ISSN 1096-7192 Volume 126 , Issue 2 , February 2019

Available online at www.sciencedirect.com Molecular Genetics and Metabolism 126 (2019) S1

Contents lists available at ScienceDirect

Molecular Genetics and Metabolism

journal homepage: www.elsevier.com/locate/ymgme

Program and Abstracts WORLDSymposium 2019* 15th Annual Research Meeting

Hyatt Regency Orlando 9801 International Drive, Orlando, Florida, USA February 4 – 7, 2019

Organizing Committee Chester B. Whitley, PhD, MD, Chair Lalitha R. Belur, PhD** Philip J. Brooks, PhD** Amber R. Brown, CPP Brenda M. Diethelm-Okita, MPA Yoshikatsu Eto, MD, PhD** Amy Gaviglio, MS, LCGC** Stephen C. Groft, PharmD** Jeanine R. Jarnes, PharmD Priya S. Kishnani, MD, MBBS** Walter C. Low, PhD** R. Scott McIvor, PhD** Jill A. Morris, PhD** Li Ou, PhD** Anne R. Pariser, MD** Marc C. Patterson, MD** David A. Pearce, PhD** Mark S. Sands, PhD** Dawn C. Saterdalen, RN, MBA Danilo A. Tagle, PhD** Cynthia J. Tiﬀt, MD, PhD** Tiina K. Urv, PhD** Steven U. Walkley, DVM, PhD

* WORLD is the acronym for We’re Organizing Research for Lysosomal Diseases ** Special thanks is given to members of the Program Committee responsible for abstract review and scoring for platform presentation. https://doi.org/10.1016/j.ymgme.2018.12.013

Available online 29 December 2018 1096-7192 Molecular Genetics and Metabolism 126 (2019) S2–S6

Contents lists available at ScienceDirect

Molecular Genetics and Metabolism

journal homepage: www.elsevier.com/locate/ymgme

WORLDSymposium™ 2019 Introduction

Overview category of Basic Science was expanded to include 2 new subcategories: 1) Disease Mechanisms, Pathology, and Biomarkers of Lysosomal Now in the 15th year, WORLDSymposium™ is celebrating numerous Diseases, and 2) Developing Therapeutic Approaches for Lysosomal milestones in research for lysosomal diseases. WORLDSymposium is a Diseases in the Laboratory. The category of Translational Research was multidisciplinary forum and research meeting with the primary purpose subdivided into 3 areas: 1) Gene Therapy, 2) Newborn Screening, and of presenting and discussing the latest advances in basic science, 3) Clinical Trial Readiness – Preclinical Trial Methods and Studies for translational studies, and clinical applications for lysosomes and lyso- Lysosomal Diseases. Finally, Clinical Applications was expanded to somal diseases. WORLD is the acronym for We're Organizing Research for address the separate areas of 1) Clinical Trials for Registration, and 2) Lysosomal Diseases, and WORLDSymposium represents the single lar- Clinical Trials: Improving Outcomes Post-Registration. gest annual global gathering of lysosomal disease experts. WORLDSymposium™ 2019 is scheduled from February 4-7, 2019, at the Poster Sessions Hyatt Regency Orlando in Orlando, Florida, USA. After consecutive record-breaking years of attendance since 2010, attendance at the 2019 In addition to the 390 abstracts submitted before the October 1 meeting is expected to exceed 1,900 participants. deadline, every year additional ‘late breaking’ abstracts can be sub- At the start of WORLDSymposium 2019, two important meetings mitted until December 1. The late-breaking abstracts may be presented help kick-off the 15th annual meeting. On Monday, February 4, 2019, in the poster sessions, but do not appear in this issue of Molecular the Lysosomal Disease Network (LDN) Council of Patient Advocates Genetics and Metabolism, nor are they systematically reviewed for a (COPA) meets in the morning to provide patient-oriented input into the platform presentation. All accepted abstracts are assigned to a specific future of the LDN. On Monday afternoon, Emerging Trends: State-of-the- time for display and discussion during the poster sessions on Tuesday Art for Experts provides updates on the basic science, diagnosis, and and Wednesday after the general session is adjourned. Abstracts are treatment of lysosomal diseases. The content provides comprehensive distributed into two separate poster sessions. First authors with last information on lysosomal diseases, but does not overlap or replace the names from A-L will present their posters on Tuesday, February 5, scientific data being presented during WORLDSymposium™ 2019. 2019, from 4:30–6:30 PM. On Wednesday, February 6, 2019, first au- The program of new research presentations runs Tuesday through thors with last names M-Z and all late-breaking abstracts will be pre- Thursday. The first day of the program is focused on Basic Science, the sented from 4:30–6:30 PM in the Exhibit Hall. second day on Translational Research, and the third on Clinical Applications. This day-by-day progression through the three stages of Molecular Genetics and Metabolism research is a signature feature of WORLDSymposium™. The main emphasis is to assess the mechanisms and obstacles for taking laboratory To enhance awareness of lysosomal diseases and advance research discoveries to actual treatments and options for patients. A number of designed to provide better therapies, the entire February 2019 special satellite and ancillary meetings — either directly associated with issue of Molecular Genetics and Metabolism is focused on lysosomes and WORLDSymposium™, or independently organized — will be held in lysosomal diseases. In addition to abstracts being presented, the issue is adjacent areas when the meeting is not in session. The week closes with filled with the highest quality, peer-reviewed research articles, some the Council of Research Experts (CORE) NIH-funded LDN Investigators' derived from the Lysosomal Disease Network™ (LDN) studies, and nu- Meeting, which occurs on Thursday evening, February 7, 2019. The merous other organized studies and observations not directly associated Program, and more information about the Symposium and lysosomal with the LDN. diseases, are available at www.WORLDSymposia.org. Nomenclature Platform Presentations The abstracts are minimally edited for clarity and continuity. For The platform program is derived from submitted research abstracts example, it is the editorial practice of WORLDSymposium™ to eliminate judged by a review panel to be of broadest interest, and featured pre- the term “storage” in the context of the name “lysosomal storage dis- sentations by invited speakers. Abstracts submitted at www. ease”. This practice focuses attention on the growing body of knowl- WORLDSymposia.org by the regular October 1 deadline are edge indicating that ‘storage’ may not be the primary common me- considered for platform presentation. For the 2019 program, 390 chanism of pathobiology in these conditions. While the physical scientific abstracts were submitted for consideration for the 74 accumulation of undegradable macromolecules might be important in platform presentations in 7 newly expanded categories. The overall disease manifestations such as skeletal deformation, other processes https://doi.org/10.1016/j.ymgme.2018.12.014

Available online 31 December 2018 1096-7192/ © 2018 Published by Elsevier Inc. Molecular Genetics and Metabolism 126 (2019) S2–S6 such as inflammation and regulation of cell activity through the mTOR health care practitioners who are interested in being current on recent pathway, are increasingly recognized as more critical factors in the advances in the basic science, diagnosis, and treatment of lysosomal underlying pathology. diseases. This course is taught at the postgraduate level, e.g., those with Additionally, disease names and abbreviations have been edited to a PhD, MD, PharmD, DDS, MS, MPH, etc. the original format of Victor McKusick. Although more recent additions such as those made to his Online Inheritance in Man (OMIM, www. Opening Reception and Awards OMIM.org) have accumulated newer names and acronyms from the literature, WORLDSymposium™ chooses to encourage uniformity with a Monday evening will include the recognition of 10 researchers who small number of selected, clinically and historically based, standard were selected for the 2019 WORLDSymposium™ Young Investigator names. Additionally, the choice of disease names also seeks to respect Awards. The award ceremony will occur at 5:15 PM in the Regency grammatically correct language, and to endorse a limited set of Foyer, followed by the WORLDSymposium™ Opening Reception in the abbreviations, e.g., MPS I but not MPS1-H. The possessive form of Exhibit Hall at 5:30 PM. eponyms has been eliminated, e.g., Parkinson disease is used instead of Parkinson's disease and Hurler-Scheie syndrome is used instead of Young Investigator Awards “Hurler's-Scheie's syndrome”. These editorial practices are implemented as an ongoing practice of the WORLDSymposium™ and the Lysosomal Of the abstracts submitted to the meeting for review, 24 in- Disease Network to enhance progress in the field, and are also applied vestigators-in-training who submitted an abstract in the ‘Basic in development of the Official List of Lysosomal Diseases available at Research’ category also applied for the 2019 Young Investigator Award. https://www.worldsymposia.org/official-list-of-lysosomal-diseases/ The WORLDSymposium™ Planning Committee reviewed all applications and 10 individuals were selected to receive the 2019 awards. The in- Monday, February 4, 2019 vestigators listed below are recognized with the 2019 Young Investigator Award and a scholarship in partial support of their parti- Council of Patient Advocates (COPA) WORLDFair™ cipation in the meeting:

On Monday morning, WORLDSymposium™ brings together the • Rhea Ashmead, University of Victoria, Victoria, Canada Lysosomal Disease Network's Council of Patient Advocates (COPA) to • Hojun Choi, Korea Advanced Institute of Science and Technology discuss how patient advocates are partnering with investigators in the (KAIST), Daedeok Innopolis, Daejeon, South Korea, (Republic of Lysosomal Disease Network™ (LDN) to facilitate, design, and implement Korea) patient-focused studies and clinical trials aimed at the issues most im- • Chloe Christensen, University of Victoria, Victoria, Canada portant to those aﬀected by lysosomal disease. This session, • Jenny Do, National Institutes of Health (NIH), Bethesda, Maryland, WORLDFair™ is intended to encourage patients and patient advocate USA group leaders to engage in a high-level discussion to plan for the future • Areian Eghbali, National Institutes of Health (NIH), Bethesda, of lysosomal disease research under the new LDN structure. (At the time Maryland, USA of this publication, the LDN has submitted a competitive grant appli- • Eric Joshua Garcia, National Institutes of Health (NIH), Bethesda, cation to the NIH Rare Disease Clinical Research Network program, Maryland, USA with a comprehensive plan for the future of funding and research for • Francyne Kubaski, Hospital de Clínicas de Porto Alegre (HCPA), lysosomal diseases.) Porto Alegre, Brazil • Murtaza Nagree, University of Toronto, Toronto, Canada Yanyan Peng, Cincinnati Children's Hospital Medical Center, Emerging Trends: State-of-the-Art for Experts • Cincinnati, OH, USA Hiroyuki Yamakawa, Keio University School of Medicine, Tokyo, In the afternoon, seasoned researchers provide a global review of • Japan the past years' advances, a state-of-the-art overview of lysosome biology, diseases and therapies (Table 1). This review evolves every Young Investigators are awarded certiﬁcates in recognition of their year, providing a summary of the latest research trends, new knowl- achievement, at 5:15 PM on Monday, February 4, in the Regency Foyer, edge, and other discoveries. The course is intended for researchers and just prior to the WORLDSymposium™ 2019 Opening Reception in the Exhibit Hall at 5:30 PM. Table 1 Emerging Trends 2019: State-of-the-Art for Experts. Tuesday, February 5, 2019 Title Speaker

1:00 PM Introduction and Overview of Course Chester B. Whitley, PhD, MD Roscoe O. Brady Award for Innovation and Accomplishment 1:10 PM Lysosomal Disease Phenotypes Chester B. Whitley, PhD, MD 1:35 PM Normal Lysosomal Function Steven U. Walkley, DVM, On Tuesday morning, the research program of WORLDSymposium™ PhD begins with an annual tradition, the recognition of one individual who 2:00 PM Break has made outstanding contributions to the field of lysosomal disease 2:05 PM Remarkable Cases I Marc C. Patterson, MD 2:30 PM Lysosomal Disease Pathogenesis Steven U. Walkley, DVM, research. The Roscoe O. Brady Award for Innovation and PhD Accomplishment was named in honor of our esteemed colleague, and 2:55 PM Break former Award recipient, Roscoe O. Brady, MD, who passed away in 3:10 PM Current Treatments for Lysosomal Jeanine R. Jarnes, PharmD 2016. The 2019 Roscoe O. Brady Award for Innovation and Diseases fi 3:35 PM Newborn Screening Amy Gaviglio, MS, LCGC Accomplishment in the eld of lysosomal disease research and therapy 4:00 PM Break is awarded to Danilo A. Tagle, PhD, MS. (Fig. 1). 4:05 PM Future Treatments for Lysosomal Jeanine R. Jarnes, PharmD Danilo A. Tagle, PhD, MS, is associate director for special initiatives Diseases at National Center for Advancing Translational Sciences (NCATS). He 4:30 PM Remarkable Cases II Marc C. Patterson, MD also recently served as acting director of the NCATS Office of Grants 4:55 PM Final Evaluation Chester B. Whitley, PhD, MD fi 5:00 PM Course Adjourned Management and Scienti c Review and currently serves as executive secretary to the NCATS Advisory Council and Cures Acceleration

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Fig. 1. Danilo A. Tagle, PhD, MS, recipient of the WORLDSymposium 2019 Fig. 2. Mark Dant, recipient of the WORLDSymposium 2019 Patient Advocate Roscoe O. Brady Award for Innovation and Accomplishment. Leader Award.

Network Review Board. Prior to NCATS, Dr. Tagle was a program di- innovations in technology and how they can be applied to early diag- rector for neurogenetics at the National Institute of Neurological nosis for lysosomal conditions, provide progress in newborn screening Disorders and Stroke (NINDS), where he was involved in developing and gene therapy, and exploit differences at the cellular level that may programs concerning genomics-based approaches for basic and trans- indicate early disease state. lational research in inherited brain disorders. Prior to joining NINDS in 2001, Dr. Tagle was an investigator and Poster Session I section head of molecular neurogenetics at the National Human Genome Research Institute and has been involved in the highly collaborative ef- Following a full day of platform presentations, the first half of over fort toward the positional cloning of genes for Huntington disease, 440 abstracts will be displayed in the poster session, facilitating face-to- ataxia-telangiectasia and Niemann-Pick disease type C. He has served on face presentations by the first-author investigators with last names numerous committees and advisory boards, including the editorial beginning with A-L. boards of the journals Gene and the International Journal of Biotechnology. Dr. Tagle leverages key resources and expertise through partner- Wednesday, February 6, 2019 ships with stakeholders in biomedical research, including various government agencies, nonprofits, patient advocacy groups, industry orga- Patient Advocate Leader (PAL) Award nizations, pharmaceutical corporations and others in the private sector. He leads and provides scientific and programmatic oversight and co- Each year, WORLDSymposium recognizes one individual for patient ordination to the following trans-NIH programs: advocacy leadership in the field of lysosomal disease. The 2019 Patient Advocate Leader (PAL) award will be presented to Mark Dant (Fig. 2), • Extracellular RNA Communication the current Chairman of the Board of the Washington DC-based Ev- • IH Microphysiological Systems (also known as the Tissue Chip for eryLife Foundation for Rare Diseases, a science-based advocacy orga- Drug Screening program) nization dedicated to accelerating biotech innovations for rare disease • Regulatory Science treatments through science-driven public policy. Mark is also the • Stimulating Peripheral Activity to Relieve Conditions (SPARC) founder and Volunteer Executive Director of the Ryan Foundation and former President and CEO of the National MPS Society. Dr. Tagle obtained his PhD in molecular biology and genetics from In 1992, while working as a police officer in the Dallas area, Mark Wayne State University School of Medicine. He was an NIH National and his wife Jeanne founded The Ryan Foundation with a bake sale Research Service Award postdoctoral fellow in human genetics in the after their only child, three-year old Ryan was diagnosed with MPS I. At laboratory of Francis S. Collins, MD, PhD, at the University of Michigan. that time, life expectancy for children with MPS I was less than He has authored more than 150 scientific publications and has garnered 15 years. Mark began an immediate global search for scientists and numerous awards and patents. philanthropists who might aid in finding a treatment in time to help WORLDSymposium™ will present Dr. Tagle with the 2019 Roscoe O. children and families living with MPS realize the promise of tomorrow. Brady Award for Innovation and Accomplishment at the opening of the A conversation with a research scientist at a symposium in Dusseldorf general session on Tuesday, February 5, 2019 at 7:30 AM, followed by Germany eventually led Mark to Dr. Emil Kakkis, a young researcher at Dr. Tagle's presentation: Innovations in Rare Disease Research. UCLA who was working on a project to help treat MPS, but had little funding. The Ryan Foundation partnered with Dr. Kakkis and provided Basic Research key funding for his project, which culminated in the development of laronidase, the first and only drug currently approved to treat MPS I. Following the annual Innovation award and featured presentation, Ryan, now 30 and a graduate of the University of Louisville, is the platform speakers in the morning and afternoon sessions will discuss longest treated person in the world with MPS I. Since the bake sale, the

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Thursday, February 7, 2019

Clinical Application

The third and final research day of the meeting begins with an important message from Chester B. Whitley, PhD, MD (Fig. 3) who will present the Thursday Keynote Address, The ‘new’ Lysosomal Disease Network. Dr. Whitley is Professor of Pediatrics and a Faculty Member of both the Division of Genetics and Metabolism, and the Department of Experimental and Clinical Pharmacology (ECP) at the University of Minnesota Medical School in Minneapolis, Minnesota. He is certified by the American Board of Medical Genetics and Genomics in the areas of Clinical Biochemical Genetics and Clinical Genetics. Dr. Whitley is the Principal Investigator for the Lysosomal Disease Network (LDN), a growing consortium of 13 medical centers collabor- ating since 2008 to address critical gaps in taking lysosomal disease research from the lab to clinical practice. The subject of Dr. Whitley's Keynote Address is The ‘new’ Lysosomal Disease Network, describing how past lysosomal disease research and current unmet needs, are the springboard for the next generations of therapies, and a vision for the LDN. After the Keynote Address, the entire third day of original research Fig. 3. Chester B. Whitley, PhD, MD, Keynote Speaker: The ‘new’ Lysosomal is dedicated to presentations regarding clinical applications of thera- Disease Network. pies. This includes clinical trials for registration, as well as outcomes from clinical trials post registration. The goal is to provide the latest updates on the actual application of new treatments, protocols and Ryan Foundation has funded millions of dollars in rare disease research. therapies in humans affected by these conditions. For the past 25+ years, Mark and the Ryan Foundation have partnered with numerous research scientists and universities to help Networking Reception innovative projects move toward treatment in lysosomal storage disease; to include the essential published laboratory work which provided At the close of the third day of research presentations, proof of concept for Intrathecal Enzyme delivery in lysosomal storage WORLDSymposium™ offers attendees an opportunity to debrief and disease. Mark and his family have been key advocates speaking to the network with colleagues in the Regency Foyer with a networking re- FDA and in 2009 successfully championed the US House of ception including a cash bar and hors d'oeuvres from 4:30–5:30 PM. Representatives to pass the Ryan Dant Health Care Opportunity Act, a bill designed to help those living on Medicaid assistance become Lysosomal Disease Network Council Of Research Experts (CORE) meeting gainfully employed. The Dant's journey has been documented on CBS 60 Minutes, CNN, Biography Magazine, Readers Digest in 13 languages The final meeting of WORLDSymposium is the gathering of the around the world, Golf Digest, the LA Times and numerous newspapers Lysosomal Disease Network Council Of Research Experts (CORE) and news outlets across the US. meeting. This year's CORE meeting will occur on Thursday evening from Mark retired from police work in 2016 as Assistant Chief of Police 5:30–7:30 PM, and will provide an update with critical information for after serving 32 years as a Patrol Officer, Detective, and Commander past and future NIH-funded projects investigators and their teams. leading multiple divisions and Bureaus to include Patrol, Criminal Investigators and teams aiming to be involved in future funding with new Investigations, Intel, and SWAT. Mark spends his time now volunteering proposals, are required to attend this meeting. This is one of the essential for the EveryLife Foundation, the Ryan Foundation, and numerous other mechanisms by which the Lysosomal Disease Network (LDN) reviews rare disease organizations to help empower the patient advocate through progress, plans new research, and reports to NIH Program Officers. the understanding that all of us have the power to turn action to hope and hope to reality. Mark's message is that it is not the entity, but the WORLDSymposium™ Past and Future individual who holds the responsibility to make true change. On Wednesday, February 6, 2019 at 7:30 AM, Mark will be presented Past and future WORLDSymposium™ meetings (NIH educational with the WORLDSymposium 2019 Patient Advocate Leader Award. meeting support acknowledgement):

Translational Studies • 1st WORLDSymposium, Minneapolis, MN, May 13–15, 2004 (1R13NS049950–01) The second day of the original research presentations focuses on the • 2nd WORLDSymposium, Phoenix, AZ, May 22, 2005 challenge of moving laboratory discoveries to therapy, the important (1R13NS053341–01) hurdles of translational research. This year researchers will provide • 3rd WORLDSymposium, Orlando, FL, December 7–9, 2006 two-dozen abstract presentations showcasing new data and observa- (1R13NS058236–01) tions in Translational Studies. • 4th WORLDSymposium, Las Vegas, NV, February 13–15, 2008 • 5th WORLDSymposium, San Diego, CA, February 18–20, 2009 Poster Session II (1U13NS066501–01) • 6th WORLDSymposium, Miami, FL, February 10–12, 2010 Following the platform program, there will be a second poster ses- (1R13NS070423–01) sion with opportunities to meet with the investigators of over 225 ad- • 7th WORLDSymposium, Las Vegas, NV, February 16–18, 2011 ditional abstracts from ﬁrst-authors with last names M-Z, as well as all • 8th WORLDSymposium, San Diego, CA, February 7–10, 2012 late-breaking abstracts. (1R13HD072632–01)

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• 9th WORLDSymposium, Orlando, FL, February 12–15, 2013 Amber R. Brown, CPP (1R13NS081934–01) Brenda M. Diethelm-Okita, MPA • 10th WORLDSymposium, San Diego, CA, February 10–13, 2014 Yoshikatsu Eto, MD, PhD* (1R13NS086124–01). Amy Gaviglio, MS, LCGC* • 11th WORLDSymposium, Orlando, FL, February 9–12, 2015 Stephen C. Groft, PharmD* • 12th WORLDSymposium, San Diego, CA, February 29–March 4, 2016 Jeanine R. Jarnes, PharmD • 13th WORLDSymposium, San Diego, CA, February 13–17, 2017 Priya S. Kishnani, MD, MBBS* • 14th WORLDSymposium, San Diego, CA, February 5–9, 2018 Walter C. Low, PhD* • 15th WORLDSymposium, Orlando, FL, February 4–7, 2019 R. Scott McIvor, PhD* Jill A. Morris, PhD* The 16th annual meeting will be: WORLDSymposium 2020 Li Ou, PhD* February 10: Lysosomal Disease Network (LDN) Council of Patient Anne R. Pariser, MD* Advocates (COPA) Marc C. Patterson, MD* Emerging Trends: State-of-the-Art for Experts David A. Pearce, PhD* February 11–13: Research Meeting Mark S. Sands, PhD* February 14: Lysosomal Disease Network (LDN) Council of Research Dawn C. Saterdalen, RN, MBA Experts (CORE) Danilo A. Tagle, PhD* Hyatt Regency Orlando, Orlando, FL, USA. Cynthia J. Tiﬀt, MD, PhD* Tiina K. Urv, PhD* WORLDSymposium™ 2019 Organizing Committee and Program Steven U. Walkley, DVM, PhD Committee *Special thanks is given to these members of the Program Committee responsible for abstract review and scoring for platform Chester B. Whitley, PhD, MD, Chair presentation. Platform presentations for WORLDSymposium are chosen Lalitha R. Belur, PhD* on a competitive basis from all abstracts submitted by October 1, 2018. Philip J. Brooks, PhD*

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Contents lists available at ScienceDirect

Molecular Genetics and Metabolism

journal homepage: www.elsevier.com/locate/ymgme

WORLDSymposium™ 2019 Program

Monday, February 4, 2019

9:00–12:00 Council of Patient Advocates The Lysosomal Disease Network (LDN) Annual Council of Patient Advocates (COPA) Meeting (COPA) Workshop: WORLDFair™

1:00–5:00 Emerging Trends: State-of-the-Art for Experts (Registration required)

1:00 PM Chester B. Whitley Introduction and Overview of Course University of Minnesota Minneapolis, MN, United States

1:10 PM Chester B. Whitley Lysosomal Disease Phenotypes University of Minnesota Minneapolis, MN, United States

1:35 PM Steven U. Walkley Normal Lysosomal Function Albert Einstein College of Medicine New York, NY, United States

2:00 PM Break

2:05 PM Marc C. Patterson Remarkable Cases Mayo Clinic Rochester, MN, United States

2:30 PM Steven U. Walkley Lysosomal Disease Pathogenesis Albert Einstein College of Medicine New York, NY, United States

2:55 PM Refreshment Break

3:10 PM Jeanine R. Jarnes Current Treatments for Lysosomal Diseases University of Minnesota Minneapolis, MN, United States

3:35 PM Amy Gaviglio Newborn Screening Minnesota Department of Health St. Paul, MN, United States

4:00 PM Break

4:05 PM Jeanine R. Jarnes Future Treatments for Lysosomal Diseases University of Minnesota Minneapolis, MN, United States

4:30 PM Marc C. Patterson Remarkable Cases Mayo Clinic Rochester, MN, United States

https://doi.org/10.1016/j.ymgme.2018.12.015

Available online 29 December 2018 1096-7192 Molecular Genetics and Metabolism 126 (2019) S7–S16

5:00 PM Adjourn

5:15 PM Chester B. Whitley Presentation of Young Investigator Awards University of Minnesota Minneapolis, MN, United States

5:30 PM Opening Reception Exhibit Hall

6:30 PM CME Satellite Symposium Unraveling a Therapeutic Conundrum: ASMD Niemann Pick Disease Accredited provider: EXCEL Continuing Education Supported by an educational grant from Sanoﬁ Genzyme

6:30 PM CME Satellite Symposium Bridging the Gap Between Research and Clinical Advances in Gaucher Disease Type 3 Accredited provider: MedIQ Supported by an educational grant from Sanoﬁ Genzyme

S8 Molecular Genetics and Metabolism 126 (2019) S7–S16 Tuesday, February 5, 2019

6:15 AM Satellite Symposium Monitoring Disease Burden and Assessing the Biochemical, Cellular and Clinical Response Supported by Sanoﬁ Genzyme to Therapy in Fabry Disease Non-US attendees; International participants only

6:15 AM Satellite Symposium Advances in the Diagnosis of MPS Disorders Supported by Ultragenyx Pharmaceutical

Basic Science I Co-Chairs: Danilo A. Tagle & Cynthia J. Tiﬀt Disease Mechanisms, Pathology and Biomarkers

7:30 AM Chester B. Whitley Welcome & Announcements University of Minnesota Presentation of 2019 Roscoe O. Brady Award for Innovation and Accomplishment to Danilo Minneapolis, MN, United States A. Tagle

7:45 AM Danilo A. Tagle Innovations in Rare Disease Research National Center for Advancing Translational Sciences National Institutes of Health Bethesda, MD, United States

8:15 AM Eric Joshua Garcia Methylomic and whole transcriptome analyses reveal several potential modiﬁer genes in National Institutes of Health GBA1-associated Parkinson disease Bethesda, MD, United States

8:30 AM Vera Niederkoﬂer CBE treatment of alpha-synuclein over-expressing and wildtype mice models Gaucher QPS Austria GmbH disease pathology Grambach, Austria

8:45 AM Annalisa Astolﬁ Hippo and necroptosis pathways are involved in cell growth defects in Gaucher disease University of Bologna Bologna, Italy

9:00 AM Pilar Giraldo Strain-elastography in musculoskeletal evaluation in Gaucher disease Institute of Health Research Aragón (IIS Aragón) Zaragoza, Spain

9:15 AM Amelia Ahern-Rindell Design and analysis of a CRISPR gene editing strategy in a sheep model variant of GM1- University of Portland gangliosidosis Portland, OR, United States

9:30 AM Anatalia Labilloy Proteomic proﬁling of engineered human immortalized podocyte cell model of Fabry Cincinnati Children's Hospital Medical disease Center Cincinnati, OH, United States

9:45 AM Break & Exhibits

10:15 AM Behzad Najafian A novel method for quantification of globotriaocylceramide (GL-3) inclusions in affected University of Washington podocytes in females with Fabry disease shows progressive accumulation of GL-3 in Seattle, WA, United States podocytes with age and no cross-correction between affected and non-affected podocytes

10:30 AM Jin-Song Shen Dysregulated DNA methylation in the pathogenesis of Fabry disease Baylor Research Institute Dallas, TX, United States

10:45 AM Siamak Jabbarzadeh-Tabrizi Eﬀects of genetic background on disease phenotypes in a mouse model of Fabry disease Baylor Research Institute Dallas, TX, United States

11:00 AM Pasqualina Colella Latent TGF-beta-binding protein 4 modulates disease severity in the knock-out mouse Généthon, Université Evry, Université model of Pompe disease Paris Saclay Evry, France

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11:15 AM Katie Harvey The evolving role of enzymology and metabolomics in the diagnosis of lysosomal disorders Great Ormond Street Hospital in the post genomic era London, United Kingdom

11:30 AM Lunch - on own or satellite symposia Exhibit hall is open

11:45 AM Satellite Symposium Treatment Experience in CLN2 Disease – New 3.5 Year Data and Case Review Supported by BioMarin Pharmaceutical Inc.

11:45 AM Satellite Symposium Biomarkers: Ready for Prime Time in the Clinical Management of Lysosomal Diseases? Supported by Shire

Basic Science II Co-Chairs: David A. Pearce & Tiina K. Urv Developing Therapeutic Approaches in the Laboratory

1:00 PM Virginia Kimonis Antisense oligonucleotide treatment targeting glycogen synthase (GYS1) in a mouse model University of California - Irvine of Pompe disease Orange, CA, United States

1:15 PM Kohji Itoh In vivo gene therapy for Tay-Sachs and Sandhoﬀ diseases by utilizing AAV9 vector encoding Tokushima University modiﬁed HEXB Tokushima, Japan

1:30 PM Kazuki Sawamoto Development of AAV gene therapy for Morquio syndrome type A Nemours/Alfred I. duPont Hospital for Children Wilmington, DE, United States

1:45 PM Saida Ortolano Functional evaluation of an AAV9 based vector expressing alpha-galactosidase A for Instituto de Investigación Sanitaria potential gene therapy of Fabry disease Galicia Sur Vigo (Pontevedra), Spain

2:00 PM Xin Chen Therapeutic eﬃcacy and safety of scAAV9/AGA gene therapy in aspartylglucosaminuria University of Texas Southwest Medical mice Center Dallas, TX, United States

2:15 PM Murtaza S. Nagree Lentiviral-modiﬁed T Rapa cells as ‘micropharmacies’ for lysosomal diseases University of Toronto Toronto, ON, Canada

2:30 PM Hojun Choi Exosome-mediated delivery of active glucocerebrosidase to Gaucher model cells Korea Advanced Institute of Science and Technology (KAIST) Daejeon, Korea

2:45 PM Break & Exhibits

3:15 PM Ibane Abasolo Targeted nanoliposomes for the treatment of Fabry disease Vall d'Hebron Institute of Research Barcelona, Spain

3:30 PM Cristin Davidson Improved disease amelioration with combination therapy for Niemann-Pick disease type C1 National Institutes of Health Bethesda, MD, United States

3:45 PM Yanyan Peng Intravenous infusion of iPSC-derived neural progenitors expressing GCase ameliorates Cincinnati Children's Hospital Medical alpha-synuclein aggregates in a mouse model of Gaucher disease Center Cincinnati, OH, United States

4:00 PM Jeanine R. Jarnes Chitotriosidase as a biomarker for central nervous system inﬂammation in the gang- University of Minnesota liosidosis diseases Minneapolis, MN, United States

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4:15 PM Chloe L. Christensen Delivery of adenine base editors to patient-derived induced pluripotent stem cells in vitro:A University of Victoria putative treatment for mucopolysaccharidosis type IIIB Victoria, BC, Canada

4:30 PM Poster Reception in Exhibit Hall

6:30 PM Satellite Symposium Sphingolipids: Opening the Black Box Supported by Sanoﬁ Genzyme

6:30 PM Satellite Symposium An Evening of Stories with Friends: How Academics, Patients, and Industry Collaborate to Supported by Amicus Therapeutics, Inc. Develop New Therapies for Pompe Disease and Other Lysosomal Disorders

S11 Molecular Genetics and Metabolism 126 (2019) S7–S16 Wednesday, February 6, 2019

6:15 AM Satellite Symposium The Case for a First-Line, Long-Term Oral Treatment Option for Eligible Adult Gaucher Supported by Sanoﬁ Genzyme Type 1 Patients

6:15 AM Satellite Symposium Title to be announced

Translational Research IA Co-Chairs: R. Scott McIvor & Mark S. Sands Gene Therapy

7:30 AM Chester B. Whitley 2019 Patient Advocate Leader Announcement and Presentation to Mark Dant University of Minnesota Minneapolis, MN, United States

7:45 AM Steven J. Gray Intrathecal and intravenous combination gene therapy in the mouse model of infantile University of Texas Southwestern Medical neuronal ceroid lipofuscinosis extends lifespan and improves behavioral outcomes in Center moderately aﬀected mice Dallas, TX, United States

8:00 AM Raymond Y. Wang Intra-articular AAV9 α-iduronidase gene therapy in the canine model of mucopolysac- Children's Hospital of Orange County charidosis type I results in rapid synovial and cartilage iduronidase expression, clearance Children's Specialists of heparan sulfate, and high serum α-iduronidase levels Orange, CA, United States

8:15 AM Roselena S. Schuh Newborn genome editing improves phenotype, cardiovascular, respiratory, and bone Universidade Federal do Rio Grande do disease in mucopolysaccharidosis type I mice Sul Porto Alegre, Brazil

8:30 AM Brian Bigger Brain targeted stem cell gene therapy provides long-term correction of mucopolysac- University of Manchester charidosis type II Manchester, United Kingdom

8:45 AM Giuseppe Ronzitti Safety and eﬃcacy evaluation of investigational liver gene transfer for secretable GAA in Généthon, Université of Evry, Université the treatment of Pompe disease Paris-Saclay Evry, France

9:00 AM Jeﬀrey A. Medin FACTs Fabry gene therapy clinical trial: Two year data Medical College of Wisconsin Milwaukee, WI, United States

9:15 AM Kevin M. Flanigan Phase 1/2 clinical trial of systemic gene transfer of scAAV9.U1a.hSGSH for MPS IIIA Center for Gene Therapy, Nationwide demonstrates 2 years of safety, tolerability, and biopotency Children's Hospital Columbus, OH, United States

9:30 AM Cassie Bebout Analysis of the eﬀect of intravenous gene therapy on brain and peripheral disease in a Auburn University feline model of GM1-gangliosidosis Auburn, AL, United States

9:45 AM Break & Exhibits

Translational Research IB Co-Chairs: Amy Gaviglio & Priya Kishnani Implementation and Impact of Newborn Screening

10:15 AM Chia-Feng Yang Very early treatment for infantile-onset Pompe disease contributes to better outcomes: Taipei Veterans General Hospital 10-year experience of nationwide NBS in Taiwan Taipei City, Taiwan

10:30 AM Barbara K. Burton Newborn screening for mucopolysaccharidosis type II (MPS II) in Illinois: The ﬁrst year's Ann & Robert H. Lurie Children's Hospital experience Chicago, IL, United States

10:45 AM Elizabeth Braunlin Hematopoietic cell transplantation for severe MPS I in the ﬁrst six months of life: The University of Minnesota heart of the matter Minneapolis, MN, United States

S12 Molecular Genetics and Metabolism 126 (2019) S7–S16

11:00 AM Adam Guenzel Improved diﬀerentiation between Krabbe disease variants, carrier status, and pseudo Mayo Clinic deﬁciency by measurement of psychosine Rochester, MN, United States

11:15 AM Francisco J. del Castillo NGS-based, 107-gene resequencing panel as ﬁrst-line screening test for lysosomal diseases Hospital Universitario Ramón y Cajal, IRYCIS Madrid, Spain

11:30 AM Lunch - on own or satellite symposia Exhibit Hall is open

11:45 AM Satellite Symposium Precision Medicine in Fabry Disease: A New Approach to Treat the Classic Phenotype Supported by Amicus Therapeutics, Inc. Patient

11:45 AM Satellite Symposium Moving Forward to Optimal Management of MPS IVA and VI Patients Supported by BioMarin Pharmaceutical Inc.

Translational Research II Co-Chairs: Lalitha R. Belur & Philip J. Brooks Clinical Trial Readiness: Pre-Clinical Trial Methods and Studies

1:00 PM Laura Adang Clinical presentation of metachromatic leukodystrophy Children's Hospital of Philadelphia Philadelphia, PA, United States

1:15 PM Cara O'Neill The natural history of facial features observed in Sanﬁlippo syndrome (MPS IIIB) using a Cure Sanﬁlippo Foundation next generation phenotyping tool Columbia, SC, United States

1:30 PM Rebecca Ahrens-Nicklas A natural history study of multiple sulfatase deﬁciency The Children's Hospital of Philadelphia Philadelphia, PA, United States

1:45 PM Lynda E. Polgreen Exploring surrogate biomarkers of skeletal and joint disease progression in mucopoly- Los Angeles Biomedical Research saccharidosis type I Institute at Harbor-UCLA Torrance, CA, United States

2:00 PM Shunji Tomatsu Eﬀect of enzyme replacement therapy on the growth of patients with Morquio syndrome Nemours/Alfred I. duPont Hospital for type A Children Wilmington, DE, United States

2:15 PM Ankit K. Desai Changing the clinical course of infantile Pompe disease with immune modulation Duke University strategies: 12 years of experience Durham, NC, United States

2:30 PM Simon Heales Urinary glucose tetrasaccharide, a useful prognostic biomarker for Pompe disease? Great Ormond Street Hospital/UCL London, United Kingdom

2:45 PM Break & Exhibits

3:15 PM Quoc-Hung Nguyen Fetal enzyme replacement and stem cell transplantation in murine Sly syndrome University of California – San Francisco targeting microglia San Francisco, CA, United States.

3:30 PM Igor Nestrasil Discovery of brain MRI signatures in infants with severe form of MPS I in the pre-HSCT University of Minnesota and post-HSCT stages Minneapolis, MN, United States

3:45 PM Adeline Vanderver Intrathecally administered recombinant human arylsulfatase A in patients with late- Children's Hospital of Philadelphia infantile metachromatic leukodystrophy: Phase 2b clinical trial design Philadelphia, PA, United States

4:00 PM Reena V. Kartha Preliminary N-acetylcysteine results for LDN 6722, Role of oxidative stress and University of Minnesota inﬂammation in Gaucher disease type 1: Potential use of antioxidant anti-inﬂammatory Minneapolis, MN, United States medications

S13 Molecular Genetics and Metabolism 126 (2019) S7–S16

4:15 PM Eric K.W. Hui Preclinical studies of a brain penetrating IgG trojan horse-arylsulfatase fusion protein in ArmaGen Inc. the metachromatic leukodystrophy mouse Calabasas, CA, United States

4:30 PM Poster Reception in Exhibit Hall

6:30 PM Satellite Symposium Hand in Hand: A Patient and Physician Journey in Pompe Disease Supported by Sanoﬁ Genzyme

6:30 PM Satellite Symposium TOO RARE? Facing Diagnostic and Clinical Development Challenges in Alpha-mannosi- Supported by Chiesi Farmaceutici S.p.A. dosis Non-US attendees; International participants only

S14 Molecular Genetics and Metabolism 126 (2019) S7–S16 Thursday, February 7, 2019

6:15 AM Satellite Symposium Fabry Disease: Treatment Guidelines to Optimize Patient Outcomes Supported by Sanoﬁ Genzyme

6:15 AM Satellite Symposium Title to be announced

Clinical Trials I Co-Chairs: Stephen C. Groft & Anne R. Pariser Clinical Trials for Registration

7:30 AM Chester B. Whitley Keynote Address: The ‘new’ Lysosomal Disease Network University of Minnesota Minneapolis, MN, United States

8:00 AM Raphael Schiﬀmann Venglustat in adult Gaucher disease type 3: Preliminary safety, pharmacology, and Baylor Research Institute exploratory eﬃcacy from a phase 2 trial in combination with imiglucerase (LEAP) Dallas, TX, United States

8:15 AM Ronald G. Crystal Design and rationale of the LYS-SAF302 gene therapy study in mucopolysacchar- Weill Cornell Medicine idosis type IIIA (MPS IIIA) children New York, NY, United States

8:30 AM Myrl D. Holida Once every 4 weeks - 2 mg/kg of pegunigalsidase alfa for treating Fabry disease; University of Iowa Hospitals and Clinics Preliminary results of a phase 3 study Iowa City, IA, United States

8:45 AM Ulla Feldt-Rasmussen Oral pharmacological chaperone migalastat compared with enzyme replacement Rigshospitalet, Copenhagen University therapy in Fabry disease: 30-month results from the randomized phase 3 ATTRACT Hospital study Copenhagen, Denmark

9:00 AM Priya S. Kishnani Safety and eﬃcacy of VAL-1221, a novel fusion protein targeting cytoplasmic Duke University glycogen, in patients with late-onset Pompe disease Durham, NC, United States

9:15 AM Paula R. Clemens Eﬃcacy and safety of AT-GAA (ATB200/AT2221) in ERT-switch nonambulatory University of Pittsburgh and Department of patients with Pompe disease: Preliminary results from the ATB200–02 trial Veterans Aﬀairs Medical Center Pittsburgh, PA, United States

9:30 AM Loren Pena NEO1 and NEO-EXT studies: Long-term safety of repeat avalglucosidase alfa dosing University of Cincinnati College of Medicine for 4.5 years in late-onset Pompe disease patients Cincinnati, OH, United States

9:45 AM Refreshment Break

10:15 AM Nathalie Guffon The first study investigating safety and efficacy of velmanase alfa (human Hôpital Femme Mère Enfant recombinant alpha mannosidase) in alpha-mannosidosis patients below six years of Lyon, France age

10:30 AM Kara Woolgar Intravenous 2-hydroxypropyl-beta-cyclodextrin for a Niemann-Pick disease type C1 Phoenix Children's Hospital infant with liver cirrhosis Phoenix, AZ, United States

10:45 AM Paul J. Orchard Preliminary results demonstrate engraftment with minimal neutropenia with University of Minnesota MGTA-456, a CD34+ expanded cord blood (CB) product in patients transplanted for Minneapolis, MN, United States inherited metabolic disorders (IMD)

11:00 AM Joseph Muenzer CHAMPIONS: A phase 1/2 clinical trial with dose escalation of SB-913 ZFN- University of North Carolina, Chapel Hill mediated in vivo human genome editing for treatment of MPS II (Hunter syndrome) Chapel Hill, NC, United States

11:15 AM Paul Harmatz EMPOWERS: A phase 1/2 clinical trial of SB-318 ZFN-mediated in vivo human UCSF Benioﬀ Children's Hospital Oakland genome editing for treatment of MPS I (Hurler syndrome) Oakland, CA, United States

S15 Molecular Genetics and Metabolism 126 (2019) S7–S16

11:30 AM Lunch - on own or satellite symposia

11:45 AM Satellite Symposium Managing Fabry Disease – Current Real World Evidence and Future Advances Supported by Amicus Therapeutics, Inc. Non-US attendees; International participants only

11:45 AM Satellite Symposium Growth and Skeletal Complications in MPS: Future Directions Supported by Sanoﬁ Genzyme

Clinical Trials II Co-Chairs: Yoshikatsu Eto, Jill Morris & Marc C. Patterson Clinical Outcomes

1:00 PM Maureen Cleary ICV-administered tralesinidase alfa (BMN 250; NAGLU-IGF2) is well-tolerated and Great Ormond Street Hospital reduces heparan sulfate accumulation in the CNS of subjects with Sanﬁlippo London, United Kingdom syndrome type B (MPS IIIB)

1:15 PM Maria L. Escolar Long-term neurodevelopmental outcomes of hematopoietic stem cell transplanta- Children's Hospital of Pittsburgh tion for late-infantile Krabbe disease Pittsburgh, PA, United States

1:30 PM Karolina M. Stepien Hormonal dysfunction in adult patients with mucopolysaccharidosis type I post Salford Royal NHS Foundation Trust hematopoietic stem cell transplantation Salford, United Kingdom

1:45 PM Ashish Gupta Allogeneic hematopoietic stem cell transplant improves outcomes in fucosidosis University of Minnesota Minneapolis, MN, United States

2:00 PM Mark Roberts Preliminary patient-reported outcomes and safety of AT-GAA (ATB200/AT2221) in Salford Royal NHS Foundation Trust Salford, patients with Pompe disease from the ATB200–02 trial United Kingdom

2:15 PM Edwin Chavez-Cintora Insights into Sanﬁlippo syndrome provided by the ConnectMPS worldwide online BioMarin Pharmaceutical Inc. registry Novato, CA, United States

2:30 PM Troy Lund Predicting intelligence in MPS IH with biomarkers University of Minnesota Minneapolis, MN, United States

2:45 PM Refreshment Break

3:15 PM Brianna Glase Robust clinical outcome measures for patients with juvenile onset GM1-gang- National Institutes of Health liosidosis Bethesda, MD, United States

3:30 PM Angela Schulz Persistent treatment eﬀect of cerliponase alfa in children with CLN2 disease: A University Medical Center Hamburg-Eppendorf 3 year update from an ongoing multicenter extension study Hamburg, Germany

3:45 PM Tama Dinur Long-term follow-up of 103 untreated adult patients with type 1 Gaucher disease Shaare Zedek Medical Center, Hadassah Medical Center, The Hebrew University Jerusalem, Israel

4:00 PM Uma Ramaswami Migalastat: Single centre experience of adult patients with Fabry disease from the Royal Free London NHS Foundation Trust Royal Free London NHS Foundation Trust, UK University College London London, United Kingdom

4:15 PM Christian J. Hendriksz Evidence-based, expert-agreed recommendations for the management of patients Steve Biko Academic Hospital with MPS IVA/VI: Recommendations to replace the speciﬁc missing enzyme Pretoria, South Africa

4:30 PM Adjourn

4:30 PM Networking Reception in Foyer

5:30 PM Council of Research Experts (CORE) The Lysosomal Disease Network (LDN) annual Council of Research Experts (CORE) Meeting meeting for NIH-funded investigators

S16 Molecular Genetics and Metabolism 126 (2019) S17–S156

Contents lists available at SciVerse ScienceDirect Molecular Genetics and Metabolism

journal homepage: www.elsevier.com/locate/ymgme

Annual WORLDSymposiumTM Abstracts 1 Targeted nanoliposomes for the treatment of Fabry disease could be a useful platform to improve the enzyme replacement therapy in Fabry patients. Ibane Abasoloa,b, Anna Lechadoa,b, Natalia García-Arandaa,b, Dolores Buenoc,b, David Ortegaa,b, Elisabeth González-Mirac,b, Solène Pas- doi:10.1016/j.ymgme.2018.12.017 semardc,b, Zamira Vanessa Díaz-Riascosa,b, Rosa Mendozad,b, José Luis Corcherod,b, Marc Moltó-Abada, Edgar Cristóbal-Lecinae,b, e,b fi e,f e,b Daniel Pulido , Jose na Casas , Miriam Royo , Guillem Pintos- 2 g c,b c,b a,b a Morell , Jaume Veciana , Nora Ventosa , Simó Schwartz , Vall Characterization of epilepsy in a large Egyptian Gaucher disease b d'Hebron Institute of Research, Barcelona, Spain, Networking Re- type 3 (GD3) cohort: A 12-year prospective study search Center on Bioengineering, Biomaterials and Nanomedicine c (CIBER-BBN), Barcelona, Spain, Institut de Ciència de Materials de a b c a d Magy Abdelwahab , Nehal Hassib , Raphael Schiffmann , Cairo Barcelona (ICMAB-CSIC), Bellaterra, Spain, Institut de Biotecnologia i b University Pediatric Hospital, Cairo, Egypt, National Research Institute, de Biomedicina (IBB), Bellaterra, Spain, eInstitute of Advanced Cairo, Egypt, cInstitute of Metabolic Disease, Baylor Research Institute, Chemistry of Catalonia (IQAC-CSIC), Barcelona, Spain, fNetworking Dallas, TX, United States Research Center on Hepatic and Digestive Diseases (CIBEREHD), g Barcelona, Spain, Vall d'Hebron Institute of Research & University There are no previous reports characterizing the epilepsy in GD3 Hospital, Barcelona, Spain patients on long-term enzyme replacement therapy even in countries known with the highest prevalence of GD3 such as Egypt Fabry disease is caused by the deficiency of α-galactosidase (GLA) and Japan. Myoclonic epilepsy is usually described in GD3. It cannot that leads to a predominant accumulation of globotriaosylceramide always be established whether the epilepsy is related to GD3. In our (Gb3) in the vascular endothelium and smooth muscle cells, Egyptian cohort, 35 of 100 [age range: 2 to 22 years 1 D409H/D409H, eventually causing an early death at the fourth or fifth decade of 30 L444P/L444P, 4 unknown) GD3 patients developed seizures. All life. Current treatment of Fabry disease consists in the bi-weekly patients had typical oculomotor palsy. None of those with D409H/ injection of recombinant GLA (algasidase alfa or algasidase beta), but L444P developed seizures. Twenty-five percent of splenectomised its effectiveness is known to be limited, at least in part, due to the patients developed epilepsy. All patients were of consanguineous short half-life and poor biodistribution of the protein. In order to marriage from 30 families with no known family history of epilepsy improve the pharmacokinetics and the efficacy of the GLA, nano- except in one family an older sibling developed epilepsy. Four liposomes containing the recombinant enzyme were prepared patients had siblings with GD3 who did not develop epilepsy whilst following the DELOS-SUSP methodology. Moreover, c(RGDfK) pep- 5 patients had GD3 siblings that developed a terminal seizure tide was incorporated in the membrane of the vesicles to enhance (Sudden Unexplained Death in Epilepsy, SUDEP) usually at a younger the uptake by endothelial cells. Using an in-house produced GLA and age. According to the initial dose of ERT received, patients were in vitro cell cultures, we previously demonstrated that GLA- divided into 2 groups, 20-30IU/kg/2 weeks (Group A) and 60 IU/kg/2 nanoformulations reduced lysosomal Gb3 deposits more efficiently weeks (Group B). Fifteen patients developed a terminal seizure than the free enzyme. Here, we report the in vivo pharmacokinetics, (SUDEP), and 20 patients had a chronic seizure disorder with the biodistribution and efficacy of nanoliposomes containing algasidase former usually occurring in Group A during infancy. Twenty-six beta. In detail, GLA knock-out Fabry mice were administered with 1 patients developed generalized tonic-clonic seizures, 5 developed mg/Kg algasidase beta or the nanoformulated version of the enzyme. complex partial seizures, and 4 patients had myoclonic seizures with Results demonstrate that nanoliposome increases the plasma half- a frequency of several times/day to once/2-4 weeks. Abnormal life and AUC of the free enzyme. Indeed, 30 min post-adminstration behaviour often preceded seizures. Interictal surface EEG usually only 0.002% of the naked algasidase beta remained in circulation, showed slow background activity commonly without a recorded whereas ten times more enzyme was available in mice receiving the epileptogenic activity. Epilepsy was refractory and required treat- nanoliposomal formulation. Biodistribution of the enzyme was also ment with 2 antiepileptic drugs in 6/20 (30%). We conclude that improved by the use of nanoliposomes, since liver sequestration of epilepsy in Egyptian GD3 patients has unique features and natural the enzyme was significantly reduced without affecting the accu- history. Epilepsy should be excluded as comorbidity in a highly mulation of the enzyme in the kidney. Finally, a significant reduction consanguineous GD3 population even in absence of family history of plasma Gb3 and LysoGb3 levels, comparable to those seen with the free enzyme, was recorded in mice treated with algasidase beta- doi:10.1016/j.ymgme.2018.12.018 nanoliposomes. Overall, our results indicate that nanoliposomes

1096-7192/$ – see front matter S18 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

3 mobility, adaptive behavior, cognitive behavior, physical character- Natural history of Sanfilippo syndrome istics, sensory-motor skills, speech, and language skills. Indicators of MPS IIIA were categorized into four progressive stages. Stage 1 is Alyssa T. Aburachis, Michele Poe, Maria L. Escolar, University of characterized by recurrent infections and developmental delays. Pittsburgh, Pittsburgh, PA, United States Stage 2 is characterized by the emergence of behavioral problems along with regression in motor and speech skills. Stage 3 is Mucopolysaccharidosis type III (MPS III), also known as Sanfilippo characterized by severe cognitive decline and loss of ambulation. Syndrome, is a rare lysosomal storage disorder characterized by low Stage 4 is characterized by frequent pneumonias, seizures, and death. production of enzymes necessary for heparan sulfate metabolism. Of For the four stages, the median ages at evaluation are 2.25 years, 4.46 the four subtypes, subtypes A and B are more common, present at years, 6.58 years, and 11.84 years respectively. Patients with MPS IIIA earlier ages, and show faster disease progression as compared to show similar patterns in disease progression that allow for subtypes C and D. This study aims to describe and compare the subdivision into four distinct stages. This staging system can be natural histories of these four subtypes. A retrospective analysis of 79 useful in guiding physicians and families as to how and when the Sanfilippo patients was performed. 50 patients had subtype A, 21 disease will progress in the absence of treatment. Additionally, this patients had subtype B, 6 patients had subtype C, and 2 patients had study can be used in developing novel therapies targeted to the subtype D. Neurodevelopmental evaluations assessed signs and problems faced at various points in disease progression. symptoms, growth, mobility, adaptive behavior, cognitive behavior, physical characteristics, sensory-motor skills, speech, and language skills. Subtype A presented at the earliest age (median=1.25 years, doi:10.1016/j.ymgme.2018.12.020 range=0-3 years, n=50) whereas subtype D presented at the oldest age (median=16.25 years, n=2). Language delays were the most common initial symptom for subtypes A and B (59% and 63% 5 respectively). Chronic ear infections, coarse facial features, and Understanding Fabry in families: Preliminary findings from a chronic irritability were the most common initial symptoms for global survey subtype C (all 50%). Cognitive decline was the initial symptom presented for subtype D (100%). Subtypes A and B showed higher proportions of patients with hearing loss (63% and 29% respectively), Jacqueline Adam, MPS Commercial, Amersham, United Kingdom whereas subtypes C and D showed no presentation of hearing loss ‘ ’ (0%). All four subtypes presented with motor and cognitive deficits, Understanding Fabry in families was designed to identify gaps in though the age at which these deficits emerged varied by subtype. pathway to diagnosis and the accessibility of pedigree testing and All subtypes showed physical attributes characteristic of the disease genetic counselling across the area FIN (Fabry International Net- ≥ along with severe cognitive and motor deficits, though this emerged work). The survey was open to any individual ( 18 years), or a at different ages. Understanding the early symptoms can reduce the parent/carer of a child, with a diagnosis of Fabry disease. The survey time delay between emergence of symptoms and diagnosis. In was sent to all 52 FIN member countries via individual patient addition, this study can be used as a comparator when designing or organisations. In total 717 responses were received from 41 evaluating the effects of novel therapies. countries. Overall, the mean age at diagnosis with Fabry disease (± standard deviation [SD]) was 33.0 ± 15.7 years (range 0-71 years), which, in the majority, followed a family member’s diagnosis (54%). doi:10.1016/j.ymgme.2018.12.019 Of those experiencing symptoms, the majority (43%) reported pain and/or discomfort in their hands/feet led to their initial visit to a doctor. Mean time to diagnosis (± SD) was 9.5 ± 13.5 years (range 0-63 years). Diagnosis was mostly within a specialist centre setting 4 (34%), and made by a genetic (31%) or Fabry specialist (24%) for the Staging of Sanfilippo syndrome type A majority. Respondents travelled far to be diagnosed mean (± SD) distance travelled to place of diagnosis was 230.5 ± 698.3 km (range Alyssa T. Aburachis, Michele Poe, Maria L. Escolar, University of 0-6000 km). Approximately two-thirds of respondents (67%) had Pittsburgh, Pittsburgh, PA, United States undergone pedigree analysis. Just over half (53%) had been offered genetic counselling. Most (92%) had received an explanation Mucopolysaccharidosis type III A (MPS IIIA), also known as regarding the inheritance patterns of Fabry. Explanations generally Sanfilippo Syndrome type A, is a rare lysosomal storage disorder came from a geneticist (35%) or a hospital specialist (32%) and caused by low production of heparan-N-sulfatase, an enzyme occurred within 6 months of diagnosis for the majority (85%). Of necessary for heparan sulfate metabolism. Accumulation of heparan those who had an explanation, 57% received take home information sulfate correlates with the progressive loss of cognitive and motor to consolidate this. Working with patient organisations across FIN, abilities. This study aims to classify the progression of MPS IIIA into this study achieved a large sample and provided global insights into four distinct stages. A retrospective analysis of 50 Sanfilippo Type A life with Fabry. Further analysis of the data collected at a regional patients was performed. 19 patients went on to have hematopoietic and a country level may identify areas that could benefit from stem cell transplantation, 10 patients who were not transplanted additional support. were followed longitudinally, and 21 patients were lost to follow-up. For patients who underwent hematopoietic stem cell transplantation, only data collected prior to transplant was included in this doi:10.1016/j.ymgme.2018.12.021 study. Neurodevelopmental evaluations assessed symptoms, growth, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S19

6 patient reported outcomes (PROs) using standard tools: MPS HAQ, Impact of two-year elosulfase alfa treatment on patient-reported EQ-5D-5L, Adolescent Paediatric Pain Tool, Brief Pain Index and the outcomes in patients with Morquio syndrome type A: Results Beck Depression Inventory. While collecting these PROs, we have from an English managed access agreement undertaken our own research to capture changes reported by patients. Patients (or parents for under 18’s) are asked to describe, Jacqueline Adama, Maureen Clearyb, Christian Hendrikszc, Tarekegn in their own words, any changes they have noticed after 4, 8 and 12 Geberhiwotd, Derralynn Hughese, Simon A. Jonesf, Ana Jovanovicg, months on treatment and then yearly thereafter (ERT naïve patients) Alexandra Morrisona, Elaine Murphyh, Suresh Vijayi, aMPS Commercial, or yearly from entry to the MAA (patients who previously received Amersham, United Kingdom, bGreat Ormond Street Hospital, London, ERT on a clinical trial). Testimonies were collected for 42 patients United Kingdom, cUniversity of Pretoria School of Medicine, Pretoria, aged between 1—58 years. The most common changes noticed, South Africa, dNew Queen Elizabeth Hospital, Birmingham, United presented as a percentage of the total number of reported changes, Kingdom, eRoyal Free London NHS FT & UC, London, United Kingdom, are given here. The most commonly reported changes for treatment fUniversity of Manchester, Manchester, United Kingdom, gSalford Royal naïve patients were at 4 months: energy levels (10/49, 20%), NHS Foundation Trust, Salford, United Kingdom, hNational Hospital for walking/movement (9/49, 18%) and pain (8/49, 16%) at 8 months: Neurology and Neurosurgery, London, United Kingdom, iBirmingham general health (22/79, 28%), walking/movement (17/79, 22%) and Children’s Hospital NHS Foundation Trust, Birmingham, United Kingdom energy levels (13/79,16%) at 12 months: general health (24/95, 25%), energy levels (19/95, 20%) and sleep/tiredness (16/95, 17%). The Morquio syndrome type A may significantly impact patients’ overall most commonly reported changes for clinical trial patients were at quality of life (QoL) and activities of daily living (ADL). Elosulfase alfa is 12 months: energy levels (11/53, 21%), general health (10/53, 19%) the only approved treatment for Morquio syndrome type A. In England, and specific health benefits and walking/movement (both 7/53, 13%) access to treatment is granted on a conditional basis through a managed at 24 months: energy levels (4/19, 21%), walking/movement, sleep/ access agreement (MAA). One criteria to continue treatment is tiredness, thinking/learning and general health (all 3/19, 16%). This stabilization/improvement in QoL/ADL assessed by patient-reported study highlights a range of outcomes that are important to patients’ outcomes (PROs). PROs assessed QoL/ADL [EQ-5D-5L and/or MPS-HAQ lives that may not be collected via current standard PRO tools. These caregiver domain (MPS-HAQ CD)], depression [Beck Depression Inven- findings may act as a guide to the selection of suitable tools or the tory (BDI)], and pain [Adolescent and Pediatric Pain Tool (APPT) or Brief development of disease specific measures for use in future studies. Pain Inventory (BPI)]. Patients with MAA baseline and 24-month assessments (20-month for BDI) were evaluated and reported. As of March 2018, 36 Morquio A patients had received treatment in the MAA doi:10.1016/j.ymgme.2018.12.023 for ≥2 years (26 initiated treatment in clinical trials, 10 initiated in MAA). Mean(SD) treatment duration was 7(1.4) and 2.1(0.3) years for each group, respectively (range: clinical trial patients 4.1-8.9 years, patients 8 initiating treatment in MAA 0.2-2.2 years). For clinical trial patients, EQ- Clinical presentation of metachromatic leukodystrophy 5D-5L (n=22) and MPS-HAQ CD (n=24) remained unchanged from baseline-24 months [EQ-5D-5L: mean(SD) change: -0.04(0.29) MPS- HAQ CD: 0.38(6.27)]. Patients initiating treatment in the MAA who Laura Adang, Akshilkumar Patel, Omar Sherbini, Julia Kramer- completed EQ-5D-5L (n=8) and MPS-HAQ CD (n=10) also remained Golinkoff, Justine Shults, Adeline Vanderver, Children's Hospital of unchanged [EQ-5D-5L: 0.08(0.53) MPS-HAQ CD: -1.3(16.79)]. From Philadelphia, Philadelphia, PA, United States. baseline-20 months, BDI remained unchanged for clinical trial patients [n=19-0.47(2.34)] while those initiating treatment in MAA improved Metachromatic leukodystrophy (MLD) is a rare, lysosomal storage [n=4-5.75(6.85)]. Clinical trial patients (n=11) and those initiating disorder caused by decreased enzymatic activity of arylsulfatase A. treatment in MAA (n=9) assessed on APPT improved from baseline-24 This can be the result of mutations in the ASA gene, or in rare cases months [-3.27(1.62) and -2.33(2.83), respectively]. Only 6 clinical trial PSAP. Historically, MLD has been subdivided into 3 forms based on age patients were assessed on BPI results remained unchanged [-0.17(2.23)]. of onset: late infantile, juvenile, and adult. These subtypes were fi No patients discontinued treatment due to intolerability or lack of de ned decades ago, prior to the appreciation of the full clinical efficacy. In the MAA, patients receiving elosulfase alfa for ≥2yearsshowed spectrum of this lysosomal storage disorder and the advent of genetic fi stable or improved PRO outcomes, in line with previously reported testing. As a consequence, these empiric age-based historical de ni- stabilization/improvement of clinical outcomes with elosulfase alfa. tions do not fully account for the spectrum of disease and are not founded in evidence-based analysis of phenotypic cohorts. Addition- fi doi:10.1016/j.ymgme.2018.12.022 ally, the antiquated de nitions do not fully predict presenting features or disease course, and they fail to stratify outcomes in the few therapies currently available to treat this disease. As novel targeted therapeutics are developed, it is essential to have a clear understand- 7 ing of the clinical presentation and natural history of MLD. Without Patient reported outcomes in MPS IVA patients receiving enzyme properly defined sub-populations, it is difficult to design a therapeutic replacement therapy clinical trial that can demonstrate efficacy in a heterogeneous group. In this project, we collected the retrospective natural history of over 50 Jacqueline Adam, Alexandra Morrison, Samantha Wiseman, MPS individuals from around the world. We created an electronic database Commercial, Amersham, United Kingdom in REDCap to able to longitudinally collect clinical information. Using this retrospective natural history approach to understanding the Individuals with MPS IVA in England have access to enzyme disease course of individuals affected by MLD, we were able to replacement therapy (ERT) via a managed access agreement (MAA). characterize age of onset, delay to diagnosis, and common presenting Under the agreement, patients are monitored clinically and via features. Our results suggest distinct clinical phenotypic subgroups. S20 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

With a better understanding of the natural history of MLD, we will be 0.06nmol/L to 137.06nmol/L) however, it was significantly higher in able to better counsel families and to design clinical trials with more males (38.41nmol/L compared to 7.00nmol/L in females, pb0.001) and coherent cohorts and more appropriate clinical endpoints. patients with classical phenotype (27.39nmol/L compared to 5.18nmol/ L in patients with attenuated phenotype, pb0.001). Moreover, the diagnostic accuracy of lyso-Gb3 to identify patients with mutations doi:10.1016/j.ymgme.2018.12.024 associated with classical phenotype was very good (area under the curve [AUC] 0.99 and 0.95 for males and females, respectively). For the entire cohort, there was a significant correlation between lyso-Gb3 and albuminuria, left ventricular mass (LVM) or Mainz Severity Score Index 9 (MSSI), but not between lyso-Gb3 and estimated glomerular filtration The role of human iPS-derived microglia-like cells in Gaucher and rate however, grouping patients by gender and phenotype, there was Parkinson disease only a significant correlation between lyso-Gb3 and LVM or MSSI in classically affected males or females, respectively. Our findings suggest Elma Aflaki, NIH/NHGRI, Bethesda, MD, United States that plasma lyso-Gb3 seems an excellent biomarker distinguishing between classical and attenuated phenotypes in FD however, after fl Chronic in ammation mediated by microglia cells is a crucial adjusting lyso-Gb3 for gender and phenotype (main contributors for its process involving to the cell death of dopaminergic neurons, which plasma values), it presents weak correlations with clinical manifesta- fl leads to in ammatory response in the brain. The gene encoding tions. These results support the need for biomarkers related to FD Glucocerebrosidase (GBA) is an important risk factor for Parkinson manifestations in each affected organ. disease. Moreover, excessive neuroinflammation was observed in neuropathic form of Gaucher disease (type 2 and type 3). Therefore, doi:10.1016/j.ymgme.2018.12.026 studying microglia the only lifelong resident immune cells in brain, is very crucial. Differentiating iPSCs into microglia has been very challenging due to controversy regarding their germline origin. In this study, we successfully differentiated iPSCs into microglia-like cells derived from 11 fibroblasts from patients with types 2 Gaucher disease as well as type 1 MicroRNAs in Fabry disease: Distinguishing between phenotypes Gaucher disease with and without Parkinson disease. The type 1, iPSCs and correlations with organ involvement with genotype N370S/N370S were corrected using CRISPR and TALEN gene editing technology. These CD11b+ cells,expressIba1,CXCR3,PU.1 Patricio Aguiara, Marina C. Costab, Olga Azevedoc, Manuela Fiúzaa, and TMEM11, markers which differentiate microglia (tissue specific Jacira Marinod, José Luís Ducla Soaresa, Derralynn Hughese, Francisco macrophages in brain) from other macrophages. Functional assays for Enguitab, aCentro Hospitalar Lisboa Norte, Lisbon, Portugal, bInstituto de chemotaxis and phagocytosis were studied in these microglia. Studies of Medicina Molecular-Lisbon Medicine Faculty, Lisbon, Portugal, cHospital neuroinflammation were then be performed by co-culturing microglia Senhora da Oliveira, Guimarães, Portugal, dRoyal Free London NHS and dopaminergic neurons derived from the same individual, enabling Foundation Trust, London, United Kingdom, eRoyal Free London NHS the evaluation of the role of microglia in neuronal synapses. Foundation Trust and University College London, London, United Kingdom. doi:10.1016/j.ymgme.2018.12.025 MicroRNAs (miRNAs) are short noncoding RNAs, controlling gene expression at the post-transcriptional level and may regulate several pathways involved in FD pathophysiology. In a previous study evaluating the entire miRNAnome in FD we found that 31 miRNAs 10 were significantly differentially expressed between FD patients and Plasma lyso-Gb3 in Fabry disease: Helpful distinguishing pheno- controls, remarkably all down-regulated. Here, we present the results of types, but not as predictor of organ involvement these miRNAs evaluation in a larger cohort of FD patients. In this multicentre, prospective, cross-sectional and diagnostic test study, we Patricio Aguiara, Olga Azevedob, Jacira Marinoc, José Luís Ducla determined the 31 differentially expressed miRNAs in a large cohort of Soaresa, Derralynn Hughesd, aCentro Hospitalar Lisboa Norte, Lisbon, FD patients. MiRNAs were quantified by reverse quantitative PCR using Portugal, bHospital Senhora da Oliveira, Guimarães, Portugal, cRoyal specific LNA primers (Exiqon). The diagnostic accuracy of miRNAs to Free London NHS Foundation Trust, London, United Kingdom, dRoyal identify patients with classical phenotype and its correlation with Free London NHS Foundation Trust and University College London, clinical variables was determined. In a cohort of 95 FD patients (49.5% London, Portugal males, mean age 51.2years old, 84.2% in ERT) and 25 controls. There was a remarkable difference in the expression profile of the 31 miRNAs Fabry disease (FD) is clinically heterogeneous in terms of clinical between patients with classical phenotype, patients with attenuated manifestations and rate of progression. A substrate-related biomarker, phenotype and healthy controls, with the second group presenting an lyso-Gb3, has been stated as a hallmark of FD however, the magnitude of intermediate expression. Moreover, the diagnostic accuracy of miRNAs its correlation with clinical manifestations and its prognostic value is not to identify patients with classical phenotype was very good (area under well established, with studies showing conflicting results. Therefore, the the curve [AUC] N0.7336 and p value b0.0001 for nine of the evaluated prognostic value of lyso-Gb3 needs to be further evaluated in large miRNAs). Two out of the 31 miRNAs showed a significant direct cohorts of patients. In this multicentre, prospective, cross-sectional and correlation with estimated glomerular filtration rate (eGFR) stronger diagnostic test study, a cohort of 73 FD patients (39.7% males, mean age than the inverse correlation between albuminuria and eGFR, mainly in 51.0±14.8years, 68.5% on ERT) was consecutively recruited. The classical phenotype. MiRNAs may help to distinguish classical and accuracy of plasma lyso-Gb3 to predict the phenotype of the patients attenuated phenotypes and may overcome albuminuria in the evalua- and their clinical manifestations has been determined. In the entire tion of FD nephropathy. Moreover, confirming that miRNAs down- cohort of FD patients, mean lyso-Gb3 was 19.48nmol/L (ranging from regulation is associated with more advanced disease, it further supports Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S21 that impaired exocytosis of the miRNAs (due to disruption of the Golgi/ therapy (ERT) is effective to only type-1 Gaucher patients, based on endosomal/lysosomal trafficking in an environment of lysosomal intravenous administration of engineered glucocerebrosidase, sub- dysfunction) may be an alternative pathophysiological pathway in FD. strate reduction therapy (SRT) is an alternative treatment strategy that involves partial inhibition of GL-1 synthesis, expanding the clinical indication to type-3 Gaucher patients. Currently 2nd genera- doi:10.1016/j.ymgme.2018.12.027 tion SRT drug, ibiglustat has been shown to effectively treat neuropathic disease such as type-3 GD and Fabry disease with improved brain penetration in phase 2 clinical study, compared with the 1st generation SRT drug, eliglustat tartrate which is limited due to 12 the lack of brain penetration. Therefore, much higher brain Design and analysis of a CRISPR gene editing strategy in a sheep penetration is still considered as the key factors to obtain better in model variant of GM1 gangliosidosis vivo efficacy in neuropathic lysosomal storage disorder and development of novel glucosylceramide synthase (GCS) inhibitor with Amelia Ahern-Rindell, David Wynne, University of Portland, Portland, increased brain penetration is urgently needed. Here, we present the OR, United States discovery and optimization of lead candidates with pharmacological and blood-brain penetration (BBB) profiles that have potential to An appropriate step in the process of identifying a disorder-causing treat type-3 GD. The series of compounds from six different scaffolds mutation is to determine if a candidate DNA sequence difference can were synthesized and evaluated for their BBB penetration and trigger the phenotypic abnormality associated with the disorder. The inhibitory activity against GCS. Among the bicyclic derivatives fi gene-editing technique known as CRISPR is an ideal tool to con rm the studied in this work, we identified promising compounds with validity of any suspected disorder-causing mutation. We report on a highly potent GCS inhibitory activity (IC50 = 5 ~ 20 nM), good preliminary study to design a CRISPR-based strategy to revert a metabolic stability and no DDI consideration from CYP inhibition. In reported-on sequence alteration, and the likely disorder-causing addition, the area under the curve (AUC) based brain to plasma ratio mutation, of an ovine variant of GM1 Gangliosidosis (GM1). GM1 is a of the selected compounds was highly excellent BBB penetration in fi lysosomal disorder (LSD) caused by a de ciency of the lysosomal mice (Kp N5.0), suggesting excellent in vivo GL-1 lowering efficacy in enzyme ß-galactosidase (ß-gal). A sheep model of GM1 has been the neuropathic disease animal models. These findings indicate the fi fi identi ed and characterized with a dual de ciency of ß-gal and alpha- series of bicyclic derivatives are promising as a potent novel α fi neuraminidase ( -neur). A sequence variation has been identi ed in therapeutic for the treatment of type-3 GD through further β the ovine GLB1 gene in exon 6, a critical coding region for the -gal development of current lead compounds. protein and where human GM1-causing mutations have been documented to occur. We identified a potential target site for CRISPR close to doi:10.1016/j.ymgme.2018.12.029 this missense mutation and designed a validation assay to sort through the mixed population of DNA sequences one would expect after utilizing the CRISPR gene editing protocol. We took advantage of an existing restriction enzyme site located at the specific DNA base 14 fi mutation by employing a restriction digest of the edited DNA. Although A natural history study of multiple sulfatase de ciency a promising approach, the restriction enzyme did not consistently cut the DNA sequence as needed regardless of whether the source of DNA Rebecca Ahrens-Nicklasa, Laura Adanga, Omar Sherbinia, Kimberly template used was artificially synthesized or isolated from sheep tissue Goodspeedb, Samuel Hughesb, Alan Finglasc, Amber Olsend, Lars in the form of genomic DNA. This research continues in order to design a Schlotawae, aThe Children's Hospital of Philadelphia, Philadelphia, PA, suitable screening process to verify CRISPR edits and continue with United States, bUniversity of Texas Southwestern Medical Center, Dallas, efforts to utilize the sheep model to help create a treatment therapy for TX, United States, cMSD Action Foundation, Dublin, Ireland, dUnited MSD GM1, a devastating and fatal neurological disorder. Foundation, Ocean Springs, MS, United States, eUniversity Medical Center Göttingen, Göttingen, Germany.

fi doi:10.1016/j.ymgme.2018.12.028 Multiple sulfatase de ciency (MSD) is an ultra-rare lysosomal storage disorder affecting approximately 1:1,400,000 live births. This disease results from mutations in the SUMF1 gene, which encodes the formylglycine generating enzyme (FGE). FGE catalyzes the post- 13 translational modification of a cysteine residue in the active site of all Development of a novel glucosylceramide synthase (GCS) inhib- 17 known human sulfatases. While loss of FGE activity globally itor with increased blood-brain barrier penetration for treatment reduces sulfatase activity, residual function of each enzyme is of Gaucher disease variable. Accordingly, we hypothesize that the broad phenotypic spectrum of MSD arises from the relative reduction of individual Sujin Ahna, Younggyu Kongb, Jung-Hwan Kimb, Donghoon Kima, sulfatases. To date, the rare and heterogeneous nature of MSD has Seulki Kwona, Jong-Suk Parka, Sooyong Chunga, Wonhui Chunga, limited our ability to prognosticate outcomes and will impede the Tae-Dong Hana, Se-Woong Oha, Soongyu Choia, aYuhan R&D Institute, design of future clinical trials with appropriate endpoint selection. To Yuhan Corporation, Seoul, Republic of Korea, bMogam Biotechnology address these issues, in collaboration with patient advocacy groups, Institute, Green Cross Corporation, Yongin-si, Gyeonggi-do, Republic of we have designed a pilot retrospective study to capture the Korea longitudinal natural history of this ultra-rare, systemic disorder. We designed an electronic database using REDCap to capture key clinical Gaucher disease (GD) is a genetic disorder caused by a deficiency components, including genotype, residual sulfatase activity, age of of glucocerebrosidase and the consequent accumulation of and clinical features at presentation, clinical trajectory, and impor- glucocerebroside (GL-1) in the lysosome. While enzyme replacement tant clinical endpoints. These endpoints include developmental S22 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 milestones, hospitalizations, and relevant surgeries as well as Frutosa, Izaskun Arenaza, Jesus Villarrubiad, Abelardo Bareze, Horacio disease-specific measures, such as the onset of ichthyosis. The Canof, Maria Teresa Molerog, Lucia Villalonh, Pilar Giraldoa, aInsituto preliminary results from this pilot study identified a novel subset of Investigacion Sanitaria Aragon, FEETEG, GEEDL, Zaragoza, Spain, affected individuals with milder disease who live into adulthood, bHospital Universitario Miguel Servet, FEETEG, Zaragoza, Spain, cCentro adding to our overall understanding of this heterogeneous genetic de Diagnostico por Imagen CEMEDI, FEETEG, Zaragoza, Spain, dHospital disorder. As a next step, we will expand our retrospective natural Universitario Ramon y Cajal, GEEDL, Madrid, Spain, eComplejo history study and anticipate the launch of a companion prospective Asistencial de Ávila, GEEDL, Avila, Spain, fHospital Virgen de la Arrixaca, study. Collectively, we anticipate that the results of these studies will GEEDL, Murcia, Spain, gHU Dr. Negrin, GEEDL, Las Palmas de Gran facilitate the design of future therapeutic trials and improve our Canaria, Spain, hHUFA, GEEDL, Alcorcon, Madrid, Spain ability to provide appropriate anticipatory clinical care. The Spanish Registry of Gaucher Disease (REsEG) was stablished within the Spanish Foundation for the Study and Therapeutics of GD doi:10.1016/j.ymgme.2018.12.030 (FEETEG), in 1993 to provide support to all those involved in the management of GD patients. This work reviews the characteristics at diagnosis of type 1 adult GD patients (GD1) diagnosed outside early diagnosis programs, focussing on leading diagnostic symptoms in 15 order to answer the current challenges in patient suspicions. Patients Chondrocytes and cardiomyocytes derived from Morquio syn- 18+ years old included in the REsEG were selected. Demographic, drome type A induced pluripotent stem cells (iPCS) genetic, clinical, biomarkers and other information were analyzed. Platelets b140x109/L defined thrombocytopenia and hemoglobin b12 Carlos J. Almeciga-Diaza, Alexander Rodriguez-Lopeza,Amanda g/dL/b13 g/dL in males/females defined anemia. To analyze the Baskfieldb,RongLib, Wei Zhengb, aPontificia Universidad Javeriana, Bogota, evolution in the diagnosis a cut-off according the year of diagnosis, Colombia, bNational Institutes of Health, Bethesda, MD, United States b2000 or 2000+, was used. 223 adults GD1 patients were included. 50.7% (113) were females, mean age at diagnosis 38.5 (18-87) y.o. Morquio syndrome type A is a lysosomal storage disease The leading symptoms were: splenomegaly 71.1%, (3 during produced by deficiency of lysosomal enzyme N- pregnancy work out), thrombocytopenia 83.3%, anemia 39.0%, acetylgalactosamine-6-sulfate sulfatase (GALNS). Currently, Morquio pancytopenia 8.5%, hepatomegaly 22.5%, bone pain: 57.9% (bone syndrome type A patients are treated by enzyme replacement crisis 48.8%, fractures 3 cases), diathesis (ecchymosis, epistaxis): therapy (ERT). Many other therapeutic strategies are under devel- 20.6%, peripartum bleeding: 3 cases, family study: 19.3%, Parkinson's opment including gene therapy, small molecules, recombinant study 3 cases, MGUS 4 cases, hyperferritinemia 7 cases. Bone marrow enzymes produced in various microorganisms and new DNA/protein aspiration/biopsy was the first test in the diagnosis process in about delivery systems. Therefore, cell-based disease models are important 80% of cases. Before year 2000 (group A) 131 (58.7%) patients were for evaluation of therapeutic candidates. In this study, we describe diagnosed, mean age 36 (18-87) y.o and 92 (41.3%) later (group B). the generation of induced pluripotent stem cells (iPSC) from mean age 41.8 (18-79) y.o. For General characteristics and genotype Morquio syndrome type A skin fibroblasts, and their differentiation see tables 1 and 2. Concerning patient’s profile, group A shows a to chondrocytes and cardiomyocytes. Three patient iPSC lines were more aggressive behavior, the incidence of thrombocytopenia was generated, showing normal karyotypes and iPS cell morphology. The similar, however, the severity were higher in group A (69.8 16-129 cardiomyocytes and chondrocytes differentiated from these iPSCs x109/L) vs group B (83.4 14-131 x109/L) (p=0.006), also anemia, expressed the expected biomarkers. They also showed increased bone pain and visceral enlargement were worse in group A. Also 7 lysosome size as observed by the LysoTracker staining. These patient cases developed Parkinson's disease and 21 neoplasms among other derived cardiomyocytes and chondrocytes were used to evaluate a interesting findings. recombinant GALNS produced in P. pastoris and four small molecules that led to significant reduction of Lysotracker staining in the patient fi cells. We also observed an increased ef cacy to the treatments in doi:10.1016/j.ymgme.2018.12.032 these cells compared to the patient fibroblasts, indicating that disease relevant cells are critical for evaluating drug efficacy. In summary, we reported the first iPSCs for Morquio syndrome type A diseases, and differentiated cardiomyocytes and chondrocytes for 17 evaluation of potential therapeutics. These cells might be used also in Prospective multi-center national study to standardize the the study of the disease pathophysiology, as well as in the design of follow-up of type 1 Gaucher disease patients treated with new therapeutic alternatives. eliglustat under standard of care practice: TRAZELGA project

Marcio M. Andrade-Camposa, Jorge J. Cebollab, Laura Lopez de doi:10.1016/j.ymgme.2018.12.031 Frutosb, Pilar Irunb, Maria Soledad Noyac, S. Nietod, Lucia Villalone, A.I. Arribasf, Abelardo Barezg, Javier Garcia-Fradeh, N. Hermosini, Jose Maria Hernandez Rivasj, Daiana Ibarretxek, M.L. Lozano Almelal, Maria T. Molerom, M. Moradon, M.A. Perez Saenzo, Jesus Villarrubiap, 16 Pilar Giraldoa, aInstituto Investigación Sanitaria Aragon, FEETEG, GEEDL, ﬁ Twenty- ve years diagnosing Gaucher disease in Spain: What we Zaragoza, Spain, bInstituto Investigación Sanitaria Aragon, FEETEG, have learned? Zaragoza, Spain, cCHU A Coruña, GEEDL, A Coruña, Spain, dH Vega Lorenzo Guirao de Cieza, GEEDL, Cieza, Murcia, Spain, eHUFA, GEEDL, a a b Marcio M. Andrade-Campos , Jose Luis Capablo , Jesus J. Fraile , Madrid, Spain, fH. La Axarquia, GEEDL, Malaga, Spain, gComplejo c a a Mercedes Roca , Pilar Irun , Jorge Javier Cebolla , Laura Lopez de Asistencial de Ávila, GEEDL, Avila, Spain, hHU. Rio Hortega, GEEDL, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S23

Valladolid, Spain, iHospital Univeritario de Jerez, GEEDL, Jerez de la 3 years on treatment. A retrospective chart review of 258 patients Frontera, Cadiz, Spain, jHCU Salamanca, GEEDL, Salamanca, Spain, with GD1 seen at the NYU Lysosomal Storage Disorders Program kIISPV Pere Virgili Health Research Institute, Reus, Tarragona, Spain, lHU over the past 30 years was conducted to assess bone density changes Morales Meseguer, GEEDL, Murcia, Spain, mHU Dr Negrín, GEEDL, Las in the lumbar spine, femoral neck, and total hip as measured by dual- Palmas de Gran Canaria, Spain, nHU La Paz, GEEDL, Madrid, Spain, oHU energy X-ray absorptiometry (DXA). Only patients with GD1 who Fund. Jimenez Diaz, GEEDL, Madrid, Spain, pHU Ramon y Cajal, GEEDL, had a DXA performed at two separate time points were evaluated Madrid, Spain and analyzed with regards to ERT treatment dose and duration. Here we report on the effects of ERT on bone density in patients with GD1, The new substrate inhibitor eliglustat (ELI), available in Spain stratified by dose. Baseline and follow up demographics including since January 2017, an effective therapy decreasing the accumulation genotype, sex, ethnic background, age of diagnosis, age of treatment of substrate by a selective inhibition of glucosylceramide synthase. It initiation, prior/present use of osteoporotic therapies, vitamin D is indicated in GD1 extensive, intermediate or poor metabolizers of supplementation, and significant co-morbid diseases are reported the cytochrome CYP2D6. The clinical trials have demonstrated and were included in the analysis. improvement and stabilization of the parameters in naïve and switched from enzyme replacement therapy patients. In this work we present the design of a prospective follow-up post-authorization doi:10.1016/j.ymgme.2018.12.034 study (GEE-ELI-2017-01) to assure the traceability of eliglustat therapy in Spanish GD1patients (TRAZELGA). A national multi-center study was designed to follow-up and evaluate the response to ELI therapy during one year, analyzing clinical parameters, biomarkers, 19 changes in medullary infiltration quantified by MRI and DEXA, Non-biliary gastrointestinal symptoms as the initial presenting compliance and side effects. A quality of life sub-study is included as symptom of type 1 Gaucher disease: A case series well as an exploratory study to analyze immune system activation markers (cytokine profile, ferritin, lipocalin, gamma globulins, Kara Anstett, Heather A. Lau, NYU Langone Health, New York, NY, oxidative stress markers). All the current recommendations previous United States ELI therapy have been reinforced, including cardiac, hepatic and renal function assessment and suitability according concomitant Gastrointestinal (GI) symptoms of gallbladder dysfunction and medications. An electronic tool was designed to help physicians to hepatosplenomegaly are well known features of type 1 Gaucher easily registry and update the patient information. All the current disease (GD1). We present a series of young adults of Ashkenazi needs for data protection were taken in account. 45 GD1 patients Jewish ancestry initially evaluated by gastroenterology for non- have started ELI. Here we provide results of 35 patients whom biliary GI symptoms, later diagnosed with GD1. Patient 1 had failure fulfilled N6 months of follow-up (median age: 43.8 years (23-75), to thrive, initially attributed to celiac disease, and was later 47% males). The median follow-up is 9 (9-18) months. Genotype: diagnosed with GD1 at 19 years on family screening and noted to c.1226ANG in homozygosity (29.4%), c.1226ANG/c.1448TNC (41.2%), have hepatosplenomegaly and thrombocytopenia. Patient 2 pre- other double heterozygotes with c.1226ANG (29.4%) metabolism of sented to gastroenterology at 22 years with chronic belching, CYP2D6 (12% poor, 64.5% intermediate and 33.5% extensive bloating, reflux, early satiety and constipation. After extensive metabolizers), 23 patients were switched from ERT and 9 from workup, but no abdominal imaging, she was diagnosed with irritable miclustat, see baseline characteristics in table1. Other features and bowel syndrome. A fourth opinion led to abdominal imaging that characteristics included two splenectomized patients, 3 patients with revealed massive hepatosplenomegaly and lymphadenopathy bone palpable splenomegaly, 6 patients with multimorbidities and poly- marrow biopsy then confirmed GD1. Patient 3 developed diarrhea medications and 6 patients whom complained fatigue as the main and weight loss at 18 years, which worsened despite dietary symptom. restriction. Evaluations with GI were nondiagnostic, but did not include imaging. At age 21, she was diagnosed with GD1 on carrier screening, and found to have mild hepatosplenomegaly and doi:10.1016/j.ymgme.2018.12.033 thrombocytopenia. Patient 4 presented at 24 years with chronic abdominal pain and reflux. Pancytopenia and moderate hepatosplenomegaly resulted in a diagnosis of CMV. Two years later, he was diagnosed with GD1 through community screening. Patient 5 18 had history of constipation with acute on chronic abdominal pain. Bone density and treatment response in a large cohort of patients Imaging was suspicious for appendicitis and revealed “incidental” with type 1 Gaucher disease splenomegaly, bloodwork revealed moderate thrombocytopenia. She underwent appendectomy one year later, GD1 was diagnosed on Kara Anstett, Heather A. Lau, NYU Langone Health, New York, NY, family screening. This case series illustrates that non-biliary GI United States symptoms may represent early features of GD1 in Ashkenazi Jewish patients. These patients initially presented to gastroenterology as The progressive bone manifestations of type 1 Gaucher disease their first contact with medical care and demonstrated other typical (GD1), including bone pain, osteonecrosis, fractures, osteopenia, and features of GD1, including organomegaly and hematological de- osteoporosis have been well documented and shown to have a rangements. Physician awareness of GD1 in high-risk populations significant impact on morbidity and mortality. Initiation of recom- would help shorten the diagnostic journey for patients. binant enzyme replacement therapy (ERT) has been shown to decrease bone crises, decrease GD1 associated bone marrow infiltration and increase bone density in adults, however, quantifi- doi:10.1016/j.ymgme.2018.12.035 able improvements in bone health may not be observed until after 2- S24 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

20 improvement of cardiac, respiratory, and muscle phenotypes in the The changing role of the clinical nurse specialist for lysosomal Gaa‑/‑ mouse model of Pompe disease. We show that secretable GAA diseases: Suggestions for support in the role outperformed native GAA, even at low doses, as evidenced by enhanced survival and muscle function in Pompe mice. Moreover, Mathilda Antonini, Mel McSweeney, Maureen Cleary, Great Ormond secretable GAA vectors demonstrated superior efficacy in restoring ‑/‑ Street Hospital for Children NHS Foundation Trust, London, United Kingdom Gaa mouse muscle strength when compared to the standard of care regimen of 20 mg/kg ERT. In an effort toward clinical translation, we Lysosomal Storage Disorders (LSDs) are rare diseases managed by further optimized the expression cassette and selected a highly specialist teams ideally including a Clinical Nurse Specialist (CNS). The hepatotropic capsid. A single infusion of the leading clinical candi- role of the CNS continues to expand as diagnosis improves and new date, SPK-GAA, an investigational liver-targeted gene therapy for the therapies emerge. The main aim of the role is to support the family in treatment of Pompe disease, in non-human primates at three managing the condition. Some roles performed by the LSD CNS are ascending doses demonstrated dose-dependent expression of GAA in common to all CNS posts such as: support at diagnosis follow-up plasma. While pre-clinical efficacy and safety studies are ongoing, no informal support with medical information and telephone liaison with toxicity has been observed in any of the studies to date. These results the family and local services. The CNS often assists families with support moving SPK-GAA into clinical development. assessments, information of disease-specific support groups, written information, liaising with school/nursery and answering ongoing queries. However, the LSD nurse role has developed specific additional doi:10.1016/j.ymgme.2018.12.037 responsibilities including: supporting homecare nurses delivering ERT (available for the majority of therapies in the UK) the management of infusion-related reactions knowledge of clinical trials and novel 22 therapies and addressing the patients’ changing needs with disease A novel antibody-enzyme fusion (AEF) platform for treating progression. As LSDs affect multiple systems of the body, it is vital that glycogen storage disorders the LSD CNS is able to support the family in liaising with the multiple specialities involved, and has a better understanding of the expected complications so they can be anticipated (by regular assessments) and Dustin Armstrong, Tracy McKnight, Nadine Aziz, Valerion managed in a timely manner. This added challenge requires good Therapeutics, Concord, MA, United States multi-disciplinary working. Regular MDT clinics and meetings can allow opportunities for learning and ensure close communication. Pompe disease is a lysosomal storage disorder characterized by a fi α Forming professional relationships within the MDT can allow the LSD de ciency of the acid -glucosidase (GAA) enzyme. Also referred to CNS to feel supported within the centre. There is a lack of structured as glycogen storage disease II (GSDII), loss of GAA activity impairs educational support for the LSD CNS so it is important they utilise other the breakdown of glycogen, which can advance to organelle rupture resources such as conferences, in house teaching sessions and ask and accumulation of extra-lysosomal glycogen (XLG). Unfortunately, questions of colleagues. Support structures to allow newer LSD CNSs to traditional enzyme-replacement therapy (ERT) does not address XLG provide peer support to each other and for senior CNSs to share their because recombinant human GAA (rhGAA) is exclusively directed to wealth of experience could also be useful. lysosomes via mannose-6-phosphate receptor (M6PR) mediated uptake. To address Pompe, GSDs, and other lysosomal disorders with XLG, Valerion Therapeutics has developed a novel antibody- doi:10.1016/j.ymgme.2018.12.036 enzyme fusion (AEF) platform by genetically-fusing a proprietary, humanized, and cell-penetrating antibody fragment to enzymes capable of degrading intracellular glycogen. The lead AEF, Fab-GAA 21 (VAL-1221), is engineered for both antibody- and M6PR-mediated Safety and efficacy evaluation of investigational liver gene intracellular delivery and is thus designed to clear XLG and lysosomal transfer for secretable GAA in the treatment of Pompe disease glycogen. Fab-GAA retains significant enzyme function at neutral pH, has enhanced activities over rhGAA alone, and can degrade XLG in Sean M. Armoura, Jayme Nordina, Francesco Puzzob, Umut Caginb, multiple in vitro and in vivo models. As such, VAL-1221 is currently Marco Crosariola, Fanny Collaudb, Helena Costa Verderab, Haley being clinically tested in Pompe disease (Clinical Trials.gov Identifier: Hanbya, Virginia Haurigota, Giuseppe Ronzittib, Pasqualina Colellab, NCT02898753) and preliminary study results will be highlighted Xavier Anguelaa, Federico Mingozzia, aSpark Therapeutics, Inc., separately at this meeting (Landy, et al.). In further support of the Philadelphia, PA, United States, bGenethon, Inserm U951 Integrare, AEF platform, a separate fusion involving human pancreatic α- University of Evry, Université Paris-Saclay, Evry, France amylase, Fab-AMY (VAL-0417) also exhibits efficacy against XLG. Unlike GAA, amylase-alone lacks M6PR-binding and has no endog- Pompe disease is a lysosomal storage disease caused by loss of enous mechanism for cell internalization making, the crucial function mutations in the acid alpha-glucosidase (GAA) gene, which functionality conferred by antibody-delivery especially evident with lead to significant accumulation of glycogen in many tissues and Fab-Amy activity. The combined preclinical data presented herein results in multi-system pathology. Enzyme replacement therapy (ERT) describes novel assays designed to detect, model, and delineate the increases survival, slows disease progression, and is the current unique glycogen-degrading capabilities of the AEF platform. To- standard of care for Pompe disease patients. However, ERT has several gether, these studies strongly support an advanced functionality of significant drawbacks such as limited efficacy, immunogenicity of the AEF over traditional GSD-focused ERT. recombinant GAA, high cost, and need for frequent lengthy infusions. We have shown that investigational liver-directed adeno-associated viral (AAV) gene therapy expressing a novel secretable GAA transgene doi:10.1016/j.ymgme.2018.12.038 results in decreased glycogen accumulation, increased survival, and Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S25

23 challenges this has entailed. A retrospective review of all patients Crossing biological membranes using PTD4: Implications for included in early phase trials receiving drugs directly into the central treatment of MPS IIIB through enzyme replacement therapy nervous system, within one paediatric Clinical Research Facility was carried out. This looked at the number of overall sedation regimes used Rhea E. Ashmead, Chloe L. Christensen, Francis Y.M. Choy, University as well as the number of AEs and SAEs related to these. There have been of Victoria, Victoria, BC, Canada a total of 8 trials involving IT or ICV drugs including a total of 13 patients over a period of four years. These patients have had a total of 9 Mucopolysaccharidosis III type B (MPS IIIB) is a neuronopathic sedation regimes with one particularly challenging patient requiring 8 lysosomal storage disorder (LSD) that results from deleterious muta- different sedation regimes over a period of 18 months. One new drug, tions in the NAGLU gene. This results in the deficient activity of its dexmedatomidine, was introduced during this time. Delivering drugs encoding enzyme, alpha-N-acetylglucosaminidase (Naglu), and leads to directly into the central nervous system of children with neurodegen- the cytotoxic accumulation of heparan sulfate in the lysosome. MPS IIIB erative disorders is a complex area. However, with continued severely affects the central nervous system: clinical manifestations experience, and the introduction of new sedative agents it has been include developmental delays, behavioral issues, and cognitive deteri- possible to successfully dose all children enrolled on such trials. oration. Traditional approaches to treating LSDs, like enzyme replacement therapy (ERT), cannot effectively target and distribute within the CNS due to the restrictive nature of the blood-brain barrier (BBB). As a doi:10.1016/j.ymgme.2018.12.040 result, there is no effective treatment for MPS IIIB. Cell-penetrating peptides, like PTD4, present a potential approach to crossing the BBB for delivery of therapeutic agents. PTD4 is a synthetic protein transduction 25 domain, which has previously demonstrated the capability of facilitat- Hippo and necroptosis pathways are involved in cell growth ing the delivery of its fusion partners–including active enzymes–across defects in Gaucher disease the BBB. We have fused PTD4 to Naglu and expressed this recombinant fusion enzyme in Spodoptera frugiperda with expression levels of fi 1.01mg/litre. Naglu-PTD4 was purified to homogeneity using three- Annalisa Astol , Daria Messelodi, Silvia Strocchi, Salvatore N. step column chromatography with a yield of 26%. Preliminary uptake Bertuccio, Davide Leardini, Alberto Taddia, Annalisa Pession, Daniela studies with Naglu-PTD4 showed a detectable increase in active Naglu Grifoni, Andrea Pession, University of Bologna, Bologna, Italy within the fibroblasts: the activity of Naglu in control fibroblasts was β approximately 1.3 U/mg in MPS IIIB fibroblasts less than 0.3 U/mg and in Gaucher disease (GD) is a monogenic disorder characterized by - fi MPS IIIB fibroblasts treated with Naglu-PTD4 approximately 5.3 U/mg. glucocerebrosidase enzyme de ciency. Typical features of the disease fl Subsequent immunofluorescence uptake studies in MPS IIIB fibroblasts are the unrestrained activation of in ammatory mechanisms and will be conducted, along with blood-brain barrier penetration studies in neuronal cell death in neuronopathic conditions, whose molecular mice, to further examine the ability of PTD4 to deliver functional Naglu pathways are still not fully understood. Recent evidences show that fl across biological membranes, and more specifically the blood-brain upregulation of the Hippo pathway is associated with neuroin amma- barrier. If this approach to CNS delivery is successful, it has the potential tion and neuronal cell death. We analyzed the Hippo pathway activity to improve the effectiveness of ERT as a treatment for MPS IIIB. Funded taking advantage of a Drosophila Gaucher-like model characterized by KO by the Sanfilippo Children Research and Rare Disease Foundations dGBA1b knock-out (GBA1b ). We found deregulation of CycE, dIAP and MYC, direct targets of the pathway both in Drosophila and mammals, which were severely reduced at transcript and protein level, suggesting a doi:10.1016/j.ymgme.2018.12.039 cell growth impairment. Analyzing upstream components we found that Fat, an atypical cadherin upstream of the kinase complex, was upregulated. Moreover the glypicans Dally and Dally-like, known to be negatively regulated by Fat and involved in glial and synapse 24 development, were found downregulated in the GBA1bKO background, One centre’s experience of sedation regimes for ICV and IT drug supporting a role in neurological damage. On the other hand, the delivery in early phase paediatric clinical trials aberrant inflammatory condition of the hematological compartment was investigated in the human model of induced Pluripotent Stem Cells Helen Ashton, Jose Rodriguez Ciancio, Maureen Cleary, Paul Gissen, (iPSC), differentiated into hematopoietic progenitors. While GD iPSCs Anupam Chakrapani, Great Ormond street Hospital, London, United were able to differentiate into CD34+/CD43+/CD45+ progenitors, they Kingdom showed a decreased proliferative potential compared to healthy donor iPSC either in semisolid and liquid culture, therefore exhibiting a growth Delivering clinical trial treatments to children with neurodegener- impairment. This effect was coupled with the activation of necroptosis ative disorders has proven to be a challenging area with limited pathway with a significant upregulation of RIPK3 and MLKL both in the expertise. The need to administer drugs directly into the central pluripotent and differentiated state. Correction of the mutation to rescue nervous system requires insertion of permanent intracerebroventric- the phenotype is ongoing, taking advantage of AAVS1 editing approach ular, or intrathecal devices that must be accessed with a sterile in order to develop a fully comparable cell model useful in a cell therapy technique. During these procedures the cooperation of the patient is perspective. These data obtained with suitable GD models support the essential to minimize the risk of adverse events. The nature of the role of Hippo and necroptosis effectors in GD, amenable of further conditions being treated means the child may struggle to sit still and consideration as potential therapeutic targets. cooperate with the procedures. They may have a level of ability that makes traditional play therapy of limited benefit. This study looks at one centres experience of delivering clinical trial drugs via such doi:10.1016/j.ymgme.2018.12.041 devices, focusing on the varying sedation regimes trialled and the S26 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

26 with keratanase II after an extraction procedure with a NH4OH 0.01 M Clinical, biochemical and molecular characteristics of five pa- solution. The detection of two KS-related disaccharides/creatinine plus tients with late infantile neuronal ceroide lipofucsinosis type internal standards was performed using a UPLC-MS/MS (Xevo TQ-S (CLN2 disease) phenotype clasical and atypical Micro, Waters) 4 minute-run. The assay showed good intraday and interday accuracy and precision (BIAS and RSDs less than 15%). Urine Nora G. Atanacioa, Consuelo Duranda, Mercedes Villanuevaa, specimens collected on filter paper were stable for at least 6 weeks. In a Francisca Masllorensb, Joaquin Frabasila, Andrea Schenonea, Mariana test-trial, all Morquio syndrome type A patients presented abnormal Loosc, aLaboratorio de Neuroquimica Dr Chamoles, Buenos Aires, results pre-treatment compared to reference values. Urine samples Argentina, bHospital Nacional Prof Alejandro Posadas, Buenos Aires, dried on filter paper facilitate the collection by parents and low shipping Argentina, cHospital Nacional de Pediatria P. Garrahan, Buenos Aires, costs of samples by regular postal service to the screening laboratory. Argentina We plan to implement this approach for mass urine screening of 70 000 newborns as part of an evaluative research project in Quebec. Informed IIntroduction: CLN2 disease is a rare progressive disorder caused by a consent will be required from parents. This methodology will also deficiency of the lysosomal enzyme tripeptidyl peptidasa 1(TPP1). CLN2 facilitate monitoring/follow-up of treated patients. most commonly presents with seizures and/or ataxia in the late- infantile period (age 2-4), often in combination whit a history of language delay, followed by progressive childhood dementia, motor and doi:10.1016/j.ymgme.2018.12.043 visual deterioration, and early death. Atypical phenotypes are characterized by later onset and in some instances longer life expectancies. Materials and methods: A retrospective medical record review was 28 conducted at our center for this case series of 5 CLN2 patients diagnosed Early-onset of symptoms and clinical course of Pompe disease between years 2016 and 2018 at Laboratorio de Neuroquímica Dr. N.A. associated with the c.-32-13TNG variant Chamoles. We analyzed the clinical, biochemical and molecular findings. Results: 3 patients presented with the classical phenotype and two patients with the atypical phenotype. We detected low TPP1 Stephanie Austin, Mrudu Herbert, Laura Case, Mugdha Rairikar, Heidi activity in dry blood spots in 5 patients and confirm it in leukocytes in 4 Cope, Lauren Flueckinger, Priya Kishnani, Duke University, Durham, patients. All 5 patients have molecular results confirming the diagnosis. NC, United States Two patients are receiving intraverebroventricular enzyme replacement therapy and outcome has been favourable so far. Discussion: CLN2 Individuals with late-onset Pompe disease (LOPD) and the common N is a rapidly neurodegenerative disorder, Age at onset and clinical c.-32-13T G variant are widely thought to have milder, adult onset features distinguishes typical from atypical forms of CLN2 disease. We disease. Previous reports of LOPD in children do not include description present 2 atypical patients from a total of 5 patients. We think it is very of the early-onset phenotype. This description of subtle signs and fi important to be aware of these atypical and less common forms of the symptoms is important to facilitate prompt identi cation and appropri- disease to ensure early diagnosis, optimal patient care, prompt ate treatment in symptomatic children. Retrospective chart review of a N treatment, multidisciplinary planning and family support. cohort of 84 LOPD patients with the c.-32-13T G variant was conducted to identify patients diagnosed clinically (as opposed to through newborn screening) who had clinically documented symptom-onset within the doi:10.1016/j.ymgme.2018.12.042 first two years of life. Four patients had early onset of symptoms, with age at onset ranging from 10 days to 20 months. Initial symptoms included gross motor delay, swallow/feeding difficulties, and sleep 27 apnea. Early deviations in skeletal muscle posture and movement where Newborn mass urine screening for Morquio syndrome type A identified in all subjects. Age at diagnosis ranged from 10 months to 26 patients using an innovative UPLC-MS/MS approach months. Median age at enzyme replacement therapy (ERT) initiation was 23.5 months. Despite ERT, progression of musculoskeletal involve- Christiane Auray-Blais, Bruno Maranda, Pamela Lavoie, Université de ment and residual muscle weakness was evident in all patients, as Sherbrooke, Sherbrooke, QC, Canada evidenced by ptosis, myopathic facies, scoliosis, lumbar lordosis, scapular winging, and trunk/lower extremity weakness. Standardized Morquio syndrome type A is an autosomal recessive lysosomal functional assessments showed gross motor function below age level as storage disorder caused by a defect of the enzyme N- measured by the Alberta Infant Motor Scales, the Peabody Developmen- acetylgalactosamine-6-sulfatase, which is involved in the catabolism tal Motor Scales-2, the Bruininks-Oseretsky Test of Motor Proficiency, of keratan sulfate (KS). This panethnic disease has a high prevalence in Second Edition, and the six minute walk test. Onset of symptoms the Province of Quebec, Canada. Enzyme replacement therapy treat- including gross-motor delay, swallow/feeding difficulties, and sleep ment has been approved for Canadian patients since 2014. Early apnea in the first two years of life is not uncommon in individuals with detection and efficient monitoring of patients under therapy might lead LOPD and the c.-32-13TNG variant. Careful evaluation for specificgross to better outcomes. Currently, quantitative KS assays are not widely motor posture and movement in patients with this variant is necessary available in biochemical genetics laboratories. The aim of this study was to identify those who have early onset of disease. Increased awareness of to devise and validate a multiplex assay for the quantitation of KS the early-onset signs and symptoms will enable early identification of normalized to creatinine using dried urine filter paper sample to disease onset in children who are diagnosed through newborn eventually perform a feasibility study for mass urine screening and early screening. detection of Morquio syndrome type A patients at 21 days of age. The infrastructure for the Mass Urine Screening Program is already established for early detection of other inborn errors of metabolism in doi:10.1016/j.ymgme.2018.12.044 Sherbrooke, QC. The KS method involves a 3-hour enzymatic digestion Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S27

29 enrolled in 2013. Presented are baseline findings for patients enrolled Development of scAAV9/SUMF1 gene therapy for multiple sulfa- through August 28, 2018 baseline is the date of the first dose of tase deficiency sebelipase alfa for ever-treated patients or enrollment date for never- treated patients. Of 149 patients enrolled, 38 were excluded from Rachel M. Baileya, Maximiliano Presab, Catherine Lutzb, Steven J. analysis (reasons: LAL-D carrier, deceased, unconfirmed LAL-D diagno- Graya, aUniversity of Texas Southwestern Medical Center, Dallas, TX, sis, treatment start date unknown) 111 were included in the analysis. United States, bThe Jackson Laboratory, Bar Harbor, ME, United States Data were available on LAL enzyme testing for 108 patients (95% were tested), genetic testing for 96 patients (91% were tested), age at LAL-D Multiple Sulfatase Deficiency (MSD OMIM #272200) is a rare, onset for 90%, medical history for 93%, and abdominal imaging for 68%. autosomal recessive pediatric lysosomal storage disease. Currently no Baseline serum aminotransferase and lipid values were available for specific treatments exist for this disorder and patients have progres- ≥65% and ≥57% of patients, respectively (≥76% and ≥78%, respectively, sive neurologic dysfunction with multisystem involvement that is among adults). Of the 111 patients, 56% were b18 years of age, 54% male, eventually fatal. The disease pathology is due to homozygous loss-of- and 86% white 5 patients (4.5%) were aged b6 months at diagnosis. function mutations in the SUMF1 gene. The SUMF1 gene encodes Median age at clinical onset was 5.3 years (range, 0.0-41.3) and median formylglycine-generating enzyme, which is required for post-transla- age at diagnosis was 10.6 years (range, -0.1 to 53.6). LAL-D manifesta- tional modification and activation of sulfatase enzymes. As such, tions were reported in 103 patients 65% had hepatomegaly, ≥60% had pathogenic mutations in the SUMF1 gene impact the function of all 17 elevated serum aminotransferase levels, 42% had high LDL-cholesterol human sulfatase enzymes, leading to a broad phenotypic spectrum levels, and 28% had low HDL-cholesterol levels. Of 43 patients with that overlaps with known inherited sulfatase disorders. Gene therapy available liver biopsy, 31 (72%) had microvesicular steatosis and 20 may provide a meaningful and long term therapeutic benefit for MSD (47%) had lobular inflammation. Nine (8%) had cirrhosis (determined patients by delivering a functional copy of the SUMF1 gene to patient by clinical history, abdominal imaging, and/or liver biopsy). Registry cells. We have developed a novel self-complementary vector encoding data of N100 LAL-D patients demonstrate diverse clinical manifestations a codon-optimized human SUMF1 gene (scAAV9/SUMF1), the unal- of the disease and a significant diagnostic delay, particularly in adults. tered design of which could be appropriate for human use. Proof-of- Funded by Alexion Pharmaceuticals, Inc. concept studies were conducted in a SUMF1 knock-out mouse model using intracerebroventricular (ICV) delivery in neonates. ICV-delivered scAAV9/SUMF1 significantly improved survival and improved doi:10.1016/j.ymgme.2018.12.046 gross motor function as assessed by open field and accelerating rotarod tests. Additionally, the safety profile of the MSD vector has been excellent with no adverse effects noted even in treated normal 31 mice. While encouraging, the translation of this approach in humans is Outcomes of 19 unplanned pregnancies in women participating complicated by the reduced relative biodistribution of the vector after in phase 2 or 3 eliglustat clinical trials and 18 pregnancies in the ICV administration when moving from a neonatal rodent to human partners of men who participated in these trials brain, and it is unknown what effect will be achieved in older animals. To test the boundaries and limitations of a gene therapy for MSD, we a b c d have intrathecally-injected scAAV9/SUMF1 in juvenile SUMF1 knock- Manisha Balwani , Elena Lukina , Nadia Belmatoug , Nora Watman , e f f out mice and studies are ongoing to assess the therapeutic benefit. Derralynn Hughes , Sebastiaan J.M. Gaemers , Yestle Kim , Mererdith f f a Overall, our preclinical results attest to the safety of scAAV9/SUMF1 Foster , M. Judith Peterschmitt , Icahn School of Medicine at Mount b delivery and predict a benefit of this treatment to MSD patients. Sinai, New York, NY, United States, National Research Center for Hematology, Moscow, Russian Federation, cUniversity Hospital Paris Nord Val de Seine, Paris, France, dHospital Ramos Mejia, Buenos Aires, e doi:10.1016/j.ymgme.2018.12.045 Argentina, Royal Free London NHS Foundation Trust, University College London, London, United Kingdom, fSanofi Genzyme, Cambridge, MA, United States

30 Gaucher disease type 1 (GD1) is an inherited lysosomal storage Clinical manifestations of LAL-D: The international lysosomal acid disorder caused by deficient enzymatic activity of acid β-glucosidase, lipase deficiency registry resulting in accumulation of its substrate glucosylceramide, leading to debilitating visceral, hematologic, and skeletal manifestations. Women Manisha Balwania, William Balistrerib, Lorenzo D'Antigac, Shona with GD1 are at increased risk for complications during pregnancy, Fangd, Simon A. Jonese, Emilio Rosf, Florian Abeld, Don P. Wilsong, delivery, and postpartum. Treatment with enzyme replacement therapy aIcahn School of Medicine at Mount Sinai Hospital, New York, NY, (ERT) is generally recommended before and during pregnancy to United States, bCincinnati Children’s Hospital Medical Center, Cincinnati, reduce risks. Eliglustat, an oral substrate reduction therapy, is a first-line OH, United States, cAzienda Ospedaliera Papa Giovanni XXIII, Bergamo, treatment for adult GD1 patients who have extensive, intermediate or Italy, dAlexion Pharmaceuticals, Inc., Boston, MA, United States, poor CYP2D6 metabolizer phenotypes (N90% of patients). Eliglustat is eManchester University Hospital NHS Trust, Manchester, United King- not recommended in pregnancy due to limited safety data. As women of dom, fHospital Clinic Barcelona, Barcelona, Spain, gCook Children’s childbearing age are an important subgroup of GD1, there is interest in Medical Center, Fort Worth, TX, United States pregnancy outcomes of women with short-term eliglustat exposure during unplanned pregnancies in clinical trials. In 4 eliglustat trials An international registry (NCT01633489) was established to better (Sanofi Genzyme) of 393 patients (202 women) with GD1 (1400 understand the natural history of lysosomal acid lipase deficiency (LAL- patient-years of exposure), women of childbearing potential were D), evaluate long-term effectiveness of treatments, and improve patient asked to use contraception and have monthly pregnancy tests. Pregnant care through evidence-based management. The first patient was women had to discontinue eliglustat promptly to limit exposure S28 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 duration. Among 145 women of childbearing age (16 to b45 years), 18 33 women had 19 pregnancies, resulting in 14 healthy infants from 13 Is there any difference in GBA1 allele frequencies depending on pregnancies (1 set of twins), 3 elective terminations, 1 spontaneous the region of Brazil? abortion, 1 tubal pregnancy, and 1 in-utero death in a previously untreated patient with severe GD1. Median time of eliglustat exposure Suelen P. Basgaluppa, Vivian Altmanna, Filippo P. Vairob, Ida V.D. during pregnancy was 38.5 days based on known date of last menstrual Schwartza, Marina Siebertc, aUniversidade Federal do Rio Grande do period. Most patients (14/18) received eliglustat after prior ERT, 4/18 Sul, Porto Alegre, Brazil, bMayo Clinic, Rochester, MN, United States, were splenectomized, and 12/18 had a genotype with an L444P allele. cHospital de Clínicas de Porto Alegre, Porto Alegre, Brazil Among partners of 16 eliglustat-treated GD1 men, there were 18 pregnancies, all resulting in healthy infants. As eliglustat is not Gaucher disease (GD) is an autosomal recessive lysosomal recommended in pregnancy, pregnant women should immediately disorder caused by pathogenic variants in GBA1 which results in discontinue eliglustat and be monitored closely to better understand glucocerebrosidase deficiency. Up to date, over 500 pathogenic the potential impact of drug exposure. variants have been reported. Given the extensive Brazilian territory, and because the interethnic ancestral crosses between Amerindians, Europeans, Jews and Africans were different depending on its doi:10.1016/j.ymgme.2018.12.047 regions, we hypothesize the most prevalent GD causing mutations vary throughout Brazil. The aim of this study was to compare the GBA1 variants frequencies in GD unrelated patients from South Brazil (SB, the region with the highest European ancestry) to patients from 32 different regions of Brazil (others). Thirty-seven GD patients from SB, Migalastat reduces globotriaosylceramide (GL-3) inclusions in and 21 GD patients from the other regions were included. DNA was renal peritubular capillaries in patients with Fabry disease and extracted from EDTA peripheral blood samples, and coding regions migalastat-amenable mutations: Post hoc analyses from FACETS as well as exon/intron junctions of the GBA1, were sequenced by Sanger and/or next-generation sequencing (Ion Torrent PGM). a b c Laura M. Barisoni , J. Charles Jennette , Nina Skuban , Jeffery P. Twenty-five different pathogenic mutations were found in the whole c d a Castelli , Robert B. Colvin , University of Miami, Miller School of sample (SB=15/74, 20% others=14/42, 33% being 4 in common). b Medicine, Miami, FL, United States, School of Medicine, University of The most common variant was N370S in SB and others (SB=31/74, c North Carolina at Chapel Hill, Chapel Hill, NC, United States, Amicus 42% others=18/42, 43% P=0.92). The second most frequent variants d Therapeutics, Inc., Cranbury, NJ, United States, Harvard Medical School, were RecNciI (SB= 14/74, 19% others=4/42, 9.5%) and L444P Massachusetts General Hospital, Boston, MA, United States (SB=14/74, 19% others=4/42, 9.5%), both with equal frequency in SB and in other regions. In SB, the 3 most frequent alleles α α Fabry disease is a rare, X-linked disorder of -galactosidase A ( -Gal corresponded to 80% (59/74) of total alleles. However, when fi A) de ciency caused by mutations in the GLA gene, leading to observing the 3 most frequent alleles in other regions, it only accumulation of the substrate globotriaosylceramide (GL-3) in many corresponded to 60% (23/42). One novel mutation was described (p. cell types, including endothelial cells of renal peritubular capillaries Gln109Argfs*9 found in a patient from Northeast). The most frequent (PTC also known as kidney interstitial capillaries [KIC]), and multi-organ genotype found in Brazil was N370S/RecNciI (SB=11/37, 29.7% fi damage. Migalastat is a rst-in-class, small molecule, oral pharmaco- others=4/21, 19%). These results suggest that there is variability in logical chaperone approved for the treatment of adults with Fabry GBA1 allele frequencies between Brazilian regions, being SB more disease and migalastat-amenable GLA mutations, as determined by an homogeneous. This could be explained by the immigration process in fi in vitro migalastat amenability assay. The primary ef cacy endpoint of which resulted in ancestry diversity between regions. Haplotype FACETS (NCT00925301), a phase 3, randomized, placebo-controlled analysis will be performed. trial of migalastat in enzyme replacement therapy-naive patients, was the average number of GL-3 inclusions per PTC, as assessed by applying BLISS, a quantitative scoring system, on whole slide images of one- doi:10.1016/j.ymgme.2018.12.049 micron plastic-embedded sections of renal cortex stained with toluidine blue. Here, we present a post hoc analysis of FACETS to evaluate the efficacy of migalastat in reducing the number of GL-3 inclusions in PTC in 17/45 patients with amenable GLA variants and higher baseline levels 34 of PTC GL-3 (≥0.3 GL-3 inclusions per PTC). After 6 months of Head circumference in individuals with MPS I compared to CDC randomized treatment, those who received migalastat (n=9) had a standard charts mean reduction of -0.88 in PTC GL-3 inclusions from baseline compared with a mean increase of 0.22 in patients who received placebo (n=8). Luisa Baya, Lorne A. Clarkeb, Nathalie Guffonc, Hillary Keenand, Joseph Differences in median change in PTC GL-3 inclusions from baseline also Muenzere, Caroline Prunetd, David H. Viskochilf, aHospital de Pediatría favored migalastat versus placebo (-0.91 versus -0.02). Seven of the 9 SAMIC Prof. Dr. Juan Pedro Garrahan, Buenos Aires, Argentina, bBritish migalastat-treated patients (78%) achieved a ≥50% reduction in PTC GL- Columbia Children's Hospital Research Institute, University of British 3 inclusions (responders) versus 2/8 of placebo-treated patients (25%). Columbia, Vancouver, BC, Canada, cCentre de Référence des Maladies These data support the therapeutic efficacy of migalastat in reducing GL- Héréditaires du Métabolisme, Hôpital Femme Mère Enfant, Lyon, France, 3 inclusions in PTCs and demonstrate substantial effects in patients with dSanofi Genzyme, Cambridge, MA, United States, eUniversity of North more severe disease at baseline. Carolina, Chapel Hill, NC, United States, fUniversity of Utah School of Medicine, Salt Lake City, UT, United States doi:10.1016/j.ymgme.2018.12.048 The natural history patterns of growth in individuals with MPS I have not been well-described. To better utilize these parameters in Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S29 patient care, head circumference (HC) data from untreated patients IgG (rho=0.66, p=0.0004), IgM (rho=0.85, p=b0.0001). sMR from the MPS I Registry were compared to CDC standard charts. correlated well with other biomarkers: chitotriosidase (rho=0.47, Longitudinal HC measurements from patients aged 0-36 months p=0.017), PARC (rho=0.66, p=0.0058), osteoactivin (rho=0.57, from the MPS I Registry were stratified by phenotype and sex and p=0.019), and MIP-1alpha (rho=0.66, p=0.005). sMR is a potential compared to CDC standards. All observations were prior to treatment new biomarker for GD and may be useful in predicting plasma cell with ERT or HSCT and included recipients of ventriculoperitoneal dyscrasiae. shunts. Data were available from 450 individuals with MPS I, 0-36 months old (severe: 205 male, 198 female attenuated: 23 male, 24 female) the majority of patients were from North America and doi:10.1016/j.ymgme.2018.12.051 Europe (97.5% of severe 93.6% of attenuated). In each phenotype, males and females showed similar trends. In patients with severe disease, HC measurements were below the CDC 95 percentile in the first 3-5 months, however, by 10 months, 50% of observations were 36 greater than the CDC 97 percentile. There were few observations for Analysis of the effect of intravenous gene therapy on brain and attenuated patients. Yet, the estimated median HC for these patients peripheral disease in a feline model of GM1 gangliosidosis were also higher than the respective CDC medians, though to a lesser extent. These data indicate that both severe and attenuated MPS I Cassie Bebout, Auburn University, Auburn, AL, United States patients have HC patterns that differ from unaffected norms. Patients with severe disease show earlier and more substantial deviation than GM1 gangliosidosis is a neurodegenerative lysosomal storage patients with attenuated MPS I. Additional observations in patients disease caused by a mutation in the GLB1 gene, resulting in a fi β with attenuated disease could be useful in defining the role of HC in de ciency of -galactosidase. In a feline model of GM1 differential phenotype assignment. Supported by Sanofi Genzyme. gangliosidosis, intravenous injection of adeno-associated viral (AAV) vectors containing a corrected GLB1 gene have proven efficacious in slowing and even halting disease progression. This doi:10.1016/j.ymgme.2018.12.050 project seeks to quantify and analyze accumulation of glycosaminoglycans (GAGs) in the urine and brain of cats treated by intravenous gene therapy, and to compare this data with that of unaffected cats and untreated GM1 cats. Previous data displays that treated cats 35 experience a decrease in urinary GAG levels to at or near normal Soluble mannose receptor is a potential new biomarker for levels, which also significantly differ from untreated cats’ urinary Gaucher disease GAG levels. Additionally, the project aims to examine any correlation between accumulation of GAGs in the brain and accumulation of Brendan Beaton, Uma Ramaswami, Atul Mehta, Derralynn Hughes, GAGs in the urine using standard colorimetric assays, isolation Royal Free Hospital NHS Foundation Trust, London, United Kingdom protocols, and methods of statistical data analysis. Strong correlation between brain and urinary GAG levels will support the validity of Gaucher Disease (GD) is an autosomal recessive lysosomal this assay as a biomarker of disease progression, and as a gauge of storage disorder caused by deficient activity of beta- treatment efficacy. Urinary data thus far has bolstered the claim that glucocerebrocidase (GBA). Clinical features of disease are heteroge- treated cats have significantly decreased storage of GAGs, but only neous as is response to treatment. Skeletal disease is complex and a after a long period of treatment, as urine collected from cats proportion of patients do not respond or progress despite Gaucher- approximately four months post-treatment contained high levels of specific therapy. Determinants of skeletal response are not known. GAGs. This finding could be used to more accurately assess drug Polyclonal gammopathy and monoclonal gammopathy of undeter- efficacy over time. Brain GAG analysis is currently underway, and mined significance (MGUS) are features of GD. Progression to these findings will provide a novel biomarker of enzymatic activity. myeloma is difficult to predict. Biomarkers currently used to assess overall disease burden are not specific to GD and do not reliably reflect individual domains of organ involvement. Soluble mannose doi:10.1016/j.ymgme.2018.12.052 receptor (sMR) has been assessed as a biomarker in various diseases where alternatively activated macrophages have been implicated, and shows promise in predicting transformation of MGUS to myeloma. It has not been assessed in GD to date. In this study, 16 37 treatment naïve type 1 GD patients, 14 with paired post-treatment Retinal thinning in Gaucher patients as a predictive test of sera and 10 non-GD controls had sMR measured by ELISA. Male: developing Parkinson disease Female 12:4, GBA mutations: 5 N370S/N370S, 11 N370S/other, 1 a a a other/other, median baseline age was 38.25 years (range: 20.8-80.4), Michal Becker Cohen , Shoshana Revel-Vilk , Ari Zimran , Tama a b a a median treatment time was 27 months (range: 11-124). Mean sMR Dinur , David Arkadir , Yishay Weill , Shaare Zedek Medical Centre, b in treatment naïve sera was 291.2 +/- 178.8 ng/ml compared to Jerusalem, Israel, Hebrew University-Hadassah Medical School, Jerusa- post-treatment 200.5 +/- 103.1 ng/ml (p = 0.01). Mean sMR of the lem, Israel control group was 182.1 +/- 48.0 ng/ml. There is good correlation with all domains of GD activity: platelet count (rho=0.56, Gaucher disease (GD) and Parkinson disease (PD) have been p=0.0004), haemoglobin (rho=0.64, p=0.0005), spleen volume known to be linked by mutations in the gene GBA. Gaucher disease (rho=0.58, p=0.011), liver volume (rho=0.43, p=0.05), bone patients and carriers are at increased risk of developing PD. Previous marrow burden score (rho=0.58, p=0.007). There was strong anecdotal publications reported thinning of the ganglion cell correlation with immunoglobulin levels: IgA (rho=0.5, p=0.012), complex (GCC), measured by optical coherence tomography (OCT), S30 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 in a small subgroup of GD patients and GBA carriers at an early stage that could potentially be used to treat MPS I-H patients who carry a of PD. This preliminary finding may serve as a biomarker of increased PTC. risk of developing PD. Diagnostic tests for the pre-motor prodromal stage of PD can assist in developing preventive treatments in the early stages of PD, preventing the neurological damage and typical doi:10.1016/j.ymgme.2018.12.054 motor symptoms. This research includes GD patients, GBA carriers and controls. Patients and controls with a neurodegenerative disease, glaucoma, high myopia (-6.0N diopter), younger than 40 or older than 75 years of age, were excluded from the research. OCT was 39 fi fi performed on all patients using swept-source domain OCT (DRI OCT- Integrative metabolic pro ling in San lippo syndrome 1 Atlantis Topcon, Tokyo, Japan). Retinal thickness and macula a a b changes data have been obtained, these will be further analysed for Soumeya Bekri , Lenaig Abily-Donval , Carlos Afonso , Stéphane a a a comparing the three groups. The results will ascertain whether Marret , Abdellah Tebani , Rouen University Hospital, Rouen, France, b retinal GCC thinning can be used as a predictive test for the Rouen University, Rouen, France prodromal phase of PD in these cohorts. Additionally, using OCT, we will assess potential macular and optic nerve changes that have Background: Metabolomics represent a valuable tool to recover fl heretofore have not been described in these unique populations. Our biological information using body uids and may help to characterize preliminary results show that the retinal nerve fiber layer in GD pathophysiological mechanisms of the studied disease. This approach patients is significantly thinner than in the matched control group. has not been widely used to explore inherited metabolic diseases. The study is ongoing, part of a larger screening to identify GBA This study investigates mucopolysaccharidosis type III (MPS III). A carriers at a significantly higher risk of PD, hence granting a research thorough and holistic understanding of metabolic remodeling in MPS window for PD preventative treatment. III may allow the development, improvement and personalization of patient care. Patient and Methods: We applied both targeted and untargeted metabolomics to urine samples obtained from a French cohort of 49 doi:10.1016/j.ymgme.2018.12.053 patients, consisting of 13 MPS IIIA, 16 MPS IIIB, 13 MPS IIIC, and 7 MPS IIID, along with 66 controls. The analytical strategy is based on ultra-high-performance liquid chromatography combined with ion 38 mobility and high-resolution mass spectrometry. Twenty-four Triamterene normalizes glycosaminoglycan accumulation in an amino acids have been assessed using tandem mass spectrometry IDUA-W402X mouse model of MPS I (Hurler syndrome) via combined with liquid chromatography. Multivariate data modeling nonsense suppression has been used for discriminant metabolite selection. Pathway analysis has been performed to retrieve metabolic pathways David Bedwella, Amna Siddiquia, Halil Dundarb, Josh Echolsa, Ming impairments. Dua, Lynn Rasmussenc, J. Robert Bostwickc, Mark Sutoc, Kim Keelinga, Results: Data analysis revealed distinct biochemical profiles. aUniversity of Alabama at Birmingham, Birmingham, AL, United States, These metabolic patterns, particularly those related to the amino bGazi University, Ankara, Turkey, cSouthern Research, Birmingham, AL, acid metabolisms, allowed the different studied groups to be United States distinguished. Pathway analysis unveiled major amino acid pathways impairments in MPS III mainly arginine–proline metabolism and Nonsense mutations introduce a premature termination codon urea cycle metabolism. (PTC) into an mRNA, which terminates translation prematurely, Conclusion: This represents one of the first metabolomics-based resulting in a truncated, non-functional protein. A PTC can also investigations of MPS III. These results may shed light on MPS III trigger nonsense-mediated mRNA decay (NMD), a pathway that pathophysiology and could help to set more targeted studies to infer degrades PTC-containing mRNAs, preventing their translation. In this the biomarkers of the affected pathways, which is crucial for rare study, a novel NanoLuc-based reporter was used to screen a library conditions such as MPS III. of 10,000 low molecular weight molecules for those that overcome the effects of PTCs by: 1) suppressing PTC termination, 2) inhibiting doi:10.1016/j.ymgme.2018.12.055 NMD, or 3) targeting both termination and NMD simultaneously. Triamterene, an FDA-approved, antihypertensive diuretic, was identified as a hit from this screen. Subsequent testing in mouse embryonic fibroblasts (MEFs) derived from Idua-W402X mice, a 40 model of mucopolysaccharidosis I-Hurler (MPS I-H), showed tri- Next generation sequencing sheds light on inherited metabolic amterene elevates α-L-iduronidase activity to levels that normalize diseases in nonimmune hydrops fetalis investigations glycosaminoglycan (GAG) storage. Our evidence suggests that this newly discovered function of triamterene operates through PTC- Soumeya Bekri, Bénédicte Sudrié-Arnaud, Abdellah Tebani, Florent dependent mechanisms that are unaffected by its ability to alter Marguet, Annie Laquerriere, Stéphane Marret, Rouen University epithelial sodium channel activity, the target of triamterene’s Hospital, Rouen, France diuretic function. Once daily oral administration of triamterene to Idua-W402X mice for two weeks showed a dose-dependent decrease Background: Hydrops fetalis is a life-threatening fetal condition, in GAG storage, with a dose of 120 mg/kg normalizing GAG levels in and 85% of all cases are classified as nonimmune hydrops fetalis all tissues examined, including tissues that are recalcitrant to (NIHF). Up to 15% of NIHF cases may be due to inborn errors of current MPS I-H therapies such as the brain, bone, and heart. These metabolism (IEM), but a large proportion of cases linked to data suggest that triamterene is a novel PTC suppression compound metabolic disorders remains undiagnosed. This lack of diagnosis Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S31 may be related to the limitations of conventional biological Conclusion: This is the largest study of nerve conduction velocity procedures, which involve sequential investigations and require in patients with MLD that was performed at one site using a multiple samples and steps. In addition, this approach is time standardized evaluation. Nerve conduction tests can be used as early consuming. We have developed a next-generation sequencing (NGS) markers of disease progression. The results demonstrate disease panel to investigate metabolic causes of NIHF, ascites, and poly- burden and can be used as a comparator for quantitating effect of hydramnios associated to another fetal abnormality. therapies such as hematopoietic stem cell transplantation, gene Patients and Methods: The hydrops fetalis (HydFet) panel was therapy, enzyme replacement and any other future therapies. designed to cover the coding regions and flanking intronic sequences of 41 genes. A retrospective study of amniotic fluid doi:10.1016/j.ymgme.2018.12.057 samples from 40 subjects was conducted. A prospective study was subsequently initiated, and six samples were analyzed using the NGS panel. Results: Five IEM diagnoses were made using the HydFet panel 42 (Niemann-Pick type C (NPC), Barth syndrome, HNF1Β deficiency, Effect of long-term migalastat treatment on plasma glo- GM1 gangliosidosis, and Gaucher disease). This analysis also botriaosylsphingosine (lyso-Gb3) levels in patients with Fabry allowed the identification of 8p sequence triplication in an disease previously treated with enzyme replacement therapy: additional case. Results from ATTRACT and open-label extension studies Conclusion: NGS combined with robust bioinformatics analyses a b b b a is a useful tool for identifying the causative variants of NIHF. Daniel Bichet , Jay A. Barth , Hadis Williams , Nina Skuban , Hôpital b Subsequent functional characterization of the protein encoded by du Sacré-Coeur, University of Montréal, Montreal, QC, Canada, Amicus the altered gene and morphological studies may confirm the Therapeutics, Inc, Cranbury, NJ, United States diagnosis. This paradigm shift allows a significant improvement of IEM diagnosis in NIHF. A national French study is ongoing to assess Fabry disease is associated with pathological accumulation of the benefit of such strategies. substrates, including globotriasosylceramide and globotriaosylsphingosine (lyso-Gb3). Although the clinical relevance of doi:10.1016/j.ymgme.2018.12.056 lyso-Gb3 as a biomarker for monitoring treatment effect in Fabry remains uncertain, it is widely used as diagnostic indicator of Fabry

disease. We examined lyso-Gb3 profiles in patients with migalastat- amenable GLA mutations in ATTRACT (NCT01218659), a randomized, 41 open-label study where enzyme replacement therapy (ERT)-experi- Nerve conduction studies as a tool in early detection of enced patients received 18 months of migalastat or ERT, followed by metachromatic leukodystrophy a 12-month open-label extension of migalastat efficacy and safety was further evaluated in a long-term open-label extension study a b b Maria L. Beltran-Quintero , Shilpa Haldal , Vince Carson , Hoda (AT1001-042, NCT02194985). Plasma lyso-Gb3 levels were measured Abdel-Hamida, Michele Poea, Maria L. Escolara, aUniversity of at baseline and at various subsequent timepoints. At baseline, Pittsburgh, Pittsburgh, PA, United States, bUPMC Children's Hospital of defined as the start of migalastat treatment in this analysis

Pittsburgh, Pittsburgh, PA, United States combining both randomized groups, the median lyso-Gb3 level was lower in females (5.8 nmol/L min, max: 0.8, 14.7 [n=26]) than in

Background: Metachromatic leukodystrophy (MLD) is an males (10.5 nmol/L min, max: 0.8, 59.1 [n=18]). Median lyso-Gb3 inherited lysosomal storage disease caused by deficient activity of levels remained low and stable with long-term migalastat treatment arylsulfatase A, an enzyme involved in the degradation of sulfatides. in males and females median changes from baseline to month 60 In MLD, the accumulation of sulfatide in myelin-producing cells leads were -2.5 nmol/L in males (n=4) and 0.4 nmol/L in females (n=6). to progressive demyelination and neurodegeneration in the periph- The lyso-Gb3 quantification assay has an assay-to-assay variability of eral and central nervous systems. MLD has a variable clinical course ~20%, which may have contributed to visit-to-visit changes. These that is classified into subtypes based on age of onset Late-infantile data validate primary trial results, but given the uncertainty in the

MLD is the most common, which presents between 1-2 years of age origin of plasma lyso-Gb3, and differences in the biodistribution and with difficulty walking and progresses to death by 5-6 years of age. mechanism of action among the current treatment options, plasma Objective: The purpose of this study is to examine the natural lyso-Gb3 should not be interpreted as a stand-alone biomarker and progression of peripheral neuropathy in patients with MLD. clinical outcomes should be considered when assessing treatment Methods: 46 patients with late-infantile (27), juvenile (13), or efficacy. Further research is warranted. adult (6) onset were evaluated from 2011 to 2018. Nerve conduction studies were all performed according to a predetermined protocol at ’ UPMC Children s Hospital of Pittsburgh. Simultaneous developmental doi:10.1016/j.ymgme.2018.12.058 assessments included growth parameters, standardized evaluations of adaptive and cognitive behavior, motor function, and speech/ language skills. All research was conducted under an approved IRB (PRO). 43 Results: There were 78 nerve conduction studies performed on Brain targeted stem cell gene therapy provides long-term the 46 MLD patients. The patients ranged from 3.5 months to 42 correction of mucopolysaccharidosis type II years at the time of testing. The sensory sural nerve velocity was the first to show abnormalities with all cases testing. No-response could Brian Bigger, Ai-Yin Liao, Helene Gleitz, Rebecca Holley, University of be detected in the 6 youngest Late-Infantile cases. Manchester, Manchester, United Kingdom S32 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Mucopolysaccharidosis type II (MPS II) is a lysosomal storage transfection of Chinese hamster ovary (CHO) cells. The identity of the disease caused by a mutation in the Ids gene, resulting in accumulation CHO derived HIRMAb-HEXA fusion protein (AGT-192) was con- of heparan and dermatan sulfate. This leads to skeletal and cardiore- firmed by hIgG1 and HEXA Western blot analyses and the purity by spiratory disease with severe neurodegeneration in two thirds of SDS-PAGE and SEC-HPLC. Fusion protein binding to the HIR was suffers. Enzyme replacement therapy is ineffective at treating saturable with an ED50 in the low nM range and comparable to the neurological disease, as intravenously-delivered IDS is unable to cross binding to the parental HIRMAb. The HEXA activity in the HIRMAb- the blood-brain barrier. Haematopoietic stem cell transplant is also HEXA fusion protein was high and comparable to recombinant HEXA. unsuccessful, presumably due to insufficient IDS enzyme production The brain uptake of HIRMAb-lysosomal enzymes in non-human from cells engrafting in the brain. We developed brain targeted primates approximates 1% of injected dose per brain. This level of hematopoietic stem cell gene therapy (HSCGT) in MPS II mice over brain uptake is able to produce therapeutic levels of HEXA enzyme both medium and long-term outcomes using a lentivirus containing activity in the brain. Successful development of a BBB penetrating IDS fused to ApoEII (LV.IDS.ApoEII) and compared to a lentivirus HIRMAb-HEXA fusion protein may be transformational for the expressing unmodified IDS (LV.IDS). As a control, a third group of MPS treatment of the brain in Tay Sachs Disease. II mice also received standard HSCT using wild-type cells. We previously reported on medium term outcomes (6 months post- transplant) and now present our findings from long-term outcomes, doi:10.1016/j.ymgme.2018.12.060 (12-14 months post-transplant). All transplant strategies resulted in significant correction of peripheral disease with enzyme levels at or above normal levels and HS and DS levels normalised in the liver. However, normalisation of brain pathology including astrocytosis and 45 microgliosis was only seen in the LV.IDS.ApoEII group, providing Against all odds: enzyme replacement therapy in non-ambulatory significantly enhanced correction compared to LV.IDS. Wild-type HSCT and ambulatory Morquio syndrome type A patients resulted in little brain IDS activity with corresponding limited correction of pathology. Interestingly, measured brain enzyme levels Pedro Paulo F. Bozzo, Amanda C. Monteiro, Daniely P. Camara, Larissa were significantly higher in LV.IDS animals compared to LV.IDS.ApoEII F. Bissoli, Charles M. Lourenco, Centro Universitario Estacio de Ribeirao despite poorer neurological outcomes in the LV.IDS group. In vitro Preto, Ribeirao Preto, Brazil. results suggested that IDS.ApoEII was more readily taken up and transcytosed more efficiently across brain endothelia than IDS, via Introduction: Mucopolysaccharidosis IVA (MPS IVA Morquio heparan sulfate/ApoE-dependent receptors and mannose-6-phos- syndrome type A) is an autosomal recessive lysosomal storage fi phate receptors. In conclusion we have shown both medium and disorder (LSD) caused by de ciency of N-acetylgalactosamine-6- fi long-term benefits from brain targeted HSCGT in MPS II mice, sulfate sulfatase (GALNS). De ciency of the enzyme leads to a demonstrating proof of principle for clinical trial development to progressive accumulation of the glycosaminoglycans (GAGs): chon- improve MPS II patient neurological function. droitin-6-sulfate (C6S) and keratan sulfate (KS). C6S and KS are synthesized primarily in cartilage, and sequentially, these GAGs are doi:10.1016/j.ymgme.2018.12.059 stored in lysosomes of chondrocytes, their associated ligaments, and the extracellular matrix (ECM). Accumulation of undegraded C6S and KS triggers progressive systemic skeletal dysplasia. Objectives: To report 4 Brazilian patients (one ambulatory male, 44 3 non-ambulatory females) under ERT for a period of 1-year with Platform technology for treatment of the brain in lysosomal elosulfase alfa. disorders: Application to Tay-Sachs disease Methods: Clinical, biochemical and radiological data retrospective analysis, including the following items: demographic characteristics, fi Ruben J. Boado, Eric K.-W. Hui, Jeff Z. Lu, Huilan Lin, William M. genetic pro le, ERT dose, and clinical manifestations. Pardridge, ArmaGen Inc., Calabasas, CA, United States Results: Enzyme Replacement Therapy (ERT) with elosulfase alfa was well tolerated in all 4 patients with no adverse events reported. The majority of lysosomal storage disorders (LSD) affect the Because of the possibility of potential hypersensitivity adverse brain. Intravenous enzyme replacement therapy does not treat the events, special attention is needed when using ERT in Morquio brain, because recombinant enzymes are large molecules that do not syndrome type A patients, in particular the ones with respiratory cross the blood-brain barrier (BBB). BBB-penetration of enzyme disorders (all infusions were performed in hospitals and medical therapeutics is enabled by re-engineering the recombinant enzyme facilities). Our patients showed respiratory improvement after as a bi-functional IgG-enzyme fusion protein, wherein the IgG starting ERT, better endurance and one out of the two non- domain is a monoclonal antibody (MAb) that targets a specific ambulatory patients started to walk again without support. No endogenous receptor-mediated transporter within the human BBB, obvious improvement was seen in hearing loss or corneal opacity in such as the human insulin receptor (HIR). A proof of concept phase II our patients. clinical trial in pediatric subjects with severe Hurler MPSI with a BBB Conclusions: ERT with elosulfase alfa is the only specif approved penetrating HIRMAb-iduronidase fusion protein (AGT-181, therapy for patients with Morquio syndrome type A. Initial clinical valanafusp alpha) produced stabilization of cognition, and reduced trials have not shown ERT experience in severely affected, non- hepatosplenomegaly. The aim of the present investigation was to ambulatory patients. Our report shows that ERT with elosulfase alfa determine if a similar BBB penetrating IgG-fusion protein can be can be used safely with appropriate multidisciplinary care in patients engineered for the treatment of Tay Sachs Disease (TSD), which is with Morquio syndrome type A. caused by mutations in the gene encoding for hexoaminidase A (HEXA). Genes encoding the heavy chain and the light chain of a doi:10.1016/j.ymgme.2018.12.061 HIRMAb-HEXA fusion protein were engineered followed by stable Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S33

46 with standard dosing protocol at 1mg/kg IV weekly. Cardiac ultra- A new research initiative amongst hematologists to address sound imaging and pulmonary function testing were performed prior current worldwide health disparities in the management and to initiation of ERT and at 6-month intervals thereafter. Six eligible treatment of Gaucher disease individuals were identified nationally: 4 declined 1 died before making a final decision. A 29 year old male, 27 years post-HCT, completed the Colm Bradleya, Sam Saleckb, Argiris Symeonidisc, aUniversity of protocol. At initiation of ERT, spirometry showed forced vital capacity Glasgow, Glasgow, United Kingdom, bUniversity of Hertfordshire, (FVC) of 1.14 L, forced expiratory volume in one second (FEV1) of 1.02 Hartfield, United Kingdom, cUniversity of Patras, Patras, Greece L, with FEV1/FVC of 0.89 which, along with the flow volume loop, was consistent with restrictive lung disease. Normal left systolic ventric- Background: Gaucher disease (GD) is a rare disease, with 75% of ular function, E/A reversal suggesting diastolic dysfunction, moderate GD patients being diagnosed and managed by hematologists. In the mitral stenosis (mean gradient 9-12 mm Hg) and mild mitral majority of cases, the appropriate diagnosis is established inciden- regurgitation were present. The mitral stenosis gradient had been tally, following a bone marrow examination, as part of the present 3 years prior to ERT. After 2 years of ERT, lung function investigation process for unexplained splenomegaly, thrombocyto- remained unchanged with a FVC of 1.12 L, FEV1 of 1.06, and FEV1/FVC penia or anemia. Despite this, many of the common causes of of 0.95. Cardiac function and mitral regurgitation remained unchanged misdiagnosis of GD when a bone marrow examination is not but mean mitral gradient decreased to 6 mm Hg. There was neither performed, are hematological in nature, with hematological malig- progression, nor improvement in pulmonary or cardiac systolic and nancy, idiopathic thrombocytopenia, and anemia of chronic disease diastolic function after 2 years of ERT in an adult MPS VI patient who being the more likely misdiagnosis. had undergone successful HCT nearly 3 decades prior. The 30-50% Aims: If we are to address possible worldwide GD health decrease in mitral valve gradient was unexpected and differs from disparities, we need to know and understand where hematologists adults with MPS VI who receive only ERT. are with regards to current treatment and care services for GD. If these services fall short, we will be able to improve their effectiveness and thus improve the quality of service that is given. doi:10.1016/j.ymgme.2018.12.063 Methods: This new research initiative can provide a real-world picture of treatment and outcomes by studying clinical characteristics, disease-management and relevant clinical outcomes to determine what is happening in the countries surveyed and pinpoint 48 fi reasons for this. It will be evidence-based and will be monitored and Hematopoietic cell transplantation for severe MPS I in the rst six evaluated. months of life: The heart of the matter Results: This future important research initiative can be used to provide a clear picture of the current worldwide management of GD Elizabeth Braunlin, Kelly Miettunen, Troy Lund, Mark Luquette, Paul by hematologists. This will help us to promote research of GD and to Orchard, University of Minnesota, Minneapolis, MN, United States develop both practical and clinical research recommendations in GD and to develop educational initiatives to allow the sharing of Hematopoietic cell transplantation (HCT) is accepted therapy for knowledge and best practice. severe mucopolysaccharidosis type I (MPS IH). With implementa- Summary/Conclusion: We aim to remove any world-wide GD tion of newborn screening (NBS) for MPS I in the US, HCT may now health disparities by building up a partnership of all sectors involved occur earlier than 1-2 years of age and it might be assumed that in GD care i.e. multi-disciplinary teams, medical societies and co-op cardiac issues will be fewer. To examine this hypothesis, we groups, politicians, the pharmaceutical industry, GD patient groups, reviewed our records for any MPS IH infant who underwent HCT ≤ the private sector and the wider community of people involved in at 6 months of age. Pre- and (most recent) post-HCT cardiac the multifaceted aspects and phases of GD care. echos and clinical courses were reviewed in all infants with MPS IH undergoing HCT at ≤ 6 months of age. 7 MPS IH infants (4 male) doi:10.1016/j.ymgme.2018.12.062 were diagnosed at median (range) (MEDRNG) of 14 (3, 22) days of life (DOL) by NBS (2) or because an older sib had MPS IH (5), and began enzyme replacement therapy at MEDRNG of 48 (7, 62) DOL. First pre-HCT echo was performed at MEDRNG of 45 (0, 88) DOL. 47 HCT (6 cord blood, 1 related) occurred at MEDRNG of 131 (105, fi Cardiopulmonary ndings with enzyme replacement therapy 183) DOL with most recent echo at MEDRNG of 408 (10, 1897) after hematopoietic cell transplantation for MPS VI days after HCT. Mitral regurgitation occurred in 2 infants before HCT and remained present thereafter PFO was common LVH Elizabeth Braunlin, Nathaniel Gaeckle, Paul Orchard, Jeanine Jarnes- occurred after HCT. One infant had severely decreased function at Utz, Chester B. Whitley, University of Minnesota, Minneapolis, MN, initial echo and required ICU management. One infant died United States unexpectedly 69 days after HCT. Newborns and young infants with severe MPS I presenting for HCT have unique medical issues related Before enzyme replacement therapy (ERT) became the presumed to their age. Early cardiac evaluation will identify features related to standard of care, hematopoietic cell transplantation (HCT) was the heart but increased awareness of the anatomic and functional performed in individuals with mucopolysaccharidosis type VI (MPS differences of all organ systems related to young age will be VI). Although HCT benefitted these patients, it was not curative a small necessary for optimal outcomes. number of long-term survivors remain available for ongoing assessment. Hypothesizing that post-HCT ERT might provide an additional fi bene cial effect, we offered a 2 year trial of ERT post-HCT to adult MPS doi:10.1016/j.ymgme.2018.12.064 VI long term survivors (NCT02156674). Galsulfase ERT was delivered S34 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

49 pharmacologically relevant levels of SOBI003 in the central nervous Consequences of newborn screening: Neuroimaging in infants system. NCT03423186 is an open-label, non-controlled, parallel, sequen- with severe MPS I ≤6 months of age tial ascending multiple-dose, multicenter phase I/II study. Patients aged 12-72 months with a development age of ≥12 months who have not Elizabeth Braunlin, Kelly Miettunen, Paul Orchard, Troy Lund, Ashish received previous treatment for MPS IIIA will be eligible to participate. Gupta, David Nascene, University of Minnesota, Minneapolis, MN, Weekly infusions of SOBI003 will be given for 24 weeks. The primary United States study objective is to evaluate the safety and tolerability of SOBI003 at 3 different dose levels. Secondary objectives include characterization of PK Newborn screening (NBS) for mucopolysaccharidosis type I (MPS I) properties, assessments of immunogenicity and pharmacodynamic allows initiation of early treatment for infants with known mutations. biomarkers as well as changes in gray matter volume and quality of life. For those with unknown mutations, treatment strategies become Further, the clinical response on neurocognition and adaptive behavior complicated as early enzyme replacement therapy (ERT) may subtly will be evaluated. Patients in the first dose group have initiated infusions alter clinical findings, preventing identification of infants with severe with good tolerability. Upon completion of the 24-week treatment mutations requiring hematopoietic cell transplantation (HCT). Recog- period, patients will be given the opportunity to receive continued nizing features of severe mutations by neuroimaging could be a useful SOBI003 treatment by participation in an extension study. tool for classifying severity. MRI images from 12 infants b 6monthsof age with known severe MPS I were reviewed for features known to be present in severe MPS I. These included: perivascular space enlargement, doi:10.1016/j.ymgme.2018.12.066 enlarged extra-axial spaces, white matter lesions, ventriculomegaly, bony abnormalities of skull and cervical spine, cervical cord stenosis, and odontoid capping. Additionally, dilated optic nerve sheaths and mastoid 51 fluid were noted, when present. Infants (6 male) were studied at a Newborn screening for mucopolysaccharidosis type II (MPS II) in median of 76 (21-185) days. Six infants had begun ERT a median of 13 Illinois: The first year's experience (range 2 - 87) days prior to imaging. No infant had a normal scan. Bony abnormalities of the skull (12/12), the presence of mastoid fluid (12/12), a,b c d and enlarged perivascular spaces (10/12) were the most common Barbara K. Burton , George E. Hoganson , Dorothy K. Grange , e f a findings. Enlarged extra-axial spaces (8/12) cervical spinal canal stenosis Stephen R. Braddock , Katherine M. Christensen , Lauren Hitchins , a g g (8/12) enlarged optic nerves (6/11) ventriculomegaly (7/12) and Rachel Hickey , Rong Shao , Khaja Basheeruddin , Khaja Bas- g a odontoid capping (7/12) occurred in N50% of infants. White matter heeruddin , Ann & Robert H. Lurie Children's Hospital, Chicago, IL, b lesions (0/12) and cortical atrophy 0/12) were not identified. Infants b6 United States, Northwestern University Feinberg School of Medicine, c monthsofagewithsevereMPSIhaveneuro-imagingfindings known to Chicago, IL, United States, University of Illinois College of Medicine, d be present in older children with MPS I, most commonly bony Chicago, IL, United States, Washington University School of Medicine, e abnormalities of the skull and enlarged perivascular spaces. Evaluation St. Louis, MO, United States, Saint Louis University, St. Louis, MO, United f of a comparable group of attenuated MPS I infants will be crucial before States, Cardinal Glennon Chilren's Hospital, St. Louis, MO, United States, g employing these findings as part of the assessment of disease severity in State of Illinois, Chicago, IL, United States unknown mutations. Newborn screening for MPS II was initiated by the Illinois Dept. of Public Health on 12/11/17. Testing was conducted by measurement doi:10.1016/j.ymgme.2018.12.065 of iduronate-2-sulfatase (I2S) activity in dried blood spots using tandem mass spectrometry. A positive test result was defined as I2S activity b 10% of the daily batch median. In the case of results N10% 50 but b13%, a repeat blood spot was requested. As of 6/30/18, a total of Safety and tolerability of SOBI003 in pediatric MPS IIIA patients - 93,219 infants had been screened. 9 (.01%) (all male) of these were key study design features of the ongoing first-in-human study referred for diagnostic evaluation. One infant was diagnosed with MPS II on the basis of deficient plasma I2S activity (3.13 nmol/4h/ml Anders Bröijerséna, Per Daléna, Fatih Ezgüb, Gunilla Huledala, Daniel normal 155-1082 MPS II 0-15), elevated urine glycosaminoglycans Lindqvista, Margareta Wikéna, Paul Harmatzc, aSwedish Orphan (GAGs) (128.2 mg/mmol creat normal 0-53) with elevated heparan Biovitrum, Stockholm, Sweden, bGazi University Faculty of Medicine, and dermatan sulfate and a known pathogenic variant in the I2S Ankara, Turkey, cChildren’s Hospital & Research Center at Oakland, gene (p.R468Q). He was started on enzyme replacement therapy at 3 Oakland, CA, United States weeks of age. Five other infants are suspected of having pseudodeficiency for I2S based on deficient plasma I2S, normal or near MPS IIIA is a rare, progressive and life-shortening autosomal recessive normal urine GAGs, a normal physical exam and the presence of a lysosomal storage disease. The disorder is caused by mutations in the previously unreported variant in the I2S gene. Three others had SGSH gene that result in the absence or deficiency of the enzyme normal plasma I2S activity ruling out MPS II. Newborn screening for sulfamidase, which will cause insufficient degradation of heparan sulfate MPS II is effective and can be implemented with a low rate of false (HS) and accumulation of HS metabolites in lysosomes. Early manifes- positive results. Pseudodeficiency for I2S does occur and appears to tations of MPS IIIA include speech and developmental delays followed by be more common than true deficiency. Further experience will be behavioral problems, sleep disturbances and progressive cognitive needed to determine whether there are recurrent alleles for decline. Patients usually die at the end of the second or beginning of the pseudodeficiency in the US population. third decade of life. SOBI003, a modified recombinant human sulfamidase, is an intravenously administered enzyme replacement therapy currently under clinical investigation (NCT03423186). The doi:10.1016/j.ymgme.2018.12.067 glycans of the enzyme have been chemically modified to facilitate Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S35

52 Enzyme replacement therapy (ERT) is the only available treatment The fruit fly Drosophila melanogaster as a model to study for subjects affected by Pompe disease, but it suffers from several Gaucher disease shortcomings, including limited efficacy and high immunogenicity. We recently reported full rescue of the Pompe phenotype in Or Cabassoa, Sumit Paula, Orly Dorota, Gali Maorb, Mina Mirzaianc, symptomatic four-month-old Gaa-/- mice 10 months after the Hans Aertsc, Mia Horowitza, aTel Aviv University, Ramat Aviv, Israel, treatment using AAV liver-directed gene therapy with secretable bHarvard Medical School, Boston, MA, United States, cLeiden University, form of GAA at a dose of 2E12 vg/kg (Puzzo, Colella et al., Sci Trans Leiden, Netherlands Med, 2017). To assess whether liver expression of secretable GAA could result in therapeutic efficacy in Gaa-/- mice with advanced Gaucher disease (GD), characterized by decreased activity of pathology, we treated nine-month-old animals with an AAV vector lysosomal acid-beta-glucocerebrosidase (GCase), is due to mutations encoding for secretable GAA and followed the animals 9 months in the GBA1 gene. As a result, there is accumulation of thereafter. AAV-treated Gaa-/- mice showed high and sustained glucosylceramide, mainly in cells of the reticuloendothelial system, circulating levels of GAA activity which correlated with a complete and of glucosylsphingosine, which is a secreted product. More than clearance of glycogen in cardiac and skeletal muscle, and a ~30% 800 mutations are known to date in the gene, leading to a glycogen reduction in the nervous system. Moreover, we observed an heterogeneous disorder, that has been divided to a non-neu- amelioration of respiratory symptoms and a complete normalization ronopathic form, type 1 GD, and two neurological forms, which of muscle strength in the AAV-treated cohort. In untreated mice, differ by severity, type 3 being less severe than type 2. Aiming at detailed investigation of muscle transcriptome by RNASeq showed studying the molecular and cellular abnormalities associated with defective calcium and mitochondrial homeostasis with pronounced GD, we used Drosophila Melanogaster models. The fruit fly has two oxidative stress due to increased mitochondrial content arising from GBA1 orthologs, GBA1a and GBA1b, each contains a minos element, defective autophagy. AAV treatment normalized expression of genes that leads to translation of a truncated GCase like protein. The involved in lysosomal and mitochondrial homeostasis and corrected GBA1a mutant protein has a 33 C-terminal amino-acids deletion, responsible energy sensing pathways including LKB1-Ampk. These while the GBA1b mutant protein has a 133 C-terminal amino-acids were also supported by the normalization of autophagic buildup and deletion, which includes one amino acid that stabilizes the substrate mitophagy in skeletal muscle observed at the protein level and by in the active pocket. We tested the expression and properties of both electron microscopy. These results in Gaa-/- mice with advanced mutant GBA1-encoded mutant proteins. While both are expressed, disease state shed light on the reversibility of the disease phenotype the GBA1a mutant GCase presents no activity toward artificial GCase in the Pompe mouse model via liver gene transfer of secretable GAA. substrates or toward an active site specific probe. GBA1b mutant encoded GCase has low activity toward GCase substrates and, therefore, flies homozygous for this mutant GCase accumulate doi:10.1016/j.ymgme.2018.12.069 substrate in their heads and bodies, present ERAD of the mutant protein, activation of the Unfolded Protein Response, inflammation and neuroinflammation. The loocomotor function of the mutant flies 54 and their life span are markedly reduced in comparison to those of Identifying a biomarker signature for Batten disease the heterozygous flies, carrying only one mutant allele. Use of the pharmacological chaperone ambroxol is effective. Our results a a a highlight the resemblance between the fly GBA1b model and neu- Jacob T. Cain , Tyler B. Johnson , Katherine White , Brandon a a a a ronopathic forms of GD and underlie their importance in future Meyerink , Clarissa Staton , Samantha Davis , David Sturdevant , b b a b studies of the disease as well as possible therapeutic modalities. Timo Bragge , Tuulia Huhtala , Jon Brudvig , Jussi Rytkonen , Kimmo Lehtimakib, Antti Nurmib, Antti Nurmib, Jill Weimera, aSanford Research, Sioux Falls, SD, United States, bCharles River Labs, Kuopio, doi:10.1016/j.ymgme.2018.12.068 Finland

Identifying biomarkers associated with the multiple forms of Batten disease is an integral step towards the goal of improving early 53 diagnosis, tracking progression of the disease, and monitoring Functional, biochemical and transcriptional rescue of advanced response to treatment in both clinical and research environments. Pompe disease in mice with liver expression of secretable GAA The search for a biomarker has been confounded not only by the many potential targets available, including proteins, genes, tran- Umut Cagina, Francesco Puzzoa, Manuel Jose Gomezb, Maryse Moya- scripts, secondary metabolites, and recently non-coding RNAs, but Nilgesc, Laetitia Van Wittenberghea, Nathalie Danielea, Nicolas also by the natural variability of each of those individual markers. Guercheta, Pauline Selliera, Jeremy Rouillona, Bernard Gjataa, Ideally, a biomarker should strongly predict the course of the disease, Jacomine Krijnse-Lockerc, Pasqualina Colellaa, Federico Mingozzia,d, and the efficacy of treatment, however, more often than not, the aGenethon, UMR_S951 Inserm, Univ Evry, Université Paris Saclay, Évry, inherent variability of a single biomarker is insufficient. To overcome France, bCentro Nacional de Investigaciones Cardiovasculares Carlos III this difficulty, we have decided to combine multiple approaches to (CNIC), Madrid, Spain, cPasteur Institute, Paris, France, dSpark Thera- generate a predictive biomarker signature of Batten Disease. Rather peutics, Philadelphia, PA, United States than relying a single method or metric, we have combined metrics from many different types of assays to identify a biomarker signature Pompe disease is a neuromuscular disorder caused by mutations of Batten disease using Principal Component Analysis. Rather than in the gene encoding for the lysosomal enzyme acid α-glucosidase looking at each metric individually, Principal Component Analysis (GAA) which converts lysosomal glycogen to glucose. GAA deficiency allows us to combine all of the data from all of the different methods leads to pathologic glycogen accumulation throughout the body. into a single dataset, reduce its dimensionality, and identify a set of S36 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 potential biomarkers from all of the different assays that most multisystem and progressive disease. Langerhans cells histiocytosis is strongly correlate with Batten disease. The power of this type of a rare clonal accumulation and proliferation of abnormal bone marrow analysis is that it is not reliant on the strength of a single potential derived Langerhans cells. We present a rare case of the coexistence of biomarker predictor, rather it measures the contribution of each these diseases in a 1 year old male. A 1 year old male who presented potential biomarker analyzed, and identifies a suite of markers that with a palpable mass in the left ninth rib, hepatosplenomegaly and most strongly predict Batten disease, and can serve as a Biomarker seborrheic dermatitis of the scalp resection surgery of the bone tumor signature for Batten Disease. and skin biopsy confirmed the diagnosis of Langerhans cells histiocytosis, bone scintigraphy with scapula involvement. The patient was diagnosed with multisystem Langerhans cell histiocytosis (MS- doi:10.1016/j.ymgme.2018.12.070 LCH) and initiated therapy according to the LCH-II protocol with “RISK” organ involvement. During the assessment of treatment response he presented stable disease, at 3 years old he showed increased enlargement of liver and spleen, coarse facial features, 55 hypertrichosis, stiff joints and contractures, umbilical hernia, upper- β -glucocerebrosidase activity is low in patients with multiple airway obstruction, extensive Mongolian spots, dysostosis multiplex myeloma without cognitive impairment. The measurement of enzyme activity of iduronate-2 sulfatase (b2.8 μmol/L/h) and gene sequencing with IDS a a a Raíssa P. Caldeira , Suelen P. Basgalupp , Yelena Perevalova , mutation c.880-8ANG diagnosed MPS II. Enzyme-replacement therapy a a b Fernanda Sperb-Ludwig , Tatiele Nalin , Rosane I. Bittencourt , initiated at 3 years of age with idursulfase administered 0.5 mg/kg b b b Kristiane Michelin-Tirelli , Filippo P. Vairo , Marina Siebert , Ida V. weekly, at 5 months of treatment he presented reduction in liver and a a D. Schwartz , Universidade Federal do Rio Grande do Sul, Porto Alegre, spleen volume, an increased height velocity, improvement of the b Brazil, Hospital de Clinicas de Porto Alegre, Porto Alegre, Brazil upper-airway obstruction and complete resolution of the MS-LCH. The key to altering the natural history of the MPS disorders is early and Several studies have described an increased risk of multiple accurate diagnosis. This case highlights the importance of early clinical myeloma (MM) development in Gaucher disease (GD) patients. Only features recognition of MPS II and raises the question of a possible one paper (Rosenbloom et al, 2009) searched for GBA1 mutations in association between these two pathologies. The question is if the MM patients (n=95), and the results were negative. The aim of this inflammatory and immunologic consequences of GAG tissue accumu- β study was to investigate the activity of -glucocerebrosidase (GCase) lation can result in a clonal accumulation and proliferation of cells? in MM patients, as well as to screen for GBA1 variants in this population. Sixty-seven patients with MM were included. Patient’s dried-blood spots (DBS) were used for measuring GCase and β- doi:10.1016/j.ymgme.2018.12.072 galactosidase (β-Gal) activities using fluorescent substrates. DNA was obtained from a peripheral blood sample in EDTA tubes. Ten patients had the entire coding region as well as exon/intron 57 junctions of GBA1 analyzed by next-generation sequencing (NGS) Assessment of plasma 7-ketocholesterol concentration, using the Ion Torrent PGM platform. Additionally, exons 9 and 10 of chitotriosidase activity and CCL18/PARC concentration in Spanish GBA1, where mutations N370S and L444P are located, were patients treated with human recombinant lisosomal acid lipase evaluated through Sanger sequencing in all MM patients. The mean value found for GCase activity in MM patients (5.98 ±2.82 nmol/h/ J.J. Cebollaa,P.Irúnb,C.Lahoza,I.Garcia-Jimenezc,D.GilOrtegad,J. mL) was lower than the value expected for healthy controls (9.6 Quintero Bernabeue, L. Aldámiz-Echevarriaf,J.deLasHerasf,A.Brea- ±2.60 nmol/h/mL, pb0.0001), but not the values for β-Gal Hernandog, N. Planah, D. Ibarretxeh,E.Balmaseda-Serranoi,P.Giraldoa, (p=0.213). No mutations were detected in the group of MM patients aUnidad de Investigación Traslacional, Instituto de Investigación Sanitaria analyzed by NGS. Only one patient was identified as being carrier of Aragón (IIS Aragón), Zaragoza, Spain, bCentro de Investigación Biomédica en the N370S variant (GCase activity = 6.5 nmol/h/mL). GCase activity Red de Enfermedades Hepáticas y Digestivas (CIBEREHD, ISCIII), Zaragoza, may be reduced in MM patients, even in the absence of mutations in Spain, cUnidad de Enfermedades Metabólicas Pediátricas, Hospital GBA1. The reason for this finding must be clarified. Universitario Miguel Servet, Zaragoza, Spain, dUnidad de Gastroenterología, Hepatología y Nutrición Pediátrica. Hospital Universitario Virgen de la Arrixaca, Murcia, Spain, eUnidad Funcional de Hepatología y Trasplante doi:10.1016/j.ymgme.2018.12.071 Hepático Pediátrico. Hospital Universitari Vall d'Hebron, Barcelona, Spain, fCentro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER, ISCIII). Hospital Universitario de Cruces, Barakaldo, Spain, gUnidad de 56 Lípidos. Hospital San Pedro, Logroño, Spain, hCentro de Investigación Mucopolysaccharidosis type II (Hunter syndrome) with multisys- Biomédica en Red de Diabetes y Enfermedades Metabólicas Asociadas tem Langerhans cell histiocytosis - A case report of a not (CIBERDEM, ISCIII), Reus, Spain, iUnidad de Gastroenterología y Nutrición described association Pediátrica.HospitalGeneralUniversitariodeAlbacete,Albacete,Spain.

fi Daniela Castillo-García, Magdalena Cerón-Rodríguez, Alba Valentina Lysosomal acid lipase de ciency (LALD, MIM#278000) is an Amici-García, Hospital Infantil de México Federico Gómez, Mexico City, inborn error of lysosomal lipid metabolism, triggering accumulation Mexico of cholesteryl esters and/or triglycerides. Historically, LALD comprises two phenotypes (Wolman disease [WD] and cholesteryl esters Hunter syndrome is a rare, X-linked disorder caused by the storage disease [CESD]) despite of the disease presents across an age deficiency of iduronate-2-sulphatase leading to progressive accumu- continuum, from infants to adults. The European Medicines Agency lation of dermatan and heparan sulfate in tissues that contribute to approved in 2015 the use of the first human recombinant enzyme Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S37

(rhLAL) for replacement therapy (ERT) in LALD patients. the 59 monitoring criteria of the therapy in these patients do not Potential role of stromal collagen in cystine crystallization in contemplate the measurement of plasma lysosomal related bio- cystinosis patients markers. The goal of this study was assessed the plasma concentrations of 7-ketocholesterol (7KC), CCL18/PARC and chitotriosidase Anuj Chauhana, Phillip Dixonb, aColorado School of Mines, Golden, CO, activity (ChT) in Spanish LALD patients under ERT with rhLAL. Seven United States, bUniversity of Florida, Gainesville, FL, United States CESD patients (median age=12 y.o.) and one WD patient (2 y.o.) were followed-up plasma concentrations of 7KC (LC/MS-MS), CCL18/ Cystinosis is a genetic disease that leads to the accumulation of PARC (immunoquantification) and ChT activity (fluorescence assay) intracellular cystine crystals in all organs including cornea due to the during 3 -27 months (median=12 months). Only the decrease of loss of cystine efflux transporters in the lysosome of the cells. While 7KC concentrations was significant in all patients (p=0.002). the mechanism for formation of intracellular cystine is well Otherwise CESD patients managed to reduce 7KC concentrations understood for most organs, it does not explain many observations (p=0.003) and ChT activity (p=0.02) as well as the normalization of for crystal accumulation in the cornea. First, the crystals in cornea are the levels of these. These results support the utility of monitoring extracellular and needle-like with several hundred microns length plasma 7KC concentration in all LALD patients, as well as ChT activity which is in sharp contrast with the rectangular or hexagonal crystals only in CESD patients, under ERT with rhLAL. found throughout other organs. Second, these crystals are arranged parallel to the stromal collagen, which is a unique to the cornea. Third, crystal growth in the cornea reaches a saturation point after doi:10.1016/j.ymgme.2018.12.073 where no further crystallization occurs. We propose a hypothesis supported by in vitro and ex vivo data to explain these observations. We hypothesize that the needle-shaped stroma crystals form extracellularly due to the ionic interactions between the cystine 58 and collagen fibrils present in the stroma. We examine cystine Identification of a novel GLA mutation (V269L) in a Mexican 2- crystal growth both with in vitro polymer solutions and ex vivo in year-old male with Fabry nephropathy: A case report rabbit cadaver eyes to show that negatively charged polymers lead to the formation of increased cystine precipitation in aqueous solution. Magdalena Cerón-Rodríguez, Daniela Castillo-García, Alba Valentina Furthermore, needle-like cystine crystal formation can be observed Amici-García, Hospital Infantil de México Federico Gómez, Mexico City, only in presence of certain polyelectrolytes including collagen. The Mexico structure of the crystals that form in vitro and ex vivo is similar to that in vivo. The stroma keratocytes likely drive formation of the Fabry disease is caused by mutations in the GLA gene that lower polygonal crystals, which become extracellular due to apoptosis α -galactosidase A activity affecting glycosphingolipid metabolism. during corneal repair. The polygonal crystals then dissolve and form fi Several GLA variants have been identi ed that are associated with needle shaped crystals in the presence of collagen. This proposed relatively elevated residual enzyme activity the challenge is to mechanism explains many of the yet unanswered questions but it determine which GLA variants can cause clinical manifestations. We needs further support from in vivo studies. The improved under- N report a novel missense hemizygous mutation, c.805G C (p. standing could lead to improved treatment of corneal cystinosis. Val269Leu) which caused Fabry nephropathy in a 2 year old male. We describe a new GLA mutation in a 2 year old male who was born in Mexico. His uncle was diagnosed with Fabry nephropathy at 28 doi:10.1016/j.ymgme.2018.12.075 years of age and his mother was carrier. The αGal-A activity in DBS was 0.39 nmol/ml/hr (mean value controls 1.04-5.9), plasma GL-3, urine GL-3 and Lyso-Gb3 were within the normal values. The gene sequencing identified a missense hemizygous mutation in exon 6 of 60 GLA, c.805GNC p.Val269Leu (V269L), the same mutation was Insights into Sanfilippo syndrome provided by the ConnectMPS identified in the uncle and mother of the patient. During the clinical worldwide online registry assessment he presented acroparesthesias and the laboratory workup showed proteinuria. We performed a kidney biopsy that Edwin Chavez-Cintoraa, Chen Yua, Paul Humphreya, Jill Woodb,Jo showed significant GL3 accumulation in several types of kidney cells, Ann Vidalc, Vanessa Rangel Millerc, Stephen M. Maricicha, aBioMarin electron microscopy showed numerous electron-dense multi-lamel- Pharmaceutical Inc., Novato, CA, United States, bJonah's Just Begun, lar inclusions in the epithelial cytoplasm, typical of Fabry disease. Levittown, NY, United States, cInvitae, San Francisco, CA, United States Enzyme replacement therapy with agalsidase beta was initiated at 1 mg/kg every other week during the course of treatment the patient Sanfilippo syndrome (mucopolysaccharidosis type III) is a group presented a diminished frequency of acroparesthesias. A novel GLA of four lysosomal storage diseases (types A, B, C, D) caused by mutation, c.805GNC (p.Val269Leu) was found in a Mexican family different enzyme deficiencies that lead to heparan sulfate accumu- with early renal manifestations of Fabry disease, the substrate lation in the central nervous system and visceral body tissues. Unlike accumulation was demonstrated with kidney biopsy that showed other MPS disorders, Sanfilippo syndrome is characterized by GL-3 deposits in a 2 year old male. The identification of this novel neurological symptoms including developmental delay, behavioral mutation will enable clinicians and genetic counselors to have better problems and progressive neurodegeneration. The rarity of these clinical understanding of Fabry disease. disorders presents a major impediment to collecting comprehensive information about how diseases like Sanfilippo syndrome present and affect multiple aspects of patient health. One method that has doi:10.1016/j.ymgme.2018.12.074 been used to circumvent this problem is creation of patient registries allowing wide-scale collection and aggregation of information about S38 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 rare diseases like Sanfilippo syndrome. In particular, online registries life, visceromegaly and hematological parameters. Huge unmet need have the advantage of being readily accessible to a large number of causes significant mortality in untreated children patients and their families around the world. To better understand how Sanfilippo syndrome affects patients and their families, we extended doi:10.1016/j.ymgme.2018.12.077 ConnectMPS, an online patient registry represented by 27 advocacy organizations, to collect additional data regarding Sanfilippo syndrome. Survey questions were designed to collect (1) demographic, disease 62 history and treatment data, (2) responses to the Sanfilippo Behavior Therapeutic efficacy and safety of scAAV9/AGA gene therapy in Rating Scale (SBRS), an inventory of questions designed to query aspartylglucosaminuria mice specific characteristics of the disorder, and (3) laboratory and other diagnostic data to better characterize medical aspects of the disease. The Xin Chena, Sarah Snanoudj-Verberb, Laura Pollardc, Sara Catheyc, surveys and enrollment information on the website have been Steven J. Graya, aUTSW Medical Center, Dallas, TX, United States, translated into a number of languages to allow access to as many bUniversité Paris Diderot, Paris, France, cGreenwood Genetics Center, families as possible. Participants were invited to complete the survey Greenwood, SC, United States. starting in January 2017 to date, 448 Sanfilippo A, B, C and D patients living in more than 40 countries have contributed information to the Aspartylglucosaminuria (AGU) is an autosomal recessive lyso- ConnectMPS registry. This presentation will summarize our findings to somal storage disease caused by the loss of functional enzyme date and discuss how the collected data informs a better understanding aspartylglucosaminidase (AGA), resulting in the accumulation of its of how Sanfilippo syndrome affects patients around the world. substrate (GlcNAc-Asn) in tissues and body fluids. AGU patients have a slow but progressive neurodegenerative disease course characterized doi:10.1016/j.ymgme.2018.12.076 by intellectual disability, skeletal and motor abnormalities, and early mortality. There is no approved treatment for AGU patients. To test our hypothesis that scAAV9/AGA gene therapy might provide a long-term therapeutic benefit, we carried out a comprehensive efficacy experi- 61 ment in which AGA-/- (AGU) mice were administered intravenously Clinical characteristics, genotype and outcome of Gaucher disease (IV) or intrathecally (IT) either high (1x1012 or 1x1011 vg/mouse for IV in Pakistani children or IT, respectively) or low (2x1011 or 2x1010 vg/mouse for IV or IT, respectively) doses of scAAV9/AGA at 6-months (early-symptomatic Huma A. Cheema, Anjum Saeed, Muhammad Nadeem Anjum, Nadia cohorts) or at 2-months (pre-symptomatic cohorts). In our 6-month- Waheed, Zafar Fayyaz, The Children's Hospital, Lahore, Lahore, old early-symptomatic cohorts, scAAV9/AGA administration led to: 1) Pakistan. dose-dependent increase and sustained AGA activity in serum and peripheral tissues to a supra physiological level 2) dose-dependent Aims/Objectives: We aimed to study the clinical characteristics, elimination of GlcNAc-Asn substrate in both central and peripheral genotype and outcome of Gaucher disease in Pakistani children. tissues and body fluids 3) significantly reduced substrate in the brain Methodology: Children’s Hospital Lahore holds the central as assessed by in vivo magnetic resonance spectroscopy 4) signifi- registry for LSD’s since 2013. 239 children with GD have been cantly restored movement in open field tests for high dose cohorts and registered. Demographic data clinical features, laboratory parameters 5) dose-dependent Purkinje cell preservation in the cerebellum. Our 2- and outcome were analyzed for 126 children who were on regular month-old pre-symptomatic cohorts are still under analysis and follow up at our center. available data have shown similar therapeutic benefit. In parallel, a Results: 126 children with confirmed GD are on regular follow-up safety experiment in AGU mice was conducted to evaluate a at Children’s hospital, Lahore. GD1 (50%) was the commonest type supraphysiological dose of scAAV/AGA (1x1012 vg/mouse, IT). Neither followed by GD3(45.9%) and GD2 (4%). There were 81 (64.2%) males abnormal neurological symptoms nor body weight loss have been with mean±SD age of 2.6±2.4 years. Majority of children (87, 69%) observed. Taken together, these results demonstrate that treatment of from all the three types presentedb2 years of age with severe disease. AGU mice with AAV9/AGA is both effective and safe, providing strong The most common clinical presentations and physical findings were proof-of-concept evidence that scAAV9/AGA gene therapy should be pallor, frequent transfusions and huge organomegaly (100%, spleenN considered for human translation. liver). Developmental delay and neurological deficit of varying degrees was prominent clinical presentation in GD2. Enzyme analysis was the doi:10.1016/j.ymgme.2018.12.078 main diagnostic tool in addition to genetic testing. Eight-six (68%) children had mutational analysis and commonest was L444P followed by few novel mutations. Only forty (31.7%) children were able to get ERT and 38 (95%) had a good response in terms of regression in 63 visceromegaly and improvement in hematological parameters. The Exosome-mediated delivery of active glucocerebrosidase to rest of the children are on follow up without treatment and suffering Gaucher model cells complications of disease. 35 (29.3%) children unfortunately died without treatment with FTT, recurrent infections and bleeding Hojun Choi, Kyungsun Choi, Chulhee Choi, KAIST, Daejeon, Republic of diathesis. 2 children of GD3 died of pulmonary disease on ERT. Korea Conclusion: GD is not uncommon in Pakistani children and has early and severe presentation with huge organomegaly. L444P mutation is Introduction: Exosomes are cell-derived extracellular vesicles the frequently seen mutation in Pakistani children with variable with a diameter of 40-150 nm that participate in intercellular phenotypic presentation even within the same family. ERT is available communication. Currently, exosomes are gaining attention for its to a limited number and has excellent results in improving quality of potential as a drug delivery system. Although enzyme replacement Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S39 therapy (ERT) is widely used for the treatment of patients with 65 Gaucher disease, ERT shows the limitations low delivery efficacy of Delivery of adenine base editors to patient-derived induced the recombinant enzyme to target cells, a poor half-life in the plasma pluripotent stem cells in vitro: A putative treatment for muco- and hypersensitivity. To overcome these drawbacks, we propose the polysaccharidosis type IIIB treatment of Gaucher disease via exosome-mediated active GCase replacement. We developed opto-genetically engineered exosome to Chloe L. Christensena, Rhea E. Ashmeada, Samuel S. Chub, Francis Y.M. load functional GCase, and demonstrated its effect in enzyme Choya, aUniversity of Victoria, Victoria, BC, Canada, bUniversity of British delivery to target cells, further establishing its potential as a Columbia, Vancouver, BC, Canada therapeutic vesicle for treatment of Gaucher disease. Methods: In the present study, we have incorporated GCase into Mucopolysaccharidosis (MPS) IIIB is an autosomal recessive the engineered exosomes by fusion with optically controlled PPI lysosomal storage disease (LSD) resulting from mutations in NAGLU, module. GCase -loaded exosomes were tested for protein loading which encodes α-N-acetylglucosaminidase (NAGLU). A lack of active efficiency by immunoblot and ELISA, and its enzymatic activity. NAGLU leads to the build up of heparan sulfate, and subsequent Patient-derived fibroblasts and Gaucher disease mouse-derived dysregulation of cellular functioning. Patients with MPS IIIB experi- neurons were used for delivery of active GCase by GCase-loaded ence sleep apnea, respiratory issues, hepatosplenomegaly, and exosomes. progressive neurodegeneration. Common treatments for LSDs in- Results: We were able to load active GCase into engineered clude recombinant enzyme replacement therapy (ERT) however, ERT exosomes and further demonstrated the intracellular delivery of fails to ameliorate neurodegeneration due to the blood-brain barrier GCase as functional proteins in the Gaucher model cells. blocking access to the brain. A large number of disease-causing Conclusion: These results clearly indicate the potential of mutations in NAGLU are missense or nonsense mutations (N114 exosome-mediated delivery of active GCase for treatment of Gaucher identified), which are suitable targets for base editing (BE), a novel disease. gene editing technology. Base editors include a Cas9-nickase with a deaminase component and are able to target a sequence of interest doi:10.1016/j.ymgme.2018.12.079 to deaminate cytidine or adenine within a predictable 5-bp window. Induced pluripotent stem cells (iPSCs) corrected for MPS IIIB mutations may be differentiated to neural precursor cells, delivered 64 intracranially, and used as a putative cell therapy. We have fi Enzyme replacement therapy for mucopolysaccharidosis type IIID generated iPSCs from MPS IIIB human skin broblasts using non- integrating Sendai viral vectors, transiently expressing transcription factors c-Myc, Klf4, Oct3/4, and Sox2. A resulting iPS cell line Tsui-Fen Choua, Derek Moenb, Chelsee Saunib, Shih-hsin Kana, Feng (homozygous c.457 GNA) was electroporated with plasmids Wanga, Xiaoyi Zhanga, Steven Lea, Jill Woodb, Sean Ekinsb, Patricia I. encoding adenine deaminase BE and guide RNA (Addgene Dicksona, aHarbor-UCLA and LA BioMed, Torrance, CA, United States, 11210143860, with insert guide sequence). Delivery of these bPhoenix Nest Inc, Brooklyn, NY, United States plasmids was confirmed through fluorescence imaging. Further analyses are being conducted to determine if electroporated iPSCs Mucopolysaccharidosis type IIID (MPS IIID Sanfilippo D) is a have incorporated a single base change, reflecting the normal devastating pediatric neurodegenerative disorder with no cure or sequence. Analyses through droplet digital PCR, sequencing, and effective treatment available. The fundamental cause of MPS IIID is an NAGLU enzyme activity assays will be conducted. In addition, we inherited mutation in glucosamine (N-acetyl)-6-Sulfatase (GNS) have outlined mutations causative for six other MPS diseases that are required to catabolize heparan sulfate (HS). Without functional GNS, putative targets for BE. This method of gene editing in iPSCs is a glycosaminoglycan accumulates in lysosomes. As enzyme replace- putative regenerative therapy for patients with LSDs affected by ment therapy (ERT) is an effective treatment for other lysosomal neurodegeneration. Funding sources: Sanfilippo Children's Research diseases, we propose to deliver recombinant human GNS (rhGNS) into & Rare Disease Foundations the patient to treat MPS IIID. The symptoms of MPS IIID are largely localized to the brain, and therefore, our strategy proposes to deliver doi:10.1016/j.ymgme.2018.12.081 recombinant human rhGNS to the brain in order to effectively treat the underlying cause. From 700 mL expression media of a CHO cell line stably expressed GNS, we can purify ~200 ug of secreted rhGNS with specific activity ~10,000 nmol/h/mg. We have demonstrated stability 66 over one month at 4°C in artificial cerebrospinal fluid. Additionally, we Development and validation of a novel multiplex LC-MS/MS assay observed intracellular enzymatic activity of rhGNS in MPS IIID human of globotriaosylceramide and globotriaosylsphingosine in human fibroblasts when rhGNS is added to the media and confirmed plasma radiolabelled HS is diminished to WT levels in MPS IIID fibroblasts treated with rhGNS. In vivo efficacy of rhGNS was evaluated by Wei-Lien Chuang, Joshua Pacheco, Felipe Lopes, Richard Yee, Crystal intracerebroventricular injection of rhGNS in neonatal mice at day 1 Sung, Sanofi, Framingham, MA, United States demonstrated return of GNS activity and statistically significant reduction of lysosomal storage markers throughout the brain.Funding Fabry disease (FD) is an X-chromosome linked lysosomal storage support NIH: 1R41NS89061-01, 2R42NS089061-02 disease, resulting from a deficiency of α-galactosidase A [1], leading to the accumulation of globotriaosylceramide (GL-3) in patient tissues. Such accumulation eventually results in impairment of the doi:10.1016/j.ymgme.2018.12.080 proper function of affected organs, including cardiac and renal failure. The relationship between clinical manifestation of Fabry S40 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 disease and plasma GL-3 levels is not well understood [2]. Studies treatment in 7/7 treated subjects. Abdominal MRI imaging have also shown that a deacylated form of GL-3, glo- demonstrates that liver and spleen volumes normalize after 24 botriaosylsphingosine (Lyso-GL-3), is also elevated in patient plasma weeks of treatment in 9/9 and 7/9 subjects, respectively. and could correlate better with disease manifestation [3]. We Stabilization in developmental quotient (DQ) was observed in 5/ recently developed and validated a novel liquid chromatograph- 7 treated subjects. Safety data indicate that ICV-administered tandem mass spectrometry (LC-MS/MS) based method to simulta- tralesinidase alfa is generally well-tolerated. These findings neously quantify both GL-3 and Lyso-GL-3 in human plasma samples. demonstrate that tralesinidase alfa can be safely administered The method incorporates a single-phase extraction system to into brain ventricles via isovolumetric bolus infusion and that this eliminate the need for solvent exchange prior to LC-MS/MS analysis. treatment approach leads to a marked pharmacodynamic re- This novel multiplex approach significantly improves inter-assay bias sponse in the CNS and visceral organs of Sanfilippo B patients. between GL-3 and Lyso-GL-3 assays as well as assay turnaround time. References: 1) Aerts et. al. Biomarkers in the diagnosis of lysosomal storage disorders: proteins, lipids, and inhibodies. J Inherit doi:10.1016/j.ymgme.2018.12.083 Metab Dis. 2011 Jun;34(3):605-19. 2) Lobato et. al. Biomarkers in Lysosomal Storage Diseases. Diseases. 2016 Dec 4(4): 40. 3) Rombach et. al. Plasma globotriaosylsphingosine: diagnostic value and relation to clinical manifestations of Fabry disease. Biochim 68 fi α Biophys Acta. 2010 Sep;1802(9):741-8. Safety and ef cacy of advanced and targeted acid -glucosidase (AT-GAA) (ATB200/AT2221) in ERT-switch nonambulatory patients with Pompe disease: preliminary results from the ATB200- 02 trial doi:10.1016/j.ymgme.2018.12.082

Paula R. Clemensa, Tahseen Mozaffarb, Benedikt Schoserc,Drago Bratkovicd, Barry J. Byrnee,OzlemGoker-Alpanf,MarkRobertsg, 67 Peter Schwenkreish, Kumaraswamy Sivakumari,AnsT.vander ICV-administered tralesinidase alfa (BMN 250 NAGLU-IGF2) is Ploegj,JacquelynWrightk, Sheela Sitaramank,JayA.Barthk, well-tolerated and reduces heparan sulfate accumulation in the Hjalmar Lagastk, Priya Kishnanil,XueMingm, aUniversity of CNS of subjects with Sanﬁlippo syndrome type B (MPS IIIB) Pittsburgh and Department of Veterans Affairs Medical Center, Pittsburgh, PA, United States, bUniversity of California, Irvine, CA, Maureen Clearya, Nicole Muscholb, Maria Luz Coucec, Paul Harmatzd, United States, cFriedrich-Baur-Institut, Neurologische Klinik, Ludwig- Joy Leee, Shuan-Pei Linf, Ilyas Okurg, Fatih Ezgug, Heidi Peterse, Maximilians-Universität München, Munich, Germany, dPARC Re- Martha Solano Villarrealh, Adam J. Shaywitzi, Heather Cahani, Anita search Clinic, Royal Adelaide Hospital, Adelaide, SA, Australia, Groveri, Andrew Meltoni, Lynn Smithi, Stephen M. Maricichi, Maria J. eUniversity of Florida, Gainesville, FL, United States, fO&O Alpan de Castro Lopezc, aGreat Ormond Street Hospital, London, United LLC, Fairfax, VA, United States, gSalford Royal NHS Foundation Trust, Kingdom, bUniversity Medical Center Hamburg-Eppendorf, Hamburg, Salford, United Kingdom, hNeurologische Klinik und Poliklinik des Germany, cHospital Clínico Universitario de Santiago, A Coruña, Spain, Berufsgenossenschaftlichen, Universitätklinikum Bergmannsheil, Bo- dUCSF Benioff Children’s Hospital Oakland, Oakland, CA, United States, chum, Germany, iNeuromuscular Research Center, Phoenix, AZ, eRoyal Children’s Hospital, Melbourne, Australia, fMackay Memorial United States, jErasmus Medical Center, Rotterdam, Netherlands, Hospital, Taipei, Taiwan, gGazi University Hospital, Ankara, Turkey, kAmicus Therapeutics, Inc., Cranbury, NJ, United States, lDuke hUniversidad del Rosario, Bogota, Colombia, iBioMarin Pharmaceutical University Medical Center, Durham, NC, United States, mRutgers Inc., Novato, CA, United States New Jersey Medical School, Newark, NJ, United States

Sanfilippo Syndrome type B (MPS IIIB) is a lysosomal storage Late-onset Pompe disease (LOPD) is a rare, inherited mul- disorder caused by deficiency of the α-N-acetylglucosaminidase tisystemic metabolic disorder characterized by progressive muscle (NAGLU) enzyme and subsequent heparan sulfate (HS) accumu- weakness and respiratory impairmentthatcanresultinpatients lation in the brain. Sanfilippo B patients display progressive losing the ability to walk. Currently, recombinant human acid α- neurocognitive decline and typically do not live past the second glucosidase (GAA) enzyme replacement therapy (rhGAA ERT or third decades of life. Tralesinidase alfa (BMN 250 NAGLU-IGF2) alglucosidase alfa) is the only FDA-approved treatment. AT-GAA is a novel enzyme replacement therapy (ERT) for Sanfilippo B (ATB200/AT2221), a next-generation rhGAA ERT/pharmacological consisting of NAGLU enzyme fused to insulin-like-growth factor 2 chaperone regimen, is being evaluated for adults with LOPD in a (IGF2) to enhance lysosomal targeting. This report presents first-in-human, open-label, phase 1/2 trial ATB200-02 preliminary results from the first human study of tralesinidase (NCT02675465). The study enrolled 20 patients in 3 cohorts: ERT- alfa (NCT02754076). NCT02754076 is a phase 1/2, open-label switch ambulatory (n=11), ERT-switch nonambulatory (wheel- study with two parts. Part 1 consisted of 3 dose-escalation periods chair-bound and unable to walk unassisted) (n=4), and ERT-naive (≥4 weeks) of intracerebroventricular (ICV) tralesinidase alfa ambulatory (n=5). This analysis evaluates the 12-month interim administered as a weekly isovolumetric bolus infusion. Subjects data for the ERT-switch nonambulatory patients, including safety from Part 1 and from an ongoing observational study of Sanfilippo assessments, muscle strength test for upper limbs, pulmonary B patients (NCT02493998) continue to Part 2, a 48-week function tests, and patient-reported outcomes. Mean GAA protein treatment period to examine efficacy/safety at the maximum exposure (protein and activity) was higher with AT2221 coadmin- tolerated tested dose (300 mg). For enrollment into Part 1, 1-10 istration. Increases in upper extremity strength, assessed by year old subjects with Sanfilippo B must have deficient NAGLU quantitative and manual muscle testing, were observed at months activity at screening. Cerebrospinal fluid (CSF) total HS levels 6 and 12. Reductions from baseline in muscle enzymes (CK, ALT, show sustained normalization after 1-3 weeks of tralesinidase alfa AST) and disease substrate (urine Hex4) were observed and Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S41 maintained for up to 12 months. Improvements from baseline 70 (mean [SD]) were observed in the Rasch-built Pompe-specific Latent TGF-beta-binding protein 4 modulates disease severity in Activity Scale (month 6, 1.5 [2.4] month 12, 1.0 [2.0]), the the knock-out mouse model of Pompe disease Rotterdam Handicap Scale (month 6, 1.5 [5.1] month 12, 0.5 [3.9]), and the Fatigue Severity Scale. All patients reported Pasqualina Colellaa, Pauline Selliera, Umut Cagina, Maria Grazia improvement in overall physical well-being (Subject Global Biferib, Nicolas Guercheta, Guillaume Tannioua, Laetitia van Impression of Change). Twenty treatment-emergent adverse Wittenberghea, Bernard Gjataa, Federico Mingozzia,b,c, aGenethon, events were reported, all mild and transient and none occurring UMR_S951 Inserm, Univ Evry, Université Paris Saclay, EPHE, EVRY, in N1 patient. One patient experienced an infusion-associated France, bSorbonne University and INSERM_U974, Paris, France, cSpark reaction (skin discoloration). These data demonstrate tolerability Therapeutics, Philadelphia, PA, United States and clinical benefit of AT-GAA in ERT-switch nonambulatory patients who have been on ERT for an average of ~9 years, Pompe disease (PD) is a neuromuscular disorder caused by the suggesting that AT-GAA (ATB200/AT2221) could be an important deficiency of the lysosomal enzyme acid-alpha-glucosidase (GAA), treatment option for nonambulatory patients with LOPD. Updated leading to pathologic glycogen accumulation. The severe infantile- data will be included in the presentation. onset form (IOPD) presents in newborns with premature death due to cardio-respiratory failure. The late-onset form (LOPD) presents in children and adults usually with milder but progressive muscular and doi:10.1016/j.ymgme.2018.12.084 respiratory impairment. Gaa knock-out miceB6;129-Gaatm1Rabn/J (Gaa-/- ), have been extremely useful to model PD, however they are limited by the late-onset of muscular myopathy (7-9 months of age) and a very mild respiratory phenotype in normoxic conditions. It has been 69 described that modulation of the genetic background may alter the Tandem promoter design confers tolerogenic and persistent severity of muscular disease. In particular, the Latent TGF-beta- transgene expression to AAV gene therapy in neonate Pompe mice binding protein 4 (Ltbp4) gene has been reported to be a strong genetic modifier of muscular dystrophy in humans and mouse models. a a a Pasqualina Colella , Pauline Sellier , Helena Costa-Verdera , Francesco Here, we developed a novel mouse model of PD by crossing the original a a a Puzzo , Laetitia van Wittenberghe , Nicolas Guerchet , Guillaume GaaB6;129-Gaatm1Rabn/J mice, naturally carrying the Ltbp4+36/+36 allele, a a b a Tanniou , Bernard Gjata , Solenne Marmier , Severine Charles , with DBA2/J mice carrying the Ltbp4Δ36/Δ36 allele, that has been a a a Marcelo Simon-Sola , Fanny Collaud , Christian Leborgne , Federico shown to enhance myopathy in mouse models of muscular dystrophy. a,b,c a Mingozzi , Genethon, UMR_S951 Inserm, Univ Evry, Université Paris Compared to the Gaa-/- Ltbp4+36/+36mice, Gaa-/-Ltbp4Δ36/Δ36 mice b Saclay, EPHE, Evry, France, Sorbonne University and INSERM_U974, Paris, presented the appearance of key disease signs at 2 months of age, the c France, Spark Therapeutics., Philadelphia, PA, United States earliest time point tested, including respiratory impairment and muscle weakness. Consequent to the worsened phenotype, male Hepatocyte-restricted, AAV-mediated gene transfer is being used to Gaa-/-Ltbp4Δ36/Δ36 animals also showed reduced survival (50% at 5 provide sustained, tolerogenic transgene expression in gene therapy. months of age, pb0.01 vs. Gaa+/+Ltbp4+36/+36). Current work is However, given the episomal status of the AAV genome, this approach focused on the molecular characterization of the model and on the cannot be applied to pediatric disorders where hepatocyte proliferation rescue of the severe phenotype with gene transfer. In conclusion, our fi fi may result in signi cant loss of therapeutic ef cacy over time. In work shows that Ltbp4 modulates the severity of Pompe phenotype addition, many multi-systemic diseases require widespread expression and provides a novel mouse model to study pathophysiology and test of the therapeutic transgene that when provided with ubiquitous or efficacy of novel therapeutic approaches to treat the disease. tissue-specific non-hepatic promoters often results in anti-transgene immunity. Here we have developed tandem promoter monocistronic expression cassettes that, packaged in AAV, provide combined hepatic doi:10.1016/j.ymgme.2018.12.086 and extra-hepatic, tissue-specific transgene expression and hepatic- mediated transgene immune tolerance. We validated our approach in infantile Pompe disease, a prototype disease caused by the lack the ubiquitous enzyme acid-alpha-glucosidase (GAA), presenting multi- 71 systemic manifestations and detrimental anti-GAA immunity. We Healthcare resource use in severe mucopolysaccharidosis type I showed that systemic AAV gene therapy in immunocompetent Gaa-/- post-transplant children via parent survey mice using the tandem promoter design confers sustained immune tolerance to the GAA transgene product. Combined with the persistent Therese Connera, Francesca Cooka, Vivian Fernandeza, Vanessa transgene expression in non-dividing extra-hepatic tissues, this re- Rangel-Millerb, Karen Rascatic, aREGENXBIO, Rockville, MD, United sulted in whole-body therapeutic efficacy upon vector delivery in States, bInvitae, San Francisco, CA, United States, cUniversity of Texas at neonate mice. We demonstrated rescue of muscle and CNS disease in Austin, Austin, TX, United States neonate Gaa-/- mice with tandem promoter GAA expression cassettes packaged in either AAV8 or AAV9. In conclusion, the tandem promoter Severe mucopolysaccharidosis (MPS) type I (or Hurler syndrome) is design overcomes important limitations of AAV-mediated gene transfer a rare, recessive genetic disorder caused by enzyme deficiency and is and can be beneficial when treating multi-systemic pediatric conditions treated with hematopoietic stem cell transplant and/or enzyme requiring persistent, tolerogenic transgene expression. replacement therapy (ERT). In the United States, little is known about a child's healthcare needs after transplant. An online survey, modified from the BURQOL-RD, collected information about MPS I children via doi:10.1016/j.ymgme.2018.12.085 parent report. ConnectMPS, a patient registry, invited its US-based members to complete the survey. This study presents utilization of S42 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 healthcare services among 32 severe MPS I children who were at least 73 one year post-transplant. Among the 32 children, 53% were female and Results of a Canadian survey on the family burden of illness in at the time of survey mean age was 9.4 (st dev=4.9) years. Nineteen severe mucopolysaccharidosis type I (61%) were covered solely by private insurance, 3 (10%) solely by Medicaid, 6 (19%) by both private payer and Medicaid, and the rest Therese Connera, Francesca Cooka, Vivian Fernandeza, Kim Angelb, (10%) by other insurers or other combinations (1 missing response). All aREGENXBIO, Rockville, MD, United States, bCandian MPS Society, children had received transplant by age 3. Eight of the children had been Vancouver, BC, Canada hospitalized at least once in the last year. Fourteen (44%) children had received ERT in the last month. In the last six months, 26 (81%) children Severe mucopolysaccharidosis (MPS) type I is a rare recessive had seen a cardiologist, 25 (78%) had seen an orthopedist, and 22 (69%) genetic disorder that is treated with hematopoietic stell cell had seen an endocrinologist at least once. The children averaged 1.8 transplant and/or enzyme replacement therapy (ERT). In Canada, occupational, 2.2 physical, and 1.5 speech therapy visits per month. The little is known about the family burden of this disease. An online results of this survey indicate that families with a severe MPS I child survey, modified from the BURQOL-RD, collected data from parents require continued medical support after transplant. Further research is of severe MPS I children, including caregiver work productivity and needed to identify therapies that reduce morbidity and mortality. the child's educational and medical needs. The Canadian MPS Society recruited 10 families to complete the survey. Nine parents said they doi:10.1016/j.ymgme.2018.12.087 were the child's primary caregiver, and six (67%) of the nine were also employed. Among these six, two had reduced their work hours in the last six months to care for their child and four reported that they had difficulty performing work tasks. All six reported they had 72 taken 1+ day off from work in the last six months to care for their Results of an online survey on family burden of illness in severe child, and two of these had taken off the entire six months during mucopolysaccharidosis type II their child's transplant. Among the 10 children, 50% were female and age at time of survey was 7 years, 8 months. Among children 5 a a a Therese Conner , Francesca Cook , Vivian Fernandez , Vanessa +years, 83% required special support to attend school or attended a b a b Rangel-Miller , REGENXBIO, Rockville, MD, United States, Invitae, special program. Eight (80%) of all respondents indicated their child San Francisco, CA, United States had received a transplant before the child's 3rd birthday. Of these, six had received transplant more than a year ago. However, two (33%) of Mucopolysaccharidosis (MPS) type II is a rare x-linked genetic the six had been hospitalized in the last year, one for hip surgery and fi disorder that is caused by a de ciency of the lysosomal enzyme one for a sleep study, and three (50%) had day surgeries for hernia iduronate-2-sulfatase (I2S). The purpose of this study was to assess repair, medical device removal, and orthopedic knee repair. The caregiver burden in families with severe MPS II children, including results of this survey indicate that families in Canada experience work productivity, time caring for the child, and access to healthcare signficant burden related to caregiver work productivity, and resources. ConnectMPS, a patient registry, invited its US-based continued educational and medical needs of the child. Further fi members to complete an online survey modi ed from the BURQOL- treatment efforts are needed that reduce the family burden of severe RD. A total of 74 surveys were completed, mostly by mothers who MPS I. reported they were the primary caregiver (80%) of their child. The average age of the child at the time of survey was 9.8 years, and age at diagnosis was 2.3 years. Twenty-four (32%) children had received intrathecal (IT) and 55 (74%) had received intravenous (IV) enzyme doi:10.1016/j.ymgme.2018.12.089 replacement therapy (ERT) in the previous month. Among primary caregivers, 70% were not working outside the home. Fifty-seven percent who were working reported that they had taken 7+ days off from work in the last six months to care for their child, and 76% 74 reported they had difficulties performing work tasks due to their Clinical and biochemical study of Brazilian patients with meta- child's disorder. Time to perform various daily activities did not chromatic leukodystrophy appear to differ by age of the child, indicating that caregivers of older a a b children spent similar time providing care as did caregivers of Maria Julia G. Costa , Frederico Mendes Borges , Maira M. Burin , b a a younger children. While 75% of parents reported that they did not Roberto M. Giugliani , Charles M. Lourenco , Faculdade de Medicina - have challenges obtaining home health supplies for their child, 32% Centro Universitario Estacio de Ribeirao Preto, Ribeirao Preto, Brazil, b spent $500+ per month out-of-pocket for home health supplies. Federal University of Rio Grande do Sul, Porto Alegre, Brazil. Supplies least likely to be completely covered by insurance (and thus costing families) included spirometry tests, breathing apparatuses, Background: Metachromatic leukodystrophy (MLD) is a lyso- urinary catheters, over-the-counter products, humidifiers and special somal disorder affecting mainly peripheral and central nervous fi beds. The results of this study indicate that families with severe MPS system, caused by arylsulfatase A (ARSA) de ciency. Usually It is fi II children experience burden in terms of work productivity, time, classi ed into three forms according to the age of onset of symptoms and costs associated with caring for their child at home. (late infantile, juvenile, and adult). Material and Methods: Retrospective clinical, biochemical and neuroimaging data analysis of 15 MLD patients (9 male, 6 females). doi:10.1016/j.ymgme.2018.12.088 Results: Mean ages at onset of symptoms and at biochemical diagnosis were, respectively, 19 and 24 months (late infantile form). The most frequently reported first clinical symptom/sign of the disease was gait disturbance and other motor abnormalities (late Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S43 infantile form) and behavioral and cognitive alterations (adult form). due to adverse events considered drug-related. In summary, bone- Diagnosis was done only after 2-3 years of symptoms onset (late related parameters showed improvement in treatment-naïve pa- infantile form). Leukocyte ARSA activity level did not present tients and stability or improvement in switch patients. significant correlation with the age of onset of symptoms. All patients had high levels of sulfatides in urine (chromatography studies). Early dementia was the main feature of the two adult doi:10.1016/j.ymgme.2018.12.091 patients (with no motor feature associated). Discussion: Our results suggest that there is a considerable delay between the age of onset of signs and symptoms and the diagnosis of MLD in Brazil. Correlation between ARSA activity and MLD clinical 76 form was not found. Further understanding of MLD natural history, Osteoblast and adipose differentiation of Gaucher mesenchymal especially with larger samples, are necessary, in particular with the stem cells availability of specific treatments in a near future. Andrea N. Crivaroa, Constanza M. Bondara, Juan M. Muccia, doi:10.1016/j.ymgme.2018.12.090 Maximiliano Ormazabala, Ricardo A. Feldmanb, Victoria Delpinoc, Paula Rozenfelda, aInstitute of Immunological and Pathophysiological Studies, La Plata, Argentina, bDepartment of Microbiology and Immu- nology, University of Maryland School of Medicine, Baltimore, MD, 75 United States, cInstitute of Immunology, Genetics and Metabolism, Effects of oral eliglustat on skeletal manifestations in patients Autonomous City of Buenos Aires, Argentina with type 1 Gaucher disease: Results from four completed clinical trials after long-term treatment Gaucher disease (GD) is caused by mutations in GBA gene that encodes the lysosomal enzyme glucocerebrosidase. Type I GD (GD1) Timothy M. Coxa, Joel Charrowb, Elena Lukinac, Pramod Mistryd, patients present visceral, hematological and bone problems. Up to Theodore Marinakise, Meredith Fosterf, Sebastiaan J.M. Gaemersf,M. now, specific treatment for GD cannot completely reverse bone Judith Peterschmittf, aUniversity of Cambridge, Addenbrooke's Hospital, problems. Bone is a dynamic tissue that is continuously remodeling Cambridge, United Kingdom, bNorthwestern University Feinberg School in order to maintain proper structure. It is composed by hematopoi- of Medicine, Ann and Robert H. Lurie Children's Hospital of Chicago, etic cells as machrophages and non hematopoietic cells as mesen- Chicago, IL, United States, cNational Research Center for Hematology, chymal stem cells (MSCs). The first group is able to differentiate into Moscow, Russian Federation, dYale University School of Medicine, New osteoclasts in presence of MCS-F and RANKL, and the other one give Haven, CT, United States, eGeneral Hospital of Athens “G. Gennimatas”, rise to chondrocytes, osteoblasts and adipocytes, so an imbalance Athens, Greece, fSanofi Genzyme, Cambridge, MA, United States between these cells could lead to bone disease. The aim of this work is to evaluate the potential of MSCs from GD and control patients to Debilitating bone complications are common among patients differentiate towards osteoblast (GDOb) and adipocyte (GDAd) with Gaucher disease type 1 (GD1). We analyzed changes in bone lineages, as well as its effect in osteoclast formation. We observed parameters among 393 GD1 patients treated with oral eliglustat for reduced mineralization, collagen deposition and alkaline phospha- 4-8 years in 4 Sanofi Genzyme-sponsored clinical trials (Phase 2/ tase activity when MSCs from GD were cultured in osteogenic NCT00358150 [N=26] Phase 3: ENGAGE/NCT00891202 [N=40] differentiation media. Moreover, the expression of osteoblastic ENCORE/NCT00943111 [N=157] EDGE/NCT01074944 [N=170]. In differentiation markers ColA1, ALP, BMP-2 and Runx2 was lower in treatment-naïve patients (Phase 2/ENGAGE), mean spine T-scores GD cells. On the other hand, we cultured THP-1 cells with MCS-F and moved from the osteopenic range at baseline to the healthy range conditioned media obtained from Control and GDOb to evaluate the after 2-3 years. Mean±SEM spine T‑score increased from ‑1.55±0.28 induction of osteoclast formation. We observed an increase number to ‑0.59±0.34 (n=14) after 8 years in Phase 2, and increased to ‑0.53 of osteoclasts number when GDOb conditioned media was used ±0.27 in ENGAGE after 4.5 years (n=9). In both trials, spine Z-scores compared to Control. Furthermore, adipogenic differentiation re- improved and femur T- and Z-scores remained normal. In ENCORE vealed that GDAd produced lower levels of lipid droplets than (patients stabilized after a mean of 10 years of enzyme replacement Control Ad. In conclusion, our results show an alteration in GD MSCs therapy [ERT]), T- and Z-scores remained in reference ranges for ≤4 differentiation process towards osteoblast and adipose lineages, years least-square mean spine Z-scores improved by 0.29 being BMP-2 pathway implicated in the altered osteoblast differen- (Pb0.0001). In EDGE (mostly ERT switch), patients stabilized during tiation. Besides, GDOb demonstrate to be able to induce osteoclas- the Lead-In maintained normal T- and Z-scores through the dose- togenesis. These changes in GD MSCs could contribute to skeletal regimen and extension periods. Mean total bone marrow burden imbalance in GD. scores improved from marked-to-severe to moderate in ENGAGE and remained stable (moderate) throughout ENCORE and EDGE. Propor- tions of patients experiencing bone pain decreased after treatment in doi:10.1016/j.ymgme.2018.12.092 all trials reported pain became less severe. Bone crises were infrequent, with none in Phase 2 or ENGAGE patients and in 3/157 (1.9%) patients in ENCORE and 7/170 (4.1%) in EDGE (4 of whom had bone crises before trial entry). Median plasma Gaucher-related 77 biomarkers decreased substantially in Phase 2 (chitotriosidase, 91% Prevalence of mucopolysaccharidoses in samples sent to the glucosylsphingosine, 92%) and in ENGAGE (chitotriosidase, 82% laboratory of inborn errors of metabolism, Sao Paulo, Brazil glucosylsphingosine, 84%). Eliglustat was generally well-tolerated. Overall, 97% of adverse events were mild/moderate and 86% Vania D. Almeida, Ariane Souza, Joyce U.S. Yamamoto, Ana M. considered unrelated to eliglustat 9 (2.3% overall) patients withdrew Martins, Universidade Federal de São Paulo, Sao Paulo, Brazil S44 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Mucopolysaccharidoses (MPS) are a subgroup of lysosomal reproducibility in the activity and sensitivity to the test during the storage diseases caused by deficiency in enzymes involved in the period. Funding: CNPq, AFIP and IGEIM. degradation of glycosaminoglycans, which leads to the accumulation of them within the cells, tissues and organs, causing multiple symptoms. MPS are classified into eleven types according to the doi:10.1016/j.ymgme.2018.12.094 lack or defect in a specific lysosomal enzyme, being: MPS I, MPS II, MPS III (A, B, C, D), MPS IV (A and B), MPS VI, MPS VII and MPS IX. Our group performed, since 2010, diagnoses for MPS (type I, II, IVA, VI and, recently, MPS VII) in samples sent from all regions of Brazil. 79 fi Analyzes of the enzymatic activity were performed in dried blood 6-sulfatoxymelatonin daily pro le in Fabry disease patients: spot (DBS), with the exception of MPS IV, which was carried out in Relationship to disease variants leukocytes. In relation to MPS I, 1505 DBS samples were received with this suspicion and, 50 were diagnosed as positive, correspond- Vania D. Almeida, Julia R.S. Vallim, Fernanda G. Amaral, Universidade ing to 3.32% of the total. For MPS II, 836 DBS suspicion samples were Federal de São Paulo, Sao Paulo, Brazil received and 52 obtained a positive result, corresponding to 6.22%. For MPS IVA, 579 leukocytes samples were received in our laboratory Fabry disease is a Lysosomal Storage Disease caused by a fi for analysis and 56 presented a positive result, corresponding to mutation in GLA gene and the enzymatic de ciency of alpha 9.67% of the total samples evaluated. In relation to MPS VI 1349 DBS galactosidase A. Depending on type of mutation, enzyme activity fi samples were received, 108 presented a positive result, 8% of the and age of symptoms onset, patients can be classi ed into classic and total samples for this MPS. Until this moment, 217 samples were nonclassic variants. Fabry patients report changes in sleep-wake evaluated for MPS VII and none of them presented a positive result. rhythms, a behavior related to melatonin concentrations. Central fi Therefore, our laboratory evaluated in the last 10 years 4486 samples biosynthesis of melatonin can be assessed by quanti cation of its with suspicion of MPS, being 266 positive, that is, 5.93% of the total metabolite in urine, 6-sulfatoxymelatonin. The metabolic impair- samples evaluated, being MPS IVA the more frequent among them. ment in Fabry disease could have an impact on patient circadian Funding: CNPq, AFIP and IGEIM. rhythms, mainly daily melatonin excretion and, due to the different clinical manifestations of the variants, these changes could differ in classic and nonclassic patients. Therefore, we aimed to evaluate the doi:10.1016/j.ymgme.2018.12.093 excretion of 6-sulfatoxymelatonin in Fabry’s disease patients. Sixteen patients with biochemical and molecular diagnosis of Fabry's disease were classified in classic (N=104 women mean ±sd = 38.9 ± 3.6 years, range: 17-54 years) or nonclassic (N=65 women 46.2 ± 4.9 78 years, range: 27-62 years) variants. Control group was matched by Differential diagnosis for mucopolysaccharidoses: Evaluation of age and gender (N = 10, 7 women, 42.4 ±3.2 years, range: 23-59 β -glucuronidase activity years). Volunteers collected urine during the day (the first urine in the morning until 7 p.m.) and night (7 p.m. until the first urine the Vania D. Almeida, Ariane Souza, Ananda Esteves, Joyce U.S. next morning). As expected, we observed in controls an oscillatory Yamamoto, Ana M. Martins, Universidade Federal de São Paulo, Sao profile of 6-sulfatoxymelatonin, with greater excretion at night Paulo, Brazil (pb0.05 d=7.80). In classic patients this oscillation was absent, without difference between day and night excretion (pN0.05). Mucopolysaccharidoses (MPS) are a group of diseases caused by Nonclassic patients presented a phase shift, with greater excretion deficiencies in enzymes involved in the degradation of glycosami- during the day (pb0.05 d=3.85). We conclude that 6-sul- noglycans (GAGs) in lysosomes, which leads to the accumulation of fatoxymelatonin excretion, and probably melatonin biosynthesis, is GAGs causing various symptoms. There are eleven types of MPS, altered in Fabry disease and this modification depends on disease classified according to the enzymatic deficiency. MPS VII, named Sly phenotypes. This alteration could impact the sleep-wake cycle of Syndrome, is caused by a mutation in the GUSBgene encoding the β- Fabry patients. Financial support: CNPq, CAPES, AFIP, IGEIM. glucuronidase enzyme. Deficiency of this enzyme triggers an accumulation of three GAGs: dermatan sulfate, heparan sulfate and chondroitin sulfate in the lysosomes of many tissues triggering doi:10.1016/j.ymgme.2018.12.095 impairment of cellular functions. The diagnosis for MPS can be made by biochemical or molecular evaluation. The biochemical evaluation consists in the enzymatic activity measurement by fluorimetric assay, which could be performed on dried blood spot samples. Since 80 MPS are chronic, progressive and multisystemic diseases that present Nurse-led clinics for Lysosomal Storage Disorders Units (LSDU) - similar signs and symptoms, the differential diagnosis is necessary. In are we prepared? A scope of advanced nursing practice in the UK this study, a differential diagnosis was performed for MPS VII in centres samples that were referenced for other types of MPS evaluation during the last year (n = 208) in our laboratory. For positive control Amanda V. Daniel, Mark A. Mckie, Derralynn A. Hughes, The Royal for the disease, two MPS VII samples were used in each assay. The Free Hospital, London, United Kingdom normal range for β-glucuronidase at our laboratory was set above 6.0 nmol/mL/h. None of the samples tested showed a positive result, Nurse-led clinics are an important part of UK healthcare and which could be justified by the low prevalence of the disease. We although there is inconsistency in defining the role, it involves also verified β-glucuronidase activity stability on DBS samples from nurses’ managing a caseload of patients utilising skills such as four healthy subjects during six months and the results showed that physical assessment and independent non-medical prescribing the enzyme presented good stability on filter paper, with good following an educational programme at masters level. Although Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S45 research is needed around the effectiveness of nurse-led clinics correlate with residual disease activity as evidenced by bone marrow inLStorage D LSD, measured health outcomes in other areas appear involvement, decreased hematologic and bone density values. to be equivalent to medical run clinics. At the Royal Free Hospital, However, in GD patients with advanced disease including complica- London, there has been an overall increase of 39% in LSD patient tions such as history of osteonecrosis or gammopathy, no significant referrals in the last 5 years resulting in increasing numbers correlation was observed. In 15% of treated subjects, there was presenting to clinic. Rebalancing the workforce through nurse ongoing immune activation with increased CHITO levels but management of less complex patients, compensates for shortage in undetectable peripheral substrate accumulation. 10% of these medical personnel, reduces delays, improves patient experience and subjects had comorbidities or late GD complications. These results allows focus of medical time on challenging issues. There is also indicate that the immune response against the stored substrate in significant expertise amongst LSD nurses which could be harnessed GD is a self-propagating event with continued downstream effects. more effectively. Embracing advanced nursing skills in practice has Even as the substrate levels were normalized, disease activity may resulted in increased autonomy, greater recognition and improved continue to worsen. inter-professional working in addition to improved quality of care for patients. However, there needs to be clear criteria for which patients can be seen by nurses and when patients need to be referred. This doi:10.1016/j.ymgme.2018.12.097 study aims to review the national status and preparedness for nurse led clinics within LSD. A questionnaire sent to all the UK LSD centres evaluated whether centres are planning to, or have introduced such schemes the benefits and risks and whether any evaluation has 82 occurred. The second part reviews the current status of nursing Evaluation of daily activity patterns using a wearable device in advanced practice including available training, clinical governance Pompe disease and support for nurses in these roles. The study results identify the a b readiness for nurse-led clinics and suggest how they can be Pronabesh DasMahapatra , Christopher Curran , Eileen Mack- b a a a implemented. The results also highlight gaps in education and we Thorley , Anureet Pabla , Alaa Hamed , Sanofi Genzyme, Cambridge, b make recommendations for a unified approach for nurse education MA, United States, PatientsLikeMe, Cambridge, MA, United States in this field. Late-onset Pompe disease (LOPD) is a rare metabolic disorder caused by deficiency of lysosomal acid alpha-glucosidase (GAA) with ≥ doi:10.1016/j.ymgme.2018.12.096 age of onset 1 year. Patients experience progressive muscle weakness, leading to decreased mobility and respiratory weakness. Wearable activity trackers offer insights into real-world health and wellness in patients. Analyses were conducted to detect patterns of 81 activity and sleep from a prior study that monitored LOPD patients Evaluation of disease burden and therapy modifications using remotely with a Fitbit One device. Eligible patients (≥ 18 years, U.S. glucosylsphingosine (lyso-GL1) in Gaucher disease residents, diagnosed LOPD, ambulatory with/without aid, no invasive ventilation) were recruited from the Acid Maltase Deficiency Julia Dao, Ozlem Goker-Alpan, Renuka Limgala, Lysosomal and Rare Association. Patients received a wearable tracker (Fitbit One) for Disorders Research and Treatment Center, Fairfax, VA, United States remote monitoring after creating a PatientsLikeMe profile and were asked to complete self-assessment questionnaires at baseline, during, Gaucher Disease (GD) is an autosomal recessive disorder that and study exit. Distribution based method (mean, median, quartiles) results from mutations in the GBA gene encoding the lysosomal was used to characterize daily physical activity in the study enzyme acid β-glucosidase, resulting in accumulation of the population. Twenty-nine LOPD patients shared wearable data substrates, glucosylceramide (GL1) and its deacetylated form, patients were on average 43 years of age, 90% female, 93% diagnosed glucosylsphingosine (lyso-GL1). Build-up of these lipids in lysosomes in adulthood, and 86% on enzyme replacement therapy. The number of macrophages is associated with immune dysregulation and of days of available wearable data ranged from 13 - 112 days. Visual chronic inflammation. Although GL1 is the primary substrate of inspection of diurnal activity heat map showed increase in activity GCase, lyso-GL1 has been shown to correlate well with disease around hour 05:00, peaking around hours 10:00 and 18:00, and burden and hence plasma lyso-GL1 is used as a valuable biomarker declining again around hour 23:00. Median mobility levels between to monitor disease progression. In current study, 36 GD subjects 05:00 to 23:00 hours were 2,703 steps per day (lowest quartile b (10M/26F) were analyzed for plasma lyso-GL1 and correlation with 1,552, highest quartile ≥ 4,460) and 21 steps per active minute biomarkers for macrophage activation and disease involvement. The (lowest quartile b 11, highest quartile ≥ 37). Activity was lower on cohort included 10 pediatric (1-16 yr, mean: 7yr) and 26 adult Sundays (median: 2,307 steps) compared to other days of the week. patients (19-72 yr, mean: 41 yr). Plasma lyso-GL1 levels showed Exploratory correlations between weather and activity will be significant correlation with Chitotriosidase (CHITO) as well as CCL18 presented. Sleep analyses were limited due to imprecision in levels (p=0.002 and 0.01 respectively) demonstrating the relation- wearable measured sleep. Characterization of real-world mobility in ship between substrate accumulation and macrophage activation. In LOPD is important to understand the progression of disease and untreated patients (naïve or with prolonged treatment interrup- potential benefits of new therapies. Sponsor: Sanofi Genzyme tions), lyso-GL1 correlates with disease load and severity. The highest levels (up to 200 fold elevations) were observed in GD2 and GD3 patients. In a subset of GD patients undergoing treatment, doi:10.1016/j.ymgme.2018.12.098 modest elevations from baseline were still observed and may S46 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

83 involvement translates as a progressive loss of intellectual and motor Agalsidase beta delays the progression to kidney disease in Fabry function for NPC patients, who eventually succumb to NPC disease. patients: Results from an individual patient data meta-analysis At present, the NPC community has 3 distinct clinical trials in different phases of development. While very exciting, the expected Pronabesh DasMahapatraa, Steve Kantersb, Eugene Poggioc, Manish approval of one or even two compounds begs the question of Maskia, Elvira Poncea, Mario Aguiara, Rachel Goldgrubb, Dieter Ayersb, whether combination therapy will provide even greater benefit. To Jeroen Jansenb, Alaa Hameda, aSanofi Genzyme, Cambridge, MA, United address this important question, two combination therapy studies States, bPrecision Xtract, Bethesda, MD, United States, cBiostatistical were carried out in NPC1 mice: 2-hydroxypropyl-beta-cyclodextrin Consulting Incorporated, Lexington, MA, United States (HPBCD) + miglustat and HPBCD + gene therapy (AAV9.EF1a(s). hNPC1). Individually, all 3 therapies have shown significant disease Fabry disease (FD) is a rare, genetic disease that, if untreated, amelioration in animal studies. Mice were enrolled in the two studies progresses to irreversible and life-threatening renal, cardiac, and and efficacy of combination treatment was evaluated. Assays cerebrovascular events. Preventing organ damage is a critical included behavioral testing, weight monitoring, survival, disease treatment goal in FD. A meta-analysis was therefore conducted to pathology, and gene copy number. All data suggested greater disease estimate the effect of agalsidase β (agal-β) treatment on the amelioration with combination therapy as compared to individual reduction of chronic kidney disease (CKD) progression in FD. mono-therapies. These results are particularly enticing given the Individual patient data (IPD) were combined from phase III and IV recent surge of companies expressing interest in development of clinical trials, a natural history study and 6 studies identified from a therapies for NPC disease and even more so with respect for systematic literature review. Eligible patients were ≥ 16 years, had improving the quality of life for NPC patients. Funding generously the classic phenotype, a baseline serum creatinine below 3 mg/dl, provided by: NIH, SOAR-NPC, Ara Parseghian Medical Research Fund were not previously on FD specific therapy, and did not have prior at Notre Dame, NHGRI IRP, and Liferay Foundation. kidney transplantation. There were 315 eligible patients (161 treated). Data were analyzed using a robust 2-step approach: 1) change in eGFR over time was modeled using linear regression for doi:10.1016/j.ymgme.2018.12.100 each patient and 2) resulting annualized eGFR slopes were modeled using weighted quantile regression adjusting for imbalances in proteinuria and sex. The resulting equation was applied to project 85 the effect of agal-β on time to CKD progression. In males aged 16 2 Carpal tunnel syndrome in mucopolysaccharidosis type I Hurler- years, with baseline eGFR of 100 ml/min/1.73m and unchanged Scheie/Scheie and effect of enzyme replacement therapy negative/trace proteinuria over a lifetime, immediate initiation of agal-β delays both the progression to stage 3 CKD (40 vs. 12 years) and to stage 5 CKD (84 vs. 24 years) by three-fold relative to natural James E. Davison, Laura Guilder, Maureen A. Cleary, Matthew Pitt, history. Immediate initiation of agal-β also delays the progression to Benan Dala-Ali, Deborah Eastwood, Great Ormond Street Hospital, stage 3 CKD by two-fold (40 vs. 18 years) relative to patients London, United Kingdom initiating agal-β at age 25 years. Limitations include heterogeneity in fi annualized eGFR slopes across studies and extrapolation of projec- Neurophysiologially de ned carpal tunnel syndrome (CTS) is a tions beyond the 5-year follow-up. Robust analyses on a large recognised feature of mucopolysaccharidosis type I Hurler-Scheie evidence base suggest that early treatment with agal-β helps FD (MPS I-HS) affecting 39% of patients in a recent registry study. The patients conserve their renal function over much longer time periods effect of enzyme replacement therapy (ERT) on CTS in paediatric than in patients delaying treatment. patients is not well documented. This retrospective single-centre review reports the incidence of CTS determined by serial neuro- physiological studies and surgical interventions in a cohort of paediatric MPS I-HS patients treated with ERT and MPS I-S patients doi:10.1016/j.ymgme.2018.12.099 not receiving ERT. Nerve conduction studies were performed to determine median and ulnar sensory palm-wrist onset latency, amplitude and conduction velocity. Prolonged median:ulnar signal 84 latency indicated CTS with latency differential indicating severity of Improved disease amelioration with combination therapy for CTS. 11 children with MPS I-HS and 3 with MPS I-S were identified, Niemann-Pick disease type C1 median age diagnosis 4.75 years (range 2.72-9.71). All MPS I-HS patients commenced ERT. At the point of diagnosis, 5/11 MPS I-HS Cristin Davidsona, Alana Gibsona, Tansy Gua, Nafeeza Alib, Randy and 1/3 MPS I-S patients had evidence of severe CTS requiring Chandlera, Charles Vendittia, Steven U. Walkleyb, William Pavana, surgical intervention, 2/11 MPS I-HS and 2/3 MPS I-S had normal or aNational Institutes of Health, Bethesda, MD, United States, bAlbert mild CTS, and 1/11 MPS I-HS had moderate CTS. Of those patients Einstein College of Medicine, Bronx, NY, United States with normal, mild or moderate CTS at point of diagnosis, all remained stable or improved and none deteriorated over time or Niemann-Pick C (NPC) disease is a fatal, inherited metabolic required subsequent surgical intervention (median follow up disorder resulting from defects in one of two genes: NPC1 or NPC2. 4.2years, range 0.74-14.95). One patient who had surgical interven- Both of these proteins are believed central to a cell’s ability to traffic tion had subsequent evidence of unilateral deterioration of CTS. CTS cholesterol out of the lysosome to other cellular locations. A defect in occurred in 43% of the cohort and was evident in all these patients either the NPC1 or NPC2 protein results in significant lipid storage with a severe defect at the point of diagnosis. No patient without within late endosomal/lysosomal compartments, impeding normal severe CTS at diagnosis developed significant CTS during the follow cell function and eventually leading to cell death. Particularly hard up period. This finding is in contrast to the reported 57% of patients hit are neurons within the central nervous system. Clinically, the CNS with attenuated MPS I who developed CTS after initiating treatment Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S47

(Viskochil 2017). Further, there was no correlation between urine appropriate monitoring and management critical. Spinal bone glycosaminoglycans concentration, iduronidase activity or genotype density is a well-recognized independent risk factor for fracture with the occurrence of CTS. however, utilizing a widely available set of clinical and demographic characteristics may also provide a useful GD-oriented risk assessment tool. We developed a fracture risk score combining the 5 doi:10.1016/j.ymgme.2018.12.101 following factors derived from previous research on fracture in GD1 patients: sex, age at treatment initiation (ATI), time interval between diagnosis and treatment initiation, splenectomy status and prior bone crisis at treatment initiation. GD1 patients in the International 86 Collaborative Gaucher Group (ICGG) Gaucher Registry Hypogammaglobulinemia, impaired vaccine response and recur- (NCT00358943) who initially received alglucerase/imiglucerase rent infections in mucolipidosis type II treatment and had known splenectomy status and at least one skeletal assessment were included in the analyses (3143 of 6204 a b a James E. Davison , Francesco Saettini , Maureen A. Cleary , Adrian patients) (data-download: 01JUN2018). Data were analyzed sepa- a a a Thrasher , Claire Booth , Great Ormond Street Hospital, London, rately by ATI group (b18, 18 to b50, or N=50 years of age) using Cox b United Kingdom, University of Milan-Bicocca, Monza, Italy proportional hazards regression with all 5 risk factors included in the multivariable model. A risk score was calculated by summing the Mucolipidosis (ML) II is a rare and fatal autosomal recessive disease contribution of each parameter weighted by the strength of its fi affecting lysosomal enzyme traf cking. Immunological abnormalities association with fracture risk. Patients were followed from the date have not been extensively studied in this patient population but of treatment initiation (or age 18 for patients if treatment started research suggests the immune system may be affected. Here we earlier) to the date of first adult fracture (n=275 first fracture evaluated the immune status of MLII patients at our institution. 6 endpoints), death, or end of follow-up. Across all 3 ATI groups, the fi fi patients with con rmed MLII (age 7 months-7 years) were identi ed composite risk score was associated with a 2.7-fold increased and clinical data on frequency of infection and results of any (pb0.01) risk of adult fracture for each one-point increase in score. immunological investigations were collated. Recurrent respiratory These analyses provide the clinician with a GD-oriented tool to be infections requiring multiple admissions were common. Viral respira- used in conjunction with bone density and other non-GD-specific tory infections can be severe (two patients experienced H1N1/VZV and risk factors (e.g. smoking, alcohol, frailty) to better assess age- ADV/RSV infection) proving fatal in one. Antibiotic prophylaxis was specific fracture risk and to help guide patient monitoring, use of initiated in 5/6 patients with no reduction in admission rate. One preventative treatments and appropriate life style modification. patient commenced immunoglobulin replacement therapy resulting Sponsored by Sanofi-Genzyme. in a reduction in infection frequency. Although white cell counts were variable, severe B-cell lymphopaenia was evident in one patient and B doi:10.1016/j.ymgme.2018.12.103 lymphocytosis in another one. Memory B cells were decreased in 2 patients. Hypogammaglobulinemia, particularly affecting IgM was present in 3/4 patients with panhypogammaglobulinaemia in one. 3 of 5 patients showed impaired vaccine responses (to tetanus or 88 fi pneumococcus). Interestingly one patient, after VZV pneumonia, NGS-based, 107-gene resequencing panel as rst-line screening failed to produce VZV IgG. Analysis of T cell compartment showed test for lysosomal diseases increased T cell absolute count (3/4 CD3+, 2/4 CD4+, 4/4 CD8+, 2/4 a a a CD4+/CD8+). Hypogammaglobulinemia and impaired vaccine re- Francisco J. del Castillo , Gloria Muñoz , Crina Ciubotariu , Marta a a a b sponses are evident in MLII and should be evaluated in patients, who Gandía , Juan M. Rosa , Miguel Piris , Luis G. Gutiérrez-Solana , María c d e may benefit from prophylactic strategies. Further work is required to Teresa García-Silva , Katrin Palk , Antonio González Meneses , Josep f g g h h understand the mechanism underlying these immune abnormalities. M. Grau , N. Fdil , Es Sabir , Nadia Slitine , Imane Ait Sab , Aicha Bourrahouath, Noureddine Radah, Mohammed Bouskraouih, Luis J. Aldámiz-Echevari, Jesús Villarrubiaa, aHospital Universitario Ramón y b doi:10.1016/j.ymgme.2018.12.102 Cajal, IRYCIS, Madrid, Spain, Hospital Infantil Universitario Niño Jesús, Madrid, Spain, cHospital Universitario Doce de Octubre, Madrid, Spain, dNorth Estonia Medical Centre, Tallinn, Estonia, eHospital Universitario Virgen del Rocío, Sevilla, Spain, fHospital Clinic, Barcelona, Spain, gCadi 87 Ayyad University, Marrakech, Morocco, hCHU Mohammed VI, Marra- A composite fracture risk score for assessing adult fracture risk in kech, Morocco, iHospital Universitario de Cruces, Barakaldo, Spain imiglucerase-treated type 1 Gaucher disease patients using data from the International Collaborative Gaucher Group (ICGG) Determination of lysosomal enzyme activity in leucocytes or Gaucher Registry fibroblasts is currently considered the gold standard for the diagnosis of lysosomal storage diseases (LSDs). However, two problems Patrick Deegana, Aneal Khanb, José S. Cameloc, Julie Batistad, Vipin hamper obtaining an early and accurate diagnosis: (1) there are Aggarwald, Neal Weinrebe, aAddenbrooke's Hospital, Cambridge, not assays for every lysosomal protein underlying a LSD and (2) the United Kingdom, bUniversity of Calgary, Calgary, AB, Canada, relative unspecificity of early symptoms and signs in many LSDs does cUniversity of São Paulo, São Paulo, Brazil, dSanofi Genzyme, Cambridge, not provide clues on which particular enzyme tests should be MA, United States, eUniversity of Miami Miller School of Medicine, considered first. Those problems are compounded by the fact that Miami, FL, United States tests for LSDs are usually performed at reference laboratories and most clinical services resort for a first screening to sending dried Fractures in Gaucher disease type 1 (GD1) patients can lead to blood spot (DBS) samples, with lesser sensitivity and specificity than significant morbidity in both treated and untreated patients making the gold standard. We have tested the efficiency of using a next- S48 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 generation sequencing (NGS)-based gene panel as a first screening 90 test for LSDs. We devised and validated a 107-gene resequencing Changing the clinical course of infantile Pompe disease with panel that interrogates 47 of the genes underlying LSDs plus the immune modulation strategies: 12 years of experience genes involved in their differential diagnoses, covering in all 207 different OMIM phenotypes. We analyzed genomic DNA samples Ankit K. Desaia, Zoheb B. Kazia, Deeksha S. Balia, Catherine W. from 24 unrelated patients referred to our unit from hospitals in Rehdera, Susan Richardsb, Amy S. Rosenbergc, Priya S. Kishnania, Spain, Estonia and Morocco. Among them, we identified mutations aDuke University, Durham, NC, United States, bSanofi Genzyme, underlying 11 cases of Hurler syndrome/MPS Ih (IDUA), 2 cases of Framingham, MA, United States, cUS FDA, Bethesda, MD, United States Sanfilippo disease 3A/MPS IIIA (SGSH), and single cases of Hunter syndrome/MPS II (IDS), Tay-Sachs disease/GM2 gangliosidosis Cross-reactive immunologic material (CRIM)-negative infantile (HEXA), Sandhoff disease (HEXB) and thrombocythemia-2 (MPL), Pompe disease (IPD) patients develop high and sustained antibody for a diagnostic rate of just over 70% (17/24 cases). Our experience titers (HSAT ≥51,200) and sustained intermediate titers (SIT ≥12,800 indicates that when our NGS panel results are backed up with and b51,200) within 12 months on enzyme replacement therapy subsequent enzyme tests and discussed within a committee of (ERT) resulting in clinical decline and ultimately ventilator depen- clinical and genetic experts, an accurate diagnosis can be reached in dence or death by 27.1 months. Approximately 40% of CRIM-positive a very short time (usually under one month). Funding: This work IPD also develop HSAT and SIT with outcomes similar to CRIM- was supported by Sanofi Genzyme. negative. Prophylactic immune tolerance induction (ITI) with rituximab, methotrexate, and/or IVIG and transient low-dose methotrexate (TLD-MTX) have helped in achieving immune tolerance in doi:10.1016/j.ymgme.2018.12.104 CRIM-negative and CRIM-positive patients. We present long-term experience on 43 CRIM-negative and CRIM-positive IPD patients on various ITI regimens. Data on 29 (22 CRIM-negative and 7 CRIM- positive) patients who received this ITI protocol was assessed for 89 long-term safety and efficacy. Overall, the median antibody titer at Severe cardiac involvement: Management in a homozygous final assessment for 22 CRIM-negative was 100 (range 0-25,600) at D409H Gaucher patient under enzyme replacement therapy median time since ERT initiation of 69 weeks (range 11-285 weeks). Of the 22 CRIM-negative patients, 50% (n=11) did not seroconvert. Mireia del Toro, Antonio Segura, Clara Carnicer, Ana Felipe, Guillem Five CRIM-negative patients required more than one cycle of ITI. Pintos, Marc Moltó, Jose A. Barrabes, Hospital Vall d'Hebron, Barcelona, Sixteen CRIM-negative patients were alive, with median age of 65 Spain months (range 7-133). CRIM-positive patients (n=7) did not seroconvert or maintained low titers (LT) with highest observed Gaucher disease (GD) type 3 with homozygous D409H mutation has antibody titer of 200. An attenuated TLD-MTX protocol has been been associated with a particular phenotype which includes oculomotor implemented in 13 CRIM-positive and 1 CRIM-negative patients. In fi apraxia and cardiac valve calci cations in late childhood. We report a TLD-MTX group, 86% (12/14) patients maintained LT, 7% (1/14) patient with type 3 D409H phenotype and severe cardiac involvement patient developed SIT, and 7% (1/14) patient developed HSAT. fi treated with enzyme replacement therapy (ERT) since the rst year of Fourteen TLD-MTX recipients had a median last titer of 150 (range life. A 22-year-old patient was diagnosed with GD at the age of 45 days 0-51,200) at median time since ERT initiation of 83 weeks (range 36- because of massive hepatosplenomegaly and showed to be homozygous 122 weeks). Overall, both these protocols were safely tolerated. This for D409H mutation. ERT was started at the age of 2 months with represents the largest cohort of IPD patients (n=43) treated with alglucerase and switched to imiglucerase at the age of 2 years, which has prophylactic ITI. These data suggest that implementation of prophy- been well tolerated. Clinically he developed an oculomotor apraxia at lactic ITI is safe and efficacious at eliminating or minimizing the the age of 6 and corneal opacities at the age of 14. At the age of 13, a deleterious immune response to ERT. routine echocardiographic exam was strictly normal. However, 2 years later he was re-evaluated for chest pain and was diagnosed of severe aortic stenosis and valve calcifications with coronary artery disease. At doi:10.1016/j.ymgme.2018.12.106 the age of 15 he underwent aortic valve replacement and left anterior descending artery (LAD) revascularization with a internal mammary artery graft. He required several admissions for chest pain showing progression of the lesions and he underwent several percutaneous 91 revascularization procedures with implantation of stents. At the age of Quantitative muscle MRI in Pompe disease: A 4 years follow-up 16, he was admitted for acute myocardial infarction. A cardiac MRI exam study showed a subendocardial infarction in LAD and LCx territories with a left ventricular ejection fraction of 45%. From then on he has required Jordi Díaz-Manera, Claudia Nuñez-Peralta, Sonia Segovia, Izaskun several percutaneous revascularization procedures for restenosis. He is Belmonte, Irene Pedrosa, Elena Montiel, Alicia Alonso-Jimenez, JOrge now on medical management, with NYHA class II dyspnea, sporadic Alonso-Pérez, Jaume Llauger, Ana Carrasco-Rozas, Esther Fernández- chest pain and LVEF of 40%, waiting for cardiac transplant. To our Simon, Xavier Suarez-Calvet, Eduard Gallardo, Isabel Illa, Hospital de knowledge this is the oldest D409H homozygous patient treated with la Santa Creu i Sant Pau, Barcelona, Spain ERT from infancy. Treatment hasn’t prevented the development of rapidly progressive cardiac involvement. Late onset Pompe disease is characterized by progressive weakness of skeletal and respiratory muscles. Enzymatic replacement therapy (ERT) changes natural history of the disease. A recently doi:10.1016/j.ymgme.2018.12.105 published long term follow-up study suggests that patients continue worsening despite the treatment. However, this fact has not been Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S49 confirmed in other cohorts. We have followed-up a long cohort of 37 years, and this observation beyond saving millions of dollars, allows LOPD for a period of 4 years. We have visited them once per year treatment-free life, which in the case of ERT is associated with a life-time performing the following procedures: clinical interview, muscle dependency on bi-weekly intravenous treatment, and in the case of SRT function tests (MRC scale, hand-held dynamometry, time to walk 10 with the need for a daily or twice daily use of a capsule, with non-trivial meters, 6 minutes walking test, timed up&go test, MFM-20), quantita- safety profile. tive muscle MRI (qMRI) using 3-point Dixon and daily live activity scales (Activlim and rPact). 29 patients were symptomatic while 8 patients had no muscle weakness at baseline visit. All symptomatic patients doi:10.1016/j.ymgme.2018.12.108 were treated with ERT. In the case of symptomatic patients, we identified a significant decrease in forced vital capacity (FVC) after 4 years of follow up while muscle function tests remained stable. qMRI demonstrated a significant increase on fat present in muscles of the 93 ’ thighs and trunk during the follow-up, that ranged from 5 to 15%. In the A3-UTR variant in SCARB2 modulates LIMP2 in patients with case of asymptomatic patients, we did not identify significant changes in Gaucher disease and myoclonic epilepsy the muscle tests and spirometry despite we observed a significant increase on fat present in paraspinal muscles. In summary, muscle fat Jenny Do, Sam Gary, Barbara Stubblefield, Emory Ryan, Grisel Lopez, replacement increased in trunk and thighs muscles in symptomatic Ellen Sidransky, Nahid Tayebi, National Institutes of Health, Bethesda, LOPD patients using qMRI despite being treated, although most of MD, United States muscle function tests remained unchanged except for a decrease in the FVC. Therefore, qMRI seems to be more precise to follow-up muscle Lysosomal integral membrane protein 2 (LIMP2) is essential for fi degeneration than conventional muscle function tests. These results lysosomal targeting of the enzyme glucocerebrosidase (GCase). De - support the inclusion of qMRI in clinical trials and daily clinics for the ciency in GCase causes one of the most common lysosomal storage follow-up of LOPD patients. disorders, Gaucher disease (GD). SCARB2, the gene encoding LIMP2, has been implicated as a genetic modifier in patients with GD who develop progressive myoclonic epilepsy (PME). Sequencing of SCARB2 in three fi doi:10.1016/j.ymgme.2018.12.107 patients with GD/PME identi ed two heterozygous coding mutations in two of the patients. However, a heterozygote, single-nucleotide deletion at the 3’-UTR (c. 2611delT) was present in all three patients. Quantitative PCR showed that SCARB2 RNA expression in the patients with GD/PME 92 was significantly reduced compared to two age-matched controls. Long-term follow-up of 103 untreated adult patients with type 1 Western blots of GCase and LIMP2 revealed significantly lower GCase Gaucher disease and LIMP2 protein levels when compared to controls. Additionally, GCase activity assay yielded significantly lower GCase activity in the probands. Tama Dinura, Ari Zimrana, Michal Becker Cohena, David Arkadirb, We hypothesized that the sequence encompassing the SCARB2 3’-UTR Claudia Cozmac, Marina Hovakimyanc, Sebastian Oppermannc, Arndt variant found in the patients with PME might be a target site for miRNA Rolfsc, Shoshana Revel-Vilka, aShaare Zedek Medical Center, Hadassah binding, thereby downregulating SCARB2 expression. Using the Human Medical Center, The Hebrew University, Jerusalem, Israel, bHadassah TargetSearch algorithmic website, we selected candidate miRNA based Medical Center, The Hebrew University, Jerusalem, Israel, cCentogene, upon their sequence homology to the mutated SCARB2 3’ UTR site. Four Rostock, Germany miRNA sequences, miR-935, miR-153-5p, miR-9-3p and miR-182-5p were identified. These miRNAs are known to regulate post-transcrip- The introduction of disease specific therapy for patients with adult- tional gene expression, and are reported to be associated with epilepsy type Gaucher disease (GD1) was a revolution in the management of and neural development. A TaqMan MicroRNA assay will be used to patients, but not without cost. Thus, the management of mildly effected quantify and validate the miRNAs in patient RNA and plasma samples. patients is still debated. In this study, we report a long-term follow up of Further studies in additional patients with PME with and without GD are 103 (45 males) GD1 patients at a mean (range) age of 43.48 (18-84) being performed. Identifying miRNAs that regulate SCARB2 expression years, who have never received any specific therapy (enzyme replace- will aid our understanding of how miRNAs contribute to mRNA export, ment therapy (ERT) and substrate reduction therapy (SRT)). Nine translation of LIMP2, and LIMP2 protein stability, as well as how they may patients who were unwilling/unable to receive treatment, but fulfilled serveasbiomarkersincaseswithPME. the criteria for ERT in Israel, were excluded. Of 103 patietns,92 were homozygous for the N370S mutation and 11 N370S were compound heterozygotes. The mean (range) age at diagnosis and time from doi:10.1016/j.ymgme.2018.12.109 diagnosis were 23.89 (0-63) years and 18.4 (0.3-58) years, respectively. At the time of last follow-up, the mean (range) lysol-Gb1 value, platelet count and hemoglobin levels were 129.9 (7.1-465) ng/ml, 163 (56-408) 10^3/ul and 13.30 (7.9-16.8) g/dl, respectively. The mean (range) spleen 94 and liver size (multiples of normal values MN) 6.12 (0-13.71) and 1.25 From birth to the sixth decade - A natural history study of 42 (0.74-4.0), respectively. Nine patients underwent splenectomy at a mean patients with neuronopathic Gaucher disease (range) of 28.77 (3-41) years prior to last analysis. 4 patients developed a a b Parkinson Disease (PD) at a mean (range) age of diagnosis of 49.75 (43- Aimee Donald , Simon A. Jones , Ashok Vellodi , Gaucherite Con- c d a 61) years. No episodes of avascular necrosis or other significant bone sortium , Timothy Cox , St Marys Hospital, Manchester, United b complications occurred during the entire follow-up period. The mean Kingdom, Great Ormond Street Hospital, London, United Kingdom, c d (range) lumber spine T score was -0.86 (-2.9 to 2.8). This rather large University of Cambridge, Cambridge, United Kingdom, Univeristy of cohort confirms that mildly affected patients may remain stable for many Cambridge, Cambridge, United Kingdom S50 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Neuronopathic Gaucher Disease (nGD) is subtype of the lysosomal pathogenic variant in GLA, consistent with Fabry disease. He reported a storage disorder resulting from deficiency of the enzyme history of hypohidrosis with an inability to run in hot weather, Glucocerebrosidase. Disease is as a result of autosomal inheritance of acroparesthesias, tinnitus with decreased hearing, and angiokeratomas. mutations of the GBA1 gene. Enzyme Replacement Therapy has He has sinus bradycardia on EKG. He has begun treatment with enzyme revolutionised the way Lysosomal Storage Disorders are managed replacement therapy. His brother and mother were also identified to and improved morbidity and mortality outcomes significantly. ERT have Fabry disease. In summary, we report three individuals with LSDs fails to penetrate the blood brain barrier and modify disease for identified by ECS. Three additional relatives have also been identified. To patients with nGD, as such, these patients experience disease our knowledge, this is the first report of clinical features in those progression. A series of therapeutic approaches are being employed identified. We expect to see increasing numbers of individuals with to address this unmet need and trials are in varying levels of LSDs identified by ECS as its use expands. development a fundamental limitation to trial design is the lack of clinical outcome measures due to inadequate natural history data. We present the natural history of nGD following detailed review of 42 doi:10.1016/j.ymgme.2018.12.111 patients from the observational study GAUCHERITE, in the UK. All living patients (n=32) were reviewed prospectively over a three-year period with serial examination by a single examiner. Parallel retrospective review of all case-notes and investigations available 96 was undertaken to generate natural history data spanning paediatric RVT-801, a developmental enzyme replacement therapy for and adulthood periods. When analysed in genotypic groups, clear Farber disease, ameliorates characteristic features of the disease consistency of phenotype and rate of disease progression was seen phenotype in a Farber mouse model suggesting genotype-phenotype correlation. Patients with genotype a b a c L444P/L444P demonstrated a slow consistent progression of disease Katie S. Duke , Christine M. Coquery , Eric J. Gaukel , Xingxuan He , c d e over time with non-neurological features contributing significantly to Edward H. Schuchman , Melissa Rhodes , Alexander Solyom , Brante a a b morbidity. Although significant difference in rate of disease progres- P. Sampey , Enzyvant, Durham, NC, United States, Immunovant, c sion by genotypic group was identifiable (as measured by mSST score) Durham, NC, United States, Ichan School of Medicine at Mount Sinai, d the pattern of clinical progression was consistent across all groups. New York, NY, United States, Altavant, Durham, NC, United States, e This natural history description is unique in the depth of detail and the Enzyvant, Basel, Switzerland duration of follow up up to the sixth decade. We summarise the experience of disease progression for this large cohort of patients with Farber disease is a progressive ultra-rare lysosomal storage a view to suggest the best clinical outcomes for use in trial design. disorder with a broad phenotypic spectrum, resulting from loss-of- function mutations in the acid ceramidase (AC) gene, ASAH1. Farber doi:10.1016/j.ymgme.2018.12.110 disease is characterized by intracellular accumulation of the AC substrate, ceramide, leading to increased monocyte chemoattractant protein (MCP)-1, and monocytic tissue infiltration resulting in painful nodules, granulomas, and joint pathology with progressive 95 symptoms and most often premature death. RVT-801 is a novel Identification of lysosomal diseases by expanded carrier human recombinant AC in development as an enzyme replacement screening therapy to treat patients with Farber disease. A severe Farber disease “knock-in” mouse model, characterized by ceramide accumulation, Theodore G. Drivasa, Stephanie B. Asherb, Maria Bonannib, Staci increased MCP-1 and monocytic infiltration of multiple tissues, was Kallishb, aThe Children's Hospital of Philadelphia, Philadelphia, PA, dosed intraperitoneally with 0, 0.1, 1, 3 or 10 mg/kg RVT-801 once United States, bHospital of the University of Pennsylvania, Philadelphia, weekly for 6 weeks. Endpoints assessed were joint, bone and major PA, United States organ histopathology, tissue ceramide and sphingosine levels, plasma MCP-1, body weight, and organ weight. Here we report, for Use of expanded carrier screening (ECS) has become more the first time, the identification and characterization of histiocytic widespread and the number of disorders included has increased with granulomatous infiltration of the soft tissues of the femoro-tibial time. As expected, those affected with the disorders included in the joint. We also report novel amelioration of joint pathology resem- screening may be identified. One study of ECS in more than 23,000 bling clinical nodules and granulomas, including normalized bone individuals (GIM 2013) found 78 who were homozygous or compound structure in Farber mice following repeat dose RVT-801 treatment. heterozygous for one of the included conditions, including 4 with Moreover, RVT-801 administration produced a dose-dependent Gaucher disease, 1 with hexosaminidase A deficiency, 1 with Niemann decrease in plasma MCP-1 and ceramide levels in the liver, spleen, Pick C disease, and 1 with Pompe disease. It was not reported if these kidney, heart, brain and muscle, increased sphingosine in a subset of diagnoses were previously known to those identified or if these tissues, improved body weight gain, and resolved splenomegaly. individuals were symptomatic. Our center has evaluated 3 individuals Systemic, repeat dose administration of RVT-801 in Farber mice was with LSDs identified by ECS, 2 with Pompe disease and 1 with Fabry well tolerated and impacted hallmark features of Farber disease, disease. Two women were identified with late-onset Pompe disease including amelioration of bone, joint and soft tissue lesions and both homozygous for the c.-32-13TNG splice site pathogenic variant. reducing levels of MCP-1. RVT-801 therapeutic effects in the severe One was identified during pre-pregnancy ECS she was asymptomatic. Farber mouse model indicate its potential as an enzyme replacement She has an identical twin, also asymptomatic, who has not presented to therapy for human Farber disease. care. The other woman was identified by ECS during pregnancy. She reports poor coordination from childhood and fatigue (increasing during pregnancy) initial evaluation revealed low-normal GAA enzyme doi:10.1016/j.ymgme.2018.12.112 activity and normal urine Hex4. ECS also identified a male patient with a Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S51

97 patients. In particular, the impact of co-existing pathologies and co- Mucopolysaccharidosis type VII: Clinical and biochemical data of morbidities on renal dysfunction was examined. Patient demo- 8 patients from Argentina graphics, measured GFR, urine protein, renal histology where available and clinical co-morbidities were recorded. Three groupings Consuelo Duranda, Joaquín Frabasila, Juan Politeia, Nora Atanacioa, of patients were identified: Group A, 21 patients in whom renal Sebastian Fortinib, Romina Floresc, Georgina Mateosd, Andrea histology was available from renal biopsy, nephrectomy, or post- Chirinoe, Andrea B. Schenonea, aLaboratorio de Neuroquimica Dr. N.A. mortem. Group B, 10 patients with end stage renal failure of whom 7 Chamoles, Ciudad Autónoma de Buenos Aires, Argentina, bHospital del had biopsy data Group C, 18 patients with CKD stage 3 and 4 in Niño Jesús, San Miguel de Tucuman, Argentina, cHospital Churruca whom no histological data available. GFR at the time of analysis in Visca, Ciudad Autónoma de Buenos Aires, Argentina, dex Hospital Dr. patients in whom renal biopsy had been performed ranged b15- Guillermo Rawson, San Juan, Argentina, eClinica y Maternidad del Sol, 61ml/min and proteinuria b0.15-1.75g/24 hour Analysis of clinical Cordoba, Argentina and histological data revealed a significantly higher proportion of patients with co-existing renal pathologies in late onset compared to Background: MPS VII is a very rare and variable disorder due to β- classic patients with CKD3-5 (p=0.0362) and CKD3-4 (p=0.0318). Glucuronidase deficiency. Patients’ phenotypes vary from severe Co-existent pathologies included diabetic nephropathy, IGA ne- forms with non immune hydrops fetalis (NIHF), skeletal dysplasia phropathy and focal segmental glomerulosclerosis. The study and mental retardation to milder forms with fewer manifestations. highlights the need to consider additional explanations for renal Methods: A retrospective medical record review was conducted at impairment in females and later onset patients with FD with a low our center for this case series of 8 patients with MPS VII. We threshold for renal biopsy. analyzed the biochemical and clinical data including: family history, age of onset, signs and symptoms and natural progression of the disease. Results: We collected information on 8 patients from doi:10.1016/j.ymgme.2018.12.114 Argentina from 2001 to 2018. Two patients had NIHF and died in the neonatal period they had family history of NIHF. Six patients had a clinical disease similar to MPS I, including the same degree of 99 clinical heterogeneity. None of the patients has undergone bone How do we explain very discordant phenotypes among three marrow transplantation or enzyme replacement therapy yet. We siblings with neuronopathic Gaucher disease? Whole exome performed β-Glucuronidase activity in DBS and confirmed it in sequencing and transcriptome analyses leukocytes. Discussion: MPS VII is a lysosomal storage disease with variable phenotype. As NIHF is a distinguishing clinical feature of MPSVII, we consider that MPSVII should be part of the workout of Areian Eghbali, Eric Garcia, Yuichiro Tanima, Pankaj Sharma, Frank fi NIHF. The other clinical aspects are similar to MPSI, so we suggest Donovan, Abdel Elkahloun, Alta Steward, Barbara Stubble eld, using the same selective screening methods used to detect MPSI, to Settara Chandrasekharappa, Grisel Lopez, Ellen Sidransky, Nahid detect this rare disease. Tayebi, National Institutes of Health/National Human Genome Research Institute, Bethesda, MD, United States doi:10.1016/j.ymgme.2018.12.113 Gaucher disease (GD) is a rare, monogenic, lysosomal storage disorder due to mutations in GBA1, which lead to the deficiency of the enzyme glucocerebrosidase (GCase) and the consequent accumulation of its substrate glucocerebroside. Of the three types of GD, 98 type 3 or chronic neuronopathic GD includes a spectrum of Renal involvement in classical and late onset patients with Fabry phenotypes, encompassing manifestations including abnormal hor- disease and the role of co-existing pathologies izontal saccadic eye movements, myoclonic epilepsy, cardiac calcification, and hydrocephalus, among other abnormalities. Moreover, Hatim Y. Ebrahim, Lucia Lavalle, Brendan Beaton, Matthew C. Reed, this type is associated with learning disabilities in certain patients. Shabbir Moochhala, Uma Ramaswami, Atul Mehta, Derralynn A. We present data from a family with three affected children, Hughes, Royal Free London NHS Foundation Trust, London, United homozygous for mutation L444P, who each have different neurolog- Kingdom ical manifestations, including one boy with profound autism. DNA and RNA were extracted from fibroblast lines from the parents and Fabry Disease (FD), OMIM 301500, due to a deficiency of alpha children. Illumina’s HiSeq and Affymetrix ClariomTM S transcriptome galactosidase A, results in cellular accumulation of assays were used to investigate the whole exome sequencing (WES) globotriaosylceramide and organ dysfunction. Two presentations of and the transcriptome-level expression profile of the samples, the condition are described: classical disease which manifests early respectively. GenomeStudio, R-Studio, and Affymetrix Transcriptome with acroparasthesia, angiokeratoma and sweating abnormalities Analysis Console were employed to analyze the transcriptome data. progressing to multiple organ dysfunction and later onset disease Compound heterozygous mutations were identified in the gene associated with higher residual alpha-galactosidase A activity, and AP3D1 only in the child with autism. This gene encodes a protein single organ dysfunction most commonly cardiac. Whilst renal involved in Golgi membrane structure, and could impact protein impairment progressing to end stage renal failure is well described trafficking to lysosomes. Several other highly significant variants are in classical males it is less common in classical females and males under evaluation. From the transcriptome data, we identified several and females with late onset disease. In this study, renal involvement genes that are critical for neuronal development including NSG1, in two-hundred and fifty-five FD patients attending the Royal Free PPF1BP2, and IL1RAP, that were differentially expressed among the Hospital NHS Foundation Trust was retrospectively reviewed to children (fcN2orfcb-2). Further RNA and protein analyses of the assess features of renal presentation in classical and late onset products of these candidate genes are underway. The findings will S52 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 then be verified in other patients with type 3 GD to establish their responses to therapeutic interventions. In Gaucher disease (GD), a potential role as modifiers impacting type 3 GD phenotypes. Such pan-ethnic lysosomal disorder, there are further variables such as the studies utilizing genomic and transcriptomic strategies can shed light recently-recognized risk of co-morbid disorders such as Parkinson’s on the genetic factors underlying the diversity of phenotypes disease and some cancers, and the psychosocial overlay of chronic observed in these patients. disease with impact on significant others. A GD-specific PRO questionnaire was developed with input from 75 patients and 10 parents of patients in Israel. A qualitative interview study assessed doi:10.1016/j.ymgme.2018.12.115 content validity in 33 patients in the US, France and Israel according to FDA standards. First, a concept elicitation exercise explored patient experience of symptoms and treatments findings were used to assess conceptual coverage of the questionnaire. A cognitive 100 debriefing exercise involved patients completing the questionnaire Lipidomics in translational research and clinical relevance for the using a “think aloud” process, and specific probing questions to fi fl identi cation of biological uids sphingolipids biomarkers for explore patients’ understanding and relevance of instructions, muccopolysaccharidoses items, response scales, and recall period. Five expert clinicians and a patient advocate were engaged as advisors at key stages and Farah El Turk, John J. Mitchell, McGill University Health Center, helped inform modifications to the questionnaire. The question- Montreal, QC, Canada naire includes 15 questions with a 6-point Verbal Response Scale plus nine 0-10 Visual Analogue Scales (VAS) is intended for fi Lipids are emerging at the forefront of scienti c research due to routine patient monitoring the clinical trial version includes eight their prominence in the metabolism and pathogenesis of various of the questions and all the VAS time to respond is usually b7 fl diseases, ranging from neurodegenerative, in ammatory to meta- minutes. The objective of this tool is to serve patients and bolic disorders. With the development of mass spectrometric physicians as a quantitative and qualitative approach to a more technologies and methodologies as tools for analysis, lipidomics is complete range of GD concerns, as an impetus to dialogue fi allowing a quantitative and comprehensive analysis of both speci c especially of sensitive/intimate concerns, and of symptoms not lipid classes as well as individual lipid species within a biological assessed in generic Health-Related Quality of Life questionnaires. system, and therefore, a better understanding of lipid-driven This study and writing support were funded by Shire. mechanisms. Mucopolysaccharidoses (MPS) are a group of rare inherited recessive metabolic disorders characterised by the paucity of the necessary lysosomal enzyme(s) required for the stepwise doi:10.1016/j.ymgme.2018.12.117 breakdown of specific metabolites, the glycosaminoglycans (GAGs), leading to accumulation in tissues and biological fluids. Despite its unique clinical manifestations, diagnosis delay of MPS still occurs. 102 GAGs were proven to be inefficient biomarkers for disease outcome. The correlation between brain MRI imaging and biochemical and Hence, lack of specific biomarkers makes it challenging to monitor molecular findings in Japanese female patients with Fabry disease therapies in patients. Drastic changes in GAGs contents in patient’s organs have been shown to affect the inflammatory pathway (i.e. a b b b TLR-4, TNF-α and sphingolipids ceramides). As part of a more Kaoru Eto , Arif Hossein , Hiroko Yanagisawa , Takashi Miyajima , b b a a extensive study looking at new biomarkers in MPS, we developed Wu Chen , Yoshikatsu Eto , Satoru Nagata , Tokyo Women's Medical b two new LC-tandem MS techniques based on targeted and semi- University, Tokyo, Japan, Advanced Clinical Research Center, Institute of targeted sphingolipidomics to evaluate the level of three species of Neurological Disorders, Kanagawa, Japan sphingolipids ceramides and identify new sphingolipids biomarkers in biological fluids of MPS patients. Introduction: Fabry disease is a most common lysosomal storage disorder caused by deficiency of α-galactosidase(GLA) activity. Progressive accumulation of globotriaosylceramide (GL-3) leads to Funding acknowledgment: Eleanor Mackenzie Harpur Endow- multi-organ dysfunction caused by vascular dysfunction, vessel ment Fund occlusion and tissue ischemia. The peripheral nerve and CNS involvements in Fabry disease are characterized by small fiber doi:10.1016/j.ymgme.2018.12.116 neuropathy as well as cerebral micro and macro-angiopathy. Brain MRI/MRA shows progressive white matter lesion (WML) and basal artery tortuosity and dolichoectasia. It has been reported that about 101 4.3% female patients presented cerebrovascular disease, though the Gaucher disease (GD)-specific patient-reported outcome (PRO) CNS involvement among female patients has not been fully studied. measures for clinical monitoring and for clinical trials We evaluate the correlation between brain imaging, and biochemical and molecular manifestations among female patients with Fabry Deborah Elsteina, Martin Klemena, Charlotte Panterb, Nicola Bonnerb, disease. Chloe Johnsonb, Ari Zimranc, aShire, Zug, Switzerland, bAdelphi Values, Object: We enrolled 22 female Japanese Fabry patients (30- Macclesfield, United Kingdom, cShaare Zedek Medical Center and the 67years) to examine the correlation between clinical manifestations Hebrew University-Hadassah Medical School, Jerusalem, Israel and brain MRI/MRA imagings, GLA activity, Lyso-Gb3 and GLA genotype. Whole patients were treated with agalsidase alfa or beta. Disease-specific patient-reported outcome (PRO) measures are Results: Cardiomegaly, proteinuria and juvenile cerebral infarc- fundamental to understanding the condition/mindset and disease- tion were observed in 14, 5 and 2 patients. Brain MRI showed WML related expectations of patients with disorders of varying age-of- in 6 patients. The mean GLA enzyme activities were 5.6nmol/mg/hr onset, unpredictable phenotypic trajectories, and non-uniform in WML(+) and 6.6nmol/mg/hr in WML(-) (n=19), respectively. Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S53

The mean plasma lyso-Gb3 values in these patients were 12.19 London NHS Foundation Trust, London, United Kingdom, cMedical and 12.09nM (n=20). Molecular analysis revealed 10missense University of Vienna, Vienna, Austria, dUniversity of California, Davis mutations, 5nonsense mutations, 3deletions, 1insertion, 1 splicing MIND Institute, Sacramento, CA, United States, eCareggi University mutation and 1other. All patients with nonsense mutations did not Hospital, Florence, Italy, fUniversity of Pittsburgh Medical Center present WML, though two patients with D33Y showed WML. Children’s Hospital of Pittsburgh, Pittsburgh, PA, United States, gNational Basilar artery diameter in the patients with positive WML were Hospital for Neurology and Neurosurgery, London, United Kingdom, 2.79mm whereas with negative WML were 2.88mm. Internal hJikei University Hospital, Tokyo, Japan, iOsaka City University Graduate carotid artery diameters in these patients were 3.76mm and School of Medicine, Osaka, Japan, jAmicus Therapeutics, Inc., Cranbury, 3.87mm, respectively. NJ, United States, kRoyal Melbourne Hospital, Parkville, Australia Conclusion: Cerebrovascular findings are sometimes recognized in younger aged female Fabry patients. There was no distinct Fabry disease is a rare, devastating X-linked lysosomal storage correlation between CNS involvement and genotype, GLA activity disorder caused by GLA mutations that result in α-galactosidase A and Lyso-Gb3 in our female cases. X chromosomal inactivation deficiency. Migalastat is a first-in-class, oral pharmacological chap- including methylation and other epigenetic factors may influence erone that binds and stabilizes amenable α-galactosidase A variants, severity of clinical manifestation in female patients. restoring lysosomal trafficking. Migalastat is approved for patients with Fabry disease and amenable GLA variants and is an alternative doi:10.1016/j.ymgme.2018.12.118 to intravenous enzyme replacement therapy (ERT). ATTRACT (NCT01218659) was a phase 3, randomized, open-label, active- controlled study that included an initial randomized 18-month treatment period (migalastat HCl [150 mg, every other day] or 103 continued ERT [agalsidase alfa 0.2 mg/kg or beta 1.0 mg/kg Multiple sclerosis as a misdiagnosis of Fabry disease biweekly]), followed by a 12-month open-label extension (OLE) period, during which patients either continued migalastat Francois Eyskens, University Hospital of Antwerp, Antwerp, Belgium (migalastat-migalastat, n=31) or discontinued ERT and started migalastat (ERT-migalastat, n=15). Data are reported for patients Introduction: Fabry disease (FD) is a X-linked lysosomal storage who entered OLE. Renal function remained stable over 30 months in disorder that causes excessive accumulation of globotriaosylceramid the migalastat-migalastat arm and during the ERT (0-18 months) (GB-3) in various organs. Clinical manifestations of cerebrovascular and migalastat (OLE) periods in the ERT-migalastat arm. Among involvement in Fabry disease include ischemic stroke, transient patients with left ventricular hypertrophy at baseline, mean (95% CI) ischemic attack, intracerebral hemorrhage, subarachnoid hemor- left ventricular mass index (LVMi, g/m2) decreased only in the rhage, microbleeds, cerebral venous thrombosis, and cervical carotid migalastat-migalastat group (0-30 months [n=10]: -10.0 [-16.6, dissection. White matter hyperintensities occur in the subcortical, -3.3]) deep, and periventricular white matter. Several studies have found that 37.5-70.3% of patients with FD present MRI lesions, with a LVMi changes in the ERT-migalastat group were 3.9 (-16.7, 25.8) progressive increase in prevalence and lesion load with age: the with ERT (0-18 months [n=4]) and 0.1 (-13.0, 13.3) during the 12- misdiagnosis of Multiple Sclerosis is often made. month migalastat OLE (n=4). During 0-18 months, the composite Case presentations: A 16-year-old male presented with episodic clinical outcome incidence was 23% (n=7/31) with migalastat and headaches and a brain magnetic resonance imaging (MRI) that 40% (n=6/15) with ERT during OLE, few patients experienced a new showed multifocal punctate to patchy white matter lesions. The event: migalastat-migalastat arm (2 patients [both renal]), ERT- diagnosis of FD was made. A 69-year old female presented with migalastat arm (3 patients [2 renal, 1 cardiac]). Plasma glo- recurrent chest pain and was diagnosed with FD as well as her son, botriaosylsphingosine levels remained low throughout the study postmortem, who died of presumed ‘multiple sclerosis’ 7 years period (both treatment arms). Migalastat was generally safe and well before the diagnosis was made in his mother. The diagnosis was tolerated in the long-term treatment of Fabry disease. These missed in this male patient although he manifested a classical observations extend 18-month findings and support use of oral phenotype of FD and died of renal insufficiency. migalastat as an alternative to intravenous ERT in patients with Conclusion: FD should be searched for in patients presenting Fabry disease and amenable mutations. with white matter lesions on MRI of the brain, especially when there is an association with heart and/or renal disease in the patient or in family members. doi:10.1016/j.ymgme.2018.12.120 doi:10.1016/j.ymgme.2018.12.119

105 Prevalence of Fabry disease in the hemodialysis unit of the 104 Instituto Mexicano del Seguro Social, in Ciudad Obregon, Sonora, Oral pharmacological chaperone migalastat compared with Mexico enzyme replacement therapy in Fabry disease: 30-month results from the randomized phase 3 ATTRACT study Sergio R. Figueroa Sauceda, Ronald Gonzalez-Jaimes, Guillermo Valadez-Juvera, Juan M. Romero-Trejo, Instituto Mexicano del Seguro a b c Ulla Feldt-Rasmussen , Derralynn Hughes , Gere Sunder-Plassmann , Social, Ciudad Obregón, Mexico. Suma Shankard, Iacopo Olivottoe, Damara Ortizf, Robin H. Lachmanng, h i j j Toya Ohashi , Takashi Hamazaki , Nina Skuban , Julie Yu , Jay A. Introduction: Fabry disease (FD), given its wide range of j k a Barth , Kathleen Nicholls , Rigshospitalet, Copenhagen University nonspeciﬁc clinical manifestations and its relative rarity, the problem b Hospital, Copenhagen, Denmark, University College London, Royal Free is often underdiagnosed and, for this reason, has recently been called S54 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

"the new great impostor".1 Due to its impact at the renal level, subjects observed a 69.32% decrease (range 63.64%-75%) heparan patients on hemodialysis (HD) represent a high risk group for FD, sulfate fragments in the CSF (CSF-HS), the biomarker of disease many case reports have reported a significantly high prevalence of pathology, at month 12, which is sustained at 24 months (63.64% FD in this population. A prevalence of 0.1-1.2% of FD cases in HD decrease). Subjects in Cohort 2 (N=3, 1 X 1013 vg/kg) demonstrated a patients is estimated.2 In our hospital, the prevalence of FD has not similar decrease in CSF-HS at 12 months (63.81%, range 50%-71.43%). been reported in a high-risk population such as patients with CKD in Importantly, subjects in Cohort 3 (N=7, 3 X 1013 vg/kg) demonstrated HD. a dose dependent 81.67% decrease (range 80%-83.33%) in CSF HS at Objective: To determine the prevalence of patients with FD who month 6 post-injection. Liver volume measured by MRI demonstrated are in renal replacement therapy with HD who attend the High a fast decrease in all subjects as early as 30 days post administration of Specialty Medical Unit of Cd. Obregon, Sonora (UMAE). scAAV9.U1A.hSGSH in all three Cohorts (39.1% in Cohort 1, 39.6% in Materials and methods: Transversal-descriptive study that Cohort 2, and 49.2% in Cohort 3), that continue to decrease through included 300 patients of the HD program of the UMAE, the men Month 24 in Cohort 1 (53%, range 46.5%-59.4%), Month 12 in Cohort 2 were measured the enzymatic activity of the alpha-galactosidase, if (53.8%, range 43%-64.7%) and Month 6 in Cohort 3 (78.7%, range 77.4%- this was below the reference level, they were then performed 80.8%). We now report evidence for stabilization or improvement of molecular study and measurement of Gb3. The women underwent adaptive behavior and/or cognitive function in all three cohorts. 24- molecular study and Gb3 directly. The main antecedents of month data from Cohort 1, 12-month data from Cohort 2 and 6-month importance for CKD were analyzed, such as the time of diagnosis of data from Cohort 3 will be presented. These data demonstrate CKD, time in HD, DM and HT, as well as other causes of CKD. The significant time and dose dependent reduction of heparan sulfate percentage prevalence was determined with the number of patients and liver volume, neurological stabilization or improvement and detected with FD among the total number of patients treated in tolerability of scAAV9.U1A.SGSH. chronic HD in the UMAE at the time of the study. Results: We found 6 patients with mutation related to FD, 4 reported as a mutation variant with uncertain significance and 2 as doi:10.1016/j.ymgme.2018.12.122 pathogenic mutations. A prevalence of FD was estimated at 2% in the population studied. Conclusions: The prevalence of mutations related to FD found in HD patients in the UMAE is higher than that published 107 internationally. Evolving challenges in the era of newborn screening for Pompe disease doi:10.1016/j.ymgme.2018.12.121 Lauren B. Flueckinger, Priya S. Kishnani, Duke University Medical Center, Durham, NC, United States

106 Prior to the 2015 addition of Pompe disease to newborn Phase 1/2 clinical trial of systemic gene transfer of scAAV9.U1a. screening (NBS), the diagnosis relied on clinical identification hSGSH for MPS IIIA demonstrates 2 years of safety, tolerability, complicated by a diagnostic odyssey. Newborn screening (NBS) and biopotency allows for early intervention resulting in improved outcomes, however, challenges persist. We present two cases highlighting a a b a K.M. Flanigan , T.R. Simmons , K.L. McBride , K.L. Kunkler ,S. these evolving difficulties. Case one is a male identified by NBS with c d e f f Alyward , M.L. Couce , M.T. Oreiro , N.J.C. Smith , L. Jaensch ,M. biventricular hypertrophy and EKG changes. Patient was CRIM g h a a Fuller , J. Ruiz , K.V. Truxal , Center for Gene Therapy, Nationwide positive with previously reported pathogenic GAA gene variants. b Children's Hospital, Columbus, OH, United States, : Center for Based on presumed infantile onset Pompe with cardiac involvement Cardiovascular and Pulmonary Research, Nationwide Children's Hospi- with limited life, parents opted for two enzyme replacement therapy c tal, Columbus, OH, United States, Division of Neurology, Nationwide (ERT) infusions before discontinuation and adoption of palliative d Children's Hospital, Columbus, OH, United States, UDyTECM, Depart- care measures. At age 10 months he presented with musculoskeletal ment of Pediatrics, Hospital Clínico Universitario de Santiago de involvement, however, was weight bearing and attempting to walk. e Compostela, Santiago, Spain, Center for Gene Therapy, Nationwide A discussion of the spectrum of disease presentation within the first f Children's Hospital, Santiago, Spain, Paediatric and Adult Neuro- year of life occurred in addition to a longer lifespan and ERT was metabolic Diseases, Women’s and Children’s Hospital, Adelaide, Austra- reinitiated. This case illustrates the need to discuss the clinical g lia, SA Pathology, Women’s and Children’s Hospital, Adelaide, Australia, spectrum of disease the terminology used even in those with cardiac h Abeona Therapeutics, Cleveland, OH, United States involvement in the first year of life. Case two is a male identified with low GAA enzyme activity via NBS genetic testing identified fi San lippo syndrome type A (mucopolysaccharidosis type IIIA) is a three gene variants: a maternally inherited variant of uncertain lysosomal storage disorder caused by mutations in the SGSH gene and significance and a known pseudodeficiency allele and a paternally results in intracellular glycosaminoglycan accumulation that leads to inherited likely pathogenic variant. This molecular finding resulted in cellular dysfunction and death. It manifests with multisystem disease, confusion of whether the child was truly affected with Pompe, or including severe CNS neurodegeneration and hepatomegaly. An open- was experiencing low levels of GAA enzyme activity due to the label, dose-escalation Phase I/II gene transfer trial has utilized an combination of a pathogenic variant with the pseudodeficiency intravenous administration of a scAAV9.U1A.hSGSH vector into 13 allele. These cases highlight the importance of routine followup for subjects divided into three Cohorts to target both the CNS and somatic newborn screening diagnoses, the development of a systematic tissues. All subjects have tolerated gene transfer well through over database for clinical followup outcomes, and the need for a 6,000 days cumulative follow up. Cohort 1 (N=3, 5 X 1012 vg/kg) Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S55 consensus on terminology used to clinically describe and prognos- of life in a patient switching between treatments. The overall response to ticate often symptom free children identified via NBS. Fitbit use is positive. Our patients appear to have more control of their lifestyle and are willing to try and manage their health pro-actively. By having the wearable devise they were encouraged to be engaged in the doi:10.1016/j.ymgme.2018.12.123 data interpretation, correlating their symptoms with the Fitbit results and decision-making.

108 doi:10.1016/j.ymgme.2018.12.125 Corticobasal syndrome in a man with type 1 Gaucher disease: Expansion of the understanding of the neurological spectrum

Lauren B. Flueckingera, Kunal C. Potnisa, Stephanie M. DeArmeya, Roy 110 N. Alcalayb, Jeffrey W. Cooneya, Priya S. Kishnania, aDuke University The use of Fitbit data in monitoring the improved functioning Medical Center, Durham, NC, United States, bColumbia University, New and quality of life in a case of Fabry disease York, NY, United States Stuart Forshaw-Hulme, Mairead Jones, Gisela Wilcox, Ana Jovanovic, Gaucher disease (GD) is an autosomal recessive condition that Karolina Stepien, Salford Royal NHS Foundation Trust, Manchester, results from a deficiency of the enzyme β-glucocerebrosidase. The United Kingdom increased risk of primary Parkinsonism symptoms among individuals affected with GD and carriers for the disorder is well documented in the The wearable technology is frequently used in the management of literature. However, these risks and case reports often reflect patients medical conditions and clinical research. The data that can be collected with classical Parkinson’s disease (PD) symptoms. We report a patient from the Fitbit include heart rate, sleep pattern and step count. This with GD type 1 who was diagnosed with corticobasal syndrome (CBS), a device can monitor patient physical activity over months to years. It can form of atypical Parkinsonism, in his sixth decade of life. Our case be used to assess the impact of a disease on patient’s quality of life. We highlights the need to consider forms of atypical Parkinsonism such as present a case of a man, who used Fitbit to self-manage his condition. A CBS in addition to PD in the differential diagnosis of cognitive and motor 62-year-old man, diagnosed with Fabry disease (p.R301Q) at the age of changes in patients with GD type 1. We also recommend careful 55 was treated with enzyme replacement therapy (ERT). When the assessment and routine monitoring of cognition, mood, behavior, sleep pharmacological chaperone therapy (CT) became available, he re- patterns, olfaction, and memory in patients with GD type 1 to identify quested that his ERT is switched to oral therapy due to convenience. early symptoms indicative of neurological involvement. There were no concerns regarding his adherence, however he developed drowsiness and lethargy and after 2 months of SRT he decided to take ‘drug holiday’. After 10-day break from treatment, his doi:10.1016/j.ymgme.2018.12.124 energy level increased and drowsiness resolved. He decided to restart CT that again caused extreme lethargy. Subsequently, his ERT was restarted. The changes in his quality of life were captured during two weeks of CT, during the ‘drug holiday’ and the first two weeks of ERT. It 109 enabled the patient to self-manage and monitor CT-related side effects. Self-management using wearable technology to promote pa- The data that was collected suggested that the overall physical activity ’ tients knowledge and skills in managing their own care of the patient was higher from day 7 of ERT whereas heart rate was significantly increased when he was on CT. There was no signification Stuart Forshaw-Hulme, Diane Green, Andrew Oldham, Gisela Wilcox, difference in sleep pattern between both treatment modalities. In Karolina Stepien, Salford Royal NHS Foundation Trust, Manchester, conclusion, the Fitbit device is a sophisticated, easy-to-use activity United Kingdom tracker that encourages high level of self-monitoring among patients with rare diseases. Further study on a large cohort of patients is needed Effective self-care among patients with Lysosomal Storage Disorders to evaluate the long-term utility of the Fitbit as part of a strategy for (LSDs) is a lifelong process of knowledge and skill. NHS England has quality of life and drug side effects monitoring. acknowledged that four in ten people with long-term conditions struggle with the ability to manage their own care. We therefore aimed to assess fl the in uence of wearable technology in assisting patients in managing doi:10.1016/j.ymgme.2018.12.126 their own care. Six patients within our Lysosomal Storage Disease cohort (Fabry disease 3, MPS II 2, Pompe disease 1), who had previously expressed their interest in developing self-management skills, were provided with a Fitbit device. Each patient agreed to synchronise the 111 Fitbit with the application on the phone this would allow the up-to-date The effectiveness of vein mapping in reducing the number of data to be transferred onto the Electronic Patient Record system in our missed infusions among Fabry disease patients: One centre Trust. In order to transfer the data, consent was obtained from all experience individuals. The aim of the data collection was to assess patients’ ability to self-manage their condition, quality of life and side effects of treatment. Stuart Forshaw-Hulme, Mairead Jones, Janet Gorton, Reena Sharma, The patient compliance in uploading the data accounted for 70%. Patients Gisela Wilcox, Karolina Stepien, Salford Royal NHS Foundation Trust, felt empowered to be involved in discussions regarding their pain Manchester, United Kingdom management. Their engagement in their own medical care helped them understand their activity, sleep pattern and heart rate and correlate them Anderson-Fabry disease is a rare Lysosomal Storage Disease for with symptoms. The device enabled us to provide real time data of quality which intravenous enzyme replacement therapy (ERT) is one of S56 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 available treatment modalities. Administration of intravenous ERT by suspected. These results clearly remark the importance of ruling out the homecare delivery and is standard practice in the UK. Some MPS VII in patients with phenotype and clinical features compatible patients affected with Fabry disease and treated with ERT, having had with MPS spectrum, since the different types of MPS may have similar unsuccessful cannulation, may have their treatment cancelled or have manifestations and share some signs and symptoms. multiple cannulation attempts per visit. We therefore aimed to evaluate the efficacy of using vein mapping to assist in the cannulation of Fabry patients prior to treatment. Five patients maintained on ERT doi:10.1016/j.ymgme.2018.12.128 for more than one year, and having had numerous failed cannulation attempts, were identified for vein mapping. By using vein illumination with the AccuVein AV400, clinical photographs were taken of the patients’ veins and given to the patient for the nursing staff to use as a 113 visual guidance on their cannulation. Before vein mapping was Single intravenous dose of AAVHSC15 vector with human introduced, the average number of cannulation attempts was on phenylalanine hydroxylase transgene results in sustained correc- enu2 average 2.8 (range 1-8) post vein mapping this number was reduced to tion of phenylketonuria in the PAH mouse model the mean of 1.2 (range 1-3) attempts. Patient satisfaction and confidence in the nursing staff increased from 37% at baseline to 74% Omar L. Francone, Seemin S. Ahmed, Jeff L. Ellsworth, Deiby Faulkner, post vein mapping. Prior to the introduction of vein mapping, two Arnold Sengooba, Hillard Rubin, Serena Dollive, Diana Lamppu, patients had missed 43% of their infusions across ten nurse visits due to Teresa Wright, Albert Seymour, Homology Medicines Inc, Bedford, unsuccessful cannulation. Post vein mapping no patients missed an MA, United States infusion due to unsuccessful cannulation. The use of vein mapping is promising in reducing unsuccessful cannulation. This method should Novel recombinant clade F adeno-associated viruses, originally be used as first-line support for patients who are difficult to cannulate. isolated from normal human CD34+ hematopoietic stem cells Vein mapping is cost-effective and can reduce the need to have (AAVHSCs), show high liver tropism and potential for liver-based gene indwelling central venous access (e.g. PICC lines and port-a-caths). therapy. Phenylketonuria (PKU) is a rare metabolic disease, resulting Patient satisfaction is key to any successful treatment patients not from mutations in the hepatic phenylalanine hydroxylase (PAH) gene. satisfied with their care may jeopardize their compliance with ERT. PKU is a suitable candidate for rAAV-based gene therapy as it is an autosomal recessive, monogenic defect. There are no current treatments that address the underlying genetic defect in PKU. A missense mutation enu2 doi:10.1016/j.ymgme.2018.12.127 (F263S) in the PAH gene greatly reduces enzyme activity in PAH mice, causing a 40-fold elevation in serum phenylalanine (Phe) on a normal chow diet (containing 1% Phe), making the mice an appropriate animal model for severe PKU. The human PAH transgene, packaged in 112 AAVHSC15 and driven by a ubiquitous promoter (AAVHSC15-PAH), was Mucopolysaccharidosis type VII: Selective retrospective screening administered as a single intravenous injection in PAHenu2 mice. Mice detects 4 new cases were fed normal chow throughout the study. Serum levels of Phe and tyrosine (Tyr) [byproduct of Phe metabolism required for production of Joaquin Frabasil, Consuelo Durand, Silvia Sokn, Ricardo Carabajal, neurotransmitters] were measured weekly. Livers were harvested and Patricia Carozza, Daniela Gaggioli, Juan Politei, Andrea B. Schenone, processed to measure vector genomes, mRNA, and PAH enzyme activity. Laboratorio de Neuroquimica Dr. Chamoles, Buenos Aires, Argentina One week post-dose, Phe levels decreased from 2000μMtob150μM (pb0.0001) and were sustained out to N28 weeks post-dosing (pb0.0001) Mucopolysaccharidosis type VII (MPS VII Sly syndrome) is a highly even as mice had a dietary intake of 1% Phe. Dose-dependent increases in heterogeneous, ultra-rare lysosomal storage disorder, caused by the PAH vector genomes, mRNA, and enzymatic activity were observed. deficiency of β-Glucuronidase (βGUS). Patients with MPS VII may Serum AST/ALT levels in treated mice did not show significant changes. course with non immune hydrops fetalis, cognitive impairment, Modifying the transgene to include a liver-specificpromoterdemon- skeletal dysplasia and/or hepatosplenomegaly, but clinical presenta- strated decreased serum Phe and increased serum Tyr in PAHenu2 mice at tion and disease progression span a wide range of phenotypes: from ten-fold lower doses compared to the initial research vector. Sustained early severe multisystem manifestations to a milder late onset correction was seen out to N44weeksinmiceonnormalchow.These phenotype. Epidemiological data of this disorder is really scarce and data demonstrated that a single dose of AAVHSC15-PAH, designated underdiagnosis of this disorder is suspected. The aim of this study was HMI-102, resulted in long-term correction of PKU in PAHenu2 mice. to carry out a retrospective screening of MPS VII in patients with clinical features compatible with MPS spectrum, in whom other MPS (types I- II- IVA- VI) had been ruled out previously. For this purpose, doi:10.1016/j.ymgme.2018.12.129 we analyzed 586 dried blood spots samples from Argentinian patients with clinical suspicion of MPS, collected between 2017 and 2018, where MPS I, II, IVA and VI were previously ruled out. βGUS enzyme activity was measured by fluorometric method with deproteinization 114 β-Galactosidase activity was also measured as a control enzyme, to Safety evaluation of pentosan polysulfate for treatment of two assess quality of DBS samples. From 586 samples analyzed, we Japanese siblings with mucopolysaccharidosis type VI in a phase 2 detected 4 patients with βGUS deficiency in DBS. βGUS activities of study these patients ranged between 0 and 0.9 umol/h/l (Reference value: N a b c d 80). These results were confirmed in leukocytes. Ages of positives Mahoko Furujo , Kenji Orii , Shunji Tomatsu , Yasuyuki Suzuki , e a cases were 2, 11, 11 and 12 years old. We could detect 4 patients with Toshiyuki Fukao , National Hospital Organization Okayama Medical b βGUS deficiency in cases where others MPS but no MPS VII were Center, Okayama, Japan, Nagamori Pediatric Clinic, Gifu, Japan, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S57 cNemours/Alfred I. DuPont Hospital for Children, Wilmington, DE, United from the proband at age 56 were compared to her data from age 20, States, dGifu University, Gifu, Japan, eGraduate School of Medicine, Gifu and to the control siblings, two methylated and four demethylated University, Gifu, Japan genes involved in potassium voltage-gated channels, and lipid metabolism were identified in CpG islands from promoter regions Early initiation of enzyme replacement therapy (ERT) has (p-value b0.001). The expression level of the genes was confirmed by demonstrated clinical benefits in patients with muco- transcriptome data (fcN2orfcb-2). The differentially methylated and polysaccharidosis type VI (MPS VI). Despite treatments with ERT, expressed genes will be investigated further to explore whether the joint stiffness persists as a significant problem for patients with MPS observed changes are directly related to the development of I, II and VI. The joint problems may severely restrict mobility and Parkinsonism in this patient with GD. ability to perform activities of daily living. We previously reported that new approaches or bone-targeting treatment for MPS VI patients should be necessary. Glycosaminoglycans (GAG) storages doi:10.1016/j.ymgme.2018.12.131 in MPS cells activate toll-like receptor 4 (TLR4) signaling which is leading to the release of tumor necrosis factor-alpha (TNF-alpha) and other inflammatory cytokines. Pentosan polysulfate sodium (NaPPS) inhibits this TLR4 -TNF- alpha activation pass way. Therefore, NaPPS 116 was evaluated in young patients with MPS VI. NaPPS doses were Biochemical and molecular analysis of MPS III in 6 Mexican escalated from 0.5mg/kg up to 1.5mg/kg through 17 times subcuta- patients neously every week with ERT. Patients were evaluated at baseline and after the last administration with physical Jose E. Garciaa, Esther Lieberman-Hernandezb, Carmen Esmer- function, laboratory evaluations and several inflammatory markers. Sanchezc, Adriana Ruizd, David Cervantes-Barragane, Alberto Bravo- NaPPS was well tolerated and there were no serious adverse events. Hidalgof, Sandra C. Mendoza-Ruvalcabaa, Karol Carrillo-Sanchezg, Following clinical evaluation including one female MPS I patient are Carolina Molina-Garayg, Luis L. Flores-Lagunesg, Marco Jimenez ongoing, and improvements of joint functions (e.g. handling Olivaresg, Carmen Alaezg, aInstituto Mexicano del Seguro Social, chopsticks and sitting down on knees) were observed for the MPS I Guadalajara, Mexico, bInstituto Nacional de Pediatría, Mexico City, patient. Mexico, cHospital Infantil Teleton de Oncología, Querétaro, Mexico, dHospital Pediátrico de Alta Especialidad, León, Mexico, eHospital PEMEX Picacho, Ciudad de Mexico, Mexico, fInstituto Nacional de g doi:10.1016/j.ymgme.2018.12.130 Rehabilitacion, Ciudad de Mexico, Mexico, Instituto Nacional de Medicina Genomica, Ciudad de Mexico, Mexico

Introduction: MPS III is a lysosomal disorder caused by null or 115 reduced enzymatic activity leading to the accumulation of the Methylomic and whole transcriptome analyses reveal several heparan sulphate glycosaminoglycan in several tissues. Prevalence potential modifier genes in GBA1-associated Parkinson disease of MPS III is about 1/70,000 live newborns. The diagnosis is based in the clinical features of the patients and requires biochemical or Eric Joshua Garcia, Frank Donovan, Abdel Elkahloun, Barbara molecular analysis for confirmation by any of the following enzymes Stubblefield, Alta Steward, Sampath Arepalli, Settara and genes: SGSH NAGLU HGSNAT or GNS. Chandrasekharappa, Dena Hernandez, Grisel Lopez, Ellen Sidransky, Objective: To describe biochemical and molecular findings in 6 Nahid Tayebi, National Institutes of Health, Bethesda, MD, United cases of MPS III by 4-MU fluorometric assays and NGS gene panel. States Methods: Blood samples of 6 patients, referred by her/his physician and were recruited in a 18 months period (February 2017 – Gaucher disease (GD) is an autosomal recessive lysosomal storage September 2018). We used the 4-MU-alpha-D-N-sulpho-gluco- disorder caused by mutations in GBA1. Furthermore, mutations in saminide, 4-MU-alpha-D-N-Acetyl-glucosaminide as substrates. Re- GBA1 are the most common genetic risk factor for the more sidual activities were measured using published techniques. DNA prevalent neurological disorder Parkinson disease (PD). We hypoth- was extracted by AS1010 cartridge/Maxwell 16. Nextera Rapid esize that epigenetic mechanisms play an important role in the Capture Illumina oligonucleotides capture was used and Next association between GD and PD. We analyzed DNA methylation in Generation Secuencing was performed in MiSeq equipment. Bioin- fibroblasts collected from a patient with type 1 GD when she was 20 formatic analysis was performed in Trimmomatic, BWA and GATK years old and again at age 56 after she developed PD. To account for programs. Variant annotations were based in snpEff and interna- sex, age and familial background, fibroblast samples from her two tional literature. older sisters without PD, one with GD and the other with wildtype Results: All the patients (3 M, 3F) all of them unrelated but a pair GBA1, were also evaluated. DNA and RNA were extracted from all of sibs. 2 were positive to IIIA (reference values 0.8846-3.4657 nMol/ four fibroblast lines. To identify aberrantly methylated sites, 45 mg prot/17 h) 3 to IIIB (reference values 7.09-22.24 nMol/mg prot/ healthy females age 20 ± 5 years and 45 at age 56 ± 5 years from 17hr) and one was negative to both assays. Molecular analysis the NCBI’s GEO and GSE87571 Northern Sweden Datasets were used revealed the following results: SGSH: one case with homozygous as controls. An Illumina Human MethylationEPIC BeadChip and mutation c.1339GNA (p.Glu447Lys) and another case with Affymetrix ClariomTM S transcriptome assay were used to investigate c.1063GNA (p.Glu355Lys) and c.1080delC (p.Val361Serfs*52) the methylation of CpG dinucleotides and the transcriptome-level NAGLU: the pair of sibs (F/M) were homozygous for a novel expression profile of the samples, respectively. GenomeStudio, R- mutation c.407GNT (p.Cys136Phe) and the third case was compound Studio, Affymetrix Transcriptome Analysis Console were employed heterozygous for c.700CNT (p.Arg234Cys)/c.432GNA (p.Trp144Ter) for data analysis, and Cytoscape and GeneMANIA were used to HGSNAT: A male case with c.372-2ANG and c.1610delT (p. elucidate biological functions and pathways analysis. When samples Leu537Profs*27). S58 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Conclusions: In this sample of 6 Mexican patients with suspected 118 diagnosis of MPS III, the confirmation of diagnosis by leukocyte Screening sp2-iminosugar N-glycosides as pharmacological chap- enzymatic activity and NGS confirmed the diagnosis of MSP IIIA, IIIB erone candidates for α mannosidosis: The effect of aglycone and IIIC. nature and valency doi:10.1016/j.ymgme.2018.12.132 Jose M. Garcia Fernandeza, Rocio Rísquez-Cuadrob, Fernando Ortega- Caballerob, Eiji Nanbac, Katsumi Higakic, Carmen Ortiz Melletb, aInstituto de Investigaciones Químicas, CSIC, Sevilla, Spain, bUniversity of Sevilla, Sevilla, Spain, cTottori University, Yonago, Japan 117 Tailoring the inhibitory versus chaperoning behavior of amphiphilic α-Mannosidosis (OMIM 248500) is an ultra-rare monogenic sp2-iminosugar glycomimetics targeting β-glucocerebrosidase: disorder with an estimated incidence of 1/1,000,000 live births. It is From micromolar to picomolar chaperones for Gaucher disease an autosomal recessive disease caused by mutations in the MAN2B1 gene causing dysfunction of lysosomal α-mannosidase (LAMAN), an Jose M. Garcia Fernandeza, Teresa Mena-Barragánb, M. Isabel García- enzyme involved in glycoprotein catabolism. As a result, intra- Morenob, Alev Sevšekc, Tetsuya Okazakid, Eiji Nanbad, Katsumi lysosomal accumulation of mannose-rich oligosaccharides occurs in Higakid, Nathaniel I. Martinc, Roland J. Pietersc, Carmen Ortiz Melletb, all tissues. Long-term enzyme replacement therapy (ERT) with Yoshiyuki Suzukie, aInstituto de Investigaciones Químicas, CSIC, Sevilla, b c velmanase alfa is approved in Europe for the treatment of non- Spain, University of Sevilla, Sevilla, Spain, Utrecht University, Utrecht, α d e neurological manifestations in patients with mild to moderate - Netherlands, Tottori University, Yonago, Japan, Tokyo Metropolitan 1 mannosidosis, but an efficient therapy is missing. Competitive Institute of Medical Science, Tokyo, Japan. inhibitors of the enzyme could potentially be used as pharmacological chaperones (PCs) to enhance the endogenous mutant LAMAN Glycolipid mimicry is a powerful strategy to access competitive activity. In the iminosugar series, D-manno-DNJ showed no inhibi- inhibitors of the lysosomal glycosidases involved in tion toward human LAMAM and only L-allo-DNJ inhibited this glycosphingolipid metabolism. The structure of glycolipid mimics enzyme with an IC value of 64 μM, but no chaperone activity has can be further tuned to favor enzyme binding at the endoplasmic 50 been reported.2 Unfortunately, the better characterized LAMAM reticulum and dissociation from the corresponding enzyme:inhibitor inhibitor, swainsonine, is a lysosomotropic compound and accumu- complex at the lysosome. This property can be exploited for the lates rapidly in lysosomes of normal cells in culture to produce rescuing of lysosomal storage disease enzyme forms hosting intracellular inhibition of this enzyme, thwarting any medical use misfolding causative mutations, which is at the basis of the LAMAM inhibitors that are suitable for the treatment of α- pharmacological chaperone (PC) therapy concept. Yet, unless a mannosidosis remain to be discovered. We have now found that stimuli-sensitive component switching-off enzyme binding at the sp2-iminosugars N-glycosides3 with lipidic aglycons can behave as lysosome is incorporated in the molecular design,1 active site- effectors or inhibitors of human LAMAN in wild type fibroblast directed PCs simultaneously behave as competitive inhibitors of the depending on their mono or multivalent presentation. In a prelim- rescued enzyme, drawing a complex scenario where the inhibitory inary assay, some representatives showed LAMAM activity enhance- and chaperoning activities must be favorably balanced to be ments over 200% in wild type fibroblasts. Further evaluation in therapeutically useful if inhibition prevails, the disease will be fibroblasts from three different α-mannosidosis patients evidenced aggravated. Typically, potent competitive inhibitors of the enzyme mutation-dependent responsiveness, the best candidate affording a are good PCs at subinhibitory concentrations, meaning that correct 30-40% activity enhancement. To the best of our knowledge, this is dosing is critical. The inhibitor/chaperone balance and the nontoxic the first evidence of a potential PC therapy for α-mannosidosis. concentration range is a function of the molecular structure of the PC References: 1) M.R. Ceccarini et al., Int. J. Mol. Sci. 2018, 19, 1500. 2) depending on the basal enzyme activity levels and the disease type, a A. Kato et al. J. Med. Chem. 2005, 48, 2036-2044. 3) E. M. Sánchez- less potent but safer chaperone might be preferable. We have Fernández et al., Chem. Commun. 2016, 52, 5497-5515. previously reported that amphiphilic sp2-iminosugar glycomimetics are particularly well-suited for PC design, current examples covering Gaucher (GD), G -gangliodisosis and Fabry diseases.2 We now M1 doi:10.1016/j.ymgme.2018.12.134 show that their structure can be chemically tuned to produce PCs active on N188S/G183W mutant β-glucocerebrosidase (GCase), associated to type 3 GD, with optimal chaperoning concentrations going from 20 μM (with no inhibition detected) down to 2-20 pM 119 (with inhibition at 200 nM), an interesting feature in the optics of Tandem mass spectrometry of 15 lysosomal diseases, biotinidase 3 developing personalized chaperone therapies. References: 1) T. deficiency and galactosemia type 1 at 2.4 minutes per assay Mena-Barragán et al., Angew. Chem. Int. Ed. 2015, 40, 11696- 11700. 2) E. M. Sánchez-Fernández et al., Chem. Commun. 2016, Michael H. Gelb, Univ. of Washington, Seattle, WA, United States 52, 5497-5515. 3) T. Mena-Barragán et al., Molecules 2018, 23, 927. We have developed a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for measuring enzymatic activities doi:10.1016/j.ymgme.2018.12.133 and biomarkers in dried blood spots relevant to lysosomal storage diseases. The analysis time is 2.4 min per assay. The assay is appropriate for newborn screening or post-screening diagnosis. Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S59

Enzymatic assays for the following disorders are quantified: Batten Kingdom, bManchester University Hospitals NHS Foundation Trust, disease (CLN1, CLN2), Biotinidase deficiency, Fabry, Galactosemia Manchester, United Kingdom, cUniversity of Manchester, Manchester, type I, Gaucher, Krabbe, MPS (I, II, IIIA, IIIB, IVA, VI, and VII), United Kingdom Niemann-Pick-A/B, Pompe, and Wolman disease. The following biomarkers are also quantified in the same LC-MS/MS run: lyso- Membranous nephropathy due to an allo-immune response to Gb1 (Gaucher), lyso-Gb3 (Fabry), lyso-Sphingomyelin (Niemann- enzyme replacement therapy (ERT) has been reported in Pompe Pick-A/B), lyso-sphingomyelin-509 (Niemann-Pick C), psychosine disease and Mucopolysaccharidosis Type VI. Here we report a 3-year (Krabbe), and sulfatides (Metachromatic leukodystrophy), and C26- old boy with infantile onset lysosomal acid lipase deficiency who LPC (X-ALD). The new assay shows the power of tandem mass developed membranous nephropathy during treatment with spectrometry for quantifying all of these analytes. sebelipase alfa. He was diagnosed and commenced ERT by 2 weeks of age due to known family history. At 6 months, he developed hypertension, treated with amlodipine, and moderate proteinuria doi:10.1016/j.ymgme.2018.12.135 (48-150 mg/mmol creatinine). Due to suboptimal response to ERT and high anti-sebelipase antibody titres, he received immunomodulation with rituximab and bortezomib at 20 months, further bortezomib at 25 months, and finally haematopoietic stem cell 120 transplantation (HSCT) at 30 months. Sebelipase alfa was continued Biomarker and pathway analysis in Pompe disease but the dose was reduced from 5mg/kg to 3mg/kg at 4 months post- HSCT. At 6 months post-HSCT, he presented with clinical features of Kelly George, Rena Baek, Monica Lane, Petra Oliva, Kate Zhang, Alison nephrotic syndrome: proteinuria (5770 mg/mmol creatinine), McVie-Wylie, Sanofi Genzyme, Framingham, MA, United States hypoalbuminaemia (8 g/l) and peripheral oedema and received multiple albumin infusions and furosemide. Renal biopsy showed Pompe disease is a rare autosomal recessive lysosomal storage features consistent with immune complex mediated membranous disorder in which a lack of acid alpha glucosidase (GAA) leads to nephropathy: glomerular capillary loop spikes multiple subepithelial, glycogen accumulation within cells. Patients with Pompe disease mesangial and paramesangial deposits on electron microscopy, and initially present with muscle weakness, respiratory compromise or granular staining for IgG in capillary loops on immunofluorescence. failure, and cardiac failure. Current treatment for Pompe disease is Anti-sebelipase antibody titres were 1:191, compared to 1:260476 enzyme replacement therapy (ERT), alglucosidase alfa (Lumizyme/ pre-immunomodulation and 1:61087 pre-HSCT. Allo-immunisation Myozyme), which provides patients with exogenous recombinant against sebelipase is suspected to be the cause of disease. No other human GAA. Avalglucosidase alfa (neoGAA) is the second generation cause of immune complex disease was found, including normal anti- of ERT that is currently in Phase III trials. The current clinical PLA2R antibodies and anti-GBM antibodies. Results of immu- biomarkers in use, including glycogen content post muscle biopsy, noperoxidase staining of biopsy tissue with anti-LIPA antibodies are MRI, CRIM status, and urinary Hex4 are invasive, technically awaited. The patient was treated with furosemide and prednisolone, challenging, or more useful for monitoring the infantile form of the though proteinuria was not steroid-responsive, and prednisolone disease. Previous discovery experiments profiling biofluids from late- was weaned and stopped. Five months after presentation, protein- onset Pompe patients and normal controls identified a number of uria has improved from nephrotic range but remains elevated (370 novel biomarkers that may be useful for prognostic monitoring of mg/mmol creatinine) and lisinopril was commenced. Membranous disease progression, triggering treatment decisions, or measuring nephropathy is a potential acute complication of allo-antibodies to response to treatment. These studies also identified a number of ERT and vigilance for this is recommended. pathways that were altered in Pompe patients. Of particular note, energy metabolism was shown to be decreased with preferential fl utilization of fatty acids over glucose. In addition, in ammatory and doi:10.1016/j.ymgme.2018.12.137 coagulation pathways are also altered. Based on the discovery profiling results, targeted and unbiased mass spectrometry based assays have been developed to evaluate these candidates and pathways of interest. More sensitive assays are also being used to 122 measure proteins that are near or below the level of detection for High dose genistein aglycone in Sanfilippo syndrome: Results of a mass spectrometry. These assays have confirmed our previous randomized, double-blinded, placebo controlled clinical trial findings and have identified new points of dysregulation in these processes and others. This is a unique and rich resource of Arunabha Ghosha, Stewart Rustb, Maria Canalc, Daniel Weisbergb, information with significant clinical potential. Michelle Hepburnb, Karen Tyleea, Brian W. Biggerc, Simon A. Jonesa, aSt Mary's Hospital, Manchester, United Kingdom, bManchester Univer- sity Hospitals NHS Foundation Trust, Manchester, United Kingdom, doi:10.1016/j.ymgme.2018.12.136 cUniversity of Manchester, Manchester, United Kingdom

Genistein aglycone is an isoflavone thought to reduce glycosaminoglycan (GAG) synthesis and to cross the blood-brain barrier, 121 making it a potential substrate reduction therapy for Muco- Membranous nephropathy in a patient with infantile onset polysaccharidosis Type III (MPS III, Sanfilippo syndrome), a rare fi lysosomal acid lipase de ciency and anti-sebelipase antibodies neurodegenerative, lysosomal disorder. High doses of genistein aglycone have been shown to correct neuropathology and hyperac- Arunabha Ghosha, Gauri Batrab, Robert F. Wynnb, Paul Brenchleyc, tive behaviour in mice, but its efficacy in humans is uncertain. We Amrit Kaurb, Simon A. Jonesa, aSt Mary's Hospital, Manchester, United report the results of a single centre, double-blinded, randomised, S60 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 placebo-controlled study with open-label extension phase. Lyso-Gb1 (32.9 ng/mL 0-144). 13/19 patients (68.4%) showed Randomised participants received either 160mg/kg/day genistein tendon elasticity abnormalities, Grade 3 (10/38), Grade 2 (11/38), aglycone or placebo for 12 months subsequently all participants Grade 1 (16/38). The fibrillar pattern, tendon thickness or received genistein for 12 months. The primary outcome measure was ecostructure were normal. QoL evaluation shows a low score in heparan sulfate concentration in cerebrospinal fluid (CSF), with physical and emotional roles and general health and high in pain. secondary outcome measures including heparan sulfate in plasma There are a significant correlation between the degree of tendon and urine, total glycosaminoglycans in urine, neuropsychological, hardness and the low score on the QoL scales, CCL18 (p=.011) and behavioural and quality of life measures and actigraphy. 21 LysoGb1 levels (p=0.035). Strain-elastography is a sensitive, non- participants were randomised and 20 completed the placebo- invasive method to detect subclinical muscle-tendon dysfunction controlled phase. CSF heparan sulfate after 12 months of genistein that may contribute to chronic fatigue in GD1 and identified patients was 5.5% lower than after 12 months of placebo (adjusted for at risk for skeletal complications developing baseline values), but this was not statistically significant (p=0.26), and CSF heparan sulfate increased in both groups during the open- label extension phase. Reduction of urine glycosaminoglycans was doi:10.1016/j.ymgme.2018.12.139 significantly greater in the genistein group (32.1% lower than placebo after 12 months, p = 0.0495). Other biochemical and clinical parameters showed no significant differences between groups. No deviation from natural history was evident in neuropsy- 124 chological outcomes. High dose genistein aglycone (160mg/kg/day) Localized lymphedema in a male with classic Fabry disease was not associated with clinically meaningful reductions in CSF a b c heparan sulfate and no evidence of clinical efficacy was detected. Pilar Giraldo , MPilar Lopez-Royo , Beatriz Ordoñez , Mercedes Roca- d a b However, there was a statistically significant reduction in urine Espiau , IIS Aragon, Zaragoza, Spain, Universidad San Jorge, Zaragoza, c d glycosaminoglycans. Overall, these data do not support the use of Spain, Hospital Quironsalud, Zaragoza, Spain, Mercedes Roca, Zara- genistein aglycone therapy in Mucopolysaccharidosis Type III. goza, Spain

Fabry disease is a treatable multisystem disease caused fi doi:10.1016/j.ymgme.2018.12.138 byde ciency of the lysosomal enzyme a-galactosidase that induce a progressive accumulation of globotriaosylceramide in various organs and tissues. In some cases develop lymphatic microangiopathy that provoke peripheral lymphoedema. We have studied a male 37 years 123 old diagnosed as classical Fabry disease with leucocyte a-galactosi- Strain-elastography in musculoskeletal evaluation in Gaucher dase activity 0.1 nmol/mg prot/h, hemizygous variant p.Q279R, disease increased lyso Gb3. He started ERT at 28 years old. At present he has a mild-moderate renal damage (Creatinine 1.08 mg/dL, GFR: 77 mL/ Pilar Giraldoa, Marcio Andrade-Camposb, Blanca Medrano-Engaya, min/1.73m2, urine albumin/creatinine rate: 741 mg/g creat) Hearth MPilar Lopez-Royoc, Mercedes Roca-Espiaud, aIIS, Aragon, Zaragoza, MRI showed light-moderate left ventricular hypertrophy, with Spain, bFEETEG, Zaragoza, Spain, cUniversidad San Jorge, Zaragoza, maximum thickness at the base previous septal level aortic diameter Spain, dRadiologic Centre CEMEDI, Zaragoza, Spain 42 mm. Normal LVEF. He has developed a progressive and asymmetric inflammation in the left leg (perimeter in cm in Chronic fatigue (CFg), the most prevalent symptom in type 1 malleolar region R/L: 25/29 perimeter at 15 cm from malleolar R/L: Gaucher disease patients (GD1) (79%) persists in spite of prolonged 29/35 perimeter at upper patellar pole R/L: 36/40). We have applied therapy. Their origin is attributed to hypermetabolism or myopathy, a noninvasive techniques: MRI 0.5 and 1.5T (SE T1,T2 and WFT2* however ~70% of patients have bone abnormalities and 11% sequences, sagital, axial and coronal slices), and peripheral neuropathy that can contribute. We hypothesized that MusculoskeletalStrain-Elastography (Hitachi system EUB-8500, others factors like muscle-tendon weakness could influence the L54M transducer, frequency 6-13 MHz). Morphological changes and development of CFg. In order to evaluate, for first time, this aspect in modifications on signal in MRI sequences and MIP processing with GD patients, we study the fibrillar structure (rigidity) of muscle- evident lymphatic channels dilatations in subcutaneous tissues and tendinous tissues in Achilles tendon applying a noninvasive tech- liquid collection periaponeurotic anterolateral and posterior in the nique based on Strain-Elastography (Hitachi system EUB-8500, L54M middle third of left leg to ankle. The strain-elastography showed transducer, frequency 6-13 MHz). We combined this with bone asymmetry with reduction of elasticity in superficial muscles of left marrow burden study using Spanish-MRI score and calcaneus leg suggesting a higher fibrous content. No alterations in deep ultrasound densitometry for bone mineral density evaluation.19 muscle planes are detected. A punch deep skin biopsy was GD1 patients (mean age: 46.1 y, 18-65, 10 females) on enzymatic performed and a high number of lymphatic small vessels were replacement therapy (mean time on ERT: 13.7 years, 2-25), without observed. A planned thirty minutes daily program of active mobility history of diabetes, endocrine diseases or concomitant steroid and strength was established and physical measurements in lower therapy were selected. Correlation with biomarkers, genotype, extremities, achieving in three months a significant reduction in concomitant diseases and SF36 quality of life scale (QoL) were perimeters and increase the range of joint movement. MRI exam performed. Results: Spanish-MRI (6,8 points, 0-18), T-score (mean showed a reduction on lymphatic channels. -0.91, -2.64-1.51), Z-score (mean -0.53, -2.43-1.45), genotype N370S/L444P (9), N370S/other (7), L444P/other (3). Hematological levels were normal, Ferritin: 420 ng/mL (110-550) Chitotriosidase doi:10.1016/j.ymgme.2018.12.140 (1,813 nmol/mL/h 50-6,575). CCL18/PARC (256.3 ng/mL 55-553), Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S61

125 against the well-validated Vineland Adaptive Behavior Scales, Second The MPS I Registry - 15 years of service to the community Edition. The Vineland-II was administered by a clinical psychologist to the parent(s) of 21 juvenile GM1 patients during a neurodevelopmental Roberto Giugliania, Cinde L. Clatterbuckb, Nathalie Guffonc, Joseph assessment of their child for a total of 32 administrations. Children were Muenzerd, Maria Veronica Munoz-Rojasb, David H. Viskochile, also scored on the 6-point ambulation metric using physiatry and aUniversidade Federal Rio Grande do Sul, Porto Alegre, Brazil, bSanofi neurology assessments as source documents. We found strong negative Genzyme, Cambridge, MA, United States, cHôpital Femme Mère Enfant, correlation among patient age and Vineland domains of communica- Lyon, France, dUniversity of North Carolina, Chapel Hill, NC, United tion, socialization and daily living (pb0.0001 for each) and the States, eUniversity of Utah, Salt Lake City, UT, United States composite adaptive behavior score (pb0.0001). Our ambulation scale score strongly correlated with patient age and the Vineland gross motor The MPS I Registry (NCT00144794) was established in June 2003 age equivalent score (pb0.0001 for both) and will be an important under the leadership of an international advisory board (IAB) of expert clinical outcome measure for upcoming clinical trials. This ambulation physicians and is the largest global, observational database for this rare metric can also be applied to many other juvenile onset LSDs with gross disease By the end of 2003, nine patients were enrolled at five sites in motor and cognitive decline. five countries as of July 6th, 2018, these numbers increased to 1125 participants, 107 sites and 30 countries. The MPS I Registry holds a variety of retrospective, baseline, and follow-up data, obtained doi:10.1016/j.ymgme.2018.12.142 through routine clinical and laboratory assessments. These data are recorded in electronic case report forms (CRFs), first developed by the IAB members in 2003 and refined over the years. In 2018, following an 127 evidence-based study, the CRFs were updated and reduced by 64% to Hurler syndrome: Severe sleep apnea as initial presentation in a best reflect what and how patient data are currently collected. These 10-month-old child data are used to support the patient and scientific community through interactions with regulatory agencies, presentations at international congresses, and scientific publications. The collective data of the MPS I Stella C. Godinho, Joyce R. Bisinoto, Gabriela D. Mussi, Leila M. Registry, as analyzed in ten articles published to date, has transformed Azevedo, Charles M. Lourenco, Universitario Estacio de Ribeirao Preto, our knowledge of the natural history of MPS I, the burden of disease, Ribeirao Preto, Brazil and its complications. This allowed for the optimization of patient monitoring and clinical interventions resulting in improved outcomes Background: Sleep abnormalities have been described for all and better quality of life for patients and their families. The MPS I mucopolysaccharidoses (MPS) disorders. They can be very debilitating Registry continues to generate meaningful evidence that can be used and affect the quality of life of both patients and their families, not by physicians, patients and their families, to make decisions for patient only at night, but also during daytime, causing excessive daytime care and to answer new questions that arise from the unmet needs of fatigue, lack of sleep, affecting memory, cognition, behavior, and patients. Supported by Sanofi Genzyme. activity level. Material and Methods: Clinical, laboratory and polysomnographic retrospective data review of a patient with MPS I (Hurler syndrome). doi:10.1016/j.ymgme.2018.12.141 Case Report: Female patient, 10 months old, born to a non- consanguineous couple, was referred to evaluation for investigation of respiratory and sleep problems. According to family, patient 126 showed many episodes of apnea and snoring since 6 months of age Robust clinical outcome measures for patients with juvenile that were getting worse. During the day, patient had some daytime onset GM1 gangliosidosis sleepness and could take several “naps”. Initial laboratory screening was normal but PSG studies showed severe sleep obstructive apnea. Brianna Glase, Jean Johnston, Precilla D'Souza, Dee Adedipe, Cristan Since patient showed some coarse facial features and motor delay at Farmer, Audrey Thurm, Cynthia Tifft, National Institutes of Health, 12 months, patient was seen by a clinical genetics team and a Bethesda, MD, United States lysosomal storage disorder was suspected. Biochemical testing pointed out to MPS I diagnosis. Family declined hematopoietic stem GM1 gangliosidosis, a multisystem disorder of β-galactosidase cell transplantation (HSCT) and child was placed in compassive deficiency caused by mutations in GLB1, is characterized by GM1 program to early ERT access. ganglioside storage in lysosomes and progressive neurodegeneration Discussion: In MPS I patients with upper airway abnormalities, with no effective therapy. Three subtypes, defined by the timing of sleep-induced loss of airway muscle tone can lead to airway collapse symptom onset, display a continuum of clinical manifestations. The and obstructive sleep apnea (OSA), causing loud and disruptive systems that are most functionally impaired in Type II juvenile onset snoring. OSA is a frequent observation in Hurler patients, being patients are ambulation and speech in addition to cognitive decline. We initially associated with short recurrent events of absent or reduced report longitudinal and cross-sectional measures of disease progression ventilation due to upper airway obstruction, alternating with periods over 2 to 10 years in 21 patients with Type II juvenile onset disease. of increased ventilation between periods of obstruction. Patients with juvenile GM1 gangliosidosis have typical development for Conclusion: Long term follow-up with systematic screening for 3 to 5 years, then plateau in attaining new skills before variable but OSA is mandatory in all MPS I patients and should be performed on relentless decline. Validated instruments to track mobility such as the yearly basis and maybe earlier than 2 years of age since many Hurler Functional Mobility Scale are difficult to apply to children who are patients can show severe symptoms of OSA. rapidly losing cognitive as well as gross motor function. We have developed a 6-point scale that reflects the progressive loss of doi:10.1016/j.ymgme.2018.12.143 ambulation in a cohort of GM1 gangliosidosis patients and tested it S62 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

128 preferred source given its rapid availability and superior clinical Incidence of Fabry disease in patients with angiokeratoma outcomes. CB, however, is associated with delayed recovery and relatively poor engraftment due to limited numbers of hematopoi- Jorge Francisco Gomez Cerezoa, Fernando Dominguezb, Rafael etic stem/progenitor cells (HSPCs). An aryl hydrocarbon receptor Cristobalc, Alvaro Hermidad, Marc Moltoe, Miguel Angel Barbaf, antagonist-based culture has been shown to expand CB CD34+ cells Xavier Solanichg, Daniel Lopez Wolfh, Monserrat Moralesi, Pablo 330-fold, leading to rapid hematopoietic recovery after infusion of Garciaj, Irene Gonzalezk, aHospital Infanta Sofía, San Sebastián de los the clinical product, MGTA-456 (Wagner et al., Cell Stem Cell 2016). Reyes, Spain, bHospital Puerta de Hierro, Majadahonda, Spain, As microglia are derived from HSPCs, we hypothesized that MGTA- cHospital de Fuenlabrada, Fuenlabrada, Spain, dComplexo Hospitalario 456 might lead to enhanced microglial engraftment and enable Santiago, San Sebastián de los Reyes, Spain, eHospital Vall Hebron, reduced intensity conditioning. MGTA-456 led to an 8-fold increase Barcelona, Spain, fHospital General Albacete, Albacete, Spain, in hematopoietic engraftment and a 10-fold increase in microglial gHospital Bellvitge, Hospitalet de Llobregat, Spain, hHospital Alcorcon, engraftment (pb0.0001, n=15) in sublethally-irradiated mice, Alcorcon, Spain, iHospital 12 de Octubre, Madrid, Spain, jHospital relative to naïve CB CD34+ cells. Most donor cells localized to the Gregorio Marañon, Madrid, Spain, kHospital Ramon y Cajal, Madrid, non-perivascular region. As high-dose busulfan is critical to micro- Spain glial engraftment (Capotondo et al., PNAS 2013), we assessed engraftment with MGTA-456 after conditioning with busulfan (20 Most angiokeratomas (AK) in the general population are or 40 mg/kg). MGTA-456 led to a 60-fold increase in microglial spontaneous, not associated with other diseases. AK are a very engraftment relative to mice transplanted with unexpanded or frequent manifestation in Fabry disease (FD), and there is none vehicle-expanded CB CD34+ cells (pb0.001, n=8). Notably, trans- screening studies conducted to assess the incidence of the disease in plant of MGTA-456 into mice conditioned with low-dose busulfan these patients. Therefore, the objective of the study is to determine led to a 21-fold increase in engraftment relative to high-dose the incidence of FD in patients with AK. This is a Spanish multicenter busulfan-conditioned animals transplanted with unmanipulated CB cross-sectional observational study that includes patients with AK CD34+ cells (pb0.01, n=8). A 28-fold increase in microglial diagnosed by skin biopsy in the last 20 years, without prior diagnosis engraftment was observed as early as 2 weeks post-transplant of FD. Family history of FD was evaluated size and location of the AK (pb0.0001, n=8). Mechanistically, only CD34+CD90+ cells, but not and alpha galactosidase A activity, GLA gene sequencing and lyso- CD34+CD90- or CD34- cells, led to brain engraftment. These studies Gb3 levels were determined in DBS [GA1] kits. Patients carrying demonstrate that microglial engraftment is faster and greater in variants in the gene were studied to evaluate renal, cardiac, ocular recipients of MGTA-456, even after lower dose busulfan condition- and neurological system function. Eighty-one patients were selected, ing, supporting the use of MGTA-456 as a cell therapy for IMD of which two were carriers of genetic variants. A 55-year-old woman patients. with no family history of FD was diagnosed she had 3 AK of 0.5 cm in diameter in the right leg and acroparesthesias as the only guide symptoms. The patient is heterozygous for p.Asp313Tyr and p. doi:10.1016/j.ymgme.2018.12.145 Ala143Thr and had normal values of enzymatic activity and lyso- Gb3. The second was a 76-year-old man with no family history of FD with one AK of 1.7 cm in the right leg, and without manifestations of 130 EF, hemizygous for the variant p.Asp313Tyr, with reduced enzyme Pitfalls and potential clues in diagnosis in attenuated form of activity, and normal values of lyso-Gb3. With these results, the Hunter syndrome incidence of FD established was 1.23%. These results are similar to those found in other studies of prevalence in risk populations. In ﬁ conclusion, although studies with a larger population would be So a A.L. Goncalves, Marina S. Silva, Letícia C. Feltrin, Gabriela C. needed, the screening of FD in patients with AK can be very [GA2] Garbuio, Charles M. Lourenco, Centro Universitario Estacio de Ribeirao useful for the diagnosis. Preto, Ribeirao Preto, Brazil

Introduction: Attenuated forms of MPS can be easily overlooked doi:10.1016/j.ymgme.2018.12.144 because of paucity of clinical signs. Moreover, they are often misdiagnosed as Legg-Calve-Perthes disease or skeletal dysplasia in presence of prominent skeletal abnormalities and lack of involvement of other systems. Cardiovascular findings can be misdiagnosed 129 as valve disease due to “rheumatic fever”. Unlike most patients with MGTA-456, a first-in-class cell therapy that enables a reduced mucopolysaccharidoses (MPS), individuals with Hunter syndrome intensity conditioning regimen and enhances speed and level of (MPS type II) do not have corneal clouding and can present with human microglia engraftment in the brains of NSG mice normal intelligence (attenuated phenotype) and mild/no organomegaly. Kevin A. Goncalves, Shuping Li, Melissa L. Brooks, Sharon L. Hyzy, Case report: Here we report a 7-year-old boy with normal fi Anthony E. Boitano, Michael P. Cooke, Magenta Therapeutics, stature, mild distal ngers stiffness, normal intelligence, mild valve fi Cambridge, MA, United States disease whose rst diagnosis was rheumatic fever, although no obvious laboratory finding could confirm this hypothesis. Dermato- Allogeneic transplant can prevent or ameliorate neurological logical evaluation for pebbling of the skin over the scapulae also complications associated with selected inherited metabolic disorders failed in lead to a diagnosis. Clinical evaluation in medical genetics (IMDs). Donor-derived microglial cells are protective, limiting service pointed out to suspicion of mild Hunter syndrome, later neurological disease progression. For IMD patients who lack an confirmed by enzyme assay, GAGs chromatography and molecular HLA matched, non-carrier related donor, cord blood (CB) is the analysis of IDS gene. Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S63

Discussion: In addition, relatively late onset of the disease and 132 absent/minimal clinical signs in the attenuated forms pose great Adherence to a pharmacological chaperone therapy among difficulties for the clinicians to suspect such condition. Furthermore, patients with Fabry disease: One centre experience the attenuated form is more rare than the severe, which occurs 3-4 times more frequently. Urinary spot test, though cheap and Janet Gorton, Stuart Forshaw-Hulme, Mairead Jones, Ana Jovanovic, commonly used in resource-restricted setting, has poor sensitivity. Karolina Stepien, Salford Royal NHS Foundation Trust, Manchester, So, appropriate patients should ideally be screened either with United Kingdom quantitative measurement of total urinary GAG or qualitative analysis by electrophoresis or chromatography to identify the Fabry disease is an X-linked metabolic disorder resulting in specific GAG and specif enzyme assay. reduced alpha-galactosidase activity. This causes Conclusion: Healthcare providers across specialties should have globotriaosylceramide accumulation resulting in a multi-systemic in account that mild Hunter patients can present with non-classical clinical presentation. Two treatment modalities are available: features as mild valve disease or a dermatological condition. Early Enzyme Replacement Therapy (ERT) and a pharmacological chaper- diagnosis and early treatment significantly improve the clinical one therapy (CT). While adherence to ERT, available in an outcome and activity of daily living in such patients. intravenous form, is monitored by homecare companies, the adherence to CT is difficult to control. Due to its pharmacokinetics, doi:10.1016/j.ymgme.2018.12.146 it is required to take it before meals and within a twelve-hour window. We therefore aimed to evaluate the compliance of oral CT in our centre. Patients were educated on how to self-manage before the therapy with CT was commenced. They were given a welcome 131 pack with patient information sheet/booklet. At a follow-up clinic Spine instability in patients with mucopolysaccharidosis (MPS) visit, they were given a questionnaire to test their knowledge and type VII adherence to treatment. A standard questionnaire was used to collect relevant information such as: frequency of missed tablets and Antonio González-Meneses, María del Amor Bueno Delgado, Javier methods patients use to remember to take medication in a timely Márquez Rivas, Mónica Rivero, Hospital Universitario Virgen del Rocío, manner. Among 320 patients affected with Fabry disease, 63 are Sevilla, Spain treated with CT (20%) for the mean period of 12 months (range, 6- 12). Among these 63 patients, twenty questionnaires from our Fabry MPS VII is an ultra-rare, autosomal recessive, lysosomal disease in patients were completed. 41% of patients reported they forgot to take which patients are deficient in the beta-glucuronidase enzyme, with CT at least twice, 24% of them took the medication without fasting fewer than 100 patients published worldwide. MPS VII is character- and only 35% where aware that there was a twelve-hour window ized by accumulation of glycosaminoglycans which causes progres- when it is safe to take the tablet. In conclusion, patient education can sive organ dysfunction including skeletal complications such as be enhanced by educational tools such as booklets, welcome pack kyphosis, scoliosis and gibbus. Although spine instability seems more and online access to the materials. A regular monthly telephone common in MPS IV (Morquio disease), MPS V (Maroteaux -Lamy), consultation with a nurse proved to be useful in monitoring patient MPS I (Hurler) and MPS II (Hunter), very few published reports have tolerance and adherence to the new therapy. To improve patient highlighted this feature in MPS VII. Here, we present two patients compliance we plan to develop a leaflet as a quick reference guide with MPS VII with early childhood-onset spine instability: Case 1: A used in our clinics. 4 year-old female with dysostosis multiplex was diagnosed with MPS VII. Spinal instability assessment, performed at age 4, showed several canal protrusions at the cervical and lumbar vertebrae level. Cervical doi:10.1016/j.ymgme.2018.12.148 spine was successfully stabilized surgically, with no further complications. Treatment with vestronidase alfa for MPS VII was started at 5 years. Case 2: A 2 year-old male with dysostosis multiplex was diagnosed with MPS VII. Patient had thoracolumbar kyphosis and 133 sternal protrusion. Treatment with vestronidase alfa for MPS VII was Intrathecal and intravenous combination gene therapy in the started at 5 years. At 7 years, during hernia surgery, patient mouse model of infantile neuronal ceroid lipofuscinosis extends developed severe cervical compression, possibly due to neck lifespan and improves behavioral outcomes in moderately hyperextension resulting in tetraplegia. Spine and neuroimaging affected mice studies showed vertebral instability in cervical, thoracic and lumbar spine. These two cases highlight the importance of evaluating spine Steven J. Graya, Alejandra Rozenbergb, Erik Lykkena, Linas instability in patients with MPS VII as early as possible, with regular Padegimasc, Timothy Millerc, aUniversity of Texas Southwestern follow up. Awareness of this manifestation, careful monitoring, and Medical Center, Dallas, TX, United States, bUniversity of North Carolinat use of preventative measures may help avoid more serious at Chapel Hill, Chapel Hill, NC, United States, cAbeona Therapeutics, complications. Appropriate surgical intervention for spine instability Cleveland, OH, United States. may be necessary for patients with MPS VII, whether receiving vestronidase alfa treatment or not. Infantile neuronal ceroid lipofuscinosis (INCL), also known as infantile Batten or CLN1 disease, is a severe neurodegenerative lysosomal storage disorder affecting the CNS and peripheral organs. doi:10.1016/j.ymgme.2018.12.147 The INCL mouse model (CLN1-KO) recapitulates the major features of the disease, with neurological deficits appearing at 4.5 months and S64 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 a median survival of 8 months of age. We explored dosing 1 month predominantly females, alternative mechanisms such as diet, anxiety old “presymptomatic” mice and 6 month old “symptomatic” mice, or lifestyle should be explored as symptoms may not be linked to with a single intrathecal (IT) injection of 7x1010 vg in the lumbar Fabry disease. cistern of a self-complementary adeno-associated virus serotype 9 carrying the human CLN1 gene (scAAV9-hCLN1). Our results showed significantly increased survival when treating at 1 month, but no doi:10.1016/j.ymgme.2018.12.150 apparent benefits when treating at 6 months. We next explored the treatment at different ages (representing the different stages of the disease) and higher doses. We were able to establish that: 1) early treatment is better and 2) a higher dose is better. Only modest 135 survival effects were seen with the high IT dose (IT-HD, 7x1011 vg) Pharmacological chaperone therapy using migalastat: 1 year administered at 6 months. In contrast, treatment with the IT-HD at experience of starting a new therapy as reported by Fabry 4.5 months (early symptomatic) showed significant survival benefits, patients at a single UK centre delayed onset of symptoms, and improved behavioral performance. We next explored combining the IT dose with an intravenous (IV) Giuseppina Grillo, LSDysosomal isease Unit, Royal Free Hospital, dose, to increase the efficacy in these symptomatic mice. Adminis- London, United Kingdom tering scAAV9-hCLN1 IV in conjunction with IT delivery allows for dose escalation and potentially transduces regions of the brain not Fabry disease is an x-linked disorder in which there is an enzyme fi α adequately treated by IT alone. A low dose IT along with a low, de ciency of - galactosidase. For several years Fabry disease medium or high IV dose did not provide greater therapeutic efficacy patients have been treated effectively with enzyme replacement over the IT-HD alone in early symptomatic mice. However, treatment therapy (ERT) which is administered every 2 weeks as an with an IT-HD and IV-HD combination provided significant benefits intravenous infusion. Migalastat (Galafold®) is an oral chaperone in this same age cohort (currently 17 months old with improved therapy and was approved by the National Institute of Health and behavioral phenotypes). Our preliminary results in mice suggest the Care Excellence in the UK in 2017 for patients with an amenable fi possibility of treating moderately affected patients. mutation. The rst cohort of patients in the Lysosomal Storage DisorderDs Unit (LSDU) at the Royal Free Hospital received migalastat in July 2017 and currently more than 80 patients have doi:10.1016/j.ymgme.2018.12.149 been treated with migalastat. In order to evaluate the patient experience of starting a new therapy an anonymous questionnaire was sent via post or given in clinic to all patients who commenced migalastat since July 2017. The aim of the questionnaire was to gain 134 an insight into the type of information patients received before A review of gastrointestinal symptoms among patients affected starting a new therapy and to evaluate the support they were given. with Fabry disease- One centre experience In addition, the questionnaire may identify any barriers to taking the medication, whether there were issues with taking every other day Diane Green, Hannah Ellis, Karolina Stepien, Ana Jovanovic, Reena therapy and any effects of food restrictions on adherence. This Sharma, Gisela Wilcox, Lorraine Thompson, Salford Royal Hospital information will help to inform staff on how to further counsel and NHS Trust, Salford, United Kingdom advise patients in the future. Early results indicate patients are overall very satisfied with the information received and have Fabry disease is an X-linked lysosomal storage disorder caused by adapted well to an oral therapy and report it as a positive experience. fi α abnormalities in the GLA gene, resulting in a de ciency in - Full results of the questionnaire will be presented. galactosidase A. The consequent abnormal accumulation of glycosphingolipids results in a variety of clinical manifestations. Gastrointestinal (GI) disturbance is the second most common doi:10.1016/j.ymgme.2018.12.151 symptom with 19-52% reporting symptoms include nausea, vomiting, abdominal pain and diarrhoea (Zar-Kessler et al, 2016). GI problems are not life threatening however they have a substantial impact on their life. The Birmingham Irritable Bowel Syndrome (IBS) 136 Symptom Questionnaire was sent out randomly to patients with Improved differentiation between Krabbe disease variants, carrier Fabry disease who attend our Adult Metabolic Medicine Clinic. Out of status, and pseudo deficiency by measurement of psychosine 60 sent out questionnaires, 48 were returned and analysed (20 males/28 females). The patient age range was 28-82 years. 7/ 48 Adam Guenzel, Coleman Turgeon, Amy White, Dawn Peck, Gessi patients had the missense mutation p.N215S of whom 6 received ERT Bentz Pino, April Studinski Jones, Devin Oglesbee, Dimitar Gavrilov, and reported moderate to severe diarrhoea. Of the total cohort, Silivia Tortorelli, Piero Rinaldo, Dietrich Matern, Mayo Clinic, diarrhoea was reported in 23 (45%), flatulence in 31 (66%) and pain Rochester, MN, United States in 22 (46%) patients. 33/48 (88%) patients experienced symptoms while on ERT. There was no difference in frequency of GI symptoms Newborn screening for Krabbe disease (KD) was initially between patients on Enzyme Replacement Therapy (ERT) (median implemented in the state of New York and is currently performed 108 months of treatment, range 6-192 months) and non-treated in six US States. High frequency of pseudodeficiency alleles in the ones. Among 64% of females pain was a prevalent symptom. GI galactocerebrosidase (GALC) gene has led to a large number of false symptoms are very common in our Fabry cohort with incidence positive cases, resulting in unnecessary clinical follow up. In addition higher than the general population. ERT has been shown to be to galactosylceramide, psychosine (PSY) is a substrate of GALC that is effective in managing GI symptoms in children and in adult males. As expected to be elevated in the circulation of KD patients and is ERT does not appear to improve these symptoms in our patients, assumed to play a significant role in the disease pathology. Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S65

Measurement of PSY in dried blood spots (DBS) as a second tier test children of all ages, rhLAMAN-08 is the first study investigating has been implemented in our newborn screening program with the safety and efficacy of VA in paediatric patients of such young age. aim to avoid false positive results while identifying those with infantile onset disease for early treatment. Additional refinement of the assay’s sensitivity allowed for better differentiation of infantile doi:10.1016/j.ymgme.2018.12.153 from later-onset KD. To date, results in DBS are as follows: normal controls (n=209) 0.76 nM (range: 0.31-1.70) GALC pseudodeficiency and mutation carriers (n=41) 0.79 nM (range: 0.32- 1.57) infantile KD (n=23) 57.92 nM (range: 24-112.78) LOKD (n=8) 138 4.99 nM (range: 1.97-8.6). Testing of PSY in erythrocytes has also Long-term visceral and hematologic outcomes of enzyme re- been added to allow for better monitoring of patients before and placement therapy in a pediatric cohort of type 1 and type 3 after bone marrow transplantation because PSY concentrations can Gaucher disease: A single center experience be normalized to hemoglobin and specimen collection is not affected by collection artifacts as is frequent in DBS. Ersin Gumus, Asuman Nur Karhan, Hulya Demir, Hasan Ozen, Inci Nur Saltik Temizel, Serap Dokmeci Emre, Aysel Yuce, Hacettepe University, Ankara, Turkey doi:10.1016/j.ymgme.2018.12.152 Enzyme replacement therapy (ERT) has shown to be effective in reversing visceral and hematologic manifestations of Gaucher disease (GD). We present a single center experience on long-term visceral and 137 hematologic outcomes of ERT with imiglucerase. Available data from 35 fi fi The rst study investigating safety and ef cacy of velmanase alfa children diagnosed with GD at our institution in the last 20 years were (human recombinant alpha mannosidase) in alpha-mannosidosis analyzed. Median age at diagnosis was 42 months. The interval from patients below six years of age diagnosis to first infusion was 1-86 months with a median of 1.5 months. The most common genotype was L444P/L444P (31%) followed a b c Nathalie Guffon , Nicole M. Muschol , Vassiliki Konstantopoulou , by D409H/D409H (17%). Fourteen patients (40%) had at least one d d e Federica Cattaneo , Elisabetta Gelmetti , Albina Tummolo , Irene N370S allele, with 2 (6%) being homozygous. Two patients with f g h g Bruno , Line Borgwardt , Julia B. Hennermann , Allan M. Lund , splenectomy excluded from the outcome analysis. Both types of disease a b Hôpital Femme Mère Enfant, Lyon, France, University Medical Center were presented equally (GD1/GD3=15/18). Mean hemoglobin level c Hamburg-Eppendorf, Hamburg, Germany, Medical University of Vi- increased from 9.89±1.75 g/dL (n=33) at baseline to 12.21±1.05 g/dL d e enna, Vienna, Austria, Chiesi Farmaceutici S.p.A., Parma, Italy, Children (n=32) in first year (pb0.0001) reaching 12.53±0.72 g/dL (n=18) after f Hospital Giovanni XXIII, Bari, Italy, IRCCS Burlo Garofolo, Trieste, Italy, 5 years. While 60.6% of patients were anemic at baseline, the percent of g Copenhagen University Hospital Rigshospitalet, Copenhagen, Denmark, patients with anemia decreased significantly to 5.5% at year 5. Patients h University Medical Center, Mainz, Germany had a mean platelet count of 107.5×103/mm3 at baseline, which improved to a mean of 207.9×103/mm3 in first year (pb0.0001) and Alpha-mannosidosis (AM) is a lysosomal storage disorder caused 236.3×103/mm3 after 5 years. The percent of patients with moderate to by the decreased or absent activity of the enzyme alpha-man- severe thrombocytopenia was decreased dramatically from 60.6% to nosidase, leading to accumulation of mannose-rich oligosaccharides 16.6% after 5 years. Significant improvements in liver (from 2.2 to 1.6 within the lysosomes of all tissues. Velmanase alfa (human multiples of normal) and spleen (from 15 to 7.6 MN) volumes were recombinant alpha-mannosidase VA) is an enzyme replacement observed in the first year of ERT (pb0.0001). One third of patients had therapy approved in Europe for the treatment of non-neurological severe hepatomegaly while half of the patients had severe splenomeg- manifestations in mild to moderate AM patients. rhLAMAN-08 is an aly at baseline. After 5 years of treatment, only 6% of patients had severe ongoing, 24-month, multi-centre, open-label trial investigating the splenomegaly and no patient had severe hepatomegaly. In conclusion, fi safety and ef cacy of repeated VA administrations in AM patients ERT with imiglucerase improved visceral and hematological parameters below 6 years of age at the start of treatment. Patients receive in a group of patients with GD1 and GD3. weekly infusion of VA at a dosage of 1 mg/kg of body weight. Five patients are participating in the study. The primary endpoints are safety and tolerability of VA based on adverse events (including doi:10.1016/j.ymgme.2018.12.154 infusion related reactions), vital signs and clinical laboratory parameters. The secondary endpoints include the changes from baseline at 24 months of a series of parameters and assessments: serum oligosaccharides (biomarker), functional assessments such as 139 Peabody Developmental Motor Scale - 2nd edition (PDMS-2) scores Type 3 Gaucher disease presented with cardiac manifestations and Mullen’s Scale of Early Learning (MSEL) scores, endurance (3- minute or 2-minute stair climbing test depending on patient’s age), Ersin Gumusa,KursadTokelb,AsumanNurKarhana,HulyaDemira, Hasan hearing, immunological profile and immunophenotype (CD3/CD4/ Ozena, Inci Nur Saltik Temizela, Lale Olcayb,AyselYucea, aHacettepe CD8 for T-lymphocytes, CD19/CD20 for B-lymphocytes), quality of University, Ankara, Turkey, bBaskent University, Ankara, Turkey. life and disability questionnaires. Patients that had undergone hematopoietic stem cell transplant are excluded from the study. Cardiovascular involvement with ocular and neurological manifes- The visit schedule foresees evaluations of both safety and efficacy at tations has been reported to be related with a very rare subgroup of baseline, 6, 12, 18 and 24 months. Due to the small sample size, data Gaucher disease type IIIc. The aim of this study is to emphasize the will be analysed and presented at individual patient level. The last cardiovascular involvement by presenting two cases of Gaucher disease patient last visit is planned for the first quarter of 2020. Despite the type IIIc. A 15-year-old girl was presented with pneumonia. Cardiac fact that VA is already licensed in Europe for the treatment of examination revealed apical pansystolic murmur radiating to axilla. S66 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Severe mitral valve stenosis, fourth-degree mitral regurgitation and EKGs post-HCT. This experience suggests early HCT is a feasible option mitral valve thickening with mitral annular calcification were detected and can slow down the decline in function in patients with fucosidosis. on echocardiography. When calcific valvular heart disease was taken Underlying pulmonary issues need to be monitored closely to together with other findings including parental consanguinity, minimize post-HCT morbidity and mortality. hepatosplenomegaly (splenomegaly being more prominent) and bicytopenia (anemia and thrombocytopenia) Gaucher disease was suspected. Bone marrow aspiration revealed Gaucher cells. The doi:10.1016/j.ymgme.2018.12.156 diagnosis was further confirmed by decreased enzyme activity and molecular analysis. The patient was homozygous for D409H. Family screening led to the diagnosis of the older sibling with the same genetic defect. She also had cardiac valvular involvement with increased 141 echogenecity and thickening of the aortic valve indicating aortic Detecting a variant in the GLA gene in multiple family members fi valvular calcification. Both patients had hazy cornea on slit-lamp as an incidental nding examination and some degree of neurological findings including severe headache, syncope, vertical gaze palsy, dysartria and dystonic tremor. Punita Gupta, Lorien Tambini-King, Alejandra Gomez, Leah Fried, St. Patients were put on enzyme replacement therapy. Although marked Joseph's Regional Medical Center, Paterson, NJ, United States improvement in visceral and hematologic manifestations was observed, neurological symptoms and corneal involvement did not benefitfrom Our patient is an eight-year old male who was seen by the fi enzyme replacement therapy. Mitral valve replacement surgery was neurologist for autism and learning dif culties. An institutional performed successfully in the index patient with an uneventful genetic panel which tests for approximately 200 conditions was fi N recovery. Gaucher disease should be kept in mind in young patients performed, detecting a variant of unknown signi cance c.239G Ain with left sided calcific valvular heart disease especially in patients with the GLA gene. Subsequent enzyme analysis was found to be decreased accompanying visceral, ocular and neurological findings. in the range of affected for Fabry disease. Maternal testing detected the same variant in her. Both patient and his mother were referred to our clinic for further evaluation and work up. Pertinent history and doi:10.1016/j.ymgme.2018.12.155 physical exam on both the eight-year boy and his mother were negative for any gastrointestinal symptoms, acroparesthesias, temperature dysregulation, hearing loss, eye findings or skin findings. Urine was negative for microalbumin. GL3 and lysoGL3 results are 140 pending. We were subsequently notified of two male siblings, 13 and Allogeneic hematopoietic stem cell transplant improves out- 18 years old, who also carried the same variant as well as reduced comes in fucosidosis enzyme activity- in the affected range. Both denied any symptoms and had normal urine microalbumin levels. Review of databases revealed Ashish Gupta, Troy Lund, Nicole Anderson, Kelly Miettunen, Julie that this variant has been previously reported in two patients with Eisengart, Elizabeth Braunlin, Paul Orchard, University of Minnesota, suspected phenotype as unaffected or type II Fabry disease. This brings Minneapolis, MN, United States to light the variability in the expression of phenotypes and challenge in deciding when to initiate treatment. While this has been a topic of Fucosidosis is a rare autosomal recessive lysosomal storage discussion with relation to the implementation of lysosomal storage fi disorder (LSD) associated with de ciency of enzyme alpha-L- disorders in newborn screening, it also presents itself as an incidental fucosidase, leading to accumulation of glycolipids and glycoproteins finding in large panel testing or whole exome/genome sequencing for in various organs including brain, lungs, bones, liver, heart and skeletal various indications. The diagnosis of a rare disorder with a variable fi muscle. Mutations in the FUCA1 gene result in de cient enzyme phenotype and unpredictable presentation, which now has two production and impaired cleavage of fucose, which is an important treatment options available, can both be a burden or a boon to the component of glycoproteins and glycolipids. Based on the principal of patient and family members as well as a constant reminder to the metabolic cross-correction with hematopoietic stem cell transplant physician to treat not just the disease but the patient as a whole. (HCT) in LSDs, we report the outcomes of three patients with fucosidosis transplanted within the last 3 years at the University of Minnesota. Patient 1 presented at 5 years of age with global doi:10.1016/j.ymgme.2018.12.157 developmental delay, hypotonia, multiple pulmonary and ear infections underwent matched sibling donor HCT at age 6. Engraftment was successfully achieved, and 6-months post-HCT, myeloid cell lysate fucosidase level and urine oligosaccharides are normal. Patient 2 had a 142 similar presentation at 4 years of age and underwent HCT with an Understanding Sanfilippo syndrome signs, symptoms and physi- umbilical cord blood (UCB) stem cell graft, did not achieve engraft- cian testing patterns: Insights from the Simply Test for MPS™ ment, and was re-transplanted using haploidentical donor bone enzyme-panel program (ST4MPS) marrow stem cells. Patient 2 is now 14 months post-HCT with full donor engraftment, normal enzyme levels and urine oligosaccharides. Stephanie Gurnona, Wayne Pana, Dorna Chua, Christine Lya, Tim There is also significant improvement on pulmonary imaging. Patient 3 Woodb, Laura Pollardb, aBioMarin Pharmaceutical Inc., Novato, CA, presented at 7 months of age and underwent UCB HCT at 11 months of United States, bGreenwood Genetics Laboratory, Greenwood, SC, United age. Patient engrafted completely but developed pulmonary compli- States cations and died about 6 months post-HCT. All three patients had normal cardiac examinations, echos and EKGs pre-HCT. One patient Sanfilippo syndrome is a rare mucopolysaccharidosis (MPS) developed symptomatic pericardial effusion requiring drainage after disorder caused by enzyme deficiencies leading to heparan sulfate the first HCT. The two survivors had normal examinations, echos and aggregation in the central nervous system and visceral body tissues. Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S67

Heparan sulfate accumulation in turn causes developmental delay, correlations, exploratory factor analysis and item-response theory behavioral abnormalities and deteriorating neurologic function. models suggested that a total symptom score (TSS) could be Diagnosis of Sanfilippo is typically delayed because early signs/ calculated. Due to redundancy among items, a neuropathy parcel symptoms of the disorder are indistinguishable from more common and an audiovisual parcel were created in generating the TSS (items causes of developmental delay. Early diagnosis is likely to improve within a parcel were averaged and treated as a single item). The quality of life, patient outcomes and caregiver burden by avoiding a sweating item did not correlate with the other items and was prolonged diagnostic odyssey and providing earlier access to excluded from the TSS. Internal consistency (Cronbach’s alpha) of anticipatory guidance. The objective of this analysis is to gain insight the TSS was ≥ 0.89 across weeks test-retest reliability (intraclass into the factors that lead clinicians to suspect and order confirmatory correlation coefficient), was ≥ 0.91. Known-groups validity showed tests for Sanfilippo syndrome. ST4MPS was initiated in the United that TSS scores incremented properly with each higher health States in February 2017 to streamline confirmatory testing for severity group significantly different from the least severe health patients suspected of having Sanfilippo and other MPS disorders. group. The TSS was correlated with clinical and patient reported Specimens are analyzed by Greenwood Genetics Laboratory. Lab assessments as expected (r N |0.4|). The study demonstrates strong requisition forms were reviewed. Brief results are reported. As of measurement properties of the FD-PRO, a novel disease specific June 18, 2018, 14 patients with enzyme levels within affected range patient-reported instrument that measures FD symptoms. (Sanfilippo A, heparan-N-sulfatase activity range: 0-1 nmol/24hr/mg protein Sanfilippo B, n-acetyl-alpha-glucosaminidase activity range: 0-2 nmol/17hr/mL plasma Sanfilippo C, acetyl CoA: glucosamine N- doi:10.1016/j.ymgme.2018.12.159 acetyltransferase activity range: 0-1.5 nmol/17h/mg protein) were identified. The ordering physicians consisted of 12 geneticists and 2 pediatric neurologists. Average patient age at time of diagnosis was 10.5 years (range: 2-45 years). Review of 12 available forms revealed 144 symptoms of developmental or intellectual delay prompted testing Salmeterol with liver depot gene enhances the skeletal muscle in 8 patients. Other reasons (forms can include multiple reasons) response in murine Pompe disease included coarse facial features (n=5), sleep disturbance (n=2), macrocephaly (n=2) and autistic behavior (n=1). These data Sang-oh Han, Songtao Li, Dwight D. Koeberl, Duke University, demonstrate that ST4MPS facilitates confirmation of Sanfilippo Durham, NC, United States syndrome diagnoses based on clinical signs/symptoms. We hypothesize that greater Sanfilippo disease awareness by pediatricians, Gene therapy for Pompe disease with adeno-associated virus behavioral specialists and other providers will lead to earlier (AAV) vectors has advanced into early phase clinical trials. However, diagnoses, which may lead to overall improved outcomes. the paucity of cation-independent mannose-6-phosphate receptor (CI-MPR) in skeletal muscle, where it is needed to take up acid α- glucosidase (GAA), has impeded the efficacy of Pompe disease gene doi:10.1016/j.ymgme.2018.12.158 therapy. Salmeterol, a long-acting selective β2 receptor agonist, enhanced the CI-MPR expression in muscle. In this study we have evaluated salmeterol in GAA knockout (KO) mice in combination with an adeno-associated virus (AAV) vector expressing human GAA 143 specifically in the liver, AAV2/8-LSPhGAA. Quadriceps GAA activity Measurement properties of the Fabry Disease Patient Reported was significantly increased by administration of the AAV vector, with Outcome (FD-PRO), a new instrument to measure symptoms in or without salmeterol, in comparison with untreated GAA-KO mice Fabry disease (pb0.05). Importantly, glycogen content of the quadriceps was reduced to its lowest level by the combination of AAV vector and a a b Alaa Hamed , Pronabesh DasMahapatra , Charlie Iaconangelo , salmeterol administration. Vector administration reduced the abnor- b c a Daniel Serrano , Chad Gwaltney , Sanofi Genzyme, Cambridge, MA, mal vacuolization and accumulation of nuclei in skeletal muscle. b c United States, Pharmerit, Bethesda, MD, United States, Gwaltney Rotarod testing revealed significant improvement following either Consulting, Westerly, RI, United States vector or salmeterol or combination therapy, in comparison with untreated mice. Thus, salmeterol could be further developed as Fabry disease (FD) is a rare, genetic disease that if left untreated, adjunctive therapy in combination with liver depot gene therapy for progresses to irreversible and life-threatening renal, cardiac, and the treatment of Pompe disease. cerebrovascular events. FD symptoms impact daily functioning and quality of life, but no disease-specific measure of these symptoms exists. The novel Fabry Disease Patient Reported Outcome (FD-PRO) doi:10.1016/j.ymgme.2018.12.160 consists of 18 items that measure neuropathic symptoms, headache, abdominal pain, heat intolerance, swelling, tinnitus, fatigue, hearing/ vision impairment, and hypohidrosis. Measurement properties of the instrument were evaluated among 139 FD diagnosed patients 145 (enzyme deficiency in males GLA genotyping in females) from 10 Fatigue in Gaucher disease: A key quality-of-life concern countries and 25 sites. Patients completed the FD-PRO daily on a handheld electronic diary for 4 weeks demographic, other patient Rosenbaum Hanna, Hematology and Gaucher Clinic Clalit Medical and clinician reported outcomes were also collected. The mean age of Consulting Center, Nazareth, Israel patients was 43 years majority (72%) was treated with enzyme replacement. Patient compliance was high ≥ 87% completed at least 4 Gaucher disease (GD) is the most common lysosomal disorder, FD-PRO entries each week (mean completion time: 171 seconds in caused by inherent deficiency of glucocerebrosidase, which is respon- week one). Empirical evaluation of item properties via inter-item sible for lysosomal glucosylceramide degradation. Glucosylceramide S68 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 accumulation in the lysosomes of macrophages, namely Gaucher cells, meters on 6MWT with a mean (SD) follow-up of 6.7 (3.20) years and an leads to hepato-splenomegaly, pancytopenia and bone manifestations, increase of 23.7 (42.05) stairs/min on 3MSCT with a follow-up of 4.3 resulting in a progressive and multi-organ disorder. GD is a panethnic (3.35) years. Pulmonary function also improved [FEV1 mean (SD) disorder classified into 3 clinical subtypes: type 1, the non-neuropathic increase 0.30 (0.53) L, n=55 FVC increase 0.39 (0.63) L, n=58] with a GD, is most common among the Ashkenazi Jewish population types 2 mean (SD) last follow-up of 6.9 (2.89) years. 79% reported ≥1adverse and 3 are associated with acute and chronic neurologic symptoms, event (AE), most frequently reported as MPS-related complications. respectively. GD is a disabling chronic disease with heterogeneous Nine (4.3%) patients experienced drug-related serious AEs, with the clinical symptoms. One of the main patient complaints is fatigue, most common being infusion-related reactions in two patients (1%). 32 resulting in functional disability and impaired quality of life. However, patients discontinued for reasons unrelated to galsulfase. No new safety fatigue does not correlate with the visceral, hematologic and skeletal concerns were observed. Galsulfase was associated with a tolerable manifestations but is often the presenting symptom of GD. Not safety profile and long-term maintenance of endurance and pulmonary infrequently, fatigue is the first and only complaint of GD patients. function, suggestive of ongoing clinical benefit. Management options for GD include enzyme replacement therapy, substrate inhibition and supportive care. Monitoring of signs, symptoms, biomarkers and organ imaging are essential for treatment doi:10.1016/j.ymgme.2018.12.162 evaluation. However, current practice guidelines do not include measurement of fatigue or therapeutic goals for fatigue and this may discourage healthcare providers from evaluating or treating this domain. Although several validated tools measure fatigue, there is no 147 scale specific for GD. The objective of this report is to demonstrate the EMPOWERS: A phase 1/2 clinical trial of SB-318 ZFN-mediated in importance of fatigue in GD through case examples. An overview of the vivo human genome editing for treatment of MPS I (Hurler regulatory process of drug approval in Israel will be presented, together syndrome) with case examples. Novel approaches to measure fatigue in GD patients a b c d should be developed to encourage healthcare authorities and treating Paul Harmatz , Heather A. Lau , Coy Heldermon , Nancy Leslie , e e f a physicians not to ignore or minimize the role of fatigue in GD. Quality of Cheryl Wong Po Foo , Sagar A. Vaidya , Chester B. Whitley , UCSF b life is frequently ignored in chronic disorders but is very important in Benioff Children’s Hospital Oakland, Oakland, CA, United States, NYU c fact, fatigue should be regarded as a criterion to initiate treatment in GD. Langone Medical Center, New York, NY, United States, University of Florida, Gainesville, FL, United States, dCincinnati Children’s Hospital e doi:10.1016/j.ymgme.2018.12.161 Medical Center, Cincinnati, OH, United States, Sangamo Therapeutics, Richmond, CA, United States, fUniversity of Minnesota, Minneapolis, MN, United States

146 Mucopolysaccharidosis type I (MPS I or Hurler syndrome), is a Enzyme replacement therapy in patients with muco- rare, progressive, autosomal genetic disorder that is caused by polysaccharidosis type VI: Updated findings from the MPS VI mutations in the human IDUA gene and is characterized by deficient clinical surveillance program activity of the lysosomal enzyme alpha-L-iduronidase (IDUA). Without functional IDUA activity, the glycosaminoglycans (GAG) Paul Harmatza, Christina Lampeb, Rossella Parinic, Reena Sharmad,Elisa dermatan sulfate and heparan sulfate, accumulate in the body and Leão-Telese, Julie Johnsonf, Debbie Sivamf,ZlatkoSisicf, aUCSF Benioff lead to widespread tissue and organ damage. MPS I patients develop Children's Hospital Oakland, Oakland, CA, United States, bCentre for Rare progressive cardiac and respiratory disease, skeletal abnormalities, Diseases, Clinic for Children and Adolescents, Helios Dr. Horst, Schmidt and vision loss, despite treatment with enzyme replacement therapy Kliniken, Wiesbaden, Germany, cCentre for Metabolic Diseases, Fondazione or bone marrow transplant. SB-318 is a new type of investigational MBBM, San Gerardo Hospital, Monza, Italy, dSalford Royal Hospital NHS treatment for MPS I. SB-318 uses zinc finger nuclease (ZFN)- Foundation Trust, Salford, United Kingdom, eSão João Hospital, Porto, mediated in vivo genome editing to insert a normal copy of the Portugal, fBioMarin Pharmaceutical Inc., Novato, CA, United States IDUA transgene into liver cells, delivered via AAV2/6 vectors. The precision and specificity of SB-318 allows for integration at a Clinical trials of galsulfase, approved for mucopolysaccharidosis VI targeted genomic location and is intended to reduce accumulation (MPS VI), have shown galsulfase as efficacious with a tolerable safety of GAG with lifelong continuous endogenous production of IDUA. profile. In 2005, MPS VI Clinical Surveillance Program (CSP) was EMPOWERS is an ongoing Phase 1/2 clinical trial to determine if SB- initiated to characterize disease progression and evaluate long-term 318 is safe and tolerable in patients with attenuated MPS I. efficacy/safety of galsulfase. Previously published interim results EMPOWERS is a multicenter, open-label, dose escalation study with corroborated clinical trial observations. Updated findings from the CSP one-time peripheral intravenous infusion of SB-318, followed by are reported. As of March 2017, 223 patients were enrolled 13 patients three years of observation. One adult subject with attenuated MPS I never received galsulfase and 48 initiated treatment in clinical trials. For has received SB-318 at a dose of 1e13 vg/kg. The infusion was all patients (n=219), median age at CSP enrollment was 12.6 years generally well-tolerated and no serious adverse events related to the (range 0.8-65.0). For those already receiving galsulfase (n=206), study drug were reported with up to 3 months of exposure. No median age at first infusion was 9.9 years (0.1-63.3) and treatment persistent transaminitis was observed. Additional analysis of the trial duration was 9.2 years (0.08-16.2). Outcomes were assessed for data is ongoing and will be presented as available. These results patients with pretreatment baseline and last follow-up assessment. support further development of SB-318 for the treatment of MPS I. From a mean (SD) last follow-up of 7.1 years (3.36), uGAG (n=132) decreased by 250.2 (262.17) μg/mg creatinine. Improvements in the six- minute walk test (6MWT n=74) and three-minute stair climb test doi:10.1016/j.ymgme.2018.12.163 (3MSCT n=40) were observed: mean (SD) increase of 48.2 (145.43) Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S69

148 and a true ketogenic diet. Mice were euthanized at 11 weeks to The evolving role of enzymology and metabolomics in the determine HS content and UHPLC-MS of brains. We report for the diagnosis of lysosomal disorders in the post-genomic era first time that global metabolomics revealed profound metabolic differences between WT and MPS IIIB mouse brains. MPS IIIB mice on Katie Harvey, Derek Burke, Simon J.R. Heales, Great Ormond Street a normal diet had 49 dysregulated metabolites out of 274 known Hospital, London, United Kingdom metabolites identified compared to WT mice on a normal diet. We observed profound disturbances in amino acid metabolism in In recent years there has been an exponential increase in genetic accordance with previous reports in urine and serum. A ketogenic testing for metabolic disorders, in particular the use of gene panels diet corrected 18/49 dysregulated metabolites in MPS IIIB mice, and whole exome/genome sequencing. With this wealth of data we including over a third of the altered amino acids. Surprisingly, have seen an increase in the finding of genetic variants of unknown despite correction of key metabolic changes, MPS IIIB mice on significance and identification of patients with a genetic diagnosis of the ketogenic diet had increased HS and other glycosaminoglycan a lysosomal storage disorder but with an atypical phenotype or a (GAG) accumulation compared to MPSIIIB mice on a normal diet. clinical diagnosis but no mutations found in the relevant gene. There Though no improvement in GAG was seen, the metabolic improve- is increasingly a need for diagnostic tests which reflect in-vivo gene ments seen with the ketogenic diet may warrant further study to function as an adjunct to molecular genetics. This is leading to a better understand the contributions of the metabolic alterations in change in the way many patients are investigated. Here we discuss MPS IIIB. recent experience in the Enzyme Unit at Great Ormond Street Hospital, of a number of cases where extensive molecular genetics have been performed but further tests have been required to doi:10.1016/j.ymgme.2018.12.165 elucidate genotype phenotype correlations and/or confirm the diagnosis. Patients discussed include 2 cases of unexpected genetic diagnosis of mucopolysaccharidosis type III from whole genome 150 testing in the ophthalmology setting, the identification variants of High α-galactosidase A over expression and/or mitochondrial unknown significance in genes for mucopolysaccharidosis type IV dysfunction may inhibit efficacy of gene therapy for Fabry disease and glycogen storage disease type III, in patients with atypical clinical presentation and clinically suspected GM2 gangliosidosis in a a a b c patient where only a single mutation was identified. In all cases, Simon Heales , Derek G. Burke , Ahad Rahim , Steven Howe , a a enzymology and the measurement of biomarkers and metabolites Jonathan Lambert , Great Ormond Street Hospital/UCL, London, United b c has helped to confirm diagnosis. These cases represent examples Kingdom, UCL School of Pharmacy, London, United Kingdom, UCL where the increased use of molecular genetics as a screening tool has Great Ormond Street Institute of Child Health, London, United Kingdom inevitably led to the identification of atypical patients. The appropriate investigation of such patients requires support from experi- Fabry disease (FD) is caused by mutations in GLA that encodes α α α enced and responsive specialised laboratories using sensitive enzyme lysosomal -galactosidase-A ( -gal-A). Loss of -gal-A leads to assays and biomarkers. glycosphingolipid storage in cells and associated pathology. A number of therapeutic approaches are available/being considered for treating FD including gene therapy using a lentivirus to integrate doi:10.1016/j.ymgme.2018.12.164 GLA into ex-vivo haematopoietic stem cells before autologous transplantation into patients for long-term enzyme expression. In this study, we further examined the efficacy of a lenti-vector previously engineered in our laboratory. Transduction of the human lympho- 149 blastic leukaemia cell-line (Jurkat) resulted in dose-dependent Effects of ketogenic diet on lysosomal storage and CNS metabo- increase of α-gal-A expression, without apparent toxicity. The lism in MPS IIIB mice enzyme produced by cells with 0.4 transgene copies per cell (vg/

cell) had comparable kinetic properties to wild type (Km for wild Kimberly E. Hawkins, Alexander P. McNally, Joseph A. DeCosmo, type enzyme 1.53+/-0.13mM. Km for enzyme in treated cells 1.54 Priya Shil, Lingbao Ai, Coy D. Heldermon, University of Florida, +/-0.10mM). Increasing exposure to 1.8vg/cell resulted in an b Gainesville, FL, United States apparent increase in Km to 2.53+/-0.29mM (p 0.01, compared to wild type enzyme). Therefore, less virus dosage may be more Mucopolysaccharidosis IIIB (MPS IIIB) is a lysosomal storage therapeutically efficient. In addition to increased intracellular disease resulting from the deficiency of a-N-acetylglucosaminidase enzyme activity, a dose and time dependent release of enzyme into (NAGLU) which is necessary for heparan sulfate (HS) degradation. the extracellular cell culture medium could be demonstrated. Accumulation of HS within lysosomes throughout the body, espe- Furthermore, uptake of the extracellular enzyme into naive Jurkat cially the CNS, results in severe neurological disorders and eventually cells could be demonstrated, thereby indicating cross-correction. death. Serum and urine metabolomic impairments in MPS IIIB mice Finally, as previous research has reported mitochondrial dysfunction, and humans have been described however, to our knowledge no we considered the possibility that this could compromise factors research is available on metabolomic differences in the CNS of such as protein expression and cross-correction. Our findings MPSIIIB mice. A ketogenic diet composed of a high fat, moderate demonstrate inhibition of respiratory chain complex I did not affect protein and minimal carbohydrate content was given to assess lentivirus transduction efficiency or uptake of extracellular α-gal-A, effects on lysosomal storage and metabolomic dysfunction in the but inhibited enzyme release into media by 37% (pb0.001). In brain of MPSIIIB mice. In this study, 58 WT and MPS IIIB mice were conclusion, lentivirus-mediated gene delivery is a promising thera- grouped and given three separate diets upon date of weaning for peutic option for FD. However, excessive enzyme generation could eight weeks a normal control mouse diet, a ketogenic control diet, result in a protein with inferior kinetic properties. Correction of the S70 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 impaired mitochondrial function associated with FD should also be glycogen. Patients with Pompe disease exhibit clinical phenotypes considered in order to promote efficacy of treatment. across a variety of tissues, including glycogen buildup in cells, deficits in cardiac, respiratory, and skeletal muscle function, and CNS pathology. Previously, we described novel multi-tissue GAA express- doi:10.1016/j.ymgme.2018.12.166 ing rAAV vectors that reduced glycogen in skeletal muscle, heart, and brain 4 weeks after treatment of GAA-/- mice. We now describe longer term follow up of animals treated with hybrid liver/muscle/ neuronal vectors. Mice were dosed with rAAV vectors at 1x1013, 151 3x1013, or 1x1014 vg/kg, and in-life measures and bioanalytical Urinary glucose tetrasaccharide, a useful prognostic biomarker endpoints were assessed relative to vehicle-treated GAA -/- or WT for Pompe disease? littermate control mice. Assessment of diaphragm, heart, and skeletal muscle tissue indicated dose-dependent increases in hGAA expres- Simon Heales, Katie Harvey, Maureen Cleary, Helen Prunty, James sion and activity, and reduced glycogen accumulation in these Davison, Derek Burke, Laura Guilder, Great Ormond Street Hospital/ tissues. Vectors with incremental expression in liver contributed to UCL, London, United Kingdom reduced antibody reactivity to hGAA, compared to mice dosed with a control vector containing a muscle-only promoter element, although Urinary glucose tetrasaccharide (Glc4) is proposed to be a useful the muscle-only promoter constructs achieved high levels of GAA fl biomarker for Pompe disease with levels suggested to re ect activity and glycogen reduction in key tissues. Finally, we observed glycogen storage. In our centre, we have established a high dose-dependent expression and activity of hGAA in nervous performance liquid chromatography (HPLC) method (reagent free system tissue and demonstrated that a significant reduction in ion chromatography with eluent generation and pulsed amperomet- glycogen levels in spinal cord could be achieved in mice dosed ric detection) that permits relatively fast turnaround and high with a systemically-administered rAAV8 vector construct. Our sample throughput. This method is now incorporated into our findings represent the first nonclinical application of engineered diagnostic algorithm for Pompe disease where it is used to hybrid promoters providing tissue-specific neuromuscular fi corroborate ndings from our dried blood spot enzyme assay service. transgene expression combined with immune-tolerizing liver ex- Whilst there is a clear role for assessment of GLc4 is the diagnostic pression and suggest key attributes for refinement of vector design workup of patients, less is known with regards to its use in for the clinical translation of an rAAV-based gene therapy for Pompe monitoring patients on treatment and predicting/correlating with disease clinical progression. To address this we now evaluate urinary Glc4 status every 3 months and relate levels to clinical parameters and laboratory findings such as antibody titres. Our preliminary data doi:10.1016/j.ymgme.2018.12.168 suggests a marked decrease in urinary Glc4 levels following commencement of enzyme replacement therapy (ERT), e.g. from 85 to 10 umol/mmol creatinine (unaffected b5 umol/mmol creatinine) 153 over a 6 month period. Furthermore, in a 3 year old patient an Increased frequency of enzyme replacement therapy in a Fabry increase in Glc4 from 66 to 112 umol/mmol creatinine was recorded disease cohort close to a cardiac arrest. Similarly, an increase in urinary Glc4 (from 10 to 70 umol/mmol creatinine) was associated with the development of anti ERT antibodies. Our results, to date, may provide Nadene D. Henderson, Joshua Barch, Kayla Segady, Megan Hoenig, evidence for a prognostic role for urinary Glc4. Work is now Damara Ortiz, UPMC Children's Hospital of Pittsburgh, Pittsburgh, PA, continuing to build and expand upon our findings to permit us to United States generate the evidence to justify continued monitoring of urinary fi Glc4 in Pompe disease. Fabry disease is a lysosomal storage disease caused by de ciency of the enzyme, alpha-galactosidase. Enzyme replacement therapy (ERT) in Fabry disease has been FDA-approved since April 2003. The doi:10.1016/j.ymgme.2018.12.167 standard dosing strategy of agalsidase beta is 1 mg/kg every two weeks. A subset of individuals treated with ERT for 9-12 years, described a cyclical pattern of feeling well after their biweekly dose followed by a variety of worsening symptoms 2-4 days before their 152 next scheduled dose. The range of symptoms reported include Continued analysis of GAA-/- mice treated with novel hybrid worsened diarrhea, acroparesthesias, abdominal pain, fatigue, car- promoter rAAV vectors expressing acid alpha-glucosidase diac findings, and a lack of mental clarity or “fogginess.” We trialed a period of more frequent dosing in which infusions were adminis- Garrett Heffnera, Lucy Jamesa, Christina Chaivorapola, Chanchal tered every 10 days with improvement in the symptomatology and Sadhua, Miranda Meia, Hui Mengb, Tim Stinchcombea, Jennifer Tona, improved perceived quality of life. We will present the collected Benedikt Schoserc, Michael Lawlorb, Suyash Prasada, John Graya, data, including laboratory measurements, cardiology evaluations, aAudentes Therapeutics, South San Francisco, CA, United States, and quality of life surveys during this period of increased frequency bMedical College of Wisconsin, Milwaukee, WI, United States, cKlinikum of dosing, which is ongoing. Munchen, Munich, Germany

Pompe disease is caused by mutations in the acid alpha- doi:10.1016/j.ymgme.2018.12.169 glucosidase gene (GAA) that is responsible for processing lysosomal Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S71

154 multidisciplinary Steering Committee (SC) of 29 MPS experts. Methodology to develop guidelines for the management of Following a systematic literature review process, the SC developed patients with neuronal ceroid lipofuscinosis type 2 disease 117 recommendation statements covering general principles, routine monitoring and assessments, interventions to treat the underlying Christian J. Hendriksza, Jeffrey Donohueb, Yanique Donohueb, Sara E. disease and surgical interventions. The statements were ratified Molec, aSteve Biko Academic Hospital, Pretoria, South Africa, bCare using a blinded modified-Delphi online survey in which they were Beyond Diagnosis Foundation, Columbia, TN, United States, cUniversity rated using a Likert scale by international MPS physicians. Consensus College London, London, United Kingdom. was reached when ≥75% respondents agreed with a given statement and following two rounds of voting, all 117 statements reached The multiple clinical manifestations and progressive nature of consensus. During this process, the MPS community voted upon the CLN2 disease, a major form of the neuronal ceroid lipofuscinoses or use of enzyme replacement therapy (ERT) and hematopoietic stem Batten disease, make the management of patients challenging. cell therapy (HSCT) to address the underlying enzyme deficiency in Although guidelines are available, the methodology used to formulate patients with MPS IVA/VI. For each treatment the rationale, evidence clinical recommendations has come under increased scrutiny, base and deliberations for management (including safety and highlighting a need for robust, independent guidance on the efficacy, monitoring response and adverse events) were considered benefits/risks of disease-modifying treatments and the medical at length. The expert consensus determined that the risk-profile interventions used to manage this condition. This method will be benefit of HSCT in patients with MPS IVA/VI is less clear than in other implemented to develop evidence-based and expert-agreed practical types of MPS, and further research, including a comparative study to recommendations for the wide range of interventions currently used better understand the long-term efficacy and safety of HSCT is to manage CLN2 disease and will be independently managed by Care required. Therefore, initiation of long-term ERT is currently recom- Beyond Diagnosis, a 501(c)3. The first part of the project was to mended as soon as possible after diagnosis for patients with MPS develop an expert mapping tool to identify two chairs, who would lead IVA/VI. Whilst HSCT is not currently recommended in patients with the project to ensure the process is transparent and unbiased. After MPS IVA due to limited evidence, HSCT may be an option for patients identifying 1,454 professionals through the KOL tool, they were with MPS VI with consideration of the associated safety risks, donor sequentially approached until two were able to commit to participa- status and institution experience. Further research is required to tion in this project. Key statements or questions will be developed and address evidence gaps and to determine the long-term outcomes of supported by a systematic review of the published evidence using ERT and HSCT in patients with MPS IVA/VI. PRISMA guidance to form the basis of an international Delphi consensus determination process which will then be conducted. Considering that the evidence-based CLN2 literature is limited, a doi:10.1016/j.ymgme.2018.12.171 series of consensus processes involving field experts will be employed via the Delphi Method. Multiple stakeholders have been identified and approached to support funding this project, including potential 156 companies developing therapeutics in this area, patient organisations The SPARKLE study: Shedding light on alpha mannosidosis and an international steering committee as recommended by the steering committee chairs. The guidelines will be assessed against the Julia B. Hennermanna, Nathalie Guffonb, Federica Cattaneoc, Stefania Appraisal of Guidelines for Research and Evaluation (AGREEII) criteria Pirondic, Line Borgwardtd, Allan M. Lundd, Mercedes Gil-Campose, and multiple tools will be developed to support their implementation Anna Tylki-Szymanskaf, Nicole M. Muscholg, aUniversity Medical and after completion, a written report will be produced. This approach Center Mainz, Mainz, Germany, bHôpital Femme Mère Enfant, Lyon, has been used successfully for development of ACMG PKU and the France, cChiesi Farmaceutici S.p.A., Parma, Italy, dCopenhagen University MSUD Guidelines, which led to publication. Hospital Rigshospitalet, Copenhagen, Denmark, eHospital Reina Sofía, Cordoba, Spain, fThe Children’s Memorial Health Institute, Warsaw, g doi:10.1016/j.ymgme.2018.12.170 Poland, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

Alpha-mannosidosis (AM) is a lysosomal storage disorder caused 155 by reduced activity of the enzyme alpha-mannosidase, leading to Evidence-based, expert-agreed recommendations for the man- intracellular accumulation of mannose-rich oligosaccharides in agement of patients with MPS IVA/VI: Recommendations to various organs. Velmanase alfa (VA) is a human recombinant replace the specific missing enzyme alpha-mannosidase approved in Europe in 2018 as enzyme replacement therapy for the treatment of non-neurological manifestations Christian Hendriksza, Roberto Giuglianib, Paul Harmatzc, Maurizio in mild to moderate AM patients. A post-authorization safety and Scarpad, aSteve Biko Academic Hospital, Pretoria, South Africa, bFederal efficacy registry called SPARKLE was designed as a multi-centre, University of Rio Grande do Sul, Porto Alegre, Brazil, cUniversity of multi-national, non-interventional, prospective cohort study. SPAR- California San Francisco Benioff Children’s Hospital Oakland, Oakland, KLE has two objectives. The first is to assess post-marketing safety CA, United States, dCenter for Rare Diseases, Helios Dr. Horst Schmidt and effectiveness of VA (product registry) the second is to gain new Kliniken, Wiesbaden, Germany understanding of the natural course of AM (disease registry). In terms of effectiveness, the primary focus is to estimate the treatment Management of mucopolysaccharidosis (MPS) is complicated, response rate based on a multi-parametric response model. The therefore robust independent guidance is required. This program effectiveness assessment includes the measurement of serum used a modified-Delphi methodology to develop evidence-based, oligosaccharides, serum immunoglobulin G levels, endurance, pul- expert-agreed recommendations for the management of patients monary function, hearing tests, incidence of infections, use of with MPS IVA/VI. The program was led by an international, antibiotics, psychotic events, questionnaires on quality of life and S72 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 burden of disease for caregivers, among others. All patients affected Liang, Joseph Duque, Hai Tran, Robert C. Wells, Giuseppe Astarita, by AM can participate into the registry, regardless of their treatment. Mark S. Dennis, Kannan Gunasekaran, Ankita Srivastava, Kirk R. Based on the current protocol, the study will be opened for Henne, Dolo Diaz, Adam P. Silverman, Ryan J. Watts, Zachary K. participation for an indefinite time and each patient will be followed Sweeney, Denali Therapeutics, South San Francisco, CA, United States up for 15 years. The protocol does not foresee a pre-specified sample size and aims at including most of the patients treated with VA in Most lysosomal storage diseases (LSDs) involve progressive clinical practice. The SPARKLE study will start in Europe in the central nervous system (CNS) impairment resulting from deficiency second half of 2019. in one or more lysosomal enzymes. Treatment of neuronopathic LSDs remains a considerable challenge, as the recombinant enzymes approved to treat these conditions are ineffective in modifying CNS doi:10.1016/j.ymgme.2018.12.172 disease because they do not effectively cross the blood–brain barrier (BBB). To address this unmet need, we developed the Enzyme Transport Vehicle (ETV), a lysosomal enzyme fused to an engineered Fc domain that binds to the transferrin receptor and enables 157 receptor-mediated transcytosis across the BBB. ETV fusions contain- Retina and optic nerve degeneration in alpha-mannosidosis ing iduronate 2-sulfatase (ETV:IDS), the lysosomal enzyme whose deficiency leads to Mucopolysaccharidosis II (MPS II), were gener- Julia B. Hennermanna, Thea Zindela, Yasmina Amraouia, Laila Arash- ated that have high intrinsic activity and effectively degrade Kapsa, Susanne Pitzb, Juliane Matlacha, aUniversity Medical Center accumulated substrates in IDS-deficient cells and in vivo mouse Mainz, Mainz, Germany, bBürgerhospital, Frankfurt, Germany models of MPS II disease. We establish that ETV:IDS, through its interactions with TfR, significantly improves delivery of IDS to the Alpha-mannosidosis is an ultra-rare autosomal-recessive lyso- brain without affecting the native properties of the enzyme somal storage disease caused by a reduced activity of the enzyme necessary for efficient lysosomal targeting and function. This alpha-mannosidase. Clinical characteristics include facial coarsening, translates to robust reductions of accumulated substrates in both intellectual, motor and hearing impairment as well as skeletal the brain and peripheral tissues of IDS-deficient mice and supports abnormalities. Opacities of the cornea or lens, strabismus, and ocular the potential of ETV:IDS as an intravenous monotherapy to treat motility disorders were reported as typical ocular manifestation in peripheral and CNS manifestations of MPS II. Further, these data, patients with alpha-mannosidosis. In contrast, retina and optic nerve together with the inherent modularity of the platform, suggest that degeneration have been rarely described. We report ocular findings the ETV can be harnessed more broadly to enhance brain delivery of of 32 patients with alpha-mannosidosis and especially concentrated additional lysosomal enzymes, yielding a new class of on retina and optic nerve abnormalities which we supported by biotherapeutics for the treatment of neuronopathic LSDs. posterior segment examination, fundus photography and Spectral- Domain optical coherence tomography (OCT). Retina degeneration with bone spicule formations in the peripheral retina or macular doi:10.1016/j.ymgme.2018.12.174 changes were seen in 3 patients (9.4%) on funduscopy. 8 retinal images could be obtained by OCT or fundus photography of these 6 showed thinning of the outer retinal layers on OCT. Optic nerve atrophy was seen in 6 patients (18.8%) of these 4 with partial 159 atrophy. 2 patients had partial optic nerve atrophy with no retinal Screening for Fabry disease and hereditary ATTR amyloidosis in abnormalities on funduscopy. Cataract was seen in 3 (9.4%), corneal idiopathic small fiber and mixed neuropathy haze in none of the patients. 3 patients (9.4%) had manifest strabismus, 4 (12.5%) with nystagmus, and in 5 patients (15.6%) Aki J. Hietaharjua, Kristin Samuelssonb, Ana Radovicc, Rayomand impaired smooth pursuit eye movements were seen. In conclusion, Pressb, Mari Auranend, Emil Ylikalliod, Henna Tyynismaae, Mikko ocular pathologies in patients with alpha-mannosidosis are not Kärppäf, Matilda Veteläinenf, Niina Peltolag, Svein I. Mellgrenh, Åse exclusively confined to opacities of the cornea or lens, strabismus or Myglandi, Chantal Tallaksenj, Henning Andersenk, Astrid J. ocular motility disorders but retina degeneration and optic nerve Terkelsenk, Freja Fontaink, aTampere University Hospital, Tampere, atrophy may be common eye pathologies in these patients. In Finland, bKarolinska Institutet, Stockholm, Sweden, cKarolinska Univer- contrast to conventional funduscopy, OCT technology provides early sity Hospital, Stockholm, Sweden, dUniversity of Helsinki, Helsinki, diagnosis of retina degeneration by showing thinning of the outer Finland, eHelsinki University Hospital, Helsinki, Finland, fUniversity of retinal layers which can progress with age and potentially leads to Oulu, Oulu, Finland, gTampere University Hospital, Faculty of Medical vision loss over time. and Life Sciences, Tampere, Finland, hUiT The Arctic University of Norway, Tromso, Norway, iSorlandet Hospital, Kristiansand, Norway, jOslo University Hospital, Faculty of Medicine, Oslo, Norway, kAarhus doi:10.1016/j.ymgme.2018.12.173 University Hospital, Aarhus, Denmark

Fabry disease (FD) and hereditary ATTR (hATTR) amyloidosis are treatable diseases known to cause small fiber neuropathy (SFN). The 158 aim of this study was to explore the value of genetic screening for Improved brain uptake and efficacy of iduronate 2-sulfatase with these 2 diseases. This was a retrospective and prospective multicen- the enzyme transport vehicle ter study involving 9 participating neurology departments from 4 Nordic countries (Denmark, Finland, Norway and Sweden). Alto- Anastasia G. Henry, Mihalis S. Kariolis, Julie C. Ullman, Jennifer A. gether, 172 patients were enrolled in this study. Of them, 17 were Getz, Annie Arguello, Cathal Mahon, Junhua Wang, Akhil Bhalla, Tina later excluded. Genetic sequencing of the GLA and TTR genes was Giese, Catherine Bedard, Do Jin Kim, Jessica Blumenfeld, Nicholas performed. A single patient, 69-year old Swedish male, had a Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S73 possible pathogenic variant, R118C, in the GLA gene, but the clinical QOL and offering an alternative regimen compared to existing investigations showed no definite signs of FD. No pathogenic treatments. mutations in the TTR gene were found, either. Our findings show that the yield of screening for FD and hATTR amyloidosis in patients with idiopathic SFN or mixed neuropathy without any clinical doi:10.1016/j.ymgme.2018.12.176 characteristics or disease-specific symptoms in a Nordic population appears to be extremely low in a clinical setting. 161 doi:10.1016/j.ymgme.2018.12.175 Renal and cardiac outcomes of young male patients with Fabry disease initiated on agalsidase beta treatment before age 30: A Fabry registry analysis

160 Robert J. Hopkina, Gustavo H. Cabrerab, John L. Jefferiesc, Eva Brandd, Once every 4 weeks - 2 mg/kg of pegunigalsidase alfa for treating Ulla Feldt-Rasmussene, Dominique P. Germainf, Nathalie Guffong, Ana Fabry disease Preliminary results of a phase 3 study Jovanovich, Ilkka Kantolai, Amel Karaaj, Ana M. Martinsk, William R. Wilcoxl, Meng Yangm, Han-Wook Yoon, Michael Mauero, aCincinnati Myrl D. Holidaa, John Bernata, Nicola Longob, Ozlem Goker-Alpanc, Children's Hospital Medical Center, University of Cincinnati College Eric Wallacea, Raphael Schiffmannd, Patrick Deegane, Nedd Khanf, Medicine, Cincinnati, OH, United States, bCentro Médico Santa María de Camilla Tøndelg, Francois Eyskensh, Derralynn Hughesi, Michael la Salud, Buenos Aires, Argentina, cUniversity of Tennessee Health Westj, Pilar Giraldok, Fatih Ezgul, Einat Almonm, Sari Alonm, Bat-chen Science Center, Methodist University of Tennessee, Memphis, TN, United Amit-Cohenm, Mali Szlaiferm, Raul Chertkoffm, William Wilcoxn, States, dUniversity Hospital Münster, Münster, Germany, eRigshospitalet, aUniversity of Iowa Hospitals and Clinics, Iowa City, IA, United States, Copenhagen University Hospital, Copenhagen, Denmark, fUniversity of bUniversity of Utah, Salt Lake City, UT, United States, cO&O Alpan LLC, Versailles, Paris-Saclay University, Montigny, France, gFemme Mère Fairfax, VA, United States, dBaylor University Medical Center, Dallas, TX, Enfant Hospital, Lyon, France, hSalford Royal NHS Foundation Trust, United States, eCambridge University, Cambridge,, United Kingdom, Salford, United Kingdom, iTurku University Hospital, Turku, Finland, fInfusion Associates, Grand Rapids, MI, United States, gHaukeland jMassachusetts General Hospital, Harvard Medical School, Boston, MA, University Hospital, Bergen, Norway, hAntwerp University Hospital, United States, kFederal University of Sao Paulo, Sao Paulo, Brazil, lEmory Antwerp, Belgium, iLSDU, Institute of Immunity and Transplantation, University School of Medicine, Atlanta, GA, United States, mSanofi Royal Free London NHS Foundation Trust, London, United Kingdom, Genzyme, Cambridge, MA, United States, nUniversity of Ulsan College of jDalhousie University, Halifax, NS, Canada, kHospital de Dia Quiron, Medicine, Asan Medical Center, Seoul, Republic of Korea, oUniversity of Zaragoza, Spain, lGazi Universitesi, Ankara, Turkey, mProtalix, Carmiel, Minnesota, Minneapolis, MN, United States. Israel, nEmory University School of Medicine, Atlanta, GA, United States Fabry disease (FD) is a progressive X-linked disorder from Pegunigalsidase alfa, a novel α-galactosidase-A enzyme for the deficiency of α-galactosidase and lysosomal accumulation of glyco- treatment of Fabry disease (FD) is a stable, homo-dimeric PEGylated lipids. This Fabry Registry (NCT00196742) analysis assessed renal protein, previously demonstrated (Hughes et al, LDN-2016) to have and cardiac outcomes of agalsidase beta treatment (1 mg/kg 2- substantially improved pharmacokinetics (PK) parameters compared weekly) for ≥2 years in male patients with age at first treatment to the two currently available enzyme replacement therapies (ERTs). (AFT) of 5-30 years. GLA variants were classic or unclassified (fabry- The ongoing phase 3 trial “BRIGHT” (PB-102-F50, NCT03180840) is database.org), which shared similar clinical features. Longitudinal an open-label switch-over study assessing safety, PK, and efficacy of post-treatment analyses included estimated glomerular filtration pegunigalsidase alfa (2 mg/kg) given every 4 weeks for 52 weeks to rate (eGFR, bedside Schwartz equation) stratified by low (LRI, urine FD patients previously treated with either agalsidase alfa or protein-to-creatinine ratio [UPCR] ≤0.5 or albumin-to-creatinine agalsidase beta. Aim of the study: to explore a more convenient ratio [UACR] ≤0.3), or high renal involvement (HRI, UPCR N0.5 or dosing regimen exploiting the improved PK characteristics of UACR N0.3), and Z-scores of cardiac interventricular septum thick- pegunigalsidase alfa, as shown by PK modelling from the Phase 1-2 ness (IVST) and left ventricular posterior wall thickness (LVPWT), study results (Warnock et al, LDN-2017). The trial will enroll up to stratified by median AFT. The 31 HRI patients had higher AFT and 30 patients without severe clinical symptoms and relatively slow lower baseline eGFR compared to 189 LRI patients (median AFT: 26.1 disease progression, as evaluated by the investigator. vs. 17.1 years eGFR: 81 vs. 92.6 ml/min/1.73m2 Pb0.01). Among Pegunigalsidase alfa plasma levels are measured at multiple time patients with repeated post-treatment assessments, eGFR declined points before, during, and up to 28±3 days post-infusion for PK more prominently among HRI patients (slopes: HRI [n=16] -2.56 LRI evaluation. Preliminary PK results, from the first 15 patients, show a [n=66] -1.29 ml/min/1.73m²/year P-interaction=0.002). Median half-life of ~80 hours and persistent plasma levels over the entire 4- AFT was 25.3 and 20.6 years, and follow-up 4.4 and 3.9 years, week dosing interval, with a mean concentration on day 28 post- respectively. Median AFT among patients with cardiac measures was infusion of 138 ± 42 ng/mL. The Area Under the Curve (AUC) of day 22.3 years (14 ≥ the median AFT, 13 b median AFT). Post-treatment 0-28 was found to be 2,126,091 ng*h/mL, of which the mean partial LVPWT and IVST Z-scores did not significantly change. Slopes of AUC during the 4th week was ~30,000 ng*h/mL confirming the study LVPWT and IVST Z-scores were -0.10 and -0.07, respectively, for the assumption. The AUC of day 0-28 is significantly greater than older group and -0.08 and -1.11 for the younger group (P- agalsidase beta (AUC0-∞ b11,000 ng*h/mL based on package insert). interaction: LVPWT: 0.25 IVST: 0.11). In conclusion, young males This interim analysis of the ongoing BRIGHT study provides with HRI had a worse baseline clinical profile and started treatment evidences of how pegunigalsidase alfa’s unique biochemical charac- later than LRI patients. After treatment, patients with HRI had a teristics translate into prolonged systemic exposure potentially greater decline in eGFR. Cardiac measures remained stable over time supporting a monthly infusion option. This feature is expected to regardless of age at treatment initiation. The limited patient numbers ease treatment-related burden for FD patients, potentially impacting and absence of untreated FD comparators warrant careful S74 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 interpretation of the results. Funding (Fabry Registry, abstract): females after sustained agalsidase beta treatment. Funding (Fabry Sanofi Genzyme. Registry, abstract): Sanofi Genzyme. doi:10.1016/j.ymgme.2018.12.177 doi:10.1016/j.ymgme.2018.12.178

162 163 Significant abdominal and acute pain improvements in young Bone/joint abnormalities in children/adolescents: A screening patients with Fabry disease initiated on agalsidase beta treatment protocol for mucopolysaccharidosis before age 30: A Fabry registry analysis Dafne D.G. Horovitza, Anneliese L. Bartha, Pedro H. Barros Mendesb, Robert J. Hopkina, Gustavo H. Cabrerab, John L. Jefferiesc, Eva Mariana Q.G. Gonzagaa, Yuri L. Chistéb, Rommel B. Medeirosb, Maria Brandd, Ulla Feldt-Rasmussene, Dominique P. Germainf, Nathalie L. Oliveirac, Fernanda B. Scalcoc, aNational Institute of Women, Children Guffong, Ana Jovanovich, Ilkka Kantolai, Amel Karaaj, Ana M. and Adolescents Health Fernandes Figueira - Fiocruz, Rio de Janeiro, Martinsk, William R. Wilcoxl, Meng Yangm, Han-Wook Yoon, Brazil, bInstituto Nacional de Traumato Ortopedia - Into, Rio de Janeiro, Michael Mauero, aCincinnati Children's Hospital Medical Center, Brazil, cLABEIM, Rio de Janeiro, Brazil University of Cincinnati College Medicine, Cincinnati, OH, United States, bCentro Médico Santa María de la Salud, Buenos Aires, Mucopolysaccharidoses (MPS) are under-diagnosed, especially Argentina, cUniversity of Tennessee Health Science Center, Methodist milder forms. A screening protocol was tested at the National Institute University of Tennessee, Memphis, TN, United States, dUniversity of Orthopedics in Rio de Janeiro - Brazil. Patients were selected from Hospital Münster, Münster, Germany, eRigshospitalet, Copenhagen Pediatric Orthopedics, Spine and Hand clinics. Inclusion criteria were University Hospital, Copenhagen, Denmark, fUniversity of Versailles, age under 18, suspected Legg-Calve-Perthes disease, scoliosis, kypho- Paris-Saclay University, Montigny, France, gFemme Mère Enfant sis, genu valgum, trigger finger, carpal tunnel syndrome, atypical Hospital, Lyon, France, hSalford Royal NHS Foundation Trust, Salford, epiphyseal dysplasia and short stature, associated or not to dysostosis United Kingdom, iTurku University Hospital, Turku, Finland, exclusion criteria were confirmed diagnosis of other genetic bone jMassachusetts General Hospital, Harvard Medical School, Boston, MA, disease and chronic disorders with bone abnormalities. Patients were United States, kFederal University of Sao Paulo, Sao Paulo, Brazil, clinically evaluated to identify other signs / symptoms of MPS, with lEmory University School of Medicine, Atlanta, GA, United States, information on infections, surgeries and other morbidities, other mSanofi Genzyme, Cambridge, MA, United States, nUniversity of Ulsan specialists seen and family history. Selected patients underwent hand/ College of Medicine, Asan Medical Center, Seoul, Republic of Korea, wrist and lateral lumbar spine X-rays and provided urine sample for oUniversity of Minnesota, Minneapolis, MN, United States GAG analyses. Clinical forms, X-rays and urinary results were reviewed by a genetics team familiar with MPS. Patients with X-ray abnormal- Fabry disease (FD) is a progressive X-linked disorder from ities suggestive of MPS and/or abnormal urinary analysis were selected deficiency of α-galactosidase. Gastrointestinal symptoms, neuro- for re-evaluation by genetics and blood collection for enzyme activity pathic pain, and attacks of excruciating pain are among the earliest studies. If necessary, additional exams (complete skeletal X-rays, symptoms. This Fabry Registry (NCT00196742) analysis assessed echocardiogram or others) were ordered. From November 2015 to changes in symptom reports from male and female patients on October 2017, 1119 pediatric patients were evaluated and 55 selected agalsidase beta with age at first treatment of 5-30 years. GLA variants for X-rays and urinary GAG analyses. Two did not complete the exams were classic or unclassified (fabry-database.org), which shared and were excluded. In the remaining 53, MPS was ruled out in 51 after similar clinical features. We compared patient-reported binary comprehensive clinical evaluation, more complete imaging and responses (yes [present]/no [not present]) for abdominal pain, enzyme activity studies in selected cases. Two patients with short diarrhea, and peripheral and acute pain between baseline and last stature, genu valgum, oar shaped ribs, abnormal spine and metacarpals follow-up ≥0.5 or ≥2.5 years on treatment separately. Ages at first had normal urinary GAGs enzyme activity studies will be ordered. So treatment and follow-up durations were expressed as group far, no MPS diagnosis was confirmed. We diagnosed a spondilo- medians. Of the early symptoms, peripheral pain was most epiphyseal dysplasia and an epiphyseal dysplasia. Despite not having frequently reported at baseline by the vast majority of the patients. MPS confirmed, we believe the protocol can be effective, although In Acute pain was least frequently reported. Compared to baseline, future studies isolated scoliosis will no longer be an inclusion criterion. significantly fewer males with ≥0.5 years follow-up reported acute Acknowledgements: Biomarin - financial support Rede MPS Brasil - pain after 2.1-year follow-up (17/78 [21.8%] vs. 7/78 [9%], P=0.012, enzyme studies. age at first treatment 17.3 years). Data was more limited for patients with ≥2.5 year responses. Among males, the significant decrease in acute pain reports remained after median 4.2-year follow-up doi:10.1016/j.ymgme.2018.12.179 (baseline 11/31 [35.3%], follow-up 1/31 [3.2%], P=0.002). Addition- ally, much fewer males and females reported abdominal pain after median 4.2- and 3.6-year follow-ups, respectively, compared to baseline (males: 21/29 [72.4%] vs. 10/29 [34.5%], P=0.002 females: 164 12/18 [66.7%] vs. 6/18 [33.3%], P=0.034, ages at first treatment were Enzyme replacement therapy in mucopolysaccharidosis type II 21.3 and 22.4 years, respectively). There were no significant changes with alternative dosing 1mg/kg idursulfase in every other week in yes/no responses for diarrhea or peripheral pain. In conclusion, in infusions young FD patients a reduction in reported acute pain in treated males was sustained over the duration of follow-up. Furthermore, Dafne D. Horovitza, Anneliese L. Bartha, Carolina F. Souzab, Sueli G. there were fewer reports of abdominal pain in both males and Canossac, Marco A. Curiatic, Sandra O. Kyosenc, Maria L. Oliveirad, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S75

Fernanda B. Scalcod, Ana M. Martinsc, aNational Institute of Women, laringomalacia (13% severe). Epiglottis was thickened in 57%, 13% with Children and Adolescents Health Fernandes Figueira - Fiocruz, Rio de total loss of organ support. Tracheal deformities were present in 53%, Janeiro, Brazil, bHospital de Clinicas de Porto Alegre, Porto Alegre, Brazil, most with diffuse involvement. Abnormal anatomy of bronchi was cCentro de Referencia em Erros Inatos do Metabolismo, São Paulo, Brazil, observed in 13,3%. 4 patients (3 MPSII and 1 MPSVI) presented dLABEIM, Rio de Janeiro, Brazil endobronchial obstruction secondary to possible GAG deposits and 2 MPSII died of respiratory complications. Abnormalities varied among Enzyme replacement therapy for mucopolysaccharidosis II (MPS II) MPS types: while the MPSI and II had supraglottic redundancy and with Idursulfase (ElapraseR) has proven effective in reducing urinary tracheomalacia, no laryngeal disease was observed in the MPS-IVA and glycosaminoglycan (uGAG) levels, liver and spleen volumes and in VI group. On the other hand, all MPSIVA, 75% of the MPSII, 64% of the increasing walking distance. During phase II/III studies, 0.5mg/kg MPSVI and 50% of the MPSI had tracheobronchial deformity, being the every other week (EOW) showed effectiveness in combined outcomes MPSIVA group the most striking and severe. Significant and impressive (6-minute walk test - 6MWT, pulmonary function and uGAG involvement of the trachea and bronchi architecture varied according to reduction), although some were not significantly effective when MPS subtype, severity of the phenotype and age of the patient. There individually analyzed. A weekly 1.5mg/kg dose was tested in 4 was a strong correlation between airway damage, patient age and patients for 48weeks and proven safe (Muenzer et al, 2007). Double duration of disease, despite adequate ERT. On the other hand, the dose EOW infusions was considered an acceptable treatment option in introduction of ERT at a very young age seemed to correlate with slightly MPS I in a dose study (Giugliani et al, 2009) and proven safe and milder lower airway disease. effective in 20 patients (Horovitz et al., 2016). Such data might suggest that double dose Idursulfase every other week infusions for MPS II doi:10.1016/j.ymgme.2018.12.181 might also be effective. A dose of 1mg/kg EOW substituted the recommended weekly dose in seven MPS II patients in three different centers in Brazil. Patients had to be under enzyme replacement therapy with Idursulfase for at least one year without adverse events. 166 Before switching to EOW thorough clinical examination, 4 week uGAG Evaluation of long-term effects by ERT for Fabry disease collection, and additional evaluations (echocardiogram, endurance biochemical and EM pictures tests, pulmonary function studies, abdominal ultrasound and quality of life questionnaires) were performed whenever possible. Upon Mohammad A. Hossaina,b, Chen Wua, Takashi Miyajimaa, Keiko completion of 48 weeks under EOW intravenous infusions of 1mg/kg Akiyamaa, Rina Itagakia, Kaoru Etoc, Takeo Iwamotob, Yoshikatsu of ERT evaluations were repeated. Six patients completed 48 weeks on Etob,a, aAdvanced Clinical Research Centre, Kawasaki, Kanagawa, Japan, EOW one patient died from acute complications of cardiac surgery bJikei University of Medical School, Tokyo, Japan, cTokyo Women’s after 4 months under the protocol. No infusion-associated adverse Medical University, Tokyo, Japan events were observed. uGAG evaluated regularly showed stable excretion, similar to when receiving the weekly dose. Patients were Fabry disease is an X-linked lysosomal storage disorder caused by a well and families very satisfied with the new dosing regimen repeated deficiency of alpha-gal A, leading to the progressive accumulation of evaluations after one year on EOW therapy showed no difference sphingolipids. Enzyme replacement therapy (ERT) is the most common when compared to the previous dose. Most patients chose to continue therapy at present which has been approved in Japan since 2004. with the EOW regimen after the protocol. Different degrees of clinical improvements by ERT were reported in numerous articles based on patients’ compliance, improved GFR, improved cardiac activity and etc. Few direct evidences of tissue biopsy doi:10.1016/j.ymgme.2018.12.180 during ERT were reported so far except for kidney biopsy. Here, we present plasma lyso-Gb3 data and electron microscopy (EM) of skin biopsy from 30 cases including male and female who are receiving ERT for almost 13 years. All the cases were diagnosed based on their family 165 history, clinical, biochemical and molecular findings. Lyso-Gb3 was Mucopolysaccharidoses and laryngeal, tracheal and bronchial measured in plasma, skin biopsy was taken for electron microscopy disease: Type-specific abnormalities and long-term implications (EM). The IgG antibody titer of the patients were measured against alpha-gal A enzyme. The male patients having severe type of mutations Dafne D. Horovitz, Paulo Pires de Mello, Mariana Pires de Mello with high antibody titers showed high plasma lyso-Gb3 and still Valente, Danielle Torres, Anneliese L. Barth, National Institute of massive accumulation of sphingolipids in the fibroblasts, exocrine Women, Children and Adolescents Health Fernandes Figueira - Fiocruz, glands and Schwan cells. The severely affected female cases also showed Rio de Janeiro, Brazil high lyso-Gb3 and sphingolipids accumulation. Morphological pictures in treated female cases showed rather different patterns from those of Airway complications are among the most common early and lethal male. The patients who were receiving ERT before the phenotype manifestations of mucopolysaccharidoses (MPS). Not much is known, appeared, had low plasma lyso-Gb3 and minimal sphingolipids however, on the anatomical characteristics of the airways in this patient accumulations in skin by EM. Several reports suggested that high group, the exact sites of involvement and variability between the plasma antibody titer minimizes the effect of the enzyme and also the different subtypes of MPS. We evaluated the lower airways of 32 MPS uptake of enzyme were varied by tissues or cell types. Our studies patients (23 males, 9 females), divided in the following MPS types: I(6), clearly indicate that EM studies in skin plus plasma lysoGb3 measure- II(10), III(2), IVA(3) and VI(11). Exams were performed with a flexible ment could be valuable for the evaluation of the efficacy of ERT to follow 3,5mm bronchoscope, trespassing the glottis and viewing the anatomy up Fabry patients. of the lower airways. Two MPSII patients transplanted before 3months with normal airways were excluded from the analysis. Laryngeal deformities were present in 37% (edema in aritenoids, thickening of doi:10.1016/j.ymgme.2018.12.182 false vocal cords, post-cricoid edema). 33% presented with S76 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

167 administrator collated the responses. In round 2, each panel member CRISPR-Cas9 generated Pompe knock-in murine model exhibits rated each indicator for importance on a 5-point Likert scale (1=not early-onset cardiac hypertrophy and motor impairment important to 5=extremely important). Indicators rated most important (a score of ≥3 assigned by N75% of respondents) were Jeffrey Y. Huanga, Anthony D. Rangela, Yunghang Chanb, Nancy reissued to the panel in round 3 for agreement rating using a 5-point Daltonb, Jon Neumannc, Raymond Y. Wanga, aChildren's Hospital of Likert scale (1=strongly agree to 5=strongly disagree). Consensus Orange County, Orange, CA, United States, bUniversity of California San was achieved for an indicator if N67% of respondents rated their Diego, La Jolla, CA, United States, cUniversity of California Irvine, Irvine, agreement ≥4. Based on responses in round 1, the panel (100% CA, United States response rate) voted on 15 renal, 15 cardiac, 13 CNS, 24 patient- reported and 16 other indicators. In round 2, 7 renal, 12 cardiac, 5 The goal of this study is to generate and characterize a knock-in CNS, 13 patient-reported and 8 other indicators were deemed most model of Pompe disease (PD) - a rare, progressive, fatal disorder important. In round 3, consensus was reached on 6 renal, 10 cardiac, primarily affecting the cardiac and musculoskeletal systems. While a 2 CNS, 6 patient-reported and 5 other indicators. At least 90% of the murine model of PD exists, it bears a Cre/loxP induced exonic panel agreed that: elevated albumin:creatinine ratio or micro- insertion of a neomycin cassette and does not completely recapitu- albuminuria histological evidence of renal involvement minor late severe human PD - displaying nonfatal hypertrophic cardiomy- abnormalities of glomerular filtration rate markers of early systolic/ opathy only late in its natural history. We therefore designed a diastolic dysfunction neuropathic pain and pain in the extremities/ CRISPR-Cas9 knock-in system targeting the Gaa gene to introduce neuropathy, were important early indicators of organ damage. the known pathogenic CRIM negative Gaa mutation c.1826insA (p. Findings from this consensus initiative may guide early appropriate Y609*). Following optimization of our knock-in strategy in cultured disease-specific therapy initiation in FD. murine myoblasts, we successfully generated a Gaac1826insA mouse model using a dual sgRNA with ssODN donor template approach. Whole genome sequencing and analysis of the Gaac1826insA murine doi:10.1016/j.ymgme.2018.12.184 model establishes that our system is highly specific for the Gaac1826 target locus and does not induce any off-target mutations or genomic rearrangements. Next, we examined GAA mRNA transcript, protein 169 expression and enzymatic activity levels in our PD knock-in mice. Clinical features of Fabry disease in patients with mutations c1826insA fi Gaa mice display signi cantly reduced levels of GAA expres- amenable and non-amenable to migalastat sion and enzymatic activity relative to wild-type mice. We performed echocardiography on Gaac1826insA mice to assess cardiac Derralynn Hughesa, Cathy Qina, De Silvaa, Lucia Lavallea, Uma structure and function. Gaac1826insA mice exhibit early-onset, pro- Ramaswamib, Atul Mehtab, aUniversity Colege London, London, United gressive cardiac hypertrophy as measured by significant increases in Kingdom, bRoyal Free London NHS Foundation Trust, London, United left ventricular wall thickness and mass index by 3 months of age. Kingdom We also conducted functional tests - grip strength, inverted screen, c1826insA gait analysis - on Gaa mice every 3 months to assess overall Alpha-galactosidase A mutations on chromosome Xq22.1 lead to a c1826insA motor performance. Gaa mice display impaired motor reduction in cognate enzyme activity, with resultant accumulation of strength and coordination relative to wild-type mice. Altogether, globotriaosylceramide, and the clinical features of Fabry disease c1826insA our results demonstrate that the Gaa murine model recapit- (OMIM 301500). Genotype-phenotype relationship is not straightfor- ulates human infantile-onset Pompe disease and is better suited for ward. Disease-specific therapy involves intravenous enzyme infusion evaluation of therapeutic strategies such as genome correction. or more recently oral pharmacological chaperone therapy (for patients with ‘amenable’ mutations, exhibiting relative increase in α-Gal A activity ≥ 1.2-fold above baseline and absolute increase in α-Gal A doi:10.1016/j.ymgme.2018.12.183 activity ≥ 3% of wild-type in HEK cells after incubation with 10 uM migalastat). Clinical phenotype was assessed in 200 consented patients with Fabry disease attending the Royal London Free NHS 168 Foundation Trust, with emphasis on features among those with an A global consensus on early indicators of organ damage in Fabry ‘amenable’ GLA mutation. Additionally, we collated published data on disease and implications for treatment initiation genotype and phenotype recorded in Fabry disease publications describing 147 separate mutations in males and 98 in females, which Derralynn Hughesa, Jack Johnsonb, Sandro Feriozzic, aThe Royal Free was used to construct heat maps of clinical features among those with Hospital, London, United Kingdom, bFabry Support & Information Group, amenable and non-amenable mutations. In the local cohort, a total of Concordia, MO, United States, cBelcolle Hospital, Viterbo, Italy 22 distinct amenable mutations were noted in 93 females and 54 males, whilst 28 females and 25 males had a total of 19 non-amenable Diagnostic delays are common in Fabry disease (FD), and mutations. The mean age-adjusted severity score of patients with initiation of disease-specific therapy is only recommended in certain amenable mutations was -0.81 (males -2.27, females 0.64) compared patients at diagnosis. Timely treatment of FD is critical to mitigate or with 5.9 in patients with non-amenable mutations (males 6.5 and prevent organ damage. Earlier treatment could be facilitated if females 4.8) pb0.001. Whilst acroparasthesia (88% versus 33% physicians knew which early indicators of organ involvement in FD p=0.04) and angiokeratoma (96% versus 26% p=0.002) were more are most predictive of later damage. In round 1 of an anonymous, common in patients with non-amenable mutations organ involvement iterative, three-round Delphi process, a panel of 21 expert physi- was comparable between the groups (male left ventricular hypertro- cians, from 15 countries, were asked independently to list early phy 56% and 59% female left ventricular hypertrophy 25% and 17% indicators of organ damage that can be readily assessed and that male CKDN3 17% and 17% respectively female CKDN3 7% and 4% might merit initiation of disease-specific therapy an independent respectively). Disease burden, as described, in end organ sites of Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S77 pathology (heart, kidneys) is comparable between those with and GalA). FD is characterized by progressive systemic accumulation of without amenable mutations. the enzyme’s substrates, globotriaosylceramide (Gb3) and lyso-Gb3, leading to renal, cardiac and/or cerebrovascular disease and culminating in premature demise. FD is treated by enzyme replacement doi:10.1016/j.ymgme.2018.12.185 therapy (ERT), however the short enzyme half-life necessitates lifelong biweekly infusions. A more effective and long-lasting treatment would benefit FD patients. An AAV-mediated liver-targeted gene therapy was evaluated in a mouse model (GLAKO) that lacks α-GalA 170 activity and accumulates high levels of Gb3/lyso-Gb3 in plasma and Preclinical studies of a brain penetrating IgG Trojan horse- tissues. This strategy employs an episomal AAV (serotype 2/6) vector arylsulfatase fusion protein in the metachromatic leukodystro- encoding human GLA cDNA (hGLA) driven by a liver-specific prophy mouse moter. One-time administration of increasing amounts of AAV-hGLA cDNA generated using a clinical scale manufacturing process resulted Eric K.-W. Hui, Jeff Z. Lu, Ruben J. Boado, William M. Pardridge, in supraphysiological expression of plasma α-GalA (over 300-fold of ArmaGen Inc., Calabasas, CA, United States WT) by study day 15, was well tolerated, and was stable for 3 months post-injection. Dose-dependent increases in α-GalA activi- Metachromatic Leukodystrophy (MLD) is a neurodegenerative ties were achieved in liver, heart and kidney with a corresponding disease caused by mutations in the lysosomal enzyme, arylsulfatase A reduction of Gb3/lyso-Gb3. This initial vector was compared to an fi (ASA). ASA de ciency leads to sulfatide glycolipid accumulation in improved cDNA vector in a 1-month study using two different AAV brain. Treatment of MLD with intravenous recombinant ASA is not doses in wild type C57BL/6 mice. The improved cDNA vector possible because this molecule does not cross the blood-brain barrier produced on average 7-fold higher levels of plasma α-GalA activity (BBB). BBB penetration by ASA was enabled by re-engineering ASA as an at study day 28 than mice administered the same dose of the initial IgG-ASA fusion protein, where the IgG domain is a chimeric monoclonal cDNA. The high levels of α-GalA activity seen in these studies, along antibody (MAb) against the mouse transferrin receptor (TfR), desig- with the concomitant marked reduction in the accumulated Gb3/ nated cTfRMAb. The IgG domain acts as a molecular Trojan horse to lyso-Gb3 in key tissues of the GLAKO mouse model, provide “proof- deliver the ASA into brain via transport on the endogenous BBB TfR. The of-concept” for AAV-mediated targeting of hepatocytes to express fi cTfRMAb-ASA fusion protein bound to the mouse TfR with high af nity therapeutic levels of human α-GalA. The clinical scale manufacturing and retained high ASA enzyme activity comparable to recombinant process developed for these studies will enable rapid development human ASA. In an intial dose ranging study, 14-week old MLD mice and production in 2019 of clinical-grade material featuring the were treated for 5 weeks with thrice-weekly intraperitoneal or improved cDNA construct. subcutaneous injections of 5 mg/kg of the cTfRMAb-ASA fusion protein. This short-term chronic administration of the cTfRMAb-ASA fusion protein was well tolerated, and no clinical injection related reactions doi:10.1016/j.ymgme.2018.12.187 (IRR) were observed. In contrast, the short-term administration of recombinant human ASA alone to MLD mice causes severe IRRs, which prevents long-term treatment (Hum Mol Genet 2005). Based on these findings, a long-term efficacy study was initiated, and MLD mice were 172 treated twice-weekly with saline, the cTfRMAb isotype control alone, or Challenges of regulatory requirements for patient registries in the cTfRMAb-ASA fusion protein intraperitoneally at 5mg/kg for up to different countries 12 months. Preliminary analysis completed at 6 months of treatment a b c c with N500 doses confirmed the safety of the cTfRMAb-ASA dosing with Jackie Imrie , Marc Patterson , Jim Green , Toni Mathieson , Shaun d a b no clinical IRRs, normal body weight and normal hematocrit and Bolton , INPDR, Bolton, United Kingdom, Mayo Clinic Children's c reticulocytes. The study demonstrates for the first time that ASA, Centre, Rochester, MN, United States, INPDR, Tyne and Wear, United d following re-engineering as an IgG fusion protein has an excellent safety Kingdom, University Hospital Birmingham, Birmingham, United profile allowing for long-term treatment of MLD mice. Kingdom

The International Niemann-Pick Disease Registry (INPDR) is a fi doi:10.1016/j.ymgme.2018.12.186 single, rare disease-speci c registry collating global Niemann-Pick Disease data. Created by professionals and patients for worldwide use, it collects clinical and patient reported data with separate datasets for ASMD and NP-C. It will replace the need for multiple 171 registries and offer a single, effective and robustly regulated data Liver-targeted AAV gene therapy vectors produced by a clinical resource for NPD. The registry is currently established in 6 countries scale manufacturing process result in high, continuous therapeu- - UK, Czech Republic, Spain, Italy, Ireland and partly in Germany. tic levels of enzyme activity and effective substrate reduction in Countries currently progressing R and D applications include mouse model of Fabry disease Germany, USA, Argentina, Brazil, Turkey, Netherlands, France and Australia. Additionally discussions have started with Israel, Taiwan Marshall W. Hustona, Makiko Yasudab,SilverePagantb, Susan St Martina, and China. The way forward for the INPDR as well as other LSD and Liching Caoa, Lillian Falesea, Kathleen Meyera, Robert J. Desnickb, rare disorder registries requires international collaboration. Our Thomas Wechslera, aSangamo Therapeutics, Richmond, CA, United States, experience to date in establishing the INPDR is that there exist bMount Sinai School of Medicine, New York, NY, United States major challenges in meeting regulatory requirements, which vary between centres, countries and supranational organisations, such as Fabry disease (FD), an X-linked lysosomal storage disease, is the European Union. We will be looking at the regulatory caused by mutations in the GLA gene encoding α-galactosidase A (α- requirements of all centres currently undergoing the approval S78 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 process to compare requirements and identify potential approval 174 barriers, initially starting with those that are a particular challenge In vivo gene therapy for Tay-Sachs and Sandhoff diseases by for various reasons: USA, France and Germany. In each of these utilizing AAV9 vector encoding modified HEXB jurisdictions, there are high standards for research compliance, which are not aligned. For example, in the United States, all human Kohji Itoha, Daisuke Tsujia, Yukiya Ohnishia, Ryo-suke Watanabeb, research must comply with regulations administered by the Katsuhito Asaic, Shin-ichi Muramatsud, aGraduate School of Pharma- Department of Health and Human Services, specifically 45 CFR 46, ceutical Sciences, Tokushima University, Tokushima, Japan, bFaculty of administered by the Office of Human Research Protections (OHRP), Pharmacy, Tokushima University, Tokushima, Japan, cGene Therapy where as in the European Union human research may be subject to Research Institute, Kawasaki, Japan, dJichi Medical University, Shimono, the General Data Projection Regulation. We will compare and Tochigi, Japan contrast, in detail, the regulatory requirements that apply to the establishment of an international rare disease registry. Tay-Sachs disease (TSD) and Sandhoff disease (SD) are autosomal recessive GM2 gangliosidoses, caused by the gene mutations of HEXA and HEXB encoding the α and β-subunits of lysosomal β- doi:10.1016/j.ymgme.2018.12.188 hexosaminidase (Hex), respectively. These incurable diseases associate with the HexA (αβ heterodimer) deficiency and excessive accumulation of GM2 gangliosides (GM2) in brains of the patients 173 and neurological manifestations. Although high incidence of TSD in Neuronal ceroid lipofuscinosis (NCL) types 1 and 2: Enzyme Ashkenazy-Jewish populations is well known, there are 12 of characteristics of PPT1 and TPP1, and their high risk and Japanese TSD patients at present. We constructed an adeno- fi newborn screenings associated viral vector (AAV9/3-modHEXB) encoding the modi ed HEXB to produce the modified HexB composed of homodimeric β- subunits carrying 9 amino acid residues substituted to those of the Rina Itagakia, Masahiro Endoa, Hiroko Yanagisawaa, Mohammad Arif α-type involving GM2 degradation. We demonstrated that the Hossaina, Keiko Akiyamaa, Takashi Miyajimaa,b, Chen Wua,b, Takeo intracerebroventricular and intravenous administration of AAV9/3- Iwamotoa,c, Junko Igarashib, Haruo Shintakud, Yoshikatsu Etoa,e, modHEXB to SD model mice could restore the GM2-degrading aAdvanced Clinical Research Center, Institute of Neurological Disorder, activity, reduce the GM2 accumulated in the brains, repress the Kanagawa, Japan, bInstitute of Rare Disease, AnGes Co., Tokyo, Japan, motor dysfunctions and prolong the life span. We also found that the cInstitute of Medical Science, Tokyo Jikei University School of Medicine, neuronal cells induced from iPS cells derived from a TSD patient Tokyo, Japan, dOsaka City University Graduate School of Medicine, secreted the human active modHexB after treatment with AAV9/3- Osaka, Japan, eJikei University School of Medicine, Tokyo, Japan modHEXB, suggesting the “cross-correction” effect. From these fi Neuronal ceroid lipofuscinosis (NCL) is lysosomal storage disease ndings, the in vivo gene therapy by utilizing AAV9/3-modHEXB will caused by accumulation of ceroid/lipofuscin in various tissues, which be expected as a novel low-antigenic treatment especially for TSD. leads to devastating brain disorders. The NCL consisted of 14 different genetic types and among these, we focused on NCL 1 and doi:10.1016/j.ymgme.2018.12.190 2. NCL 1 is caused by the deficiency of palmitoyl protein thioesterase (PPT1), whereas NCL 2 is caused by the deficiency of tripeptidylepeptidase (TPP1). Currently, intrathecal enzyme treatments/AAV gene therapy for NCL 1 & 2 are now developing. 175 Therefore, early diagnosis and treatment will be essentially necessary Effects of small molecule therapies on lysosomal function in for better clinical outcome. We studied the enzyme characteristics of Gaucher disease PPT1 and TPP1. We characterized PPT1 and TPP1 enzymes in DBS, plasma/serum, leukocytes/lymphocytes, and fibroblasts from NCL 1 Margarita Ivanova, Julia Dao, Nida Sharief, Ozlem Goker-Alpan, and 2 patients and control subjects, using fluorescent substrates Lysosomal and Rare Disorders Research and Treatment Center, Fairfax, according to Lukacs et al. The PPT1 enzyme activities in control VA, United States subjects had one acidic form, optimal pH 5.0. The PPT1 enzyme activities in NCL 1 patients showed deficient activities in acidic Gaucher disease (GD), the deficiency of the lysosomal enzyme condition, pH 5.0. Conversely, the TPP1 enzyme activities in control glucocerebrosidase (GCase), is caused by the mutations in GBA. Most cells consisted of two forms, an acidic form and a neutral form, and mutations lead to misfolding of GCase in the endoplasmic reticulum the optimal pH was 4.5 and 6.5, respectively. In NCL 2 patients, TPP1 and altered protein trafficking to the lysosome. The accumulated enzyme activities were markedly reduced in acidic conditions, pH substrate, glucosylceramide in the lysosomes is also suggested to 4.5, whereas it showed higher enzyme activities at pH 6.5-7.0. We alter autophagy-lysosome pathway (ALP) functions. Enzyme replace- also carried out high risk screening in epileptic patients with ment therapy successfully treats hematologic manifestation, how- progressive intellectual problem (about 30 patients) and newborn ever has lessor no effect on other end organs, such as brain, lungs, screening (about 2,000 DBSs from newborn infants) were carried out and bones. Small molecule therapies haves the potential for better using this new enzyme assay system. We are now continuing both tissue distribution and nervous system penetration. Eliglustat high risk and newborn screening, and we could not yet currently find hemitartrate (EGT), a substrate reducing agent, works by inhibiting out a PPT1 or TPP1 deficient patient. UDP-glucosylceramide synthase, the first enzyme that catalyzes the biosynthesis of glycosphingolipids, thus reducing the load of glucosylceramide influx into the lysosome. Ambroxol (ABX), is a doi:10.1016/j.ymgme.2018.12.189 pharmacologic chaperone shown to partially recover the mutated GCase. The effects of both molecules on ALP are yet to be studied. The Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S79 effects of EGT and ABX on autophagy-lysosomal pathways were suggests that the effect of genetic background on the phenotype examined in various primary cells derived from patients with GD. is determined by other genetic and epigenetic factors. Overall, both compounds increased GCase activity, but this effect was only observed in some patients. GCase expression analysis demonstrated elevated protein but not mRNA levels after ABX and EGT doi:10.1016/j.ymgme.2018.12.192 treatments. There was also an increase in autophagic vesicle formation and autophagosome accumulation as evidenced by changing levels of LC3A/BII. There was an increase in LysoTracker signal and mitochondria levels, but RT-PCR demonstrated no change 177 fi in LC3 and LAMP1 in primary fibroblasts, PBMC and macrophages. Distribution of chemically modi ed rhSulfamidase to CNS mon- Decreasing RNA levels of pro-inflammatory cytokines CCL17, IL10, itored by brain microdialysis and repeated CSF sampling after IFNγ were observed in macrophages following both treatments, intravenous administration in rat while ABX particularly inhibited CCL2 levels. While the small a a a a molecules, EGT and ABX have completely different mechanisms of Juliette Janson , Gudrun Andersson , Lars Bergquist , Maria Eriksson , b a action, they both lead to ALP activation and decreased pro- Joost H.A. Folgering , Swedish Orphan Biovitrum, Stockholm, Sweden, b inflammatory cytokine levels that could be among the reasons for Charles River Laboratories, Groningen, Netherlands improved GCase activity in select cell lines. Mucopolysaccharidosis type IIIA (MPS IIIA) is an autosomal recessive disorder caused by absence or deficiency of the lysosomal doi:10.1016/j.ymgme.2018.12.191 enzyme sulfamidase. Glycans of recombinant human sulfamidase were chemically modified to generate CM-rhSulfamidase with retained stability and catalytic activity, but with significantly reduced uptake in peripheral tissues. For the enzyme to modify the neuropa- 176 thology of MPS IIIA disease, it has to reach the lysosomes of cells in the Effects of genetic background on disease phenotypes in a mouse brain, hence pass the blood brain barrier (BBB). To monitor BBB model of Fabry disease passage of CM-rhSulfamidase in a time dependent manner, concentration in brain interstitial fluid was explored using push-pull a a a Siamak Jabbarzadeh-Tabrizi , Taniqua Day , Mouna Taroua , Michel microdialysis combined with CSF and serum collection. Push-pull b b a Boutin , Christiane Auray-Blais , Raphael Schiffmann , Jin-Song microdialysis is a sampling technique facilitating continuous mea- a a b Shen , Baylor Research Institute, Dallas, TX, United States, Faculty of surement of larger molecules (e.g. proteins), here applied in awake, Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC, freely-moving Sprague Dawley rats. Having established membrane Canada recovery, surgery was performed on rats to implant guide cannulas for the microdialysis probes using a stereotaxic frame to pin-point exact Knockout mouse model of Fabry disease (Ohshima, 1997) is location in the prefrontal cortex, and cannulas for repeated CSF and widely used, and has been proven an invaluable tool to investigate serum sampling in the cisterna magna and jugular vein, respectively. disease pathogenesis and therapies. However, the importance of After the animals fully recovered, CM-rhSulfamidase was dosed genetic background of this mouse line has been underestimated, intravenously and brain interstitial fluid was sampled by microdialy- and in many publications WT control mice were chosen incorrectly. sis, CSF and serum via their respective cannulas. Concentrations of CM- Our aim was to examine the effects of genetic background on rhSulfamidase were determined by an immunoassay and data were biochemical and functional abnormalities of Fabry mice. We subjected to quantitative pharmacokinetic analysis. While maximum characterized Fabry mice in B6/129 mixed background (~75% B6) serum concentration was observed shortly after administration, a and Fabry mice on a purely C57BL/6 strain (10 generations delay was observed of CM-rhSulfamidase entering CSF and brain backcrossing), compared with WT mice on an identical genetic interstitial fluid. Maximum CSF and brain interstitial fluid concentra- background. Both Fabry-B6/129 and Fabry-B6 developed progres- tions differed somewhat between rats, but after reaching the sive cardiac and renal hypertrophy and impaired thermosensation. maximum level reduction in concentration occurred at a similar rate However, Fabry-B6/129 exhibited earlier onset and more severe between individual rats. Mean CM-rhSulfamidase concentrations phenotypes than Fabry-B6. Cardiac and/or renal hypertrophy versus time relationships were similar in brain interstitial fluid and appeared at age of 6 months in Fabry-B6/129, but at 12 months CSF. Brain exposure was confirmed by determination of brain in Fabry-B6. At 12 months, kidney-to-body weight ratio increased homogenate concentrations. In conclusion, CM-rhSulfamidase bio- by 50% in Fabry-B6/129 (vs. WT), but only 15% in Fabry-B6. The distribution to CNS was quantified using a combined push-pull brain onset of the thermosensation abnormality was at age 2 months in microdialysis, serum and CSF approach. Fabry-B6/129 and 7 months in Fabry-B6. Cardiac expression of hypertrophy marker, atrial natriuretic peptide (ANP), was upregulated in Fabry-B6/129, but not in Fabry-B6. Despite these differ- doi:10.1016/j.ymgme.2018.12.193 ences, globotriaosylceramide (Gb3) and lyso-Gb3 levels in the heart, kidney and dorsal root ganglia were similar between Fabry-B6/129 and Fabry-B6. However, Gb3 level in liver (organ unaffected in Fabry disease) was twofold higher in Fabry-B6/129 compared to 178 Fabry-B6. Additionally, several parameters including body weight, Adding enzyme replacement therapy after hematopoietic stem organ weight, kidney Gb3 and cardiac ANP level were different cell transplantation results in increased metabolic correction in between WT-B6/129 and WT-B6. In conclusion: 1) genetic back- MPS VI ground has a significant impact on the severity and onset of phenotypes in Fabry mice 2) the lack of correlation between Jeanine R. Jarnes-Utz, Elizabeth Braunlin, Paul Orchard, Chester B. phenotype and Gb3/lyso-Gb3 levels in disease-relevant organs Whitley, University of Minnesota, Minneapolis, MN, United States S80 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Following allogenic bone marrow transplantation with predomi- 180 nance of donor engraftment, patients with Maroteaux-Lamy syn- Liver-directed gene therapy corrects Fabry disease in mice drome (MPS VI) demonstrate several indications of metabolic correction, such as reduction in liver size, and achieve near-normal Jey Jeyakumara, Azadeh Kiaa, Jenny McIntosha, Daniel Verhoefa, Petya levels of urine glycosaminoglycan (GAG). Thus it was somewhat Kalchevaa, Paniz Hosseinia, Rose Sheridana, Romuald Corbaua, Amit surprising that, a single infusion of galsulfase after 20 years of full Nathwania,b, aFreeline Therapeutics, Stevenage, United Kingdom, bUCL donor engraftment resulted in a further decline of GAG into the normal Cancer Institute, London, United Kingdom. range (Whitley and Jarnes-Utz, Mol Genet Metab 101(4): 346-348, 2010). Urine GAG declined from slightly high pre-treatment levels Fabry disease is an X-linked lysosomal storage disorder (LSD) (7.63 mg GAG/mmol creatinine range 7.0-8.5, N=3) progressively resulting from mutations in the gene encoding for α-galactosidase A declining below the age-specific normal range (b6.5) over 10 days to (GLA). It is characterised by the abnormal accumulation of neutral the lowest level of 4.4, with a mean post-treatment level of 5.60 glycosphingolipids (GSL), predominantly Globotriaosylceramide (n=9). Thus, we wondered whether this increased circulating enzyme (Gb3), in the lysosomes of multiple cell types including vascular levels (or an alternative route of administration) result in better endothelial cells. It is associated with early-onset stroke, cardiomy- therapy than is achieved by hematopoietic stem cell transplantation opathy, and progression to end-stage renal failure. We examined the alone, and would supplemental galsulfase enzyme replacement therapeutic efficacy of GLA gene therapy in Fabry mice using therapy improve the clinical outcome after transplant? To answer this recombinant adeno-associated virus (rAAV) vectors, directed by the question, the same patient was re-started on intravenous galsulfatase liver-specific promoter (FRE1). In this study, we showed that a single at age 27 and, over two years, show an immediate and sustained intravenous (IV) injection of rAAV8-FRE1-GLAco (2x1012 vg/kg) reduction in monthly urine GAG to ~5 mg/mmol in the normal range achieved supraphysiological levels of plasma GLA activity up to (b6.5) with increased endurance by an independent observer 1061-fold of normal with concomitant correction of lysosomal (mother). We conclude that increased enzyme, above that provide storage pathology in multiple key organs in a Fabry mouse model. by hematopoietic cell transplantation alone, provides increased Animals injected with our novel codon-optimised (“GLAco”) GLA metabolic correction as evidenced by reduction in urine GAG and construct exhibited a rapid elevation in plasma GLA activity levels increased physical endurance. (Supported by BioMarin.) reaching a peak level by 4 weeks post-injection, and this level of expression was maintained for the duration of the study (14 weeks). Furthermore, mass spectrometry (LC-MC/MS) analysis of GSL extract doi:10.1016/j.ymgme.2018.12.194 from different tissues provided proof of exposure and storage clearance in various key organs, which is indicative of metabolic cross-correction of liver-derived GLA enzyme. Gb3 levels in the 179 plasma were reduced by 91% (p=0.008) in the kidney by 64% b Fabry disease A143T genotype-phenotype investigation (p=0.045) in the heart by 98% (p 0.0001) in the spleen by 97% (pb0.0001) and in the liver by 99% (p=0.0075) relative to age matched untreated controls. Following a similar trend, Lyso-Gb3 in Susheela Jayaraman, Joel Charrow, Jill Frauenheim, Rachel Hickey, the plasma were also reduced by 98% (pb0.0001) normalising to that Shanna Widera, Ann & Robert H. Lurie Children's Hospital of Chicago, of wild type levels. Gb3 clearance was also supported by electron Chicago, IL, United States microscopy data. Collectively, these data provide strong evidence In 2014, Illinois became one of the first states to initiate a pilot that our liver-directed AAV-mediated gene therapy approach holds study for newborn screening for Fabry Disease. Since screening for considerable therapeutic potential for the treatment of Fabry disease. Fabry disease was implemented statewide in June of 2015, the Division of Genetics, Birth Defects and Metabolism at Ann & Robert doi:10.1016/j.ymgme.2018.12.196 H. Lurie Children's Hospital of Chicago has evaluated many infants with abnormal newborn screens for Fabry disease. Cascade family testing has allowed us to follow the phenotype and clinical presentation of previously undiagnosed individuals with Fabry 181 disease. Specifically, Ann & Robert H. Lurie Children's Hospital’s Migalastat pharmacokinetic (PK) exposure comparisons between cohort of patients with the A143T variant that have been identified race/ethnic groups and between males and females are similar through newborn screening and cascade familial testing. As there are many conflicting reports in the literature as to the clinical Franklin K. Johnson, Anthony Sileno, Nina Skuban, Jay A. Barth, significance of the A143T variant, there is little consensus to guide Amicus Therapeutics, Cranbury, NJ, United States clinical interpretation and management of these individuals identified to have the A143T variant. With little evidence available to guide Fabry disease is an X-linked lysosomal storage disorder caused by anticipatory guidance for newly diagnosed individuals and parents, a a mutation in the GLA gene, resulting in the functional deficiency of diagnosis of Fabry disease can be overwhelming and lead to α-galactosidase A (α-Gal A). Migalastat is a pharmacological increased anxiety. This poster will present the center’s data including chaperone that binds to and stabilizes amenable forms of α-Gal A enzyme levels, demographic, phenotypic and management data for and is approved for treating patients with Fabry disease and patients with the A143T variant. This poster will also further amenable GLA mutations. Pharmacokinetic data from 152 subjects emphasize the challenges of educating and counseling families on (105 White, 25 Black, and 22 Asian) were pooled across phase 1, 2, their prognosticated future of living with the A143T variant. and 3 clinical studies for race comparisons with the analysis of variance (ANOVA). Area under the curve (AUC) point estimates for Black (test) and Asian (test) vs White (reference) were 92.9 and doi:10.1016/j.ymgme.2018.12.195 102.7, respectively. Maximum concentration (Cmax) point estimates for Black (test) and Asian (test) vs White (reference) were 102 and Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S81

127, respectively. There was no significant difference in AUC or Cmax 183 between Black and White subjects as the lower and upper Clinical and neuropathologic findings in two long-term survivors confidence intervals (CIs) were contained within the 80% to 125% of Krabbe disease with and without umbilical cord blood boundaries. When comparing Asian and White subjects, these CIs transplantation were also contained within the 80% to 125% boundaries and indicated no significant difference for AUC however, the upper Kofler Julia, Maria L. Beltran-Quintero, Michele Poe, Maria L. Escolar, N bound CI for Cmax was 125% (143.8) due to lower mean body University of Pittsburgh, Pittsburgh, PA, United States weight of Asians. This difference is not clinically relevant to migalastat PK or PD effects. Rate of absorption (tmax), approximately Background: Krabbe disease or Globoid Cell Leukodystrophy is 3 hours, did not differ between race/ethnic groups. Collectively, these a lysosomal storage disease caused by deficiency of the lysosomal results demonstrated that there are no meaningful race/ethnic- enzyme Galactocerebrosidase. The early onset form is character- related differences in migalastat PK. Population PK modeling was ized by irritability, abnormal tone, developmental delay and performed to compare exposures between males and females in the feeding difficulties that start before 12 months and leads to phase 3 study, AT1001-011 (NCT#00925301). After accounting for premature death. Pathognomonic findings of classic infantile weight differences, models were run to show the relationship Krabbe disease are globoid cells, astrogliosis and axonal loss. between males and females on clearance and volume of distribution. Umbilical blood cord transplantation has shown to alter the Additional models were run to rule out sex-related effects on disease progression, by providing enzyme to the brain tissue. migalastat PK. The models demonstrated there are no sex-related There are no neuropathological studies comparing the natural differences in migalastat PK. progression of the disease with the brain of a treated patient. The purpose of this study is to examine the neuropathological differences between the treated and untreated patient that shared doi:10.1016/j.ymgme.2018.12.197 the same mutation known to cause the early onset form of the disease. Methods: Clinical characteristics, neurodevelopment and neu- fi 182 ropathological ndings were analyzed and compared in a patient Characterization of a novel porcine model of CLN3-Batten disease with Krabbe disease that received UBCT as a newborn and an untreated patient. Patients shared the same disease-causing mutation. Tyler B. Johnsona, David A. Sturdevanta, Katherine A. Whitea, Arlene Results: Developmental trajectories of treated and untreated V. Drackb, Sajag Bhattaraib, Chris Rogersc, Jonathan D. Cooperd, David patients differ significantly. The transplanted patient had no A. Pearcea, Jill M. Weimera, aSanford Research, Sioux Falls, SD, United evidence of active demyelination, however, there were myelination States, bUniversity of Iowa, Iowa City, IA, United States, cExemplar abnormalities due to incomplete remyelination partially related to Genetics, Coralville, IA, United States, dWashington University School of axon loss, and there was absence of globoid cells. The untreated Medicine, St. Louis, MO, United States patient had severe leukoencephalopathy with near-complete white CLN3-Batten disease is an autosomal-recessive disorder that matter rarefaction. fi results from mutations in CLN3. In the vast majority of cases, disease Conclusions: This is the rst reported pathology study in a onset occurs in early childhood, is characterized by progressive loss transplanted patient with Krabbe disease compared with an of vision, seizures and a failure in psychomotor development and is untreated patient with similar characteristics. Umbilical cord blood universally fatal by the third decade of life. Several mouse models of transplantation has positive results in the CNS demonstrated by CLN3 have been developed that have provided insight into the developmental assessments and neuropathological examination. pathological progression of this disease including early glial activa- There is evidence of successful engraftment of myeloid cells in tion followed by neuronal loss, however, many aspects of the disease the brain, clearance of globoid cells and prevention of new globoid are not recapitulated in these mouse models. Porcine models hold cell formation and prevention of progressive demyelination. the promise of a more accurate disease model given the similarities Umbilical cord blood transplantation failed to halt disease that pigs and humans share in terms of development, anatomy, and progression in the peripheral nervous system. physiology, and in particular, similarities in brain development such as perinatal growth spurt and brain structure. Furthermore, the doi:10.1016/j.ymgme.2018.12.199 shared characteristics of the human and porcine visual system make the pig a useful resource for studying macular-associated retinal diseases that are not possible in mice. Additionally, the size of the pig would allow for the use advanced neuroimaging techniques such as 184 MRI, CT and/or PET to model in vivo changes within the CNS Enzyme replacement therapy together with renin-angiotensin longitudinally and track the success of potential therapeutics. system inhibition seems to prevent kidney function decrease in Recently, we have developed a novel CLN3Δex7-8/Δex7-8 porcine model most Finnish Fabry patients treated either for 5 or 10 years of CLN3-Batten disease. Here we present initial characterization results showing behavioral, pathological, and visual deficits that Ilkka Kantolaa, Antti-Pekka Annalab, Agneta Ekstrandc, Tiina Heliöc, mirror changes seen in human patients. Aki Hietaharjud, Kristiina Kananene, Taru Kantolaa, Anne Karhuf, Jorma Kokkoneng, Johanna Kuusistoh, Päivi Pietila-Effatii, Miia Raintoa, Jukka T. Saarineni, Lorenzo Sandinij, Markku Savolainenk, doi:10.1016/j.ymgme.2018.12.198 Jyrki Tauriod, Pentti Tienaric, Olavi Ukkolak, Kati Valtolah, Susanne Wallsa, Iina Yliaskak, aTurku University Hospital, Turku, Finland, S82 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 bSeinäjoki Central Hospital, Seinäjoki, Finland, cHelsinki University disease progression. N-acetylcysteine (NAC), a FDA-approved med- Hospital, Helsinki, Finland, dTampere University Hospital, Tampere, ication with antioxidant/anti-inflammatory activity, is also available Finland, eKainuu Central Hospital, Kajaani, Finland, fPori Central as an inexpensive dietary supplement with a good safety profile. Our Hospital, Pori, Finland, gCentral Finland Central Hospital, Jyväskylä, objective is to determine whether oxidative stress and inflammation Finland, hKuopio University Hospital, Kuopio, Finland, iVaasa Central in the blood and brains of individuals with GD1 can be altered with Hospital, Vaasa, Finland, jSouth Karelia Central Hospital, Lappeenranta, orally administered NAC. In this open-label study, we have currently Finland, kOulu University Hospital, Oulu, Finland enrolled 13 patients who received NAC at 3600mg/day dose. Blood sample collection for biomarker analysis and brain imaging using Enzyme replacement therapy (ERT) has stabilized renal func- magnetic resonance spectroscopy (MRS) was done before and tion but its effect on proteinuria is inconsistent. Also following 3 months of NAC. Total glutathione in red blood cells, nephroprotective renin-angiotensin-aldosterone system (RAAS) which is a measure of oxidative stress, and plasma NAC concentra- inhibition is warranted. We calculated eGFR using CKD-EPI formula tions were measured using a validated liquid chromatography/mass and albuminuria using urine albumin/creatinine ratio (ACR) once a spectrometry method. Intracellular antioxidant enzyme activity and year in 12 Fabry patients with ERT for 10 years (8 females, 4 plasma markers for inflammation and lipid and protein modifications males, age 53.6 (15.0) years) and in 24 patients with ERT for 5 were measured using colorimetric and immunoassays. Interim years (11 females, 13 males, 53.3 (17.2) years). In the 10-year analysis of data from 10 patients revealed that NAC at this dose did group 5 patients and in the 5-year group 12 patients used RAAS not significantly alter any of the neurochemical markers. However, inhibition at the end of follow-up. General linear model for validated MRS markers related to neuronal health such as N-acetyl repeated measures and Pearson correlation were used. Two aspartate (NAA) and glutamate showed trends toward improvement. patients who got renal transplantation were excluded. EGFR Similar trends were also observed in blood antioxidant and decreased 5.0 (SD 15.3, median 5.0) ml/min/1.73 m2, slope -0.5/ inflammatory biomarkers. Pharmacokinetic analysis using serial year in the 10-year group and increased 0.7 (SD 9.2, median -2.2), samples collected 90-days after NAC regimen is underway to slope +0.14/year in the 5-year group. No statistically significant determine if a dose increase is warranted. If these favorable trends change was seen in either group. ACR increased 2.5 (SD 6.7, subsequently are shown to be significant, NAC could be a new median 0.07) mg/mmol, slope 0.25/year in the 10-year group and adjunctive therapeutic option. 1.3 (SD 4.6, median 2.2) mg/mmol, slope 0.26/year in the 5-year group. No statistically significant change was seen in either group. In the 10-year group eGFR decreased more than 15 ml/min/1.73 doi:10.1016/j.ymgme.2018.12.201 m² in 4 patients and in the 5-year group in one patient. ACR increased over 5 mg/mmol in 2 patients both in the 10-year and in the 5-year group. Neither age, age at the initiation of ERT, 186 gender, use of agalsidase alfa versus beta nor use of RAAS inhibitor An improved, novel, systemically administered AAV gene therapy correlated to change in eGFR or ACR in the 10-year group. In the for treatment of CLN3 juvenile neuronal ceroid lipofuscinosis 5-year group, higher age correlated to higher change in eGFR (p=0.024). According to our results, ERT for five years regardless Scott Kernsa, M. Arnolda, A. Aldrichb, R. Falletb, Tammy Kielianb, Linas of RAAS inhibition prevented decline in kidney function. This Padegimasa, Timothy J. MIllera, aAbeona Therapeutics, Cleveland, OH, finding was more consistent with younger age, suggesting earlier United States, bDepartment of Pathology & Microbiology, University of initiation of ERT. Nebraska Medical Center, Omaha, NE, United States doi:10.1016/j.ymgme.2018.12.200 Juvenile Neuronal Ceroid Lipofuscinosis (JNCL) is a lysosomal storage disease caused by an autosomal recessively inherited mutation in the CLN3 gene. JNCL is a progressive neurodegenerative disorder in which the central nervous system (CNS) is greatly 185 affected resulting in behavioral issues, vision loss, and other Preliminary N-acetylcysteine results for LDN 6722 - Role of cognitive disabilities. We developed a novel AAV capsid with oxidative stress and inflammation in Gaucher disease type 1: enhanced tropism for CNS tissue after systemic administration and Potential use of antioxidant anti-inflammatory medications optimized the CLN3 transgene cassette to improve biodistribution and expression in CNS and somatic tissues. AAV9 was used as a Reena V. Karthaa, James Joersa, Marcia Terluka, Paul Tuitea, Usha benchmark to assess the gene biodistribution in the Cln3Δex7/8 mice Mishraa, Kyle Rudsera, Gulin Oza, Neal J. Weinrebb, Jeanine Jarnes- model. AAV9-CLN3 and AAV214-CLN3 were compared in mice that Utza,c, James C. Cloyda, aUniversity of Minnesota, Minneapolis, MN, received 2.0 x 1013 vg/kg via intravenous administration. After 30 United States, bLeonard School of Medicine of University of Miami, days the animals were sacrificed and tissues were harvested for Miami, FL, United States, cFairview Health Systems, Minneapolis, MN, biodistribution analysis. We evaluated capsid delivery in the primary United States. regions of the CNS and the spinal cord (cervical and lumbar). In those tissues AAV214-CLN3 showed greater biodistribution efficacy Enzyme replacement and substrate reduction therapies have when compared to AAV9-CLN3 (brain=160% of AAV9, Cervical significantly improved outcomes in patients with Type 1 Gaucher SC=158% of AAV9 and lumbar SC=184% off AAV9). The sciatic disease (GD1), but do not completely return glucocerebroside levels nerve also demonstrated higher AAV214-CLN3 (744%) bio- to the normal range nor prevent continuing tissue injury. Preliminary distribution compared to AAV9. The optimization of the CLN3 gene biomarker analyses indicate a role for oxidative stress and inflam- and expression cassette along with a novel AAV vector was able to mation in GD1 pathophysiology. Thus, additional treatment strate- show greater tropism into the CNS via systemic administration. gies that target oxidative stress and inflammation have the potential Current studies assessing expression and dose response over a to ameliorate both systemic and brain symptoms and may alter longer duration suggest AAV-214 can be used for effective systemic Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S83 delivery and the ability to correct CLN3 in CNS tissues and peripheral striated musculature as well as the central nervous system (CNS), tissues. with early mortality. Enzyme replacement therapy (ERT) is currently the only FDA-approved therapy to treat Pompe and requires biweekly injections of relatively large quantities of recombinant doi:10.1016/j.ymgme.2018.12.202 protein. While ERT significantly reduces the mortality rate of infantile Pompe patients, who typically die by the age of two without therapy, it fails to completely ameliorate all symptoms of fi 187 Pompe, primarily due to its inability to ef ciently enter the CNS and AAV gene therapy for the treatment of Fabry disease: A novel resulting immune responses to the GAA protein. Gene therapy capsid with improved tropism to heart, kidney and CNS and strategies have been investigated and while many show great improved GLA expression promise in correcting the glycogen accumulation and other symptoms of Pompe, most have suffered from the severe immune response seen during GAA replacement. Previous work has demon- Brian M. Kevany, Scott Kerns, Linas Padegimas, Timothy J. Miller, strated that hepatic-specific expression can tolerize animals to the Abeona Therapeutics, Cleveland, OH, United States GAA protein and significantly reduce the humoral response. We have identified a novel adeno-associated virus (AAV) capsid with Fabry disease is an X-linked lysosomal storage disorder caused by improved tissue specificity and expression in the target tissues, adeficiency in alpha-galactosidase A (GLA) activity that results in the namely heart, muscle and CNS after intravenous administration. We accumulation of the glycolipid products, globotriaosylceramide have developed a novel codon-optimized transgene and improved (Gb3) and lyso-Gb3 in the lysosome. Disease manifestations include promoter for supraphysiological expression levels in target tissues as frequent bouts of peripheral neurotrophic pain, angiokeratomas, well as increased secretion for cross-correction to non-transduced reduced sweat production, corneal dystrophy, cardiomyopathy, renal cells. Proof of concept studies demonstrate significant protein insufficiency and cerebrovascular disease, which result in reduced expression and enzymatic activity en route to IND-enabling studies. lifespan in Fabry patients. While males are affected more severely due to the mutations in the GLA gene, female patients are also frequently symptomatic and are often misdiagnosed. Enzyme doi:10.1016/j.ymgme.2018.12.204 replacement therapy (ERT) is currently the only FDA-approved therapy to treat Fabry and requires bi-weekly injections of relatively large quantities of recombinant protein. While ERT reduces the accumulation of Gb3 in the heart, kidney and vasculature it fails to 189 completely treat all symptoms of Fabry, primarily due to its inability Non-clinical evaluation of a blood-brain barrier-penetrating to efficiently enter the CNS. Gene therapy strategies have been enzyme for the treatment of mucopolysaccharidosis type I investigated and while many show great promise in correcting the glycolipid accumulation, most have failed to efficiently enter the CNS Sachiho Kida, Masafumi Kinoshita, Satowa Tanaka, Miho Okumura, and also suffered from an immune response often seen during GLA Yuri Koshimura, Hideto Morimoto, JCR Pharmaceuticals Co., Ltd., Kobe, replacement. Previous work has demonstrated that hepatic-specific Japan expression can tolerize animals to the GLA protein and significantly reduce the humoral response but suffer from the same limitations as Treatment of central nervous system (CNS) disorders in patients ERT. We have identified novel adeno-associated virus (AAV) capsids with lysosomal storage diseases by intravenous enzyme replacement with improved tissue specificity and expression in the target tissues, therapy is still challenging because the blood-brain barrier (BBB) namely CNS, heart, and kidney after intravenous administration. interferes the drug delivery to the brain. We have developed JR-141, After identifying a lead AAV candidate, we have developed a novel, a fusion protein consisting of anti-human transferrin receptor codon-optimized transgene and enhanced promoter to provide antibody (named J-Brain Cargo) and iduronate-2-sulfatase, which supraphysiological expression levels in target tissues as well as can cross the BBB utilizing receptor-mediated transcytosis of increased secretion for cross-correction to non-transduced cells. transferrin, and are currently conducting clinical trials for Muco- Proof of concept studies demonstrate significant improvements and polysaccharidosis II (MPS II). Here we report a non-clinical enzymatic activity to facilitate IND-enabling studies. evaluation of JR-171, another J-Brain Cargo-applied enzyme for the treatment of MPS I. MPS I is caused by deficiency of a lysosomal enzyme α-L-iduronidase (IDUA) and characterized by pathological doi:10.1016/j.ymgme.2018.12.203 accumulation of heparan sulfate (HS) and dermatan sulfate (DS) in cells throughout the body, resulting in a broad spectrum of somatic and CNS symptoms. We prepared JR-171, a fusion protein consisting of J-Brain Cargo and IDUA, and examined its biodistribution in 188 cynomolgus monkeys. JR-171 was detected in the brain as well as in A novel AAV capsid with improved CNS tropism for treating peripheral tissues of monkeys after intravenous administration, Pompe disease by intravenous administration indicating its penetration of the BBB. We then evaluated the efficacy of repetitive intravenous administration of JR-171 in the reduction of Brian M. Kevany, Linas Padegimas, Timothy J. Miller, Abeona accumulated HS and DS in peripheral tissues and the brain using Therapeutics, Cleveland, OH, United States human transferrin receptor knock-in mice lacking Idua gene as an animal model of MPS I. The intact IDUA decreased HS and DS Pompe disease is a lysosomal storage disorder caused by a concentrations in peripheral tissues but failed to reduce the deficiency in acid alpha-glucosidase (GAA) activity that results in the substrates in the brain of the mice. In contrast, JR-171 markedly accumulation of glycogen in the lysosome. The disease presents as a reduced HS and DS concentrations in the brain as well. These results form of muscular dystrophy which primarily affects both smooth and demonstrate that the J-Brain Cargo-applied IDUA distributes to the S84 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 brain by crossing the BBB when administered intravenously, and insufficiency. Many patients have also exhibited decreased bone suggest that JR-171 has a potential to exert therapeutic effects on density and fractures as symptoms of Pompe Disease Our cohort CNS disorders as well as somatic symptoms in MPS I. included 15 Pompe patients ranging from 21- 74 years on ERT for variable durations. We tested the progressive impact of ERT on doi:10.1016/j.ymgme.2018.12.205 osteopenia or osteoporosis by studying bone mineral by comparing the z and t-scores of hips and spine using DXA scans. Our results demonstrated that females had a 2.17 units lower average z-score of the lumbar spine and 0.563 units lower average z-score of their femurs compared to males. Increasing age also changed the bone 190 dynamics, as a one-year increase in age of the patients led to a 0.09 A case of a 39-year-old man with novel mutation and classic unit score significant increase in lumbar z-scores (p=0.01) and a Fabry disease who showed different changes of several bio- 0.042 unit score significant increase in femur z-scores (p=0.04). markers and speckle tracking after enzyme replacement therapy Additionally every one year duration of ERT treatment resulted in a 0.15 unit score increase in z-scores (p=0.01) of the lumbar spine. However the femur z-score only showed a 0.08 unit score increase Gee-Hee Kim, St. Vincent’s Hospital, College of Medicine, The Catholic after one year of treatment (p=0.10) Results overall, denote that ERT University of Korea, Suwon-si, Republic of Korea treatment shows a positive correlation with a lower risk of fractures Fabry disease (FD) is a progressive, X-linked lysosomal storage resulting from osteoporosis. ERT was associated with an increase of disorder caused by a deficiency of α galactosidase A (GLA) activity. the z-scores of .09 of the lumbar spine. Females were at a higher risk Affected individuals accumulate glycosphingolipids in the lysosomes of developing osteoporosis compared to males. This study empha- and cytoplasm of cells throughout the body, leads to major organ sizes the importance of early management and ERT to prevent failure and premature death. Cardiac involvements include left osteopenia and bone fractures. A genotype-phenotype correlation ventricular hypertrophy, arrhythmia, angina and cardiomyopathy. and a larger study of Pompe subjects is needed to determine other However, it is unclear that changes in several biomarkers, including variables Lyso Gb-3, brain natriuretic peptide (BNP) and galectin-3 (Gal-3) in fl patient with Fabry cardiomyopathy (FC) results in in ammation and doi:10.1016/j.ymgme.2018.12.207 fibrotic changes in the heart. We present a case of a 39 years old man with classic Fabry disease and identified one hemizygous novel mutation in exon 4 of GLA, c.617TNC (p.Leu206 Pro) that was registered in the Fabry-database under my name. The clinical 192 symptoms were intermittent burning pain originating in the extrem- Safety and effectiveness of resistance exercise training in a pilot ities and a ringing in the ears in the past, angiokeratomas on the trunk, study of patients with late onset Pompe disease and cornea verticillata. In this patient, left ventricular hypertrophy (18.1mm) by Sokolow criteria with T wave inversion on V4-6 began to Virginia Kimonis, Jeet Shah, Caleb Bhatnagar, Bhumi Ramani, Vince appear on the electrocardiogram in the thirties and the PR interval Caiozzo, UC Irvine, Orange, CA, United States (120ms) showed a shortening. In 39-year-old, Magnetic Resonance Imaging findings did not show the delayed enhancement. We Pompe Disease is a rare inherited disorder associated with demonstrate the patient showed serial biomarkers and echocardio- muscle weakness and respiratory insufficiency which can affect all graphic changes for 6 months after enzyme replacement therapy ages, ethnicities, and gender. It is caused by the accumulation of (ERT). Biomarkers, such as pro-BNP and Lyso Gb-3 improved glycogen primarily in cardiac and skeletal muscle. There are two compared to before ERT. However, after ERT, global longitudinal strain forms of the disease, classic infantile form and juvenile/adult form. value which is the prognostic marker of cardiomyopathy aggregated The purpose of this study is to evaluate the effectiveness of the and Gal-3 was unchanged. This case highlights the importance of supervised resistance training program on muscle strength, changes in several biomarkers and echocardiographic parameters in functional capacity and body composition in patients with late early FC patient with novel mutation before and after ERT. onset Pompe disease. The study includes 10 patients over a 32 week study of increased resistance exercise 3 times a week in a doi:10.1016/j.ymgme.2018.12.206 gym with a personal trainer. The patients also undertook respiratory muscle exercise training daily with increasing resistance every 2 months. Each patient served as their own control (Weeks 1 - 8 baseline period) and subsequently physical strength, 191 stamina and respiratory function was tested with the Biodex and Effects of enzyme replacement therapy on bone density in late hand held dynamometers, 6 minute walk test (6MWT), maximum/ onset Pompe disease minimum inspiratory pressure (MIP/MEP), SF-36 questionnaire, muscle volume and texture change using MRI, and blood and Virginia Kimonis, Mahima Avanti, Yanjun Chen, Margaret Knight, urine collections of tetrasaccharide. Throughout the study, we Tahseen Mozaffar, UC Irvine, Orange, CA, United States found that there was an overall improvement in the patient’s muscle strength by Biodex dynamometry, MRC scores and 6MWT. Pompe Disease is a rare autosomal recessive disorder lysosomal For example, knee and elbow extension torque improved by a storage disorder caused by the deficiency of α-glucosidase (GAA). mean of 21.6% and a 41.6% respectively. The 6MWT showed an The disease results in the accumulation of glycogen, a polysaccharide average improvement of 6.58%, and total MRC scores showed an of glucose, in the smooth, cardiac, and skeletal muscles. Late Onset improvement of 2.1% from initial to final visit. There was a marked Pompe is mainlycharacterized by skeletal muscle glycogen accumu- improvement in the respiratory parameters in the majority of lation, proximal muscle weakness, and early respiratory patients the MIP showed an increase of 25% while MEP increased Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S85 by 10%. Overall, patients also had an improved sense of well being worsening intellectual impairment and regression. Disease progres- with very few adverse side effects. sion creates dramatic neurobehavioral symptoms, including hyperactivity, inattention, behavior difficulties, sleep disturbance, and others. These behaviors are highly disruptive, distressing to families, doi:10.1016/j.ymgme.2018.12.208 and significantly impact quality of life. But these symptoms have been difficult to measure. Existing pediatric behavioral assessment tools do not capture the unique constellation of symptoms, nor variability of expression. To address this problem, we are developing 193 a Hunter-specific caregiver rating scale of neurobehavioral symp- Antisense oligonucleotide treatment targeting glycogen synthase toms. The goal is to create a valid, reliable tool that detects change in (GYS1) in a mouse model of Pompe disease neurobehavioral profile, informing natural history and therapeutic response. A first step involves drafting a tool using methods of scale a a b a Virginia Kimonis , Lan Weiss , Michele Carrer , Howard Yu , Nina development for another MPS type with neurobehavioral impair- c b a b Raben , Tamar Grossman , UC Irvine, Orange, CA, United States, Ionis ment, the Sanfilippo Behavior Rating Scale. Item sets are informed by c Pharmaceuticals, Carlsbad, CA, United States, National Institutes of coded video recordings of: 1) semi-structured play in small groups of Health, Bethesda, MD, United States. affected boys and 2) focus groups with caregivers literature review and review of existing tools. Inclusion criteria include: 1) confirmed Pompe disease is a progressive myopathy resulting from the diagnosis of early progressive Hunter syndrome 2) age 4 to 9 years fi α de ciency of acid -glucosidase (GAA). The standard of care is ERT 3) able to ambulate. Findings aligned with the literature’s descrip- with recombinant human (rh) GAA. ERT works well in alleviating the tions of diverse symptoms, with considerable differences in presen- cardiomyopathy however, many patients continue to have progres- tation within-child, dependent upon internal and external factors sive muscle weakness from muscle glycogen accumulation produced (e.g., fatigue, hunger, novelty of a setting). Caregiver reports by muscle glycogen synthase (GYS1). Previous studies have shown indicated a need for a behaviorally anchored scale that assesses that inhibition of GYS1 reduced lysosomal glycogen. Knockdown of frequency of symptoms and context rather than severity. Next steps GYS1 mRNA by phosphorodiamidate morpholino oligonucleotide will involve piloting the draft scale and soliciting caregiver feedback. conjugated with a cell penetrating peptide however wasnephrotoxic. Future directions include validating the measure in a larger multi- Antisense Oligonucleotides (ASO) technology has emerged as a center study. Support: Shire pharmaceuticals, NIH U54NS065768, powerful therapeutic alternative for the treatment of genetic National MPS Society, UMN Center for Neurobehavioral Develop- fi disorders by targeting RNA. In order to impart speci city for the ment muscle variant of GYS1, we propose the use of ASO-mediated gene silencing through the RNaseH1 dependent degradation mechanism. Most recently therapy for spinal muscular atrophy has been doi:10.1016/j.ymgme.2018.12.210 successful using ASOs, and our hope is that ASO technology will be successful in Pompe disease. Over 150 ASOs were designed and screened in vitro to identify the most efficacious ASO for testing in wild type mice. The lead from the screen were validated in a dose 195 fi response study and the top 10 ASOs were screened in vivo. The 2 Safety and ef cacy of VAL-1221, a novel fusion protein targeting ASOs (GYS1 ASO#1 and GYS1 ASO#2) showed the best tolerability cytoplasmic glycogen, in patients with late-onset Pompe disease and efficacy profile leading to knock down of GYS1 mRNA by a b c approximately 50% of control. We have performed a pilot study of Priya Kishnani , Robin Lachmann , Tahseen Mozaffar , Crista a a d d c the efficacy of two GYS1 ASOs in Pompe mice as monotherapy and Walters , Laura Case , Matt Appleby , Vincenzo Libri , Manisha Kak , c e a have reduced muscle GYS1 mRNA levels versus PBS and a mismatch Marie Wencel , Hal Landy , Duke University, Durham, NC, United b ASO. ASO#2 resulted in weight loss of the mice. We will present the States, National Hospital for Neurology and Neurosurgery, London, c results of the study of the effect of the GYS1 ASOs on muscle United Kingdom, University of California, Irvine, Orange, CA, United d glycogen, histology, GYS1 RNA and muscle function in Pompe mice. States, UCL Institute of Neurology and the National Hospital for e These preliminary studies provide proof of principle for a promising Neurology and Neurosurgery, London, United Kingdom, Valerion adjunct treatment for Pompe disease. Therapeutics, LLC, Concord, MA, United States

Disease progression in Pompe Disease, a lysosomal storage doi:10.1016/j.ymgme.2018.12.209 disorder, occurs despite enzyme replacement therapy ERT, and is characterized by cytoplasmic accumulation of glycogen, presumably due to rupture or seepage of glycogen from lysosomes, defects in autophagy and the targeting of conventional ERT only to the 194 lysosome. VAL-1221 is a fusion protein comprising the Fab portion Feasibility of quantifying behavior in early progressive MPS II of a cell-penetrating antibody utilizing the nucleoside transporter ENT-2 to gain access to the cytosol, and recombinant human acid Kelly E. Kinga, Elsa G. Shapiroa, Chester B. Whitleya, Joseph Muenzerb, alpha glucosidase (rhGAA), which enters lysosomes via mannose-6- Julie B. Eisengarta, aUniversity of Minnesota, Minneapolis, MN, United phosphate receptors (M6PRs). Thus, VAL-1221 targets both cytosolic States, bUniversity of North Carolina, Chapel Hill, NC, United States and lysosomal glycogen. 30 mg of VAL-1221 contains 20 mg of rhGAA. We performed a 3-month Phase 1/2 repeat dose, dose- Hunter syndrome is a rare X-linked lysosomal disorder associated escalation study of VAL-1221 with an rhGAA (Myozyme/Lumizyme) with progressive multisystem involvement. Ages of onset vary, as do control to assess its safety and preliminary efﬁcacy. 12 adults with disease course and phenotypic severity. The early progressive late-onset Pompe disease (LOPD) treated with rhGAA for at least one phenotype involves delayed developmental milestones with year were randomized (1:3) to rhGAA (20 mg/kg qow) or to VAL- S86 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

1221 (Cohort 1: 3 mg/kg, Cohort 2: 10 mg/kg, Cohort 3: 30 mg/kg). 6, 9, and 12 as assessed by quantitative and manual muscle testing. Adverse events, infusion-associated reactions, anti-VAL-1221 anti- Forced vital capacity (% predicted) increased in ERT-naive patients (n=5, bodies and PD markers were assessed. Efficacy outcomes included mean change: month 6, 4.2 month 9, 6.2 month 12, 4.4), and was six-minute walk test (6MWT), pulmonary function tests, measures of generally stable in ambulatory ERT-switch patients (n=9, month 6, -1.3 muscle strength and function and patient-reported outcomes. There month 9, -1.7 month 12, -3.3). AT-GAA (ATB200/AT2221) was associated were no serious or unexpected adverse events all patients completed with reductions in creatine kinase and urine hexose tetrasaccharide. The the study. All but 1 patient had anti-VAL-1221 antibodies at Baseline most common treatment-emergent adverse events were upper and which increased initially but declined thereafter. Three patients, one lower abdominal pain, diarrhea, and nasopharyngitis. Three events of in each cohort, experienced IARs requiring transient reduction in infusion-associated reactions (skin discoloration, erythema, pruritus) dose. Although there were no significant changes in PD markers, occurred in ≥550 infusions. Data from this interim analysis show clinical there were largely dose-dependent improvements in nearly all benefits of AT-GAA (ATB200/AT2221) in ERT-naive patients and both efficacy outcome parameters. Control patients declined by 49 meters ambulatory and nonambulatory ERT-experienced patients, suggesting on 6MWT while VAL-1221 patients in cohorts 2 and 3 increased by that ATB200/AT2221 can be an effective and well-tolerated treatment for 54 meters. VAL-1221 is safe and well-tolerated in previously treated adults with LOPD. Updated 24-month efficacy/safety data will be patients with LOPD and shows preliminary efficacy at doses of 10–30 included in the presentation. mg/kg.

doi:10.1016/j.ymgme.2018.12.212 doi:10.1016/j.ymgme.2018.12.211

197 196 Data mining and machine learning for lysosomal disease drug First-in-human study of advanced and targeted acid α-glucosi- discovery and beyond dase (AT-GAA) (ATB200/AT2221) in patients with Pompe disease: preliminary functional assessment results from the ATB200-02 Jennifer J. Kleina, Nancy C. Bakerb, Kimberley M. Zorna, Daniel P. trial Russoa, Ana P. Rubioa, Alex M. Clarkc, Sean Ekinsa, aCollaborations Pharmaceuticals Inc., Raleigh, NC, United States, bParlezChem, Priya Kishnania, Benedikt Schoserb, Drago Bratkovicc, Barry J. Byrned, Hillsborough, NC, United States, cMolecular Materials Informatics, Inc, Paula R. Clemense, Ozlem Goker-Alpanf, Xue Mingg, Mark Robertsh, Montreal, QC, Canada Peter Schwenkreisi, Kumaraswamy Sivakumarj, Ans T. van der Ploegk, Vipul Jainl, Sheela Sitaramanl, Jay A. Barthl, Hjalmar Lagastl, Tahseen We have recently stressed the need to scale and “industrialize” Mozaffarm, aDuke University Medical Center, Durham, NC, United rare disease drug discovery. Finding information on compounds States, bFriedrich-Baur-Institut, Neurologische Klinik, Ludwig-Max- relevant to rare diseases across the hundreds of available databases imilians-Universität München, Munich, Germany, cPARC Research Clinic, is a complex challenge, even for experts and the linkage between Royal Adelaide Hospital, North Adelaide, Australia, dUniversity of targets and data is often non-existent. Disease-associated targets are Florida, Gainesville, FL, United States, eUniversity of Pittsburgh and only identified after a deep dive into the literature for a specific Department of Veterans Affairs Medical Center, Pittsburgh, PA, United disease as disease annotation is usually not included alongside States, fO&O Alpan LLC, Fairfax, VA, United States, gRutgers New Jersey publicly accessible structure activity relationship data. This would be Medical School, Newark, NJ, United States, hSalford Royal NHS valuable so that we could use public datasets for specific targets to Foundation Trust, Salford, United Kingdom, iNeurologische Klinik und build machine learning models through our Assay Central software. Poliklinik des Berufsgenossenschaftlichen, Universitätklinikum This could then help us to identify small molecules that might have Bergmannsheil, Bochum, Germany, jNeuromuscular Research Center, value as inhibitors or chaperones for testing with collaborators Phoenix, AZ, United States, kErasmus Medical University, Rotterdam, working on those diseases and move development forward more Netherlands, lAmicus Therapeutics, Inc., Cranbury, NJ, United States, quickly. To date, many of the lysosomal storage diseases (LSDs) have mUniversity of California, Irvine, Irvine, CA, United States no FDA approved treatments. As a first step we have mined PubMed and other databases such as ChEMBL to identify and understand Late-onset Pompe disease (LOPD) is a rare lysosomal storage disorder disease targets associated with LSDs and available datasets. We will characterized by progressive loss of muscle and respiratory function demonstrate selected targets and public datasets we have identified resulting from acid α-glucosidase (GAA) deficiency, for which recombi- to date for LSDs and show how they can be used to generate nant human GAA enzyme replacement therapy (rhGAA ERT: machine learning models (Bayesian, Random Forest and Deep alglucosidase alfa) is currently available. Here, we report the 12-month learning etc) that can suggest new molecules to test. We ultimately interim results from ATB200-02 (NCT02675465), a first-in-human, plan to create linkages between these models and disease targets for open-label, phase 1/2 trial evaluating a next-generation rhGAA/chaper- the 7000 rare and neglected diseases so that we can create one regimen AT-GAA (ATB200/AT2221) in adults with LOPD. The study hierarchies of protein families and disease categories. This will then enrolled 20 patients in 3 cohorts: ERT-switch ambulatory (n=11), ERT- enable the selection of specific machine learning models in Assay switch nonambulatory (n=4), and ERT-naive ambulatory (n=5). 6- Central which we can use with our collaborators. Efforts such as this minute walk test (meters) improved in ambulatory ERT-switch patients could help in dramatically scaling research and development efforts (n=10 mean change [SD]: month 6, 23.9 [52.2] month 9, 24.5 [40.8] for rare diseases. month 12, 42.2 [46.5]) and ERT-naive patients (n=5 month 6, 41.8 [29.4] month 9, 63.5 [23.1] month 12, 63.1 [29.1]). Other motor function tests were generally consistent with these results. Nonambulatory ERT-switch doi:10.1016/j.ymgme.2018.12.213 patients demonstrated increases in upper extremity strength at months Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S87

198 common. Diarrhea was more common in males constipation in Quantitative evaluation of white matter hyperintensities in the females. Additionally, discomfort, bloating and early satiety were central nervous system in infantile Pompe disease frequently noted. Except for early satiety, all these symptoms are addressed in the DIBSS-M. Interviews were conducted in 17 patients Aditi Korlimarla, Ela Stefanescu, Stephanie Austin, Steven Chen, diagnosed with FD (13 male, 4 female) ≥ 16 years old mean age of James M. Provenzale, Priya S. Kishnani, Duke University, Durham, NC, 40.9 years (15.0) in the United States. The most commonly reported United States symptoms were diarrhea (94% n=16), abdominal pain (64% n=11), bloating (47.1% n=8), and vomiting (41.2% n=7). While nausea is Pompe disease is a lysosomal storage disorder caused by not included on the DIBSS-M, 47.1% of patients found it relevant to deficiency of acid-alpha glucosidase. There are limited data on their FD. In contrast to the literature, interviewees infrequently central nervous system (CNS) involvement in survivors of infantile noted constipation (5.9% n=1). Patients reported substantial limita- Pompe disease (IPD). The objective of this cross-sectional study is to tion of social and work activities. Patients found the DIBSS-Measy to implement the Fazekas scale, a visual MRI rating scale, to quantify understand and answer, with an appropriate recall period and white matter hyperintensities (WMH) on T2-weighted-Fluid Atten- distinct options for each question. 88% of patients found the items uated Inversion Recovery (T2-FLAIR) brain MRI images and to measured on DIBSS-M to be relevant, especially those about stool determine the extent of white matter (WM) involvement in the frequency, stool consistency, urgency, and abdominal pain. Results CNS of patients with IPD. Twelve patients (aged 6.1-18.1 years, 7 support use of the DIBSS-M in clinical studies for FD with a focus on males, 5 females) with IPD and on long-term enzyme replacement improvements in abdominal pain and altered bowel functioning. therapy were included in the study. We assessed WM involvement in ten anatomical areas of the brain- juxtacortical U-fibers, subcortical WM, periventricular WM in the centrum semiovale and in the frontal doi:10.1016/j.ymgme.2018.12.215 and parietal lobes at the level of corona radiata, corpus callosum, anterior and posterior limb of the internal capsule, external capsule, corticospinal tracts at cerebral peduncles, midbrain, and pons and 200 the medullary decussation of corticospinal tracts. Based on the Identification of MPS clusters in Latin America: An opportunity severity of WMH, each area was graded on the Fazekas scale from for targeted health care programs zero (absent) to 3 (severe) and a total score (range 0-30) was obtained. Ten patients (ages 6.6-18.1 years) had WMH on brain MRI Francyne Kubaskia, Flávia Gameleirab, Ceila Perez de Ferránc, Joany (range of scores: 2-23) using the Fazekas scale. Five of the 10 Ramirezc, Francisca Jaquezc, Ana Carolina Brusius-Facchina, Sandra patients (ages 6.7-18.1 years) had scores greater than 15 (range 16- Leistner-Segala, Maira Graeff Burind, Kristiane Michelin-Tirellid, 23) indicating severe WMH and 2/10 patients (aged 6.1 years and 7.3 Simone S. Lopese, Paula F.V. de Medeirose, Angelina X. Acostaf, years) had no WM involvement with a score of zero. Our results Kiyoko A. Sandesf, Maria L. Castro Moreirag, Hector P. Quintero provide a means to quantify WM involvement in the CNS of patients Montanoh, Martha L. Solano Villareali, Heidy M. Barroso Sandovalj, with IPD. Fazekas scale could serve as a biomarker for longitudinal Rodolfo Bareirok, Gladys Cossiol, Roberto Giugliania, aUFRGS/HCPA/ follow up of WM abnormalities in patients with IPD and to better INAGEMP, Porto Alegre, Brazil, bHealth Secretary, Ouro Preto, Brazil, understand the extent of CNS involvement in IPD. [A portion of this cHospital Infantil Dr. Robert Reid Cabral, Santo Domingo, Dominican study was funded by Genzyme Sanofi] Republic, dHCPA, Porto Alegre, Brazil, eUniversidade Federal da Paraíba, Campina Grande, Brazil, fUFBA, Salvador, Brazil, gHospital Santa Casa da h doi:10.1016/j.ymgme.2018.12.214 Misericordia, Campo Grande, Brazil, Hospital Verdi Cevallos, Portoviejo, Ecuador, iFundacion Cardioinfantil de Bogota, Bogotá, Colombia, jHospital Provincial Del Puyo, Puyo, Ecuador, kHospital Adventista do Penfigo, Campo Grande, Brazil, lHospital del Niño de Panamá, Panama, 199 Panama Evaluating the content validity of the Diary of Irritable Bowel Syndrome Symptoms - Mixed (DIBSS-M) to assess gastrointestinal The mucopolysaccharidoses (MPS) include 11 rare disorders symptoms associated with Fabry disease caused by deficiency of specific lysosomal enzymes resulting in the accumulation of undegraded glycosaminoglycans (GAGs) and several Nerissa Krehera, Dawn Phillipsa, Sheri Fehnelb, T. Michelle Brownb, clinical consequences. The combined incidence for all MPS subtypes aAVROBIO Inc, Cambridge, MA, United States, bRTI Health Solutions, is approximately 1:25,000 live births. Clusters of these diseases have Research Triangle Park, NC, United States been identified in areas with high consanguinity and/or founder effect associated to endogamy. The MPS Brazil Network, associated to This study sought to identify the most important and relevant the Brazilian Institute of Population Medical Genetics, identified gastrointestinal (GI) symptoms experienced by patients with Fabry several MPS clusters in Latin America. Three clusters were confirmed disease (FD) and the best way to measure GI symptoms in clinical in Brazil, of MPS IIIC (state of Paraíba), MPS IVA (state of Paraíba) trials of new treatments for FD. A targeted literature review on GI and MPS VI (state of Bahia). Two clusters were identified in Ecuador: symptoms relevant in FD for measurement was completed. Qualita- MPS IIIB (state of Manabi) and MPS IVA (state of Pastaza). A cluster tive interviews with open-ended concept elicitation and cognitive of MPS VI was also identified in the Dominican Republic. Other debriefing were conducted in individuals with FD to explore the clusters are being investigated in Haiti (MPS VI), Panamá (MPS IVA), relevance of GI symptoms from the patient perspective and to and Brazil (MPS IIIB, Minas Gerais state). Haplotype analyses are evaluate the content validity of the Diary of Irritable Bowel underway to better characterize mutation profiles, and results Syndrome Symptoms—Mixed (DIBSS-M) in the FD population. In already available for the clusters of MPS IVA and MPS VI in Brazil the literature review, a high degree of variability of GI symptoms was indicate founder effects with common ancestors. As one example of reported abdominal pain and altered bowel function were most the benefits of cluster identification, a newborn screening program S88 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 for MPS VI was implemented in the Brazilian region in order to 202 provide early identification and treatment of new patients. In MPS Brazil Network: A summary of all mucopolysaccharidosis conclusion, examples of MPS clusters were identified in Latin type IIIB patients America and it is likely that several others are still unreported. We aim to properly identify and characterize MPS clusters in Latin Francyne Kubaskia, Marina Zambranob, Luciana Giugliania, Franciele America, to better understand how they were originated and to be Barbosa Trappa, Maira Graeff Burinb, Kristiane Michelin-Tirellib, able to offer targeted preventive and management measures to the Sandra Leistner-Segala, Ana Carolina Brusius-Facchinb, Maria Lucia affected communities (Acknowledgements to the MPS Brazil Net- Castroc, Hector P. Quintero Montanod, Roberto Giugliania, aUFRGS/ work, especially to Fernanda Bender, Fernanda Bittencourt, Diana HCPA/INAGEMP, Porto Alegre, Brazil, bHCPA, Porto Alegre, Brazil, Rojás Malaga, Aline Nemetz Bochernitsan, Jurema de Mari, and cHospital Santa Casa da Misericordia, Campo Grande, Brazil, dHospital Franciele Barbosa Trapp). Verdi Cevallos, Portoviejo, Ecuador

The MPS Brazil Network is a nonprofit organization founded in doi:10.1016/j.ymgme.2018.12.216 2004 that aims to provide information, diagnosis and treatment monitoring and management of patients with mucopolysaccharidoses (MPS). MPS type IIIB is an autossomal fi 201 recessive disorder caused by de ciency of alpha-N- Can MPS patients be identified by facial features acetylglycosaminidase leading to the accumulation of non-degraded or partially degraded heparan sulfate (HS). Due to this HS accumulation, patients present a wide spectrum of neurologic Francyne Kubaskia, Franciele Barbosa Trappa, Maria L. Castro symptoms including hyperactivity, autistic features, progressive Moreirab, Hector P. Quintero Montanoc, Martha L. Solano Villareald, cognitive decline and dementia. We aimed to provide a profile of Heidy M. Barroso Sandovale, Rodolfo Bareirof, Roberto Giugliania, all of the MPS IIIB patients identified by the MPS Brazil Network aHCPA/UFRGS/INAGEMP, Porto Alegre, Brazil, bHospital Santa Casa da within its 14 years of existence. Until now, the MPS Brazil Network Misericordia, Campo Grande, Brazil, cHospital Verdi Cevallos, Portoviejo, has identified 1,460 MPS patients, of whom 130 (9%) present MPS Ecuador, dFundacion Cardioinfantil de Bogota, Bogota, Colombia, IIIB. All patients were diagnosed by the identification of the specific eHospital Provincial Del Puyo, Puyo, Ecuador, fHospital Adventista do enzyme deficiency. From the 130 patients, 62 (48%) were females Penfigo, Campo Grande, Brazil and 68 (52% males). The average age at diagnosis was 6 years (range: The mucopolysacharidoses (MPS) have a wide spectrum of 1-28 years of age). 29 patients were from other countries (Argentina clinical manifestations. Taking this into consideration and exploring 1, Ecuador 24, Nicaragua 1, Saudi Arabia 2 and Uruguay 10). The 101 the benefits of noninvasive phenotypic characterization using frontal Brazilian patients were found widespread in all country regions, with fi pictures, we decided to test if a tool for facial recognition could no disease clusters con rmed in Brazil so far. In conclusion, MPS IIIB properly identify patients with MPS III and IV. The FDNA Face2Gene is a relatively common MPS in Brazil, and efforts should be made to algorithm extracts mathematical facial descriptors from patient’s diagnose all cases and to identify them earlier in life, as new frontal pictures and compares them with syndrome gestalts in order therapies for this condition (intra-cerebro-ventricular enzyme re- to suggest potential diagnoses. We tested the version 18.1.14 of the placement therapy, gene therapy) are being developed and should algorithm with pictures from 25 known MPS patients (MPS IIIB=16, become available in a near future. MPS IVA=9). For each patient, a de-identified case was created with frontal pictures, date of birth, height, weight and head circumference, and the phenotype was refined with the following descriptors: doi:10.1016/j.ymgme.2018.12.218 coarse facial features, hepatomegaly, short stature, intellectual disability, psychomotor deterioration, abnormal heart valve morphology, cardiomyopathy, hernia, hearing impairment, kyphosis, 203 splenomegaly, genu valgum, corneal opacity, dysostosis multiplex, Thoracolumbar kyphosis in MPS I: A natural history study and an respiratory distress and pectus carinatum, according to the clinical international consensus procedure for the development of a examination. The Face2gene scores were classified as low, medium clinical practice guideline and high similarity. For the scores, 1 (4%) did not receive an MPS score, 9 patients (36%) had the lowest, 14 (56%) medium, and 1 (4%) a a a the highest score. For the MPS IIIB patients, 15 (94%) had MPS as Gé-Ann Kuiper , Eveline J. Langereis , Stephanie C.M. NIjmeijer , b c d suggested diagnosis within the top 5 syndromes. Furthermore, 10 Sandra Breyer , Marco Carbone , Rene M. Castelein , Deborah M. e f f (62%) had MPS IIIB as one of the top 5 suggested diagnosis. For the Eastwood , Christophe Garin , Nathalie Guffon , Peter M. Van g h h i MPS IVA patients, all had MPS as suggested diagnosis however none Hasselt , Pauline Hensman , Simon A. Jones , Vladimir Kenis , Moyo d a j had MPS IV suggested. Other than MPS, the most common suggested C. Kruyt , Johanna H. Van der Lee , William G. Mackenzie , Paul J. k h l h diagnoses were: Angelman, DiGeorge, and Williams-Beuren syn- Orchard , Neil Oxborrow , Rossella Parini , Amy Robinson , Elke m n a drome. In conclusion, the algorithm was able to properly suggest Schubert Hjalmarsson , Klane K. White , Rick R. van Rijn , Frits A. a a MPS as a possible diagnosis in most patients based only on frontal Wijburg , Amsterdam University Medical Center, Amsterdam, Nether- b ’ c images with minimal clinical data and without any tests, nonetheless lands, Altonaer Children s Hospital, Hamburg, Germany, Institute for “ ” improvements are still necessary. This approach could be valuable in Maternal and Child health IRCCS Burlo Garofolo , Trieste, Italy, d e lower-income countries where access to diagnosis is limited. University Medical Center Utrecht, Utrecht, Netherlands, Great Ormond Street Hospital for Children, London, United Kingdom, fHôpital Femme-Mère-Enfant, Université Lyon, Lyon, France, gWilhelmina doi:10.1016/j.ymgme.2018.12.217 Children's Hospital, University Medical Center Utrecht, Utrecht, Nether- lands, hRoyal Manchester Children's Hospital, Central Manchester Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S89

University Hospitals NHS Foundation Trust, Manchester, United King- catheter use is a major risk factor for development of deep vein dom, iThe H. Turner institute for Children’s Orthopedics, Saint- thrombosis (DVT) which isassociated with significant morbidity, Petersburg, Russian Federation, jNemours/Alfred I. duPont Hospital for including debilitating extremity pain and swelling, pulmonary Children, Wilmington, DE, United States, kUniversity of Minnesota, embolism, superior vena cava syndrome as well as increased Minneapolis, MN, United States, lSan Gerardo University Hospital, overall mortality. Data from patients who require long-term Monza, Italy, mQueen Silvia's Children's Hospital, Göteborg, Sweden, central venous access show a prevalence of catheter related DVT nSeattle Children’s Hospital, Seattle, WA, United States with obstruction of 3-5%, with higher rates of asymptomatic non- occlusive thrombi. Here we report a series of six individuals with Dysostosis multiplex is a progressive skeletal disease in MPS II that presented acutely with severe symptomatic DVT. Of patients with mucopolysaccharidosis type I (MPS I). One of its our cohort of 18 MPS II patients, six (33.3 %) ages 12 to 27 years key features is thoracolumbar kyphosis and no guideline for presented with symptomatic DVT, five of which coinciding with surgical treatment is available yet. Therefore, we investigated the site of catheter placement. Veins affected included subclavian, natural course of kyphosis and initiated an international consen- jugular, axillary, common femoral, and brachial veins. All patients sus procedure with the aim to develop the first clinical practice were admitted to the ICU and underwent chest CT angiogram, four guideline for the management of kyphosis in MPS I patients. of which revealed the presence of remarkable diffuse tortuous Sequential radiographs (WKZ/UMC Utrecht and Amsterdam UMC, collateral veins. Coagulation profile and thrombophilia workup the Netherlands Royal Manchester Children’s Hospital, United were unremarkable in all cases. In conclusion, symptomatic DVT Kingdom San Gerardo Hospital, Italy) were obtained and mea- in Hunter patients appears to be at a higher rate than those with surements included the kyphotic angle (modified Cobb angle (CA) long-term central access for other indications. Interestingly, the and posterior angle (PA), both developed by us for measurement majority of patients with DVT showed evidence of extensive of kyphosis in dysplastic vertebrae) and spondylolisthesis. A formation of tortuous collateral vessels. The pathogenesis of these modified Delphi procedure was used to develop statements with findings remains unclear. This highlights the importance of an international expert panel including 10 spinal/orthopedic further understanding mechanisms and clinical implications of surgeons, 6 metabolic pediatricians and 3 physiotherapists, all DVT and vasculopathy as well as of adopting measures for experienced in MPS I. The procedure consisted of 3 written rounds screening and prevention of these findings in MPS II patients. and a face-to-face meeting. In the natural history study 44 patients (233 radiographs) were included. Intra- and inter-rater reliability was ≥ 0.9 for all measurements. The median modified CA was doi:10.1016/j.ymgme.2018.12.220 larger in Hurler patients (38°) compared to non-Hurler patients (9°) and an abnormal modified CA was present at a younger age. A modeled trend showed a progression of the modified CA of 1.8°/ 205 year for Hurler patients. The modified CA and the PA correlated Mucopolysaccharide quantitation in urine by LC-MS/MS strongly. Spondylolisthesis was present in 34 patients. During the modified Delphi procedure 16 statements on decision for surgery Jean M. Lacey, Dimitar K. Gavrilov, Dietrich Matern, Devin Oglesbee, and goals of treatment were developed. Consensus was reached on Kimiyo Raymond, Piero Rinaldo, Silvia Tortorelli, Mayo Clinic, all statements. In conclusion, we present an extensive radio- Rochester, MN, United States graphic assessment of thoracolumbar kyphosis in MPS I patients showing kyphosis is progressive over time. These natural history The mucopolysaccharidoses (MPSs) are a group of disorders data may be important for future studies. In addition, we present caused by a deficiency of enzymes involved in the stepwise the first clinical practice guideline for management of kyphosis in degradation of dermatan sulfate (DS), heparan sulfate (HS), keratan MPS I patients. sulfate (KS), or chondroitin sulfate (CS). We describe a LC-MS/MS method modification for routine determination of MPS which doi:10.1016/j.ymgme.2018.12.219 improves upon the existing method with respect to sample volume, sample preparation, and reduces the potential for false positive and false negative diagnosis when relying on a qualitative method. DS, HS and KS are enzymatically digested to disaccharides by the 204 addition of chondroitinase B, heparinase I, II, III, and keratanase, Deep vein thrombosis is a common life-threatening complication while CS is enzymatically digested to disaccharides by in mucopolysaccharidosis type II chondroitinase AC prior to LC-MS/MS analysis. Intra- and inter-assay precision was assessed using urine samples (N=3) of varying Anatalia Labilloy, Lisa Berry, Connie Wehmeyer, Robert J. Hopkin, concentrations. Intra-assay precision CVs ranged from 4.7 - 8.4% for Carlos E. Prada, Cincinnati Children's Hospital Medical Center, DS 4.9 - 10.4% for HS 4.1 - 9.1% for KS and 4.9 - 5.3% for CS, Cincinnati, OH, United States respectively (N=20). Inter-assay precision CVs ranged from 10.9 - 14.2% for DS 8.9 - 14.9% for HS 9.2 - 14.8% for KS and 6.9 - 8.8% for CS, Mucopolysaccharidosis type II (MPS II) is an X-linked lyso- respectively in the same specimens (N=20). Three urine specimens somal disorder caused by deficiency of iduronate 2-sulfatase spiked with MPS standard solutions at 2 levels exhibited recoveries resulting in progressive accumulation of dermatan and heparan ranging from 88% - 112%. Pediatric urine (age 1 day - b19 years) sulfate with multisystem involvement including neurological, (N=554) and adult urine controls (≥19 years (N=319) were respiratory, cardiac and skeletal. Long-term enzyme replacement analyzed for reference range determination. Clinical sensitivity was therapy for MPS II has demonstrable benefits in walking ability, 100% for MPS-I (N=26), MPS-II (N=20), MPS-III (N=17), MPS-IVA endurance and organomegaly and possibly life expectancy and is (N=17), MPS IVB (N=2) and MPS VI (N=5). Mucolipidoses (MLII/ usually delivered by a central venous port system. Intravenous III, N=6), multiple sulfatase deficiency (N=7), fucosidosis (N=3) S90 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 and GM1 gangliosidosis (N=3), may mimic any MPS disorder, but Mucopolysaccharidoses (MPS) are rare disorders associated with can potentially be further delineated by oligosaccharide analysis. enzyme deficiencies, resulting in glycosaminoglycan (GAG) accumulation in multiple organ systems. Because of improvements to the diagnosis and treatment of MPS, patients are increasingly surviving doi:10.1016/j.ymgme.2018.12.221 to adulthood, and a smooth transition into adult care is essential. We present transition challenges and resolutions through case studies and share best practice strategies in order to optimise transition for 206 MPS patients. Six cases were provided by four leading European Development of a technical colorimetric for the determination of metabolic disease centres, including patients with MPS I, II, IIIC and fi galactosis in blood VI, aged 15-18 years at rst transition visit (age at transfer to adult care: 16-19 years). Transition processes were based on national guidelines and institutional regulations. Strategies varied in duration Karima Lafhala, Mouna Cheggoura, Es-Said Sabira, Safya Sbyeaa, Laila of transition periods, healthcare providers involved and methods Chabaaa, Fatima Zahra Mouadb, Bouraghouat Aicha Bouraghouatb, used to support the patient and monitor their readiness for transfer Naima Fdila,c, aLaboratory of Biochemistry Faculty of Medicine of to adult care. At one centre, adult care was coordinated in a Marrakech, Cadi Ayad University, Marrakech, Morocco, bPediatric B paediatric setting at others, patient care was transferred from services of Mohammed VI Hospital, Marrakech, Morocco, cBiochemistry paediatric- to adult-care teams in different hospital departments or Research Team, Cadi Ayyad University, Marrakech, Morocco at different sites. Challenges included attachment to paediatricians and unwillingness to attend appointments with unfamiliar Galactosemia, a life-threatening disorder, is an inherited healthcare professionals. Limited knowledge of MPS in adult-care metabolic disease transmitted in an autosomal recessive manner. teams also concerned patients and parents. Challenges were resolved It is metabolized in humans and other species predominantly by by preparing for transfer at an early stage, regularly communicating the sequential activities of three enzymes that constitute the Leloir with and reassuring the patient and family, coordinating the process pathway. A deficiency of any one of the Leloir enzymes in humans, through MPS experts, and including multidisciplinary input from results in a galactosemia. Our study aims to develop a fast and MPS adult-care experts. These cases highlight the strategies that can accurate colorimetric technique of qualitative and quantitative be employed to manage transfer from paediatric to adult care. determination of galactose in blood. This technique involves Transition plans should be comprehensive but flexible to accommo- incubating the standard galactose and glucose ranges with orcinol date each patient’s needs. Sufficient time should be provided to allow in a strongly acidic solution to form a product capable of absorbing patients to understand the process and how the adult-care system light at both wavelengths. This allows the determination of both differs from the paediatric setting. Effective communication between sugars simultaneously. The standard range of 12.5 to 200 mg/ adult and paediatric teams, as well as between patients and carers, is linearity of y = 0.0027x - 0.004, R² = 0.9999 and y = 0.0017x + key to ensuring a smooth transition. 0.0129, R² = 0.9963 of glucose and galactose respectively, agitation of orcinol is 3 min, incubation in the dark is 30min. The maximum absorbance of galactose is 569.5 nm while that of doi:10.1016/j.ymgme.2018.12.223 glucose is 421 nm. Since galactose in whole blood is unstable over time, all the parameters of the assay have been standardized after 1 year of testing. We have modied a technique colorimètrique that can be applied to the detection of galactosemia. The target analyte 208 is total galactose. We continued to simplify this technique which is Interfamily variability in patients with classical Fabry disease fast and economical. The interference between glucose and galactose has been bypassed by a simple calculation which makes Dawn A. Laney, Emory University, Atlanta, GA, United States it possible to determine the concertration of these two sugars simultaneously. To have good results, certain factors must be Fabry disease (FD) is a X-linked lysosomal storage disorder maintained, such as time and temperature. Thanks to this method caused by pathogenic variants in the GLA gene leading to insufficient we could diagnose 8 suspected cases of which only one has this alpha-galactosidase A enzyme levels, progressive substrate accumu- disease. lation, and multisystemic medical complications. The evolving understanding of FD phenotype recognizes that symptoms present on a spectrum from the severe “Classical” presentation to the more doi:10.1016/j.ymgme.2018.12.222 variable, but still life-impacting “Nonclassical” presentations. Even within classical disease presentation, intrafamilial variability related to symptom onset and severity is present. In order to highlight this 207 intrafamilial variability, we discuss cases in the literature and report Transition from paediatric to adult care in patients with on signs, symptoms, and onset of FD in two multigenerational mucopolysaccharidosis (MPS) families with classical variants. Discussion will also review possible modification of phenotype in young family members through disease specific therapies including enzyme replacement and chaperone Christina Lampea, Briony McNellyb, Anahit Gevorkyanc, Christopher therapy. Family 1 includes 7 members (4 males, 3 females) over Hendrikszd, Tinatin Lobhanidzec, Jordi Pérez-Lópeze, Nato Vas- three generations with confirmed diagnosis of Fabry disease caused hakmadzec, Mireia del Toroe, aHelios Dr. Horst Schmidt Kliniken by the c.164ANT/c.171GNA (p.Asp55Val/Gln57Leu) GLA variants. Wiesbaden, Wiesbaden, Germany, bSalford Royal NHS Foundation Trust, Intrafamilial variability is most evident in the different presenting Salford, United Kingdom, cResearch Center for Children's Health, phenotypes in males in the family: two males in differing Moscow, Russian Federation, dUniversity of Pretoria, Pretoria, South generations with childhood onset of neuropathic pain, but not Africa, eVall d'Hebron University Hospital, Barcelona, Spain gastrointestinal issues and two additional males in differing Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S91 generations with gastrointestinal issues but not neuropathic pain. alfa can be maintained long-term in real-life clinical practice. This Family 2 includes three affected family members (1 male, 2 females) study and writing support were funded by Shire. over two generations with a confirmed diagnosis of FD caused by a c.647ANG (p.Tyr216Cys)] GLA variant. In this family, childhood symptom onset and progression is very similar between males and doi:10.1016/j.ymgme.2018.12.225 females including gastrointestinal issues and neuropathic pain in childhood. However, other childhood symptoms diverge with one female family member exhibited severe vertigo and the other family 210 members affected by more frequent and severe pain crises. This Clinical characteristics of patients with neuronopathic and non- discussion of intrafamilial phenotypic variability provides a reminder neuronopathic mucopolysaccharidosis type II: Data from the that phenotype is a spectrum even in patients in the same family Hunter Outcome Survey with the same classical GLA variant. Heather A. Laua, Paul Harmatzb, Jaco Bothac, aNew York University, New York, NY, United States, bUCSF Benioff Children’s Hospital Oakland, doi:10.1016/j.ymgme.2018.12.224 c Oakland, CA, United States, Shire, Zub, Switzerland

Patients with neuronopathic mucopolysaccharidosis type II (MPS 209 II Hunter syndrome) develop progressive cognitive impairment (CI), Long-term analysis of velaglucerase alfa-treated patients with whereas those with non-neuronopathic disease remain cognitively Gaucher disease who entered the Gaucher Outcomes Survey (GOS) intact all patients have somatic involvement, but the extent varies real-life registry widely. Cognitive decline is often apparent by age 5 years, although this also varies. Using data from the HOS registry (Shire July 2017 data), we examined the age at which absence of CI might be Heather Laua, Nadia Belmatougb, Pilar Giraldoc, Derralynn Hughesd, considered indicative of non-neuronopathic disease. Examination of Jaco Bothae, Steven Huyssee, Zoya Panahlooe, Ari Zimranf, Patrick cognitive data in HOS (answer to a “yes/no” question, based on Deegang, aNew York University, New York, NY, United States, clinical impression and/or standardized testing) by age revealed bUniversity Hospital Paris Nord Val de Seine, Clichy, France, cIIS Aragon, that changes in cognitive status were infrequent after a cut-off age Zaragoza, Spain, dUniversity College Medical School, London, United of 10 years. Using this criterion, we compared clinical characteris- Kingdom, eShire, Zug, Switzerland, fShaare Zedek Medical Center and the tics of patients with neuronopathic and non-neuronopathic MPS II. Hebrew University-Hadassah Medical School, Jerusalem, Israel, gAd- Overall, of 146 patients (two female) who had a cognitive denbrooke's Hospital, Cambridge, United Kingdom assessment available at age 10 years and ≥1knownfollow-up assessment aged ≥11-b20 years, 127 (87.0%) had no change in CI Gaucher disease (GD) is a rare, autosomal recessive condition, characterized by hepatosplenomegaly, thrombocytopenia and ane- status between these assessments. Aged 10 years, 80/127 (63.0%) patients had CI and 47/127 (37.0%) had no CI. Median (P10, P90) mia. Velaglucerase alfa resulted in improvements in these parameters in clinical studies, but there are limited data on long-term age at last visit was 13.8 (11.8, 18.0) and 14.8 (11.9, 19.5) years, respectively. Symptom onset occurred earlier in patients with CI efficacy and safety in real-life clinical practice. The Gaucher Outcome Survey (GOS), an international registry for patients with confirmed (1.5 [0.3, 3.0] versus 2.3 [0.5, 4.5] years). The most common signs/ symptoms generally had similar prevalence in each group. Although GD regardless of type or treatment status, collects data during patients’ routine care, and provides an opportunity for long-term not among the most prevalent manifestations, bladder/urinary incontinence, enlarged tongue, behavior problems and sleep apnea analysis of patients receiving velaglucerase alfa. This evaluation included 197 treatment naïve patients who initiated velaglucerase were more common in patients with CI. Number of surgeries by age 10 years (3 [1, 8] versus 4 [1, 8]), age at first surgery (2.3 [0.2, 6.8] alfa upon enrollment into GOS and continued velaglucerase alfa treatment for up to 8 years (mean 59.8 months). Hemoglobin levels, versus 2.3 [0.2, 5.1] years) and types of surgery were also similar. platelet counts, liver and spleen volume, and chitotriosidase levels Overall, our data indicate that patients without CI are still markedly were assessed. At baseline (start of velaglucerase alfa), 77.2% affected by somatic symptoms of MPS II. Shire sponsors HOS and patients were aged ≥18 years, 58.9% were female, 52.8% had a funds medical writing support. known N370S-containing genotype, and 87.8% had an intact spleen (including partial splenectomy). Patients showed stabilization or doi:10.1016/j.ymgme.2018.12.226 improvement in all measured clinical parameters during the study period. After 4 years of velaglucerase alfa treatment, mean (SD) hemoglobin levels increased by 7.5 g/L from baseline to 136.4 (12.8) g/L (n=19 patients with available data) and mean (SD) platelet 211 counts increased by 54.6 x109/L to 166.4 (57.9) x109/L (n=19). Liver AAV gene therapy LYS-SAF302 demonstrates widespread size remained stable at 1.2 MN (n=4) over 4 years, while spleen size sulfamidase distribution in primate brain and correction of decreased from 7.5 MN to 6.4 MN (n=4), a mean reduction of 1.2 disease pathology in MPS IIIA mice MN. Mean (SD) chitotriosidase values reduced by 4302.7 nmol/mL/h to 1019.1 (1907.45) nmol/mL/h at year 4 (n=14). Sixty-one (31.0%) Ralph Laufera, Michael Hocquemillera, Kim Hemsleyc, aLysogene, patients experienced 153 adverse events during velaglucerase alfa Neuilly sur Seine, Paris, France, bSAHMRI, Adelaide, Australia treatment 16 AEs in 10 patients were considered related to velaglucerase, but none were serious. This analysis suggests that Lysogene completed its program of nonclinical studies for LYS- improvements in clinical parameters achieved with velaglucerase SAF302, an optimized AAVrh.10 vector carrying the human SGSH S92 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 gene. LYS-SAF302 achieves 3-fold higher enzyme expression than LYS- result upon their second urine analysis. Of these latter patients, 3 had SAF301, which was previously tested in a phase 1/2 trial. A dose passed away when enzyme activity testing was requested. The ranging efficacy study was conducted with LYS-SAF302 in 5-week-old remaining four patients had normal enzyme activity or mutation MPS IIIA mice (15/gender/group) at three different doses (8.6E+08, analysis. In conclusion, no Fabry patients were detected in this study.

4.1E+10, and 9.0E+10 vg/animal) injected into the caudate putamen/ Having done 13% retests, this might indicate that urinary Gb3 is not striatum and thalamus. LYS-SAF302 was able to dose-dependently specific enough for high-risk screening of Fabry patients having correct MPS IIIA-related brain pathology at 12-weeks and 25-weeks chronic kidney disease. Nevertheless, larger cohorts might be neces- post-surgery. To study SGSH distribution in the brain of large animals, sary to draw definite conclusions. LYS-SAF302 (7.2E+11vg/animal) was injected into white matter of nonhuman primates (n=22 injections of 50μl per hemisphere), using an optimized injection method. Six weeks following dosing, animals doi:10.1016/j.ymgme.2018.12.228 were euthanized, the brain was sliced into 3-4 mm coronal slabs, and each slab was divided into 10 x10 mm sections. Viral DNA and SGSH enzyme activity were determined in each of these punches. In 97% 213 N (±2) of punches analysed, we observed a 20% increase in SGSH Pharmacological chaperone therapeutics for Krabbe disease activity relative to non-injected controls. In parallel, a GLP toxicolog- ical and biodistribution study conducted in 12 juvenile NHP did not Chris W. Lee, Arulmani Manavalan, Dana Clausen, Biomedical reveal toxicity following bilateral white matter administration of LYS- Research Institute of New Jersey, Cedar Knolls, NJ, United States SAF302, and confirmed the broad distribution of enzymatically active SGSH throughout the primate brain. Taken together, these data Loss-of-function mutations in the galactcerebrosidase (GALC) lead validate intraparenchymal AAV administration as a promising method to the development of Krabbe disease, a degenerative disorder to achieve widespread enzyme distribution and correction of disease characterized by progressive demyelination and neuroinflammation. pathology in MPS IIIA. An international Phase 2-3 clinical trial in MPS More than 100 mutations have been identified in Krabbe disease IIIA with LYS-SAF302 is planned. patients including deletions, frameshift mutations, nonsense mutations and missense mutations. Positive disease diagnosis is confirmed doi:10.1016/j.ymgme.2018.12.227 by low GALC enzymatic activity in blood sample and mutational analysis of the GALC gene. More than 20 pathogenic GALC mutations have been determined to be missense mutations, which impair GALC protein function by altering the catalytic site and/or folded structure, 212 which leads to protein mistrafficking. In these patients, GALC protein is High-risk screening for Fabry disease in chronic kidney disease present but inactive, or missing from the lysosome, respectively. Most patients disease-associated missense mutations are distant from GALC’s catalytic site, which suggests that the mutant proteins will retain Pamela Lavoiea, Michel Boutina, Mona Abaouia, Anne-Marie Côtéa, some catalytic activity if they can be stabilized enough to properly fold Ayub Akbarib, Adeera Levinc, Fabrice Mac-Wayd, Christiane Auray- and/or be transported to the lysosome. Pharmacological chaperone Blaisa, aCIUSSS de l’Estrie-CHUS, Université de Sherbrooke, Sherbrooke, (PC) is an emerging class of small-molecule therapeutics that can QC, Canada, bOttawa Hospital Research Institute, Ottawa, ON, Canada, stabilize protein structure and restore function in enzymes with a cSt. Paul’s Hospital, Vancouver, BC, Canada, dCHU de Québec, Hôtel-Dieu missense mutation. In 2010 we identified one of the first PC candidates de Québec, Quebec, QC, Canada for Krabbe disease, α-lobeline (LB). This natural compound was found to partially correct the function of the GALC D544N mutant protein in a Fabry disease is a multisystemic, X-linked lysosomal storage human cell-based assay. Later, it has also demonstrated by other group disorder caused by a deficit of the enzyme alpha-galactosidase A that LB can increase the activity of several other GALC mutants (G553R, which leads to the accumulation of glycosphingolipids in tissues and E130K, N295T and G57S) in patient’s fibroblasts. These results and biological fluids. The clinical presentation is heterogeneous with many others suggest that LB and other PC-based molecules may restore the atypical variants, leading to an underestimated prevalence. Enzyme function of a broad spectrum of GALC mutants. Therefore, we replacement therapy and chaperone therapy are some of the hypothesize that GALC missense mutations that cause protein mis- treatments offered. Since renal dysfunction is a common manifestation folding, but maintain catalytic potential, are amenable to functional of Fabry disease, our group elaborated a high-risk screening protocol correction by LB and other PC candidates. We will present the results for patients with chronic kidney disease of unknown etiology. of our initial effort to identify amenable mutations and new potential Recruitment of participants took place in four centers across Canada PC candidates for therapeutic development. from 2011 to 2017. Participants provided a urine specimen collected on filter paper at their first visit for globotriaosylceramide (Gb3) analysis. If abnormal, a liquid urine specimen was collected for an doi:10.1016/j.ymgme.2018.12.229 additional Gb3 and globotriaosylsphingosine (lyso-Gb3) and related analogue analyses. All biomarker analyses were performed by liquid chromatography/tandem mass spectrometry. If an abnormal Gb3 or 214 nd lyso-Gb3 and related analogue result was obtained upon the 2 Dose-dependent impact of ERT on neutralizing anti-drug anti- α analysis, the measurement of -galactosidase A activity was requested bodies and long-term outcomes in Fabry disease in males and a mutation analysis in females. A total of 402 patients were recruited. Fifty-three patients (13.2%) had an abnormal Gb 3 Malte Lendersa, Leon Paul Neußera, Michael Rudnickib, Peter result on the first analysis. Of these, 50 patients (94.3%) provided a Nordbeckc, Sima Canaan-Kühld, Albina Nowake, Markus Cybullaf, liquid urine specimen for retest, but unfortunately 2 patients were lost Boris Schmitza, Jan Lukasg, Christoph Wannerc, Stefan-Martin Branda, to follow-up and one died. Seven patients (14%) still had an abnormal Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S93

Eva Branda, aUniversity Hospital Muenster, Muenster, Germany, was performed by real-time PCR and fluorescence microscopy. bMedical University Innsbruck, Innsbruck, Austria, cUniversity Hospital Amenability to chaperone treatment was quantified by enzyme and University of Wuerzburg, Wuerzburg, Germany, dCharité, Campus activity assays and intracellular Gb3 measurements using flow Virchow-Klinikum, Berlin, Germany, eUniversity Hospital of Zurich, cytometry and compared to patients’ biochemical response. Under University of Zurich, Zurich, Switzerland, fCenter of Internal Medicine, chaperone treatment (N13 months), patients with the p.N215S Muellheim, Germany, gUniversity Rostock Medical Center, Rostock, mutation (n=5) showed a significant reduction of plasma lyso-Gb3 Germany (pb0.05). By contrast, lyso-Gb3 levels in carriers of the p.L294S Mutation (n=3) increased under treatment (pb0.05). While the Fabry disease (FD) is a rare X-linked disorder caused by a amenability assay in wildtype HEK293T cells suggested amenability deficiency of lysosomal α-galactosidase A (GLA) activity. Neutraliz- for both mutations (pb0.05), chaperone-treatment provoked in- ing anti-drug antibodies (ADA) against enzyme replacement therapy creased GLA activity (pb0.0001) and intracellular Gb3 reduction (ERT) are associated with disease progression in male patients. (pb0.05) only in immortalized p.N215S cells but not in p.L294S or Recently we demonstrated that ADAs can be supersaturated during IVS2+1 ANT cells. Amenabilities were confirmed using GLA knock-out infusions. In the current work we hypothesized that supersaturated cells for amenability assays. In vitro GLA activity assays predicted p. patients benefit from better clinical outcomes. Twenty-six male L294S as an amenable mutation for chaperone therapy. Our study with patients with FD receiving ERT (mean 98±70 months) and positive patient-specific immortalized cells revealed that the amenability of p. for ADAs were recruited. Free ADAs were isolated from patients' sera L294S might be too low, which was confirmed by the poor biochemical and the amount of ERT necessary for antibody supersaturation was response to migalastat observed in patients with p.L294S. Additional determined. Clinical outcomes including measurements of eGFR and studies are required to further analyze mutations with endogenous septum thickness of supersaturated patients were compared to non- GLA activity b5% of wildtype for their amenability to chaperone supersaturated patients. ADA titers decreased in all patients (n=26) treatment in a patient-specific Setting. during infusion (pb0.0001) and the capacity to bind ADAs was similar for agalsidase-alfa and agalsidase-beta (r²=0.9740 pb0.0001). Patients with supersaturated ADAs (n=14) presented doi:10.1016/j.ymgme.2018.12.231 with a yearly loss of eGFR (-2.7±2.2 ml/min/1.73m² p=0.0015), a stable septum thickness (p=0.4628) and decreasing lyso-Gb3 levels (-13.1±16.3 ng p=0.0099) since ERT initiation. By contrast, non- 216 supersaturated patients (n=12) suffered from a more severe yearly Generation of patient-specific human induced pluripotent stem loss of eGFR (-5.1±5.0 ml/min/1.73m² p=0.0005), an increasing cells to analyze mutation- and cell-specific pathomechanisms in septum thickness (0.3±0.4 mm p=0.0039) and worse lyso-Gb3 Fabry disease reduction (p=0.2834). Dose-escalation resulted in partially elevated ADA titers, but importantly in reduced lyso-Gb3 levels in 3 patients. Malte Lenders, Franciska Stappers, Christoph Niemietz, Hartmut We conclude that measuring neutralizing ADA titers is feasible to Schmidt, Eva Brand, University Hospital Muenster, Muenster, Germany calculate and control the required amount of enzyme to supersatu- rate ADAs during infusions. A non-supersaturated status is associated Fabry disease (FD) is an X-linked lysosomal storage disorder with a progressive loss of eGFR and ongoing cardiac hypertrophy. caused by the deficient activity of the lysosomal enzyme α- Dose escalation can result in supersaturation of ADAs and decreasing galactosidase A (GLA), leading to intracellular globotriaosylceramide lyso-Gb3 levels, but may also lead to increased ADA titers. (Gb3) accumulation. Induced human pluripotent stem cells (iPSCs) provide a powerful tool to analyze cellular pathomechanisms caused doi:10.1016/j.ymgme.2018.12.230 by a rare disease. Here we demonstrate the successful non-invasive generation of iPSCs from urinary cells of four male FD patients with individual GLA mutations. Urinary cells from 4 male patients with the GLA mutations p.S126G, p.N215S, p.L294S, and IVS2+1CNT were 215 isolated and cultivated for pluripotency induction. iPSCs were Patient-specific Fabry disease cell models as a tool to evaluate the induced by a transient triple transfection with pCXLE-hUL, pCXLE- amenability to chaperone therapy hSK, and pCXLE-hOCT3/4-shp53 (all Addgene). Q-PCR demonstrated a stable OCT3/4, NANOG, and SOX2 expression in all IPSCs over time Malte Lenders, Franciska Stappers, Christoph Niemietz, Boris (N passage 25), which was confirmed by flow cytometric analysis Schmitz, Andree Zibert, Hartmut Schmidt, Stefan-Martin Brand, Eva and fluorescence microscopy. Induction of embryoid bodies con- Brand, University Hospital Muenster, Muenster, Germany firmed pluripotency demonstrating increasing marker expression for the three germ layers of mesoderm (FOXF1, HAND1), endoderm Fabry disease (FD) is an X-linked lysosomal storage disorder caused (FOXA2, HNF4a), and ectoderm (PAX6), while marker expression for by the deficient activity of the lysosomal enzyme α-galactosidase A self-renewal (SOX2, NANOG, OCT4) decreased in parallel. Neuronal (GLA), leading to intracellular globotriaosylceramide (Gb3) accumu- induction of iPSCs to generate neural progenitor cells resulted in an lation. Patients with amenable mutations can be treated with a increased mRNA expression of NCAM and SOX1, as well as a high pharmacological chaperone (migalastat) to restore endogenous GLA protein expression for NCAM and nestin. Mesodermal induction of activity. Current tests to determine patients’ amenability are limited to early mesoderm resulted in an increased brachyury (T) and NCAM heterologous mutation expression in HEK cells. We established a new expression. FD-specific characterization of IPSCs demonstrated cell culture model based on immortalized urine cells from Fabry different endogenous GLA activities and amenability to migalastat, patients, as well as CRISPR-CAS 9-mediated HEK cells lacking depending on the existing genotype. Here we demonstrate the endogenous GLA activity. The model was used to test amenability of successful and non-invasive generation of patient- and mutation- GLA mutations p.N215S and p.L294S as well as IVS2+1 ANT nonsense specific iPSCs from urine of male Fabry patients. This approach can mutation, serving as negative control. Characterization of the cell lines now be used to analyze the cellular pathology of FD and response to S94 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 present and future therapeutically options in different primary cells Hughesi, aCharles University, Praha, Czech Republic, bThe Royal of interest in a GLA mutation-dependent manner. Melbourne Hospital, Melbourne, Australia, cDalhousie University, Hali- fax, NS, Canada, dHaukeland University Hospital, Bergen, Norway, eSalford Royal Dept of Inherited Metabolic Disease, Manchester, United doi:10.1016/j.ymgme.2018.12.232 Kingdom, fHospital de Dia Quiron, Zaragoza, Spain, gGeneral Hospital Slovenj Gradec, Gradec, Slovenia, hProtalix, Carmiel, Israel, iLSDU, Institute of Immunity and Transplantation, Royal Free London NHS Foundation Trust, London, United Kingdom. 217 Selective large scale screening for lysosomal diseases in minority Pegunigalsidase alfa is a novel, PEGylated, covalently-linked recom- groups shows higher incidence rates binant α-Galactosidase-A enzyme homodimer, for the treatment of Fabry disease (FD). The “Bridge” study (PB-102-F30 NCT03018730) an a a a Renuka P. Limgala , Erk Changsila , Vyalcheslav Furtak , Margarita M. on-going Phase III, open label, switch-over study, assessing the safety a b b Ivanova , Flyod Wilks , Marie N. Fidelia-Lambert , Marjorie C. and efficacy of pegunigalsidase alfa in FD patients previously treated b a a Gondré-Lewis , Ozlem Goker-Alpan , Lysosomal and Rare Disorders with agalsidase alfa for at least 2 years. The study enrolls up to 22 adult b Research and Treatment Center, Fairfax, VA, United States, Howard patients to be treated with pegunigalsidase alfa, 1mg/kg every other University College of Medicine, Washington, DC, United States week for 12 months. The kidney function related inclusion criteria are estimated glomerular filtration rate (eGFR) ≥ 40 ml/min/1.73m2 and no We investigated the incidence of four Lysosomal Storage Disorders or treated proteinuria. The baseline characteristics of the first 16 (LSDs) in a cohort of mostly urban-dwelling African Americans, patients (9 males and 7 females) enrolled: age 27-60 years kidney compared to previous work primarily in Caucasians (and Ashkenazi function: 8/16 are treated with ACEi or ARBs, of which 3/16 have Jews). LSDs are inherited genetic disorders that result in the functional proteinuria, mean eGFR of 75.4 and 86.0 mL/min/1.73m2 for males and absence or deficiency of specific lysosomal enzymes resulting in a range females, respectively with an annualized eGFR slope of -8.0 and -5.1mL/ of clinical symptoms manifesting in various organ systems. To highlight min/1.73m2/year, respectively mean residual leucocytes enzymatic diagnostic challenges originating from ambiguous clinical manifesta- activities of 5.9% and 27.9%, respectively, and plasma lyso-Gb3 of 53.6 ‘ ’ tions, we initiated a large scale screening for treatable forms of LSDs in and 13.8 nM, respectively. Preliminary results of the first 16 patients 5000 patients seeking healthcare for various health concerns. Of these, treated for six months with pegunigalsidase alfa show improvement in 85% reported as African American, 10% Hispanic, and 5% Caucasian or the mean annualized eGFR slope, from -6.8 mL/min/1.73m2/year while other. Dried blood spots were prepared from peripheral blood samples on agalsidase alfa, to +3.7 mL/min/1.73m2/year after switching and and fluorometric enzyme assays performed for β-glucosidase (Gaucher treated with pegunigalsidase alfa. These results suggest the potential disease), α-galactosidase (Fabry disease), α-glucosidase (Pompe dis- benefit of pegunigalsidase alfa on renal function for FD patients ease), and β-galactosidase (MPS IV B) enzymes. On samples which currently treated with the available enzyme replacement therapies. showed significantly lower enzyme activities, molecular analysis was performed using targeted sequencing. After two rounds of screening, we confirmed undetectable activity for β-glucosidase in 1 subject α- doi:10.1016/j.ymgme.2018.12.234 galactosidase in 3 subjects β-galactosidase in 1 subject and α- glucosidase in 5 subjects. In addition, approximately 0.05-0.1% showed significantly reduced enzyme activity, likely indicating carrier status. 219 These samples are being screened using Next-Gen Sequencing and Like mother, like daughter: A case report of multiple family preliminary results highlight higher incidence rates for abnormal members affected by Pompe disease enzyme levels and pathogenic mutations in the target population compared to previously published LSD screenings. The findings could be because the patient cohort was already under clinical care thus Valynne Long, Allison Foley, Stephanie Wechsler, Emory University, increasing the likelihood of a disease phenotype or due to ancestry- Atlanta, GA, United States based genotype and phenotype variations. It is highly likely that variations in genomics and resultant biochemical cascades contribute Pompe disease is a rare, lysosomal storage disease resulting from to health and disease. The study highlights the importance of such large reduced levels of alpha glucosidase (GAA) enzyme. The disease is scale screenings in minority groups traditionally not associated with a divided into two subtypes: infantile onset and late onset. Infantile high incidence of LSDs. The study is partially funded by Shire (IIR-USA- onset Pompe disease typically presents with cardiomyopathy and 000892), Pfizer (WI194030) & NCATS/NIH (#UL1-TR001409). hyptonia in the early neonatal period progressing to death by age 2 for those untreated. Late onset disease has a more generalized presentation with respiratory decline, muscle wasting and weakness, without cardiomyopathy, beginning in the proximal muscles, doi:10.1016/j.ymgme.2018.12.233 occurring typically after the first 12 months of life. The symptoms are variable and seem to correlate with the amount of residual GAA activity. The condition is inherited in an autosomal recessive fashion. 218 Due to this recessive inheritance, it is uncommon to see multiple Pegunigalsidase alfa for the treatment of Fabry disease: Prelim- generations in one family with the condition. Recessive disease can inary results from a phase III open label, switch over study from sometimes present as a pseudo-dominant condition in populations agalsidase alfa where carriers are common due to founder effect, such as Gaucher disease in the Ashkenazi Jewish population. They also occur more Ales Linharta, Kathy Nichollsb, Michael Westc, Camilla Tøndeld, Ana frequently where consanguinity occurs. We present a case with two Jovanovice, Pilar Giraldof, Bojan Vujkovacg, Einat Almonh, Sari Alonh, generations of a family affected by Pompe disease with multiple Bat-chen Amit-Cohenh, Mali Szlaiferh, Raul Chertkoffh, Derralynn family members in each generation affected, all females. In addition, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S95 there is possibly another, as yet unidentified, genetic disorder in the discern these gaps and assesses if online information can fill them. family. Our case highlights the importance of screening other family This study consists of a pre-survey, a research period, and a post- members for rare genetic diseases despite the lower probability of survey. The pre-survey collects demographic information, tests disease based on mode of inheritance. knowledge, and determines participants’ questions. Questions are split into current and initial questions (from the time of diagnosis). Participants are then directed to ThinkGenetic.com, a source for doi:10.1016/j.ymgme.2018.12.235 accurate and current genetic information, for research at their leisure. The post-survey determines if knowledge scores improved and if questions were answered. 100 participants were recruited from the Emory University Lysosomal Storage Disease Clinic. 34 220 participants achieved perfect scores (5) on both surveys. Scores Cyp2d6 allelic characterization on type 1 Gaucher disease patients changed for 34 participants, in which all but 4 increased. The median increase was 1. All decreased scores lowered by 1 point. The majority Laura López de Frutos, Pilar Alfonso, Carlos Lahoz-Gil, Pilar Irún, Pilar of initial and current questions pertained to treatment. Initial Giraldo, Instituto de Investigación Sanitaria Aragón (IIS Aragón), questions also focused on daily living and general knowledge of the Zaragoza, Spain disease. Current questions also focused on daily living. 51 participants reported finding answers to their initial questions, whereas Cytochrome p450 is the main drug metabolic pathway mostly by only 33 participants reported finding answers to their current CYPIIIA3 and CYPIID6 enzymes. CYPIID6 is the responsible for the questions. Participants reported that questions remained about clearance from 20% of most common drugs as paracetamol or codeine treatment, research studies, and daily living, indicating that this is and presented four different phenotypes: normal metabolizer (NM), where their knowledge gaps lie. The data suggests that accurate intermediate metabolizer (IM), poor metabolizer (PM) or ultra-rapid online information can answer questions at the time of diagnosis. metabolizer (UM). CYP2D6 (MIM*608902) is a highly polymorphic However, as they live longer with the disease, participants accumu- gene that encodes CYPIID6. The different metabolizer capabilities are late questions that may require further research. The small increase caused by different alleles on CYP2D6 due to entire gene deletions or in knowledge scores is attributed to the small number of knowledge duplications or 19 SNPs. This is the pathway to metabolize one of questions and the high baseline knowledge of the cohort. We will Gaucher disease (GD) treatments, which dose administration is continue this study with more LSD patients, family members, and regulated according to the metabolizer phenotype being this an caregivers. administration limitation. With this background we consider interesting stablish the metabolizer and allelic frequency from CYP2D6 on our GD population. Because of this, we select 109 type 1 GD patients doi:10.1016/j.ymgme.2018.12.237 genetically confirmed and we used the xTAG®CYP2D6 kit to identify the CYP2D6 gene alleles. As a result 87 patients were classified as normal, 14 as intermediate, 6 as poor and 2 as ultra-rapid metabolizers. 222 The allelic frequencies were 5.5% for duplication and 4.5% for deletion. Platform technology for treatment of the brain in lysosomal The most common allele was wild type and the second one was the disorders: Application to Niemann-Pick disease type A null allele *4. Alleles *6 and *9 presented a higher frequency than expected. There was no relationship between the patients’ gender or Jeff Z. Lu, Eric K.-W. Hui, Huilan Lin, Ruben J. Boado, William M. GBA1 genotype and the allelic or phenotypic frequencies on CYPIID6 Pardridge, ArmaGen Inc., Calabasas, CA, United States enzyme and CYP2D6 gene. To sum up, our Spanish GD series shows an unexpected distribution of alleles that encodes for enzyme with a The majority of lysosomal storage disorders (LSD) affect the reduced or absent activity than the results previously reported in brain. Intravenous enzyme replacement therapy does not treat the Spanish population. This is important due to the pharmacological brain, because recombinant enzymes are large molecules that do not interactions and the impact of unexpected secondary effect that can be cross the blood-brain barrier (BBB). BBB-penetration of enzyme higher on this population. This project has been partially supported by therapeutics is enabled by re-engineering the recombinant enzyme a grant from FEETEG and is submitted to its publication. as a bi-functional IgG-enzyme fusion protein, wherein the IgG domain is a monoclonal antibody (MAb) that targets a specific doi:10.1016/j.ymgme.2018.12.236 endogenous receptor-mediated transporter within the human BBB, such as the human insulin receptor (HIR). A proof of concept phase II clinical trial in pediatric subjects with severe Hurler MPSI with a BBB penetrating HIRMAb-iduronidase fusion protein (AGT-181, 221 valanafusp alpha) produced stabilization of cognition, and reduced Determining the disease-specific knowledge gaps in patients, hepatosplenomegaly. The aim of the present investigation was to family members, and caregivers living with lysosomal diseases determine if a similar BBB penetrating IgG-fusion protein can be engineered for the treatment of Niemann-Pick disease (NPD) type A, Georgia Loucopoulosa, Dawn Laneya, Morgan Simmonsa, Deepti which is caused by mutations in the gene encoding for acid Babub, Kathryn Garbera, Cecelia Bellcrossa, aEmory University, Atlanta, sphingomyelinase (ASM). Genes encoding the heavy chain and the GA, United States, bFreelance Consulting Medical Writer and Editor, light chain of a HIRMAb-PPT1 fusion protein were engineered Edmonton, AB, Canada followed by stable transfection of Chinese hamster ovary (CHO) cells. The identity of the CHO derived HIRMAb-ASM fusion protein There is a lack of information about knowledge gaps in the (AGT-189) was confirmed by hIgG1 and ASM Western blot analyses lysosomal storage disease (LSD) population, which limits the and the purity by SDS-PAGE and SEC-HPLC. Fusion protein binding to capacity for proper genetic counseling. This is the first study to the HIR was saturable with an ED50 in the low nM range and S96 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 comparable to the binding of the parental HIRMAb. The ASM activity United States, bUniversity of Colorado, Aurora, CO, United States, in the HIRMAb-ASM fusion protein was retained based on a cUniversity of Utah, Salt Lake City, UT, United States, dWashington fluorometric enzyme assay. The brain uptake of HIRMAb-lysosomal University, St. Louis, MO, United States, eSangamo Therapuetics, enzymes in non-human primates approximates 1% of injected dose Richmond, CA, United States, fLos Angeles Biomedical Research Institute, per brain. This level of brain uptake is able to produce therapeutic Los Angeles, CA, United States. levels of ASM enzyme activity in the brain. Successful development of a BBB penetrating HIRMAb-ASM fusion protein may be transfor- Abnormal cerebral spinal fluid (CSF) characteristics were previ- mational for the treatment of the brain in NPA Disease. ously reported in children with severe mucopolysaccharidosis type I (MPS IH Hurler syndrome), a progressive lysosomal storage disease associated with rapid neurocognitive decline, worsening multi- doi:10.1016/j.ymgme.2018.12.238 system organ dysfunction, and early death. The present objective was to evaluate whether intrathecal (IT) enzyme replacement therapy (ERT) added to intravenous ERT and hematopoietic cell transplantation (HCT) is associated with attenuation of CSF abnor- 223 malities and whether this change has clinical correlates. Twenty-four fl Twelve-year experience with a rapid and simple uorometric MPS IH patients received IT ERT at four time points in the peri- tripeptidyl peptidase 1 (TPP1) assay using dried blood specimens transplant period. At these times, CSF opening pressure (OP), heparin to diagnose CLN2 disease sulfate (HS), non-reducing ends (NRE I0S0 and I0S6), heparin cofactor II thrombin complex (HCIIT), and inflammatory markers Zoltan Lukacs, Miriam Nickel, Simona Murko, Paulina Nieves Cobos, were measured. Neurocognitive functioning (i.e., infant IQ) was Angela Schulz, René Santer, Alfried Kohlschütter, Hamburg University quantified at baseline, 1 year after HCT, and 2 years after HCT. There Medical Center, Hamburg, Germany were no adverse events due to the administration of IT ERT. An association between attenuated CSF biomarkers and IQ change CLN2 disease is a lysosomal storage disorder that belongs to the following transplant was examined. Significant reductions in CSF neuronal ceroid lipofuscinoses (NCL) and progressively leads to abnormalities were seen between time points 1 and 2, i.e. prior to fi dementia, blindness and early death. It is caused by a de ciency of HCT, for OP, NRE I0S0 and I0S6, and HCTIIT (p=0.036, 0.001, 0.006, the lysosomal enzyme tripeptidyl peptidase 1 (TPP1). The disease and 0.026, respectively), and across time points 1 through 4 for NRE has recently gained interest as an enzyme replacement therapy has I0S0 and I0S6, HS, and HCIIT (pb0.001 for all four biomarkers). become available. For this therapy to be effective, the diagnosis must Percent decrease in NRE from the first to fourth dose was be made early. We have investigated the diagnostic reliability of a significantly associated with a percent increase in IQ from baseline test for TPP1 deficiency in dried blood specimens (DBS), previously to 2 years post-HCT (Mean Fit Slope=0.703 [95% CI: (0.232,1.175)], developed by us, to detect CLN2 disease. During a 12-year period we p=0.003). IT ERT is associated with attenuation of abnormal CSF have received 3882 DBS for testing TPP1 activity. To check for quality biomarkers in MPS IH, with maximal attenuation following combi- of samples, we measured activities of two additional lysosomal nation of IT/IV ERT and HCT. This study offers the first evidence of a enzymes as controls, palmitoyl peptidase 1 and ß-galactosidase. Of correspondent biochemical change with a clinical manifestation of all samples, 1.7% were excluded from the study because of neurologic response, specifically IQ improvement. Clinicaltrials.gov subnormal activity of more than one enzyme. For all samples with NCT00638547 subnormal TPP1 activity and good sample quality, we reviewed clinical information. Consequently, we obtained adequate clinical and molecular genetic data for 51 patients. All of those had doubtless doi:10.1016/j.ymgme.2018.12.240 evidence of CLN2 disease (including seven atypical patients) as shown by symptoms of a progressive brain disease and presence of known pathogenic CLN2variants. The sensitivity of the test could not 225 be evaluated directly. However, as our NCL clinic is a major reference Preclinical gene therapy in a mouse model of Charcot-Marie- center for these disorders and we have never received feedback Tooth disease type 4J information that a patient with normal TPP1 activity in our DBS test was later diagnosed with CLN2 disease, this constitutes convincing a a a a circumstantial evidence of a high sensitivity. The TPP1 test on DBS as Cathleen Lutz , Maxmiliano Presa , Crystal Davis , Jennifer Cook , a a b b a used in this study was shown to be a reliable, convenient and Tara Murphy , Rob Burgess , Rachel M. Bailey , Steven J. Gray , The b inexpensive tool for a first diagnostic step in a suspected case of Jackson Laboratory, Bar Harbor, ME, United States, University of Texas CLN2 disease and, can be applied with minor modifications for mass Southwestern Medical Center, Dallas, TX, United States throughput, like newborn screening programs. Charcot-Marie-Tooth Disease Type 4J (CMT4J) is an ultra-rare, autosomal recessively inherited peripheral neuropathy, first described in 2007 and caused by mutations in the Fig4 Gene. FIG4 is doi:10.1016/j.ymgme.2018.12.239 a PIP2 phosphatase, and its loss leads to lysosomal abnormalities and “spongiform-like” vacuolization in neurons. Clinically, CMT4J is characterized by motor developmental delay, slow nerve conduction 224 velocities, rapidly progressive paralysis, quadriplegia, resulting Predicting intelligence in MPS IH with biomarkers respiratory complications and premature death. Later-onset patients have a more variable disease course, with some experiencing mild Troy Lunda, Julie Eisengarta, Rene Pierponta, Alexander Kaizerb, Kyle symptoms. There are currently no approved therapies for CMT4J Rudsera, Marzia Pasquallic, Patricia Dicksond, Weston Millere, Lynda patients. Gene therapy represents a reasonable and promising Polgreenf, Paul Orcharda, aUniversity of Minnesota, Minneapolis, MN, approach to provide a meaningful and long term therapeutic benefit Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S97 for this patient population. To explore this, we conducted preclinical studies recommended include larger, more generalizable studies for studies in the pale tremor mouse model, which lacks Fig4 expression. GD and GJH. These preclinical studies used a codon optimized human FIG4 gene, delivered by AAV9. Administering this vector directly to the nervous system by either intracerebroventricular (ICV) in early neonates or doi:10.1016/j.ymgme.2018.12.242 later, after the onset of symptoms by intrathecal (IT) injection produced benefit, as indicated by improved survival, improved gross motor function including grip strength, improved peripheral motor nerve conduction, and improved histopathological outcomes in 227 spinal cord and dorsal root ganglia, as well as other parts of the The impact of newborn screening for lysosomal disorders in a central nervous system. Importantly, these results indicate that the non-screening adjacent state benefit is greatest the earlier the FIG4-AAV9 is delivered. Preclinical study results also indicate a clear dose-response of the treatment up Samantha Marcellus, Lauren Jordan, Alpa Sidhu, Myrl Holida, John to the maximum feasible dose in mice, justifying the maximum Bernat, University of Iowa Hospitals and Clinics, Iowa City, IA, United feasible dose to provide the most effective treatment. Importantly, States there were no observable adverse effects, even when Fig4 was overexpressed in WT mice. Overall, our results predict a benefitof As other states have added Lysosomal Storage Disorderls (LSDs) this treatment to pediatric subjects, with no obvious toxicity to their newborn screening (NBS) programs, some families have fi expected, and support the further exploration of clinical trials for crossed borders and sought con rmatory testing and follow-up care CMT4J patients. in Iowa. With the addition of Pompe disease and Muco- polysaccharidosis type I (MPS I) to the Recommended Uniform Screening Panel (RUSP) in 2013 and 2015, respectively, states in the doi:10.1016/j.ymgme.2018.12.241 Midwest (including Missouri, Illinois, Minnesota, Wisconsin, and Nebraska) have begun screening for a variety of LSDs. While Iowa does not presently screen for LSDs on NBS, four children have come to our genetics clinic for second tier testing based on positive Fabry 226 disease Illinois NBS results. Three males from Illinois with A143T Pain and fatigue associated with generalized joint hypermobility GLA mutations were seen in our clinic for follow-up NBS testing. Two in Gaucher disease of these were siblings for familial mutation analysis and the third needed confirmatory testing for Fabry disease and subsequent Farrah R. Mahan, Laurie Bailey, Valentina Pilipenko, Carlos Prada, familial analysis for at-risk members. We now follow the three boys Cincinnati Children's Hospital Medical Center, Cincinnati, OH, United and five additional positive family members. Another male infant States was referred to us by his primary care provider for a presumptive positive Fabry NBS. The child and subsequently his mother were Gaucher Disease (GD) is a lysosomal storage disease character- found to have a c.1072_1074del mutation in the GLA gene. Mutation ized by hepatosplenomegaly, pulmonary disease, fatigue, and bone analysis is pending on additional family members. A Fabry family pain and crises. While most physicians show primary concern with identified by Missouri newborn screening on a male child relocated visceral and hematologic symptoms, patients report interest in to Iowa and is now in our care. Since moving to Iowa they have had decreasing fatigue and pain, which has an effect on quality of life. two additional children, both with Fabry disease. Two children with Generalized joint hypermobility (GJH) is characterized by joint laxity MPS 1 pseudodeficiencies, one confirmed and one pending confir- and increased joint range, and affect levels of both pain and fatigue. mation, were sent to us from Illinois for second tier testing. As the The relationship in patients with GD between chronic pain, residual number of LSD patients identified through NBS increases due to fatigue, and GJH was investigated. Participants with GD Type 1 surrounding states, we will adapt our clinic processes to incorporate fi performed a Beighton Score and ve-part questionnaire to meet the influx of patients and their families. This experience will prove diagnostic criteria for GJH. The Brief Pain Inventory (BPI) and Fatigue valuable if and when Iowa begins NBS for LSDs. Severity Scale (FSS) were used to assess pain and fatigue. Therapeu- tic goals and the Disease Severity Scoring System (DS3) evaluated treatment goals and disease involvement. 33% of our population (4/ doi:10.1016/j.ymgme.2018.12.243 12) met GJH criteria. Of those with GJH, 50% expressed moderate pain severity and 25% reported moderate pain interference, compared to 12.5% and 0% without GJH. 50% of both groups reported severe fatigue. There was a strong positive correlation between 228 reported values of pain severity, pain interference, and fatigue. Spanish multidisciplinary clinical practice guideline on Anderson- Participants met a median of 96.3% of Gaucher-related therapeutic Fabry disease in adults: A live guideline goals. In those with GJH versus those without GJH, 50% and 57.1%, respectively, had “mild disease” involvement. We did not see Ignacio Marin-Leona, Enrique Calderón-Sandubeteb, Eduardo significant changes in the levels of pain and fatigue between the Brionesc, Rafael Santamaría-Olmod, Montserrat Barcos-Martínezd, two groups of participants, but the study did reinforce those who Celedonio Márquez Infantea, Manuel Posada de la Paze, Pilar report higher levels of pain severity and interference also report high Giraldo-Castellanof, Jordi Cruz-Villalbag, Juan Gimeno-Blanesh, levels of fatigue, regardless of a GJH diagnosis. Someone with high Encarna Guillén-Navarroi, Paz Latre-Martínezj, Alberto Ortizk, aRocio levels of pain may need anticipatory management for fatigue. Our University Hospital, Seville, Spain, bRocio University Hospital. CIBERESP, patients were well-managed and met many therapeutic goals, which Seville, Spain, cDistrito Sevilla, CIBERESP, Seville, Spain, dReina Sofia may account for the levels of pain and fatigue reported. Future University Hospital, Cordoba, Spain, eInstituto de Salud Carlos III, S98 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Madrid, Spain, fCIBERER- ER, Zaragoza, Spain, gFEDER, Madrid, Spain, provided. The DMP aims to enroll approximately 35 patients with hSociedad Española Cardiología, Murcia, Spain, iArrixaca Universitity MPS VII worldwide. Diagnosis will be confirmed by GUSB enzyme Hospital, CIBERER, Murcia, Spain, jSEMFYC, Zaragoza, Spain, kFundacion assay and mutation analysis. Key assessments include demographics Jimenez Diaz, Madrid, Spain family, medical, and diagnostic history clinical presentation assessments of cognition, mobility, skeletal disease, and pulmonary The great variability in the clinical presentation of Fabry Disease function patient/caregiver-reported outcomes and health related (FD), the difficulties for its diagnosis, and the availability of several quality of life assessments and long-term vestronidase alfa safety and alternatives for its treatment, are a great challenge for the effectiveness data. Assessments occur at baseline, every 6 months for professionals who treat patients with FD, that justify the elaboration the first year (for patients ≥ 5 years) or first 2 years (for patients b 5 of an evidence based clinical practice guideline (CPG) to help years), then annually thereafter for up to 10 years. Data will be decision-making in the management of Fabry patients. METHODS: source verified based on GCP standards. For patients rolling over We applied both the guidances from NICE guideline development from clinical trials or compassionate use programs, efforts will be methods and GUIASALUD (Spanish NHS Guideline Methods). We made to analyze baseline pre-treatment data. By emphasizing site incorporated GRADE methodology (Grading of Recommendations selection, standardized assessments, monitoring with source docu- Assessment) for the evaluation of quality of scientific evidence and ment verification, and patient retention techniques, the DMP will the development and weighing of recommendations. For the final address the shortcomings of traditional registries and provide a elaboration of these recommendations, a structured consensus comprehensive dataset that offers a greater understanding of MPS process was carried out with a panel of 9 experts proposed by VII disease course. different scientific societies, research centers and patients' associations in two rounds using the Delphi-RAND method. Finally, the proposed guideline was reviewed by stakeholders (pharma indus- doi:10.1016/j.ymgme.2018.12.245 tries, scientific societies and patient associations). The guideline maintains a living process for annual updating. RESULTS: thirty-two PICO clinical questions were considered, gathered in 9 different steps 230 of the process of care. Tables were prepared for the synthesis and Quantification of 11-plex LSD enzyme activity using liquid evaluation of the quality of the evidence for each of the questions. chromatography-tandem mass spectrometry Regarding the quality of the evidence, balance between benefits and risks, values and preferences of patients, equity and use of resources Ryuichi Mashima, Mari Ohira, Torayuki Okuyama, National Center for were considered. The FD-Guideline contains 92 specific recommen- Child Health and Development, Tokyo, Japan dations for the management of FD including chapters on when to suspect the disease or how to support social and non-medical Lysosomal storage disorders are characterized by the accumula- outcomes for patients and families. CONCLUSIONS: An evidence tion of lipids, glycolipids, oligosaccharides, and mucopolysaccharides based CPG about Fabry disease is now available for both health care because of the pathogenic deficiency of lysosomal enzymes. Such workers involved in this disease and patients. diseases are rare thus, a multiplex assay for these disorders is effective for the identification of affected individuals during the doi:10.1016/j.ymgme.2018.12.244 asymptomatic period. Previous studies have demonstrated that such assays can be performed using LC-MS/MS with multiple reaction monitoring (MRM) detection. Thus, an assay procedure to quantify the activity of 11 enzymes associated with lysosomal storage 229 disorders was provided. First, a validation study was performed The MPS VII disease monitoring program (DMP) is a novel, using dried blood spot (DBS) samples with 100% and 5% enzyme longitudinal, cohort program with rigor beyond a traditional activity for quality control (QC). Under the assay condition, the registry analytical range, defined as the ratio of the peak area of the enzyme reaction products from the DBS for QC with 100% enzyme activity to Deborah Marsden, Robert Hostutler, Tricia Cimms, Ken Sansone, Qais that from the filter paper blank sample, was between 14 for GALN Abu Ali, Hank Mansbach, Ultragenyx Pharmaceutical Inc, Novato, CA, and 4561 for GLA. Based on these results, the distribution of the United States enzyme activity for the 11 LSD enzymes was further examined. Consistent with the previous data, the enzyme activity exhibited a MPS VII is an ultra-rare, autosomal recessive, heterogeneous, bell-shaped distribution with a single peak. The averaged enzyme debilitating, lysosomal disease in which patients are deficient in the activity for the healthy neonates was as follows: GLA, 3.80 ± 1.6 beta-glucuronidase (GUSB) enzyme. Vestronidase alfa (recombinant GAA, 10.6 ± 4.8 IDUA, 6.4 ± 2.3 ABG, 8.6 ± 3.1 ASM, 3.3 ± 1.1 GALC, human GUSB) is an approved enzyme replacement therapy for MPS 2.8 ± 1.3 ID2S, 16.7 ± 6.1 GALN, 1.2 ± 0.5 ARSB, 17.0 ± 8.7 NAGLU, VII. Given the rarity of MPS VII, there is a need to better understand 4.6 ± 1.5 and GUSB, 46.6 ± 19.0 μmol/h/L (mean ± SD, n = 200). In disease progression and evaluate long-term treatment with contrast, the enzyme activity in disease-affected individuals was vestronidase alfa. The MPS VII Disease Monitoring Program (DMP) lower than the minimum enzyme activity in healthy neonates. The is a multicenter program collecting long-term real world data on results demonstrate that the population of disease-affected individ- patients with MPS VII, regardless of treatment status. To overcome uals was distinguished from that of healthy individuals by the use of the common registry challenge of missing data, the DMP is designed LC-MS/MS. to provide standardized, consistent, focused outcomes data. Site selection is based on MPS expertise and location, to maximize the number of potential patients who enroll. To ensure enrollment and doi:10.1016/j.ymgme.2018.12.246 retention, appropriate site compensation and travel assistance are Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S99

231 and building on extensive preclinical data we generated previously, Long term biomarker analysis to assess cardiac involvement in we have launched a multi-center, phase I clinical trial in Canada Fabry disease using gene transfer to treat men with Fabry disease of the classical phenotype (clinicaltrials.gov#NCT02800070) and to determine the Lauren M. Mason, Renuka Limgala, Ozlem Goker-Alpan, Lysosomal & feasibility and safety of this procedure. The protocol involves Rare Disorders Research & Treatment Center, Fairfax, VA, United States mobilization of Fabry patient hematopoietic stem/progenitor cells (HSPCs), ex vivo recombinant lentivirus-mediated gene transfer of a In Fabry disease (FD), α-galactosidaseA deficiency leads to the codon-optimized human a-gal A cDNA into those cells, and infusion accumulation of globotriaosylceramide (GL-3), triggering a patho- of the transduced HSPCs into minimally ablated autologous recipi- logic cascade that causes progressive damage to multiple organs, ents. The first Fabry patient received a single dose of vector- including the heart. Disease heterogeneity and lack of meaningful transduced cells in January 2017. Since then two other Fabry patients short term clinical endpoints are the major obstacles in managing have also received the transduced cell product (with more patients patients with FD. While disease onset occurs in early childhood, the in the queue). With regards to safety - no serious adverse events diagnosis is typically made only after life-threatening cardiac, renal have occurred in any Fabry patient in our trial to date. We now or cerebrovascular complications in the fourth to fifth decade of life. report results for patient 1 at two years post-infusion of the Therefore, clinically significant markers are necessary for early transduced cells. We tracked a-gal A activity in leukocytes and detection of specific organ involvement and monitoring FD progres- plasma, vector copy number in peripheral blood and bone marrow sion. In current study, 80 patients with FD (33M:50F -80 yr, mean cells, antibody titers in plasma, and changes to enzyme substrates age: 36 yr) were seen between 2010-2018. 25 patients (13 M:12F 3- and catabolites in both plasma and urine. Results show durable 72 yr, mean age: 51 yr) were diagnosed with cardiomyopathy engraftment and persistence of vector-transduced HSPCs in this (LVPWd (cm)N1.0 and/or LVMI (g/m2) (M)N 81 and (F)N61), and 7 to Fabry patient that are functionally enabled to produce a-gal A have severe cardiac disease (LVPWdN1.5 and LVMI (M)N91 and (F)N continually. This protocol appears to be feasible and safe however, 69). Three females with mild/moderate involvement had extensive application to other patients and long-term effectiveness are still to cardiac fibrosis. Median treatment duration was 10 years among the be studied treated (19/25). All but one with severe cardiac disease, were treated for 10 years or more. Glycolipid biomarkers, plasma lyso-GL3, plasma GL3 and urine GL3, and cardiac specific markers, high-sensitive doi:10.1016/j.ymgme.2018.12.248 troponins, BNP and Galectin 3 levels were compared to organ involvement. Plasma lyso-GL3 was found to be significantly higher in the cohort with severe cardiac involvement (p=0.0086). This 233 difference was more prominent in females (p=0.0025). Troponin I Development of an algorithm to facilitate diagnosis of Gaucher fi and Troponin T were signi cantly elevated in all patients with Fabry disease related cardiomyopathy (pb0.0001, p=0.0062 respectively). BNP was elevated in patients with severe cardiomyopathy (pb0.0001). Atul Mehtaa, Oliver Rivero-Ariasb, Magy Abdelwahabc, Samantha This information is important clinically because it shows that organ Campbella, Annabel McMillana, David Kuterd, aRoyal Free Hospital, damage continues despite intervention later in FD. Disease progres- London, United Kingdom, bNuffield Department of Population Health, sion once triggered, continues even in patients under long-term Oxford, United Kingdom, cCairo University Paediatric Hospital, Cairo, therapy. Early treatment in FD is critical to prevent irreversible end Egypt, dMassachusetts General Hospital, Boston, MA, United States organ damage. Collating the opinions of 22 global experts, the Gaucher Earlier doi:10.1016/j.ymgme.2018.12.247 Diagnosis Consensus (GED-C) initiative determined which risk factors are potential indicators of Gaucher disease (GD). The aim is to create an algorithm based on these risk factors that can differentiate possible GD from other diseases in which such factors 232 occur. A feasibility study was conducted to assess whether patients FACTs Fabry gene therapy clinical trial: Two-year data in three disease groups (liver disease [LD] haematologic malignancy [HM] immune thrombocytopenic purpura [ITP] N=25 per group) Jeffrey A. Medina,b, Aneal Khanc, Ju Huangb, Dwayne Barberb,C. who had similar risk factors to GD patients (N=25) could inform Anthony Rupard, Christiane Auray-Blaise, Graeme Fraserf, Daniel H. development of this algorithm. Pearson's chi-squared test was used Fowlerg, Armand Keatingb, Michael L. Westh, Ronan Foleyf, aMedical in all comparisons. Family history of GD was only associated with GD College of Wisconsin, Milwaukee, WI, United States, bUniversity Health (n=8 overall, pb0.001). Jewish ancestry was more frequent with GD Network, Toronto, ON, Canada, cUniversity of Calgary, Calgary, AB, than LD (n=9 vs n=0 p=0.006) but differences between GD and Canada, dWestern University, London, ON, Canada, eUniversite de either HM (n=5) or ITP (n=4) were non-significant. More patients Sherbrooke, Sherbrooke, QC, Canada, fMcMaster University, Hamilton, with GD and LD had splenomegaly (n=17, both) than did patients ON, Canada, gRapa Therapeutics, Rockville, MD, United States, with HM (n=4) or ITP (n=0 overall, pb0.001), and significantly hDalhousie University, Halifax, NS, Canada more patients with GD had hepatomegaly (n=14) than did patients with LD (n=3), HM (n=1) or ITP (n=0 overall, pb0.001). Bone pain Fabry disease is due to a deficiency in alpha-galactosidase A (a- was more frequent with GD (n=17) than with LD (n=0), HM gal A) activity. Enzyme therapy is used to treat Fabry disease (n=2), or ITP (n=0 overall, pb0.001), and hyperferritinaemia was although the cost is high and biweekly infusions are required long- also more frequent with GD (n=14 versus LD [n=7], HM [n=5], ITP term. We have shown that when a-gal A is overexpressed in cells, a [n=5] overall, p=0.004). Thrombocytopenia affected more patients portion of the hydrolase is secreted and can be taken up and used with ITP (n=25) than with GD (n=14 p=0.002) and was more functionally by unmodified bystander cells. Based on this concept, severe with ITP than GD (pb0.001). Fewer patients with HM (n=9) S100 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 than GD were thrombocytopenic (p=0.038) the difference between therapy has emerged as a very promising avenue of treatment for LD (n=22) and GD was non-significant. More patients with LD various monogenic genetic disorders, allowing sustained levels of (n=19) than GD (n=9) had gammopathy (pb0.001) compared with transgene expression upon a single treatment. We hypothesised that GD, no other differences in gammopathy were significant, nor were liver-directed gene therapy could be used to ameliorate GD disease. For any differences in incidence of anaemia. Collectively, these between- this purpose, we used an adeno-associated viral vector (AAV) to drive disease differences suggest that development of a GD algorithm may the expression of human GBA gene in the liver of mice. Four weeks after be feasible. a single injection we observed a steady and dose-dependent elevation of GCase in the bloodstream of treated mice. Upon tissue analysis, noticeable uptake of human GCase was observed in organs affected in doi:10.1016/j.ymgme.2018.12.249 GD such as spleen, lung and bone marrow. Marker analysis performed in spleen tissue showed positive staining for human GCase in macrophages. Further improvement in our AAV constructs, obtained by 234 applying rational design and codon optimisation algorithms, has fi Influence of enzyme replacement therapy on the evolution of allowed us to achieve an additional ve-fold increase in GCase ocular manifestationsin a cohort of Fabry disease patients circulating in the bloodstream, resulting in increased uptake by macrophages. Overall, these data show that upon a single injection of a liver-expression-directed AAV vector it is possible to achieve elevation Langis Michauda, aFreeline Therapeutics, Stevenage, United Kingdom of GCase in the bloodstream that results in greater GCase bioavailability compared to current ERT. Therefore, this approach offers enhanced Purpose: To compare evolution of ocular manifestations of Fabry therapeutic potential for the treatment of GD. patients between those on ERT and those without ERT Methods: Observational study conducted between 2012 and 2017. Subjects were seen on an annual basis and ocular manifestations were assessed. doi:10.1016/j.ymgme.2018.12.251 Testing included: slit lamp (anterior segment), threshold visual field (FDT), corneal biomechanics (hysteresis and resistance factor), intra- ocular pressure, dilated fundus examination. Results: 28 patients completed the study after 5 years. Among them, 18 (10 hemizygotes 236 and 8 hétérozygotes) received algaside alpha or beta prior and during Two years of efficacy of oral eliglustat in treatment-naïve and the study. Another group of 10 heterozygotes, not taking ERT, were also switch patients enrolled in the International Collaborative included. A few clinical findings did not evolve: conjunctival, retinal and Gaucher Group Gaucher registry upper lid vessels tortuosity, and cornea verticilata. Many others increased in prevalence and/or severity: micro-aneurysms, corneal Pramod K. Mistrya, Manisha Balwanib, Joel Charrowc, Priya Kishnanid, haze, anterior and posterior sub-capsular cataracts. Hemizygotes on ERT Claus Niederaue, Monica R. McClainf, aYale University School of showed increased signs at baseline vs others. Heterozygotes on ERT Medicine, New Haven, CT, United States, bIcahn School of Medicine at showed a marked evolution of ocular manifestations over time, Mount Sinai, New York, NY, United States, cNorthwestern University especially for the presence of microaneurysms. All patients under ERT Feinberg School of Medicine, Ann and Robert H. Lurie Children's Hospital demonstrated evolution over time. Conclusion: ERT does not influence of Chicago, Chicago, IL, United States, dDuke University, Durham, NC, the prevalence and the progression of ocular manifestations over time, United States, eKatholische Kliniken Oberhausen, Oberhausen, Germany, compared with subjects not treated. Heterozygotes on ERT progressed fSanofi Genzyme, Cambridge, MA, United States more and, after 5 years, were at the same level than hemizygotes on ERT for a majority of clinical findings. Eliglustat (Cerdelga, Sanofi Genzyme) is a first-line oral substrate reduction therapy for adults with Gaucher disease type 1 (GD1) with extensive, intermediate, or poor CYP2D6-metabolizer phenotypes N doi:10.1016/j.ymgme.2018.12.250 ( 90% of patients). Clinical trials have demonstrated long-term clinical improvement in all major disease manifestations in treatment-naïve patients and long-term stability in patients switching from enzyme-replacement therapy (ERT). We report real-world data 235 from treatment-naïve and ERT-switch patients enrolled in the Liver directed AAV gene therapy to treat Gaucher disease International Collaborative Gaucher Group Gaucher (ICGG) Registry (NCT00358943/Sanofi Genzyme) who were treated with eliglustat Carlos J. Mirandaa, Jenny McIntoshb, Azadeh Kiaa, Miriam Canavesea, for ≥1 year. Of 400 eliglustat-treated patients in the Registry as of Jonathan Foleya, Doyoung Leeb, Paniz Hosseinia, Jey Jeyakumara, Rose July 2018, 238 had baseline and 2-year data (±1 year) for ≥1 key Sheridana, Romuald Corbaua, Amit C. Nathwania, aFreeline Therapeu- disease parameter: 15 treatment-naïve (0 splenectomized) and 223 tics, Stevenage, United Kingdom, bUniversity College London, London, switch (43 splenectomized). Overall, 213 were from the United United Kingdom States, the first country to approve eliglustat. In treatment-naïve patients, mean hemoglobin improved from 12.5 to 13.8 g/dL Gaucher disease (GD) is characterised by the deposition of (p=0.004 n=14) mean platelet count improved from 113 to 158 glucocerebroside in cells of the macrophage-monocyte system caused x109/L (p=0.0002 n=13) mean spleen volume decreased from 9.4 by impaired production of the enzyme beta-glucocerebrosidase to 4.2 multiples of normal (MN) (p=0.01, n=5), and mean liver (GCase). Over the past 20 years, enzymatic replacement therapy (ERT) volume was unchanged: 1.2 vs 1.1 MN (non-significant, n=5). In was developed as the standard of care for GD. However, since it requires non‑splenectomized-switch patients, mean hemoglobin remained continuous treatment (every other week infusions), it results in a high normal: 14.3 vs 14.1 g/dL (p=0.01, n=169) mean platelet count cumulative cost for this type of therapy, and more importantly in a remained normal: 181 vs 184 x109/L (non-significant, n=169) mean significant patient treatment burden. Over the same period, gene spleen volume decreased from 2.9 to 2.5 MN (p=0.002, n=63), and Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S101 mean liver volume remained stable at 0.9 MN (non-significant, Madrid, Spain, bHospital Universitario Ramón y Cajal, IRYCIS, Madrid, n=63). In splenectomized-switch patients, mean hemoglobin Spain remained stable: 13.5 to 13.4 g/dL (non-significant, n=36) mean platelet count increased from 295 to 319 x109/L (p=0.05, n=34) A 40-year-old man with a history of mild idiopathic or immune mean liver volume remained stable: 0.9 to 1 MN (non-significant, thrombocytopenia since childhood was referred for consultation due n=15). Mean chitotriosidase levels decreased in all groups, with to splenomegalia. He had mild normocytic anemia and thrombocy- significant reductions in treatment-naïve patients (1325 to 308 topenia, a markedly enlarged spleen (22 cm) with several focal nmol/mL/hr, p=0.03, n=6) and non-splenectomized-switch patients lesions without lymphadenopathy, and typical Gaucher cells were (838 to 725 nmol/mL/hr p=0.02, n=86). These real-world results observed in bone marrow smears. Analysis of genomic DNA by are consistent with those reported in eliglustat clinical trials. means of the Next-Generation Sequencing (NGS)-based Sple- nomegalia and Thrombocytopenia resequencing gene panel (STP version 1, 74 genes) and segregation analyses indicated that the doi:10.1016/j.ymgme.2018.12.252 patient was compound heterozygous for the known pathogenic mutations p.(Asn409Ser) (formerly N370S) and p.Leu483Pro (formerly L444P) at the GBA gene. Determination of beta-glucosidase activity indicated that only residual levels were present, confirming a 237 diagnostic of Gaucher disease. However, further analysis of panel Generation of iPS cells derived from skin fibroblasts of patients data revealed that the patient was also hemizygous for a pathogenic with Fabry disease using RNA-reprogramming mutation, p.(Arg24*), at the G6PD gene on chromosome X. Quanti- fication of glucose-6-phosphate dehydrogenase in blood confirmed Takashi Miyajimaa, Hiroko Yanagisawaa, Mohammad Arif Hossaina, reduced activity, well below normal levels. We did not find any Chen Wua, Takeo Iwamotob, Yoshikatsu Etoa, aAdvanced Clinical evidence of previous episodes of acute anemia, even after exposure Research Center, Kawasaki, Japan, bTokyo Jikei University School of to quinine or Vicia faba. Our results indicate that, by simultaneously Medicine, Tokyo, Japan interrogating many genes involved in lysosomal storage diseases (LSDs) and their differential diagnoses, NGS panel sequencing helps Fabry disease is a lysosomal metabolic disorder with X-linked identifying additional mutations that may modulate the phenotypic inheritance caused by a deficiency of alpha-galactosidase which leads presentation and therefore suggest different therapeutic approaches. to accumulation of globotriaosylceramide (Gb3) in various organs. Funding: This work was supported by Sanofi Genzyme. Clinical features of Fabry disease are renal, cardiac and CNS involvement. However, precise cellular pathogenesis in Fabry disease is still unknown. The iPS technology may be useful for the understanding of doi:10.1016/j.ymgme.2018.12.254 pathogenetic mechanism of lysosomal storage disease including Fabry disease. The generation of iPS cells generally have been carried out by the use of Sendai virus vector or episomal vector, but these technologies need the environmental attention and equipment for 239 handling viruses. Therefore, we focused on RNA-reprogramming CRISPR/Cas in iPSCs from sphingolipidoses patients procedure. Skin fibroblasts from control and Fabry disease were transfected with reprogramming factors (Oct4, Sox2, Klf4, cMyc, Luciana Moreira, Ana Joana Duarte, Diogo Ribeiro, Olga Amaral, Nanog, Lin28), interferon factors (B18, E3, K3), RNA and microRNA National Institute of Health R.Jorge & CECA, ICBAS-University of Porto, and formed iPS colonies were obtained after several cell passages and Porto, Portugal surveyed for morphological and biochemical studies. Using this method, no virus is used and feeder cells are not required, and also Clustered Regularly Interspaced Short Palindromic Repeats iPS cells can be efficiently obtained in a short period of time. Using this (CRISPR) were found as an immune adaptive mechanism in bacteria method, the iPS cells were successfully established from healthy and quickly were applied to various fields as a promising tool for gene human skin fibroblasts and skin fibroblasts derived from three Fabry editing. Lysosomal storage diseases (LSDs) are a group of metabolic male sibling patients in same family. Expression of surface markers disorders caused by defects in lysosomal proteins leading to accumu- and undifferentiated markers in these iPS cells showed positive, lation of undigested macromolecules within the cells. The lack of good demonstrated by immunostaining and RT-PCR. Generated iPS cells in vitro models hinders research of the pathophysiologic mechanisms from these Fabry patients were confirmed as iPS cells by various and the development of new therapies. Induced pluripotent stem cells pluripotency markers. Our RNA reprograming procedure to generate (iPSCs) are patient-specific and can be differentiated in any cell type. iPS cells could be more widely used for understanding of pathogenetic The advantage of iPSCs is to enable targeted studies in cells with the mechanism of various lysosomal diseases including Fabry disease. patient’s own background leading to more straightforward results than other models. Combining CRISPR and iPSCs is, therefore, a promising strategy. We aim to use CRISPR/Cas-mediated gene editing doi:10.1016/j.ymgme.2018.12.253 to provide more specific cellular models of disease, to correct causal mutations in LSDs and to create mutants for functional studies. In this work, we generated and characterized iPSCs from human fibroblasts obtained from Gaucher and Fabry patients (through Gaslini Institute) 238 and will edit them with a CRISP/Cas9 approach. Because both gene Unexpected genetic findings in a Gaucher disease patient analysed editing and iPSCs generation require manipulating the cell’s genome, by NGS-based panel sequencing we envisage multiple check points along the workflow. It will be useful to compare the “native” mutated cells with the corrected cells that Marta Moradoa, Gloria Muñozb, Miguel Pirisb, Alejandro Sanzb, Jesus modulate the “disease in a dish”. Gene editing is still recent and the Villarrubiab, Francisco J. del Castillob, aHospital Universitario La Paz, methods require improvement, namely increasing transfection rates S102 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 and mutagenesis efficiency with less off-targets. Nevertheless, CRISPR/ fact, current dynamics have led results obtained by several research Cas is a promising alternative to other therapies, and every result groups to be of diagnostic value, particularly for those working with contributes to the enhancement of this technology, broadening the low frequency diseases. This situation has allowed research labora- validation of CRISPR application and making it an accessible option. tories to incorporate concepts such as quality, quality assurance, certification, and accreditation, among other quality-related concepts. Therefore, promotion and implementation of quality manage- doi:10.1016/j.ymgme.2018.12.255 ment systems (QMS) has become as opportune as necessary. The international quality standards under which a QMS can be implemented are based on ISO standards. One of them is ISO/IEC17025, which establishes the requirements that must be met by testing and 240 calibration laboratories. This standard is based on ISO 9000 rules but Are we missing complex rearrangements by next generation incorporates technical requirements essential to achieve accredita- diagnostic approaches: A case report of a complex rearrangement tion. The ISO/IEC17025 standard is applied with the objective of in MPS II demonstrating that testing and calibration laboratories are technically competent and that their results are accurate. For the a b c Maria L. Moreira , Francyne Kubaski , Rodolfo Bareiro , Catarina accreditation process (NTC-ISO/IEC17025:2005-standard) our labo- d b a Pereira , Roberto Giugliani , Hospital Santa Casa da Misericordia, ratory selected 6 assays of lysosomal enzymatic activity performed in b Campo Grande, Brazil, UFRGS/HCPA/INAGEMP, Porto Alegre, Brazil, DBS. This process has been carried out in 2 stages: 1. Implementa- c d Instituto da Mão MS Hand, Campo Grande, Brazil, Centogene, Rostock, tion: a.Diagnosis: At initial evaluating laboratory status, it was found Germany compliance in 20% of the standard b.Planning: Activities were determine to fill existing gaps c.Design: Strategies to comply with fi Mucopolysaccharidosis type II (MPS II) is caused by de ciency of the requirements of the standard were defined: process map and Iduronate-2-sulfatase (IDS) that leads to accumulation of undegraded 100% of documentary support d.Implementation: Disclosure and glycosaminoglycans. So far, over 630 mutations have been described for application of the policy/objectives/documentary support were the IDS gene in the Human Gene mutation database and only 2% of these carried out e.Evaluation: An internal audit concluded that the fi are classi ed as complex rearrangements. Here, we present the effectiveness of the implementation was 80%. After solving the molecular diagnosis odyssey of an MPS II patient. Non-consanguineos nonconformities, we proceeded to stage 2. Application: The applica- fi parents had 3 children, the rst daughter died with 2.6 years with no tion was made to the ONAC. a.Documentary evaluation: ONAC’s fi con rmed diagnosis but dysmorphic features, the second was a evaluation team declared 95% compliance of documentary support. stillbirth with facial dysmorphic features at birth and the third son b.On-site evaluation: the conclusion was that efficiency of the QMS (proband) died with 6.1 years. He started walking with 2 years of age accomplished 96%. and had severly impaired speech development limited to few words. He started developing seizures at 4 years of age. Clinical examination revealed: short stature, coarse facial features, neurological impairment, doi:10.1016/j.ymgme.2018.12.257 hepatosplenomegaly, proximal muscle weakness, joint rigidity, cardi- opathy, dysostosis multiplex, macular degeneration, retinopathy, and hearing loss. The biochemical diagnosis was confirmed at 5.11 years by fluorimetry in dried blood spot. Next generation sequencing of the IDS 242 α by Illumina platform and MLPA analyses of the IDS gene did not reveal CRISPR/Cas9 mediated insertion of -L-iduronidase (IDUA) and any clinically significant variant. Sanger sequencing followed by gel anti-PE receptor in B-lymphocytes for selective activation into electrophoresis revealed a complex rearrangement within intron 3 and long-lived plasma cells for sustainable IDUA expression intron 7 of the IDS gene and the IDS2 pseudogene. This rearrangement have already been described in the literature. The patient died with 6.1 Branden Moriarity, University of Minnesota, Minneapolis, MN, United years of age due to pneumonia. In conclusion, we should be carefull States when using solely next generation sequencing approaches for the molecular diagnosis of MPS II patients or MLPA taking into consider- Mucopolysaccharidosis type I is a rare inherited lysosomal ation the limitations of these methodologies for the identification of storage disorder caused by mutations in the IDUA gene. IDUA fi complex rearrangements. The delay in diagnosis of this patient de ciency results in the accumulation of glycosaminoglycans contributed to his premature death due to the lack of treatment that (GAGs), which ranges in clinical severity from Scheie syndrome could have helped to delay disease progression if started early. (mild form) to Hurler syndrome (Severe form). At present enzyme replacement therapy (ERT) and bone marrow transplantation (BMT) are the standard of care for this disease1 . In order to develop new doi:10.1016/j.ymgme.2018.12.256 approaches to treating this disease we exploit the unique characteristic that B cells differentiate into antibody producing long-lived plasma cell. We use CRISPR/Cas9 2 mediated homologous directed repair (HDR) for site-specific insertion to co-express a therapeutic 241 gene (IDUA) and phycoerythrin (PE). CRISPR/Cas9 system will create Accreditation: A challenge for a research laboratory a double strand break at the endogenous B-cell receptor (BCR) of the B cell genome where homologous directed repair will facilitate the Patricia Moreno, Alfredo Uribe, Universidad de los Andes, Bogota, insertion of a therapeutic cassette under the expression of a Colombia synthetic promoter. A successfully engineered B cell will produce both IDUA and the PE receptor. A large quantity of IDUA will be A research laboratory focuses its efforts on the development of secreted into the extracellular environment and the PE receptor will experiments bringing benefit to the entire scientific community. In be transported to the cell membrane, which will allow us to Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S103 selectively activate the engineered B cells into long-lived plasma Benioff Children’s Hospital Oakland, Oakland, CA, United States, cells that produce a large quantity of IDUA enzyme. Transplantation dHospital Universitario Austral, Buenos Aires, Argentina, eSt Mary’s of this engineered plasma cells into an IDUA deficient mouse model Hospital, Manchester, United Kingdom, fInfant Jesus Children’s Hospital, will result in sustainable enzyme secretion into the circulation for Madrid, Spain, gNational Institute of Pediatrics, Mexico City, Mexico, systemic cross-correction of IDUA deficiency. This novel strategy will hShire, Lexington, MA, United States generate long-lived plasma cells that provide sustainable IDUA enzyme for a potential cell-based enzymotherapy for MPS I disorder. Two-thirds of patients with mucopolysaccharidosis type II (MPS II This engineering strategy has many potential applications for Hunter syndrome), a rare lysosomal disease characterized by treating various enzymopathies. iduronate-2-sulfatase deficiency, have cognitive impairment. This observational, prospective, longitudinal study (NCT01822184) assessed cognitive status and adaptive behavior in patients aged 2 doi:10.1016/j.ymgme.2018.12.258 −b18 years with MPS II patients with a General Conceptual Ability (GCA) score b55 measured by the Differential Abilities Scales-II (DAS-II) were excluded. Patients received IV idursulfase as standard- of-care throughout. Cognitive ability (DAS-II) and adaptive behavior 243 (Vineland Adaptive Behavior Scales [VABS-II]) assessments were Evaluation of PPS treatment in osteoclast-osteoblast imbalance performed at baseline, and 3-month intervals for up to 24 months. Of using in vitro models of Gaucher disease 55 enrolled patients (mean [SD] age: 5.60 [3.316] years), 32 discontinued (25 to enroll in a phase II/III-treatment study). Mean Juan M. Muccia, Constanza Bondara, Andrea Crivaroa, Maximiliano DAS-II GCA (n=44) and VABS-II Adaptive Behavior Composite (ABC) Ormazabala, Edward Schuchmanb, Calogera Simonarob, Paula scores (n=53) at baseline were 78.4 (19.11) and 83.7 (14.22), Rozenfelda, aInstitute of Immunological and Physiopathological Studies, respectively. Mean changes from baseline at months 12 and 24 were La Plata, Argentina, bIchan School of Medicine at Mount Sinai, New York, −0.9 (9.39) and −3.8 (12.71), respectively, for DAS-II GCA score NY, United States (n=27/n=20) and −2.4 (7.64) and −2.0 (8.07) for VABS-II ABC score (n=29/n=21). Changes in GCA scores over time varied widely Gaucher disease (GD) is caused by mutations in the gene in individual patients, with some experiencing rapid declines in encoding for the lysosomal enzyme glucocerebrosidase. Bone cognitive ability. Differences in the least-squares mean (LSM) alterations are the most disabling condition in Type I GD (GD1) changes from baseline in DAS-II GCA scores between patients with patients and remain a chronic issue in spite of enzyme replacement baseline GCA scores of ≤70 (low cognitive function n=18) and N70 therapy. Mechanisms leading to bone damage are poorly understood, (n=26) were −2.4 (p=0.5657) at month 12 and −7.4 (p=0.1461) but previous reports suggest that both osteoblasts and osteoclasts at month 24. Similarly, for VABS-II ABC scores, differences in LSM are involved. In the last years, several works have been published in changes from baseline between the GCA subgroups (≤70/N70) were relation to cartilage and bone pathology in mucopolysaccharidoses −1.9 (p=0.5545) at month 12 and 0.3 (p=0.9484) at month 24. where inflammation was shown to be a key factor in pathogenesis. A Overall, cognitive ability and adaptive behavior in the pediatric MPS compound called pentosan polysulphate (PPS) was proposed as II population who completed this study remained relatively stable therapy in MPS models and showed marked improvements in over 24 months. However, some patients experienced a rapid decline clinical behavior. In the present study we analyzed the effect of PPS in cognitive ability, while others retained stable, but impaired, treatment on osteoclast differentiation and osteoblast activity using cognitive function. Shire funded this study and medical writing in vitro models of GD. PPS treatment reduced osteoclast differenti- support. ation from GD patient PBMCs. Supernatants from two in vitro models of GD osteoblasts (MC3T3) and osteocytes (MLO-Y4) with CBE were obtained in the presence or absence of PPS. When osteoclast doi:10.1016/j.ymgme.2018.12.260 precursors were cultured in the presence of these supernatants, the PPS supernatants induced less osteoclast differentiation. Osteoblast activity was also improved by PPS treatment in these in vitro models. In addition, mineral deposition was increased in PPS treated 245 osteoblasts. Our results suggest that PPS should be considered as an Evaluation of the long-term treatment effects of idursulfase using alternative treatment for bone implications in GD. statistical modelling: Data from the Hunter Outcome Survey (HOS) doi:10.1016/j.ymgme.2018.12.259 Joseph Muenzera, Barbara K. Burtonb, Paul Harmatzc, Jaco Bothad, Christoph Kampmanne, aUniversity of North Carolina Chapel Hill, Chapel Hill, NC, United States, bAnn & Robert H Lurie Children’s Hospital c ’ 244 of Chicago, Chicago, IL, United States, UCSF Benioff Children s Hospital d Neurodevelopmental status and adaptive behavior of pediatric Oakland, Oakland, CA, United States, Shire, Zug, Switzerland, e patients with Hunter syndrome: A longitudinal observational Johannes-Gutenberg University, Mainz, Germany study Treatment for mucopolysaccharidosis type II (MPS II Hunter syndrome) is available in the form of intravenous enzyme replace- Joseph Muenzera, Barbara K. Burtonb, Paul Harmatzc, Hernan ment therapy (ERT) with idursulfase (Shire, Lexington, MA, USA). Amartinod, Simon A. Jonese, Luis González Gutiérrez-Solanaf, Matilde This analysis used statistical modelling to evaluate the long-term Ruiz-Garciag, Yuna Wuh, David Alexanderianh, aUniversity of North treatment effects of idursulfase on selected clinical parameters based Carolina Chapel Hill, Chapel Hill, NC, United States, bAnn & Robert H. on data from HOS, a global, observational registry (Shire, Lexington, Lurie Children’s Hospital of Chicago, Chicago, IL, United States, cUCSF MA, USA). Mixed modelling was used to analyse data from male S104 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 patients followed prospectively in HOS who had received idursulfase and one started idursulfase after BMT. Median time between for 5-8 years and information available for two or more timepoints, idursulfase start and BMT was 3.5 (−173.5, 24.3) months (n=7). of which one was pre-ERT. Data were excluded from patients with Cognitive impairment at any time was reported for 21 patients only pre-ERT information available, who had received a bone (58.3% n=36). In total, 13/36 patients died median age at death was marrow transplant or had enrolled in an idursulfase clinical trial. 8.8 (3.4, 17.6) years. The most common cause of death was graft Age at and time since ERT start were included as covariates and versus host disease/transplant complications (n=4) followed by results were modelled for up to 8 years of treatment. For the respiratory failure (n=3). These long-term data from HOS provide prediction of percent-predicted forced vital capacity (FVC), forced valuable information on patients with MPS II undergoing BMT. Shire expiratory volume in 1 second (FEV1) and the 6-minute walk test sponsors HOS and funds medical writing support. (6MWT), only data from patients aged ≥5 years and without cognitive impairment were used. A sensitivity analysis assessed doi:10.1016/j.ymgme.2018.12.262 robustness of the model using information from patients with data for five or more timepoints. The main model indicated a decrease over time in urinary glycosaminoglycan levels and palpable liver size. Left ventricular mass index was stable for up to 8 years of treatment 247 while there was a slight decrease in percent-predicted FVC and FEV1 CHAMPIONS: A phase 1/2 clinical trial with dose escalation of SB- and a gradual increase in distance walked in the 6MWT. Similar 913 ZFN-mediated in vivo human genome editing for treatment results were observed in the sensitivity analysis, indicating that this of MPS II (Hunter syndrome) model provides reliable estimates. The nature of this analysis means that these findings are descriptive only. However, our results support Joseph Muenzera, Carlos E. Pradab, Barbara Burtonc, Heather A. Laud, those of previous studies and indicate that idursulfase has a positive Can Ficicioglue, Cheryl Wong Po Foof, Sagar A. Vaidyaf, Chester B. effect on somatic manifestations of MPS II. Shire sponsors HOS and Whitleyg, Paul Harmatzh, aUniversity of North Carolina, Chapel Hill, funds medical writing support. Chapel Hill, NC, United States, bCincinnati Children’s Hospital Medical Center, Cinncinati, OH, United States, cLurie Children’s Hospital, Chicago, IL, United States, dNYU Langone Medical Center, New York, NY, United doi:10.1016/j.ymgme.2018.12.261 States, eChildren’s Hospital of Philadelphia, Philadelphia, PA, United States, fSangamo Therapeutics, Richmond, CA, United States, gUniversity of Minnesota, Minneapolis, MN, United States, hUCSF Benioff Children’s 246 Hospital Oakland, Oakland, CA, United States. Characteristics of patients with mucopolysaccharidosis type II Mucopolysaccharidosis type II (MPS II or Hunter syndrome) is a who have received a bone marrow transplant: Data from the rare, progressive, X-linked genetic disorder that is caused by Hunter Outcome Survey mutations in the human IDS gene and is characterized by deficient activity of the lysosomal enzyme iduronate-2-sulfatase (IDS). Joseph Muenzera, Barbara K. Burtonb, Christoph Kampmannc, Jaco Without functional IDS activity, the glycosaminoglycans (GAG), Bothad, Simon A. Jonese, aUniversity of North Carolina Chapel Hill, dermatan sulfate and heparan sulfate, accumulate in the body and Chapel Hill, NC, United States, bAnn & Robert H. Lurie Children's Hospital lead to widespread tissue and organ damage. Patients with MPS II of Chicago, Chicago, IL, United States, cJohannes-Gutenberg University, develop progressive respiratory and cardiac disease, skeletal abnor- Mainz, Germany, dShire, Zug, Switzerland, eSt Mary's Hospital, Man- malities and in the severe form, cognitive decline and early death, chester University NHS Foundation Trust, University of Manchester, despite treatment with enzyme replacement therapy. SB-913 is a Manchester, United Kingdom new type of investigational treatment for MPS II. SB-913 uses zinc finger nuclease (ZFN)-mediated in vivo genome editing to insert a Mucopolysaccharidosis type II (MPS II Hunter syndrome) is a normal copy of the IDS transgene into liver cells, delivered via AAV2/ rare, life-limiting, X-linked lysosomal storage disease. The Hunter 6 vectors. The precision and specificity of SB-913 allows for Outcome Survey (HOS) is a Shire-sponsored, global, observational integration at a targeted genomic location and is intended to reduce registry initiated in 2005 that collects real-world data on the natural GAG accumulation with lifelong continuous endogenous production history of MPS II and long-term treatment with enzyme replacement of IDS. CHAMPIONS is an ongoing Phase 1/2 clinical trial to therapy (ERT) with idursulfase. Patients receiving other forms of determine if dose escalation of SB-913 is safe and tolerable in pharmacological ERT are excluded from HOS but individuals who patients with MPS II, and is the first trial to attempt in vivo genome have received a bone marrow transplant (BMT) may be enrolled. editing in humans. CHAMPIONS is a multicenter, open-label, dose This analysis examined the characteristics of 36 male patients in HOS escalation study with one-time peripheral intravenous infusion of from Europe and North America who had received a BMT (March SB-913, followed by three years of observation. Six adult subjects 2018 data). In total, 22 patients (61.1%) were European and 14 with attenuated MPS II have received SB-913, with two subjects in (38.9%) were North American. Twenty-seven patients (75%) were each of three dose cohorts (5e12 vg/kg, 1e13 vg/kg, and 5e13 vg/kg). followed prospectively in HOS 9 (25%) were enrolled after their The infusions were generally well-tolerated and no serious adverse death (followed retrospectively). Median (10th percentile, 90th events related to the study drug were reported at any dose with up percentile) ages at symptom onset, diagnosis and last visit were 0.8 to 10 months of exposure. No persistent transaminitis was observed. (0.1, 3.0) years (n=26), 2.5 (0.8, 5.0) years (n=35) and 15.2 (4.9, Additional analysis of the trial data is ongoing and will be presented 28.4) years (n=27), respectively patients were aged 2.6 (0.7, 4.5) as available. These results support further development of SB-913 for years at BMT (n=10). Fourteen patients (38.9%) had received at least the treatment of MPS II. one idursulfase infusion, 16 (44.4%) had not received idursulfase idursulfase treatment status was unknown for six patients (16.7%). Of seven patients with data available on the relative timing of doi:10.1016/j.ymgme.2018.12.263 idursulfase treatment and BMT, six started idursulfase before BMT Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S105

248 Methods: We applied mathematical approaches to morphomet- Lentiviral-modified T Rapa cells as 'micropharmacies' for lyso- ric data of 91 glomeruli from 40 untreated males with FD. somal diseases We modeled the relationship between GL-3 inclusion volume density in podocyte [Vv(Inc/PC)] profiles with visible inclusions Murtaza S. Nagreea,b, Tania C. Felizardoc, Ju Huangd, Daniel H. and the probability of obtaining random profiles without inclu- Fowlerc, Jeffrey A. Medina,b,d, aUniversity of Toronto, Toronto, ON, sions () and derived an equation for estimating Vv(Inc/PC) in Canada, bMedical College of Wisconsin, Milwaukee, WI, United States, females independent of mosaicism. The model was applied to cRapa Therapeutics, Rockville, MD, United States, dUniversity Health biopsies from 10 females and 8 age-matched males with untreated Network, Toronto, ON, Canada. FD. Mean podocyte volume (VPC) and total GL-3 inclusions volume [V(Inc/PC)] were estimated in affected podocytes in Transgene-augmented cell therapies are a promising strategy for females. the treatment of amenable Lysosomal Storage Disorderls (LSDs). Results: A 4th degree polynomial equation best described the Most overexpressed lysosomal enzymes can be secreted from relationship between Vv(Inc/PC) and (R2=0.94) and passed k-fold vector-modified cells and taken up by uncorrected cells, a process cross-validation. VPC and V(Inc/PC) (r=0.87, p=0.002) were both termed “cross-correction”. Our lab is currently evaluating a increased with age in affected podocytes in females. Foot process treatment for Fabry disease using transduced CD34+ hematopoietic width correlated with VPC (r=0.89, p=0.02) and V(Inc/PC) stem and progenitor cells (HSPCs) in a phase I trial (NCT02800070). (r=0.87, p=0.02) . Comparisons with male biopsies showed no These HSPCs are modified ex vivo with a recombinant lentivirus to significant sex difference in Vv(Inc/PC) (p=0.75) or in V(Inc/PC) engineer overexpression of α-galactosidase A (α-gal A). Transduced (p=0.45). HSPCs are reinfused into the Fabry patient and differentiate into Conclusion: We developed and validated a novel method for leukocytes that secrete α-gal A long-term. However, HSPC collec- measuring GL-3 in Fabry podocytes independent of mosaicism in tion requires both a hospital stay and a drug regimen for cell females. GL-3 accumulation and injury in Fabry podocytes in mobilization. HSPC engraftment also requires a chemotherapy- females is progressive with age. Importantly, GL-3 content in based conditioning protocol (even for 'mild' myeloablation - as in Fabry-affected podocytes in females is similar to that in males, our case). An alternative hematopoietic cell population that can be consistent with absence of cross-correction between affected and readily obtained without mobilization and engrafted with less non-affected cells. This method will be of great value for conditioning may therefore be more desirable for the systemic measuring podocyte GL-3 as an outcome variable in studies of delivery of therapeutic cargo - such as lysosomal hydrolases. T cells females with FD. can be obtained from peripheral blood without mobilization and expanded exponentially in culture. T cells can also secrete high levels of protein. When treated with rapamycin in defined ex vivo doi:10.1016/j.ymgme.2018.12.265 conditions, ‘T Rapa’ cells acquire an anti-apoptotic and pro- engraftment phenotype. These T Rapa cells engraft with minimal conditioning. Autologous transduced CD4+ T Rapa cells have 250 “ potential to deliver enzyme systemically as circulating micro- Saccadic eye movements and its use as clinical endpoints in ” pharmacies . Our data shows that T Rapa cells manufactured from lysosomal disorders: A literature review Fabry patients can be stably transduced and secrete α-gal A in vitro. Secreted α-gal A can be taken up by fibroblasts. α-Gal A activity Luba Nalysnyka, Alaa Hameda, Camille Rochmanna, Liz Molenkampb, was also detected in the plasma of xenografted NOD/SCID/Fabry Tanya Fischera, Kacey Rawsonb, aSanofi Genzyme, Cambridge, MA, mice substrate was reduced. We are concomitantly testing the United States, bEvidera, Waltham, MA, United States potential for T Rapa cells to secrete enzymes for treating other LSDs, such as Gaucher disease, Farber disease, and Pompe disease. Saccadic eye movements (SEMs) are rapid eye movements that aid vision through the redirection of sight from target to target. doi:10.1016/j.ymgme.2018.12.264 Normal SEM is distinguished by a consistent relationship between peak velocity, amplitude, and duration (latency and termination). There are several disorders of the saccades that affect these parameters and are apparent in Gaucher disease type 3 (GD3) and 249 monogangliosidosis 2 (GM2). A targeted literature review was A novel method for quantification of globotriaocylceramide (GL- conducted to describe SEM disorders and their manifestations in 3) inclusions in affected podocytes in females with Fabry disease GM2 and GD3 and to assess its use as an endpoint in clinical trials. In shows progressive accumulation of GL-3 in podocytes with age both diseases, SEM disorders may include horizontal saccadic and no cross-correction between affected and non-affected initiation failure and slowed horizontal and downward saccades. podocytes SEMs play a critical role in coordination and the ability to perform activities of daily living (reading, interpretation, navigation of the Behzad Najafiana, Alexey Sokolovskiya, Michael Mauerb, aUniversity of visual environment, and social interactions). In GD3, SEM disorders Washington, Seattle, WA, United States, bUniversity of Minnesota, were associated with significant impairment in hand-eye coordina- Minneapolis, MN, United States tion and manual dexterity. In clinical trials of lysosomal storage disorders, SEMs have been assessed using change from baseline in Background: While females can suffer serious FD complications, either horizontal or vertical SEM velocities (HSEM-α and VSEM-α) most studies are limited to males to avoid confounding the results by and horizontal and vertical SEM amplitudes (HSEM-β and VSEM-β) mosaicism phenomenon. We developed a method for quantifying as a primary or secondary endpoint. The process for assessing SEM GL-3 content of Fabry-affected podocytes in females independent of and the thresholds for improvement and deterioration varied across mosaicism. the trials. A clear definition for a clinically meaningful change was S106 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 not always provided making comparisons across studies difficult. Introduction: Hurler syndrome (MPS IH), the most severe form SEM is not routinely used in clinical practice and changes cannot be of mucopolysaccharidosis (MPS) type I, is fatal within the first directly translated into clinical benefits. These aspects were decade of life if untreated. Hematopoietic stem cell transplantation highlighted by regulatory agencies in regulatory assessments of (HSCT) is the standard of care for MPS IH to treat central nervous miglustat for the treatment of progressive neurological manifesta- system disease. As new treatments are being developed and tions in Niemann-Pick Type C. Guidelines around the usefulness and newborn screening identifies patients earlier, it is critical to define acceptability of SEM endpoints in trials were not identified. More reliable and sensitive measures reflecting MPS I disease prior to HSCT data is needed to establish a clear link between early sign of change in infants. in SEMs and clinical benefits. Supported by Sanofi Methods: MRI scans including T1, T2-weighted images and diffusion tensor imaging (DTI) were acquired on clinical 3T MR doi:10.1016/j.ymgme.2018.12.266 system in 12 pre-HSCT MPSIH infants (age b 2 years of age). Brain and ventricular volumes, T1/T2 ratios, and DTI indices were evaluated. Results: Significant increase in fractional anisotropy (FA) 251 between scans prior/after-HSCT and limited FA change in the Quantitative brain MRI in patients with alpha-mannosidosis: post-HSCT follow-up scans were seen in most patients. Two Study from 3 centers patients exhibited no or minimal FA increase between pre-/after- HSCT scans but steeper FA increase on post-HSCT scans. RD Igor Nestrasila, Amy Paulsona, Jitka Jireckovab, David Nascenea, and T1/T2 ratio exhibited similar trends. Total brain volumes Manuela Vaneckovab, Katarina Jurickovac, Anna Hlavatac, Troy Lunda, increased in most subjects after the HSCT however, ventricular Paul Orcharda, Petr Dusekb, Martin Magnerb, aUniversity of Minnesota, volumes exhibited both increases and decreases within the Minneapolis, MN, United States, bFirst Faculty of Medicine, Charles population. University and General University Hospital, Prague, Czech Republic, Conclusions: The finding is suggestive of an immediate cComenius University Medical School, National Institute of Child response measured by DTI and T1/T2 ratio (within months) to Diseases, Bratislava, Slovakia the HSCT procedure in most subjects. In some subjects a delayed response (after a year) to the HSCT procedure was detected. The Introduction: Alpha-mannosidosis is a rare lysosomal storage results confirmed an increase in the white matter myelination in disease with multisystemic abnormalities and a wide clinical patients who received HSCT. Determinants of the response type to variability. The primary clinical brain MRI findings are described in the HSCT need to be identified. Similar trends are observed in the case report studies. Quantitative MRI outcomes using robust normal brain development thus, the exact trajectory and rate of automated methods as a prerequisite for clinical trials are not FA/RD and T1/T2 ratio changes need to be compared with healthy available. age- and gender-matched individuals. (Funded by NIH Methods: High-resolution T1-weighted MRI scans of 6 subjects U54NS065768, Lysosomal Disease Network Orphan Disease Center (age: range 6-27 mean ± SD 14.5±4.9 years of age) with alpha- at the University of Pennsylvania MPSI-16-003-02 Genzyme mannosidosis acquired on clinical MRI scanners were evaluated Sanofi). by automated volumetric analysis and compared to MRIs of 80 healthy controls (age: range 6-25 mean ± SD 14.0±7.7 years of age). Brain volumes were adjusted for intracranial volume and doi:10.1016/j.ymgme.2018.12.268 age. Results: Lower volumes of total cerebral grey matter (specifically putamen*, caudate**, and thalamus*) and cerebellum* (both 253 white* and grey* matter) were observed in patients. No change Proteomic profiling of engineered human immortalized podocyte was seen in volumes of cerebral cortex, corpus callosum, total cell model of Fabry disease cerebral white matter, and brainstem. (*pb0.001 **pb0.02) Conclu- sions: In the cohort of 6 alpha-mannosidosis patients brain MRI a a b atrophy of subcortical grey matter structures and cerebellum and a José Tibúrcio M. Neto , Gianna M. Kirsztajn , Ester M. Pereira , Helida c c d preservation of cerebral white matter structures were quantified M. Andrade , Ivana Helena R. Oliveira , Anatalia Labilloy , Adalberto b a b by automated volumetric analysis. These findings will be later S. da Silva , Federal University of São Paulo, São Paulo, Brazil, Federal c assessed to determine correlation to functional motor and University of Piaui, Teresina, Brazil, Federal University of Minas Gerais, d cognitive scores. Belo Horizonte, Brazil, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, United States. doi:10.1016/j.ymgme.2018.12.267 Renal involvment in Fabry disease (FD) is a frequent and early event with a significant morbidity and adverse prognosis, manifesting itself, initially, through microalbuminuria, evolving with a progressive reduction of the glomerular filtration rate and chronic 252 kidney disease (CKD). Enzyme replacement therapy has improved Discovery of brain MRI signatures in infants with severe form of life quality of FD patients, although it does not stop the progression MPS I in the pre-HSCT and post-HSCT stages to CKD. The mechanisms by which the lysosomal accumulation of glycosphingolipids, chiefly Gb3, results in cell and organic dysfunc- Igor Nestrasila, Alena Svatkovab, Rene Labouneka, Bryon Muellera, tion in FD are still unknown however, podocyte injury plays an David Nascenea, Troy Lunda, Paul Orcharda, Chester B. Whitleya, important role in the progression of this nephropathy. Podocytes are aUniversity of Minnesota, Minneapolis, MN, United States, bMedical well-differentiated and specialized kidney cells with limited capacity University of Vienna, Vienna, Austria for repair and replacement being the main glomerular cell involved Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S107 in Fabry Nephropathy (FN). Thus, elucidating the molecular mech- microglia. These experiments suggest that IUERT can penetrate the anisms of the podocyte in the FD context is important. In this study, BBB and decrease brain inflammation. IUHSCT results in microglial we have aimed to identify differentially abundant proteins in a engraftment and increasing chimerism may further improve results. model of immortalized human podocyte of FD, genetically edited These findings support in utero therapy for MPS7, for which we are with the aid of CRISPR/Cas9 technology using Two-dimensional currently developing a clinical protocol. Differential Gel Electrophoresis (2D-DIGE) and the proteins have been identified through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Our results have doi:10.1016/j.ymgme.2018.12.270 shown that proteins HSPD1, UCHL1 and Enolase 1 are downregulated in FD podocytes. These proteins take part in signaling pathways involved in processes of tubulointerstitial fibrosis and autophagy. In order to confirm pro-fibrosis pathways, we have compared the 255 expression of other proteins involved in this process by using Monoclonal gammopathies and hypergammaglobulemia in Western Blot and our data have shown overexpression of proteins Gaucher disease: An analysis from the French Gaucher disease TGF-β (1.5x), Vimentin (2.3x) and VEGF-2 (1.5x) in Fabry podocytes registry as regards the controls. Additionally, proteins involved in proliferative pathways, e.g. PI3K 110a (2.5x) and PI3K 110b (1,45x), are also Yann Nguyena, Jérôme Stirnemannb, Bengherbia Moniaa, Karima overexpressed. Therefore, the Fabry podocytes expresses a pro- Yousfia, Dalil Hamrounc, Florent Lautredouxd, Bérengère Cadord, fi fi brosis proliferative pattern. These ndings may improve our Marc Bergere, Bruno Fantina, Nadia Belmatouga, aHôpitaux knowledge of Fabry podocitopathy aiming new strategy at interven- Universitaires Paris Nord-Val-De-Seine - Hôpital Beaujon, Clichy, tion for preventing Fabry nephropathy. France, bHôpitaux Universitaires de Genève, Genève, Switzerland, cCHRU de Montpellier, Montpellier, France, dCHU de Rennes, Rennes, France, eCHU Estaing, Clermont-Ferrand, France doi:10.1016/j.ymgme.2018.12.269

Objectives: To determine the risk factors, the clinical significance, and the evolution of monoclonal gammopathy (MG) and 254 hypergammaglobulinemia (HG) in Gaucher disease (GD). Fetal enzyme replacement and stem cell transplantation in Methods: Patients from the French GD Registry with information murine Sly syndrome targeting microglia on the presence or absence of MG and HG (defined by an immunoglobulin level above 15 g/L) at baseline and/or during Quoc-Hung Nguyen, Russell Witt, Lucas Smith, Jeremy Shea, Bowen follow-up were studied retrospectively. The characteristics at fi Wang, Carlo Eikani, Saul Villeda, Tippi MacKenzie, University of diagnosis and the rst occurrence of MG, HG, bone events and California San Francisco, San Francisco, CA, United States severe thrombocytopenia during follow-up were collected. The risk factors of developing MG/HG and the association between MG/HG Mucopolysaccharidosis type 7 (MPS7) is a lysosomal storage with disease severity were analyzed using log-rank tests and disorder resulting in multi-organ dysfunction and in utero demise. multivariable Cox proportional hazards models, using MG/HG and Post-natal enzyme replacement therapy (ERT) and hematopoietic bone events/severe thrombocytopenia as events, respectively. Effect stem cell transplantation (HSCT) are unable to address neurologic of treatment on gammaglobulin levels was assessed using linear/ manifestations because of the blood-brain barrier (BBB). We study logarithmic mixed models. whether in utero ERT (IUERT) or HSCT (IUHSCT) can penetrate the Results: 59/187 patients (31.6%) had MG: 20 (33.9%) at diagnosis BBB, targeting microglia as they are enriched in the missing enzyme. of GD and 39 (66.1%) during follow-up after 19.0 ± 11.0 years Prior IUERT work showed neurologic benefit - we now aim to following diagnosis. 112/235 (47.7%) had HG: 15 (13.5%) at diagnosis analyze the extent of microglial engraftment, and to provide a of GD and 96 (86.5%) during follow-up. Mean ± SD age at diagnosis permanent enzyme source with IUHSCT. We treated MPS7 fetuses at of MG and HG were 49.7 ± 14.3 and 35.0 ± 16.5 years, respectively. E14.5 with IUERT or IUHSCT. Following injection of missing Multivariable analysis (hazard ratio 95% confidence interval) glucuronidase (GUS), performed flow cytometry for enzyme activity retained age at diagnosis of GD as the unique independent risk on brain at E18. Controls included GUS injection in adult mice. We factor for MG (1.08 1.05-1.10). MG or HG were neither associated stained brains for CD68 (to quantify microglial inflammation). We with the occurrence of bone events ([1.11 0.52-2.36] and [1.16 0.64- bred CX3CR1-GFP donor mice (engrafted microglia express CX3CR1 2.11], respectively), nor severe thrombocytopenia ([0.89 0.25-3.13] and are therefore green on histology) and transplanted fetal liver and [1.17 0.54-2.54], respectively). During follow-up, 5/187 had mononuclear cells, initially into wild-type mice. Chimeric brains then lymphomas (1 following MG and 3 following HG) and 1/187 underwent immunofluorescent staining for microglial markers. We multiple myeloma (following MG). Under treatment, immunoglob- injected 46 fetuses with enzyme or PBS control. 6 of 7 treated ulin levels significantly decreased (slope = -0.24 [95%CI, -0.28, -0.20] homozygous fetuses had detectable enzyme activity (24.6 ± 9.5% of per year, P b 0.001). microglia, some with wild-type activity levels). Accordingly, CD68 Conclusion: MG and HG are not associated with unfavorable staining showed decreased brain inflammation. In contrast, 6 adult outcome in GD but should encourage monitoring of hematologic homozygous mice were treated, and none developed detectable malignancies. brain GUS activity. We transplanted 5 litters of wild-type mice, and 9 mice engrafted (8.5±2.2% multilineage chimerism in blood and bone doi:10.1016/j.ymgme.2018.12.271 marrow). In chimeric mice at 3 weeks, transplanted stem cells co- stained with GFP and Iba1, indicating engraftment into the brain as S108 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

256 extraction method (modified from Alharbi et al, 2016) and analyzed The effects of long-term migalastat treatment in Fabry disease in both positive mode for ceramide trihexosides and negative mode patients previously treated with enzyme replacement therapy for sulfatides using MALDI-TOF mass spectrometry. Validation who have migalastat-amenable variants with low alpha-galacto- included specimens from confirmed patients affected with Fabry sidase A response in the in vitro migalastat amenability assay disease (N=16 males N=11 females), MLD (N=23), MSD (N=7), SAPB (N=2) and MLII (N=5). Fabry disease has a specific ceramide Kathleen Nichollsa, Iacopo Olivottob, Toya Ohashic, Hadis Williamsd, trihexoside profile pattern without presence of sulfatides. Overlap- Vipul Jaind, Nina Skuband, aRoyal Melbourne Hospital, University of ping sulfatide profiles, without ceramide trihexosides, for MLD and Melbourne, Melbourne, Australia, bCentro di Riferimento Car- MSD can be confirmed by glucosaminoglycan analysis. SAPB has both diomiopatie, Florence, Italy, cJikei University Hospital, Tokyo, Japan, the ceramide trihexoside profile pattern similar to Fabry disease and dAmicus Therapeutics, Inc., Cranbury, NJ, United States the sulfatide profile pattern similar to MLD and MSD, but reduced peak intensities. Overlapping profiles to MLII, which can have both Fabry disease is a rare, X-linked disorder of α-galactosidase A (α- ceramide trihexosides and sulfatides as well, can be confirmed by Gal A) deficiency caused by mutations in the GLA gene, resulting in oligosaccharide analysis. To further delineate profile patterns, peak substrate accumulation and multi-organ deterioration. Migalastat is a ratios to respective internal standards in positive and negative first-in-class, oral, small-molecule pharmacological chaperone that modes (N=30) are analyzed using a custom-made automated post- binds to and stabilizes endogenous α-Gal A, leading to increased analytical multivariate pattern recognition software, Collaborative lysosomal activity and reduced substrates in patients with amenable Laboratory Integrated Reports (CLIR https://clir.mayo.edu). This GLA variants, as determined by an in vitro migalastat amenability software generates additional ratios (N=248) between the analyte/ assay. This descriptive analysis evaluates the long-term effects of IS ratios to aid in reliable pattern recognition. migalastat on α-Gal A activity and clinical outcomes in enzyme replacement therapy (ERT)-experienced patients enrolled in the phase 3, randomized, ERT-controlled trial ATTRACT (NCT01218659) doi:10.1016/j.ymgme.2018.12.273 and the open-label extension study AT1001-041 (NCT01458119). The analysis focuses on 5 patients (3 females, 2 males) whose GLA variants, when tested with the in vitro migalastat amenability assay, 258 α resulted in a relatively low response of -Gal A activity (an increase CBE treatment of alpha-synuclein over-expressing and wildtype ≥ –b ‘ ’ ≥ of 3% 6% of wild-type) but still met the amenable criteria ( 1.2- mice models Gaucher disease pathology fold relative increase over baseline and ≥3% absolute increase of wild-type α-Gal A activity). These 5 patients, with an average of 2.2 Vera Niederkofler, Ewald Auer, David Amschl, Joerg Neddens, Birgit years of migalastat treatment, generally achieved stabilization in Hutter-Paier, QPS Austria GmbH, Grambach, Austria renal function and reduction in cardiac mass, and their globotriaosylsphingosine (lyso-Gb3) levels also remained stable, con- Gaucher disease is the most prevalent lysosomal storage α sistent with the overall population. White blood cell -Gal A activity disorder and is caused by autosomal recessive mutations in the (assessed in males only, n=2) was very low at baseline in this subset glucocerebrosidase gene. Glucocerebrosidase (GCase) hydrolyses and achieved sustained increases during migalastat treatment, the sphingolipid glucoceramide to glucose and ceramide. Deficiency maintaining absolute increases of 4.6% (1.7 fold) and 7.2% (16.9 in GCase activity leads to a multisystemic accumulation of substrate fold) of normal at last assessment, respectively. The every-other-day in lysosomes. Additionally, α-synuclein, tau, ubiquitin, APP and dosing resulted in consistent levels of migalastat exposure in these Abeta might build up in affected tissues. Most of these proteins are patients. These data suggest that in patients who have amenable also accumulated in several other rare diseases and neurodegener- α GLA variants with low -Gal A responses in the in vitro migalastat ative diseases such as α-synuclein in dementia with Lewy bodies or amenability assay, long-term treatment with migalastat generally Parkinson´s disease. In the last years several links between α- fi provides a bene t in renal function and cardiac mass. synucleinopathies and lysosomal storage diseases have been reported. We thus combined an inducible mouse Gaucher model doi:10.1016/j.ymgme.2018.12.272 with a transgenic Parkinson´s disease mouse by treating six months old alpha-syunclein transgenic mice and non-transgenic littermates with Conduritol B Epoxide (CBE) that serves as an irreversible inhibitor of β-glucosidase and is known to cause a Gaucher-like 257 phenotype. After CBE treatment, animals were tested for motor Multiplex assay for the tandem detection of ceramide tri- impairments in the Beam Walk test und brain tissue was analyzed hexosides and sulfatides: Efficient first tier screening for Fabry, for alpha-synuclein expression, activated microglia and astrocytosis MLD, MSD, and Saposin B in urine by alpha-synuclein, Iba1 and GFAP immunofluorescent labeling. Our results show that CBE treatment caused motor impairments that Kim K. Nickander, Jean M. Lacey, Dimitar K. Gavrilov, Dietrich were not affected by the transgene. Alpha-synuclein levels were Matern, Devin Oglesbee, Piero Rinaldo, Silvia Tortorelli, Kimiyo highly increased in transgenic animals compared to non-transgenic Raymond, Mayo Clinic, Rochester, MN, United States littermates and CBE treatment caused a further increase of alpha- synuclein levels. Activated microglia and astrogliosis were increased Fabry disease, metachromatic leukodystrophy (MLD), multiple in all CBE treated groups, independent of the transgene. Our data sulfatase deficiency (MSD), saposin B deficiency (SAPB) and validate motor impairments and neuroinflammation after CBE mucolipidosis II (MLII) are disorders caused by enzyme deficiencies treatment and additionally show an impact of CBE on alpha- leading to accumulation of ceramide trihexosides and/or sulfatides in synuclein levels in transgenic mice. Using alpha-synuclein trans- tissues and increased excretion in urine. Ceramide trihexosides and genic mice to induce Gaucher disease by CBE treatment might sulfatides are extracted in chloroform from urine using a simple therefore be a valuable alternative to the already established CBE Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S109 model in wildtype mice by combining pathologies observed in disorder. The disease is characterized by severe and progressive loss Gaucher and Parkinson disease. of cognitive and motor functions, behavioral deficits and eventually death in the second decade of life, although the severity and progression of the disease varies widely. MPS IIIA is caused by doi:10.1016/j.ymgme.2018.12.274 mutations in the SGSH gene that result in deficiency of the N- sulfoglucosamine sulfohydrolase enzyme and subsequent accumulation of undegraded heparan sulfate, lysosomal enlargement and 259 cellular and organ dysfunction. To model the disease, a MPS IIIA Characterization of 4L/PS-NA mice for enzyme activity, substrate mouse strain with spontaneous Sgsh mutation, resulting in an almost concentrations as well as inflammation to model Gaucher disease complete loss of N-sulfoglucosamine sulfohydrolase activity, was characterized. Male animals were analyzed at an age of 28 to 30 weeks for body weight changes, activity, social interaction and Vera Niederkoflera, Ewald Auera, Victoria Schiffera, Tina Loefflera, cognition. Afterwards, neuropathology was evaluated using a LIMP2 Joerg Neddensa, Staffan Schmidtb, Jan Kehrb, Birgit Hutter-Paiera, antibody, labeling lysosomal and endosomal membranes, and GFAP aQPS Austria GmbH, Grambach, Austria, bPronexus Analytical AB, and IBA1 antibodies to analyze neuroinflammation. MPS IIIA mice Bromma, Sweden demonstrated an increased body weight, combined with a decreased rearing behavior in the open field test. Social interaction of MPS IIIA Gaucher disease is the most common lysosomal storage disease. The mice was reduced and animals presented weak memory deficits in neuronal disease variant is characterized by aggregated protein accu- the Barnes maze test. Histological analyses revealed a strong increase mulations in the brain and associated neurological manifestations. It is in astrogliosis but only a slight increase of activated microglia. autosomal recessively inherited and modeled by 4L/PS-NA mice that Quantification of LIMP2 showed a severe signal increase in MPS IIIA express low levels of prosaposin and saposins, as well as a functionally mice compared to wildtype animals. In summary, these results impaired β-glucosidase (GCase) with a homozygous point mutation at confirm that MPS IIIA mice present a similar phenotype as observed V394L. To use this model for compound tests against Gaucher disease a in Sanfilippo syndrome type A patients. The mouse is therefore a detailed characterization of these mice is needed. Thus, we aimed to valuable model for preclinical research. analyze the 4L/PS-NA mice for GCase activity, glucosylsphingosine (GlcSph) and glucosylceramide (GlcCer) levels as well as inflammation over age. The GCase activity is currently analyzed in different tissues doi:10.1016/j.ymgme.2018.12.276 using a commercially available GCase activity assay. GlcSph and GlcCer were extracted from brain homogenates using solid-phase extraction cartridges and the levels of GlcSph and GlcCer in the brain extracts were measured by ultrahigh-performance liquid chromatography coupled to 261 tandem mass spectrometry and ultra performance liquid chromatogra- Reduction of paraprotein levels in type 1 Gaucher disease with phy with time-of-flight mass spectrometry, respectively. To explore enzyme therapy neuroinflammatory processes, in particular activated microglia and astrocytosis, in this animal model we performed immunofluorescent Graeme A.M. Nimmo, Mary Anne Patterson, Heather Goba, Dominick labeling on brain sections. Furthermore, KC/GRO (CXCL1) cytokine Amato, Mount Sinai Hospital, Toronto, ON, Canada measurement in the CSF was performed by immunosorbent assay. Finally, mouse embryonic fibroblasts(MEFs)of4L/PS-NAmicewere Individuals with type I Gaucher disease (GD) have a 25-50-fold analyzed as in vitro screening tool. Analyses of enzyme activity and increased risk of developing monoclonal gammopathy of undeter- substrate concentrations in 4L/PS-NA mice are currently ongoing. 4L/PS- mined significance (MGUS), and its post-cursor, multiple myeloma, NA mice showed strong neuroinflammation and increased CXCL1 levels. compared with the general population. Multiple myeloma is a major Analysis of MEFs revealed a strongly reduced GCase activity in the cells contributor to morbidity and mortality in patients with GD with up from 4L/PS-NA mice compared to C57Bl/6 MEFs but only a minor to 8.0% of patients progressing to a malignant gammopathy. Chronic reduction compared to 4L/PS+/+NA mice harboring wildtype prosaposin. immune activation and the pathogenic accumulation of lysolipids in 4L/PS-NA mice thus mimic the most prominent features of Gaucher GD are hypothesized to contribute to this increased risk. However, disease and are a valuable tool for in vitro and in vivo drug development. the response of MGUS to enzyme replacement therapy and/or an alteration in the risk of progression to multiple myeloma on treatment is not clear. Retrospective chart review of 85 patients doi:10.1016/j.ymgme.2018.12.275 with Type 1 GD revealed 12 patients with MGUS (incidence: 14%), including 4 who received enzyme replacement therapy (ERT). ERT resulted in an improvement in markers of disease activity (including 260 chitotriosidase, ferritin, lysoGb1) and in reduction of the monoclonal Behavioral and histological hallmarks of a mucopolysaccharidosis protein. In the majority of patients, the response was sustained. One type IIIA mouse model patient demonstrated a transient reduction in paraprotein with ERT but progressed to smoldering myeloma with an IgG-κ monoclonal component. These data suggest that MGUS may respond to ERT and Vera Niederkoflera, Ewald Auera, David Amschla, Joerg Neddensa, provide further evidence that altering the levels of lysolipid Elisabet Ohlin Sjöströmb, Susanne Gustavssonb, Birgit Hutter-Paiera, accumulation in GD individuals with MGUS may attenuate the risk aQPS Austria GmbH, Grambach, Austria, bSOBI - Swedish Orphan of progression to malignancy. Biovitrum AB, Stockholm, Sweden

ﬁ Mucopolysaccharidosis IIIA (MPS IIIA), also known as San lippo doi:10.1016/j.ymgme.2018.12.277 syndrome type A, is an autosomal recessive lysosomal storage S110 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

262 Background Fabry disease (FD), an X-linked lysosomal storage Fabry patients’ needs and expectations regarding their treatment disorder, results from α-galactosidase A gene (GLA) mutations in France: Development of a Patients’ Need Questionnaire (PNQ causing deficient α-galactosidase A (α-GalA) activity. There are two Fabry) major subtypes: early-onset Type 1 “Classic” and Type 2 “Later- Onset” phenotypes. Type 1 males have essentially no α-GalA activity, Esther Noëla, Bertrand Dussolb, Didier Lacombec, Najya Bedreddined, marked microvascular endothelial globotriaosylceramide (Gb3) Alain Fouilhouxe, Pierre Roncof, Delphine Genevazg, Soumeya Bekrih, accumulation, and childhood/adolescent onset characterized by Albert Hagègei, Frédérique Dupuis-Siméonj, Valérie Derrienk, Domi- acroparesthesias, angiokeratoma, corneal opacities, and hyp- nique P. Germainl, Olivier Lidovem, aHôpitaux Universitaires de ohidrosis. With age, progressive Gb3 deposition in microvascular Strasbourg, Strasbourg, France, bHôpital de la Conception, Marseille, endothelial cells, renal podocytes, and cardiomyocytes, leads to renal France, cCHU de Bordeaux, Hôpital Pellegrin, Bordeaux, France, insufficiency/failure, cardiomyopathy, and cerebrovascular disease. dAssociation des Patients de la Maladie de Fabry (APMF), Marsannay Type 2 males have residual α-GalA activity, no microvascular Gb3 la Côte, France, eCHU de Lyon - Hopital Civil de Lyon, Lyon, France, accumulation, lack Type 1 early manifestations, and present in fHôpital Tenon, Paris, France, gVaincre les Maladies Lysosomales (VML), adulthood with progressive cardiac and/or renal disease. Previously, Massy, France, hCHU de Rouen, Rouen, France, iHôpital Européen FD prevalence was estimated at ~1 in 40,000-119,000 males. Georges Pompidou, Paris, France, jAmicus Therapeutics, La Défense, However, the incidence is unknown it is best estimated by newborn France, kA+A Research, Lyon, France, lHôpital Raymond Poincaré, screening (NBS) of male newborns, as enzyme screening does not Garches, France, mHôpital de la Croix St Simon, Paris, France reliability identify female heterozygotes. Methods Online databases were searched for FD NBS studies those with GLA sequencing were Anderson-Fabry disease (FD) is a rare X-linked inherited lysosomal reanalyzed for pathogenic mutations and phenotype. Results In 15 disease caused by reduced activity of α-galactosidase A. Due to the NBS studies reporting mutations, 1,566,931 newborns were heterogeneity of disease presentation and evolution, patient-reported screened. Of the seven studies that reported 247,286 males, outcome (PRO) tools, such as SF-36, do not provide accurate insights 0.0546% had pathogenic mutations. Among these studies, the into patients’ daily lives and impact of specific treatments. In response incidence of Caucasian and Asian males was ~1:4,000 and ~1:1,400, to the “Shared Clinical Decision Making” report published by the respectively, with Types 1 to 2 phenotype ratios of 1:20 and 1:13, French Authority for Health, we aimed to construct a validated and FD- respectively. Conclusions The estimated FD male incidence was specific tool to appraise patients’ needs and expectations towards their markedly more frequent than the previously estimated prevalence, treatment. This endeavor was led with the help of French patient primarily since Type 2 males were previously under-diagnosed. In X- associations (APMF & VML) and dedicated expert centers. The linked diseases, expected heterozygotes number twice the newborn Questionnaire of the Expectations of Patients with Fabry Disease with males. For Type 1 males, NBS facilitates early treatment. For Type 2 respect to their treatment (PNQ Fabry) was developed according to males, NBS presents ethical issues, since manifestations develop in the FDA/EMA methodologies and best practices for the development of adulthood. Screening Types 1 and 2 at-risk family members will PRO tools in rare diseases. Our approach comprised of 3 steps, as identify older previously undiagnosed affected family members for follows: concept elicitation and item generation, item reduction medical management including enzyme replacement or pharmaco- (quantitative survey), and final validation of the questionnaire logic chaperone therapy. through a 2-stage survey. Intrinsic and extrinsic stability of the PNQ Fabry was established, using a validated benchmark questionnaire. With the invaluable help of patient associations, we recruited a doi:10.1016/j.ymgme.2018.12.279 satisfactory population in this rare disease setting, to ensure robust participation to validate our PNQ (final number of questionnaires: 76). At the end of the process, we obtained a 26-item patient-reported 264 questionnaire with excellent psychometric properties, exhibiting very Fabry disease genotype, phenotype and amenability: A full satisfactory measurement outcomes for reliability and validity. The country overview results of this initiative demonstrate that the PNQ Fabry is both acceptable to patients and accurate, as it addresses themes identified Albina Nowaka, Pierre-Alexandre Krayenbuehlb, Uyen Huynh-Doc, from patient interviews, that were further validated through statistical Frederic Barbeyd, aUniversity Hospital Zurich, Zurich, Switzerland, analyses of qualitative and quantitative surveys. An ongoing phase IV bSpital Linth, Uznach, Switzerland, cUniversity Hospital Bern, Bern, study is using this tool we believe it could be reliable and insightful in Switzerland, dHôpital Beaumont Lausanne, Lausanne, Switzerland daily practice, to improve patient management in FD. (Supported by Amicus Therapeutics.) Fabry disease (FD) is a rare X-linked lysosomal storage disorder due to mutations in the α-galactosidase A gene (GLA) that result in fi α α doi:10.1016/j.ymgme.2018.12.278 de ciency of -galactosidase A ( -GalA) enzymatic activity. As a result, the major glycosphingolipid substrates accumulate in plasma, urine and tissue lysosomes. There are two major phenotypes, classic and later-onset. Males with the more severe, classic phenotype have 263 an α-Gal A activity of b1% and suffer early symptoms including Fabry disease: Incidence of pathogenic GLA mutations estimated acroparesthesias, angiokeratoma, corneal opacities and hypohidrosis, by newborn screening studies culminating in cardiomyopathy, kidney disease, and premature strokes in adults. The less severe, later-onset phenotype is charac- Albina Nowaka, Robert J. Desnickb, aUniversity Hospital Zurich, Zurich, terized by an α-Gal A activity of N1%, asymptomatic childhood, but a Switzerland, bMount Sinai School of Medicine, New York, NY, United progressive heart or kidney disease in adult males. The two States phenotypic entities require different diagnostic and therapeutic Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S111 strategies. Results: A total of 164 patients from 44 independent threatening among MPS patients. It can also be used to monitor pedigrees, 61 affected males and 103 heterozygous individuals, have medication adherence. been diagnosed with FD in Switzerland since the year 1958. The ratio classic to later-onset phenotype was 3:1. Overall, 49% of the patients doi:10.1016/j.ymgme.2018.12.281 had amenable mutations, of those, 24% had classic and 25% later- onset phenotype. All patients with the later-onset phenotype had amenable missense mutations. While screening programs for FD have never been performed in Switzerland, mainly classic mutations 266 were diagnosed in the past, later-onset mutations only being Improving brain delivery of adeno-associated viral gene therapy increasingly diagnosed recently. Conclusions Although the later- vectors for the treatment of MPS IIIC onset phenotype naturally occurs 10-20 times more frequently than the classic one, as shown in newborns screening studies, in our Claire O'Learya, Gabriella Fortea, Nadia Mitchellb, Amir Saam population, the prevalence of classic was considerably higher than Youshania, Martin Wellbyb, Katharina Russellb, David Palmerb, Els later-onset phenotype, most likely because systematic screening for Henckaertsc, Ian Kamaly Asld, Brian Biggera, aUniversity of Manchester, FD have never been performed in Switzerland. However, an Manchester, United Kingdom, bLincoln University, Lincoln 7647, New increasing number of later-onset phenotype patients has been Zealand, cKing's College London, London, United Kingdom, dRoyal diagnosed in the recent time, due to increasing awareness of FD. Manchester Children’s Hospital, Manchester, United Kingdom Screening studies would help identify many so far undiagnosed later-onset patients. MPS IIIC is caused by mutations in the HGSNAT gene. HGSNAT deficiency affects lysosomal catabolism of heparan sulfate (HS), resulting in widespread CNS pathology in infants and children, doi:10.1016/j.ymgme.2018.12.280 leading to behavioural problems, cognitive decline and, finally, dementia and death before adulthood. Currently there are no existing treatments for MPS IIIC and the development of treatments is difficult as the deficient enzyme cannot cross the blood brain 265 barrier or diffuse between cells. We have developed an adeno- Potential benefits of Fitbit device in managing a patient with associated virus (AAV) based gene therapy for MPS IIIC which is mucopolysaccharidosis efficacious in a MPS IIIC murine model via injection into the brain parenchyma. Although effective in a small murine brain, the a a b Andrew Oldham , Karolina M. Stepien , Christian J. Hendriksz , architecture and volume is considerably different from a human a b Salford Royal Foundation Trust, Salford, United Kingdom, University brain, and the failure of previous clinical trials for AAV vectors using of Pretoria, Pretoria, South Africa intraparenchymal injection, means that large animal experiments are essential in optimising widespread distribution needed for clinical Mucopolysaccharidosis (MPS) IVa is a lysosomal storage disor- efficacy. Due to the inability of HGSNAT to cross-correct cells, der resulting from a deficiency of N-acetylgalactosamine-6-sulfatase intracerebral delivery of vector needs to target the maximum leading to progressive accumulation of glycosaminoglycans (GAGs) number of cells, and with the use of convection enhanced delivery in musculoskeletal system, patients manifest with an impaired gait (CED) this can be achieved. CED has the potential to treat diseases pattern, reduced levels of physical endurance, muscular weakness, with global pathology such as MPS IIIC as it overcomes the issues arrhythmias and upper airways abnormalities. Wearable life style with current delivery methods and can be administered to multiple technology is available and used in managing health issues. We sites. We compared different conditions to determine optimal vector describe the application of a FitBit device as part of physiotherapy spread of an AAV9-GFP vector in the sheep brain, which is 130mL in assessment of one MPS IVa patient. A 21-year-old male diagnosed volume, at clinically applicable volumes and titres whilst keeping with MPSIVa at the age of 3, started weekly infusions of Enzyme volume identical throughout. We compared 4 intraparenchymal Replacement Therapy (ERT) at the age of 21. Six months into the sites, against intraventricular injection, and compared different flow therapy, he started recording his heart rate, sleep pattern and rates at 2 single intraparenchymal sites. We found a rate of 1μL/min numbers of steps on a FitBit device. During regular clinic review, into the corpus striatum produced the greatest vector distribution, physiotherapy assessment included heart rate before and after 6- whilst intraventricular outcomes were comparatively poor. However, minute walk test (6MWT), saturation, Borg Scale and Dyspnea further studies are currently ongoing to improve this further with Scores. Number of steps/day was used as a hallmark of his physical the aim of determining the optimal delivery system for gene therapy activity and continued to increase in response to ERT. At baseline, 7, of MPS IIIC. 10 and 13 months of ERT, he showed significant improvement in 6MWT with 43, 60, 60 and 90 meters, respectively. His heart rate before 6MWT was 60-84/min and after the test 150-165/min. There doi:10.1016/j.ymgme.2018.12.282 were no changes in his sleeping pattern throughout 13 months of monitoring, with average 6-8 hours of sleep. In conclusion, FitBit device allows continuous observations concerning the impact of clinical interventions on mobility over a long period of time. It 267 improves the ability to share data with medical professionals. The Design and rationale of the LYS-SAF302 gene therapy study in wearable device promotes wellness and self-management of mucopolysaccharidosis type IIIA (MPS IIIA) children medical condition among MPS patients. It is a way of monitoring patient’s physical activity and quality of sleep. Most importantly it Sophie Oliviera, Samantha Parkera, Anupam Chakrapanib, Ronald may be an effective screening tool for arrhythmias that can be life- Crystalc, Lisa Emrickd, Benedicte Herone, Nicole Muscholf, Raymond S112 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Wangg, Chester Whitleyh, Frits Wijburgi, Lucie Roucha, aLysogene, mass of disease specific images. Earlier collaborations between Cure Neuilly sur Seine, France, bGreat Ormond Street Hospital, London, United Sanfilippo Foundation, Jonah’s Just Begun Foundation and Face2Gene Kingdom, cWeill Cornell Medicine, New York City, NY, United States, collected patient entered images and established a composite image dTexas Children's, Houston, TX, United States, eHôpital Armand Trous- for MPS IIIB. Here, we assessed the technology’s capability to seau La Roche Guyon, Paris, France, fUniversity Medical Center, characterize the unique facial phenotype of MPS III across different Hamburg, Germany, gCHOC Children's, Orange County, CA, United age groups. Through a secure patient portal, families submitted States, hUniversity of Minnesota, Minneapolis, MN, United States, patient photographs across the patients’ lifespan. Images were iAcademic Medical Center, Amsterdam, Netherlands stratified by age group and comparisons conducted between MPS IIIB age groups, between “normal” subjects and non-MPS IIIB Mucopolysaccharidosis type IIIA (MPS IIIA) (OMIM 252900) leads syndromic facies. Statistical analysis of the receiver operating to early-onset neurodegeneration and premature death. Though the characteristic (ROC) curve was performed to calculate the area involvement of multiple organ systems in MPS IIIA is recognized, under curve (AUC) assessing differences between age groups and central nervous system (CNS) manifestations predominate, in build the facial natural history. Face2Gene was able to accurately particular intellectual disability, progressive loss of acquired skills, identify MPS IIIB patient photos in the 1-3 year-old age a critical behavioral and sleep disturbance. The reduced lifespan is related to window for intervention. Development of a facial natural history for progressive neurological deterioration rather than involvement of MPS III could aid clinicians in identifying affected patients. This study other organ systems. A direct-to-brain administration is thought best further illustrates the value of patient engagement in the advance- for such diseases because of the profound CNS component and the ment of rare disease diagnosis and characterization. blood brain barrier, which limits the passage of therapies administered via peripheral routes. Long-term 5-year data in four MPS IIIA patients treated with an intracranial administration of a first- doi:10.1016/j.ymgme.2018.12.284 generation adeno-associated viral (AAV) gene therapy (LYS- SAF301) showed that direct to CNS gene therapy was well tolerated. Lysogene’s Phase 2-3 trial (NCT03612869) uses an optimized gene 269 therapy construct (LYS-SAF302). The study is a single-arm, multi- Meaningful treatment outcomes for Sanfilippo syndrome: A study center study of AAV serotype rh.10 carrying the human SGSH of caregiver preferences and prioritization (AAVrh.10-SGSH) for the treatment of MPS IIIA. The study will include 20 patients. The primary objective will be to assess the drug Cara O'Neilla, Katherine Porterb, Elise Drakea, Holly L. Peayb, aCure efficacy in improving or stabilizing the neurodevelopmental status of Sanfilippo Foundation, Columbia, SC, United States, bRTI International, patients compared to the expected evolution based on natural Research Triangle Park, NC, United States history data. Safety, tolerability, effect on behavior, sleep and quality of life are secondary endpoints. Eight clinical sites in the US and Sanfilippo syndrome (MPS III) is a rare, degenerative disorder Europe are participating in this trial. The principal inclusion criterion with no approved therapies. This study explored caregiver percep- is a cognitive developmental quotient (DQ) score N50% based on the tions of the burden of Sanfilippo syndrome on the child and family, Bayley Scales of infant and toddler development third edition (BSID- and meaningful benefits that are desired from a non-curative III). Status of the first patients treated in the study will be presented. therapy. We used an innovative mixed-method approach with 25 caregivers (biological and step-parents) of children ages 4-36 years doi:10.1016/j.ymgme.2018.12.283 old from 17 U.S. states. We conducted three focus groups comprising three activities: (1) formative validity testing of a symptom/staging survey (not described here), (2) moderated exploration of burden and meaningful treatment benefit, and (3) best-worst scaling (BWS) 268 activities to quantify the relative importance of twelve treatment The natural history of facial features observed in Sanfilippo benefits, which arose during activity 2 and thus varied across focus syndrome (MPS IIIB) using a next generation phenotyping tool groups. Thematic analysis revealed common themes and the quantitative analysis used ‘best-worst’ scoring to prioritize mean- Cara O'Neilla, Nicole Fleischerb, Jill Woodc, aCure Sanfilippo Founda- ingful benefits from most to least important. Caregivers’ experiences tion, Columbia, SC, United States, bFDNA Inc., New York, NY, United reflect uncertainty and distress related to progression and increased States, cJonah's Just Begun Foundation, Brooklyn, NY, United States family burden with progressing symptoms. Participants expressed willingness to “try anything” to slow or stop progression. Meaningful Sanfilippo syndrome (MPS III) is a rare lysosomal storage disease benefits to the family included reducing risky and perceived resulting from a deficiency in one of four enzymes needed to aggressive/impulsive behaviors and improving ability to communi- breakdown the glycosaminoglycan heparan sulfate. Excessive storage cate needs and preferences. For their children, focus group 1 of heparan sulfate leads to debilitating, progressive neurodegenera- prioritized addressing communication (relative importance tion while systemic disease is relatively less severe. Despite MPS III score=.50), pain (score=.35), and child unhappiness (score=.33) being the most prevalent form of mucopolysaccharidosis, significant group 2 prioritized communication (score=.58), frustration delays occur in diagnosis due to the less pronounced facial and (score=.33), and aggressive behavior (score=.31) and group 3 physical features. Several distinct facial features are present in nearly prioritized communication (score=.50), pain (score=.48), and sleep all patients progressive coarsening facies, prominent eyebrows, and (score=.23). Caregivers reflected on the relevance of communication frontal bossing. Face2Gene is a freely available tool for medical challenges across disease progression. Consistent with prior reports, providers, that uses facial analysis, deep learning, and artificial most caregivers identified considerable burden. Modest treatment intelligence to analyze 2D photographs and suggest possible benefits to communication limitations and problematic behaviors syndrome matches. To establish an accurate composite facial image would be highly valued, as would reduced pain-especially given for which to compare new cases, the system must contain a critical caregiver uncertainty regarding the child’s pain experience. Our next Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S113 steps include using these data to develop a larger BWS survey to Intra-venous enzyme replacement therapy has its limitations in prioritize meaningful treatment benefits across stages of MPS III Anderson-Fabry disease. Genetically amenable Anderson-Fabry pa- progression. Funding: Cure Sanfilippo Foundation, BioMarin, Lyso- tients can now be treated with a novel chaperone therapy. Migalastat gene, Sobi, Orchard Therapeutics. (Galafold) is an oral agent that stabilises mutant enzyme function without the need for enzyme replacement. Here we present real- world data from patients initiated on migalastat in a quaternary doi:10.1016/j.ymgme.2018.12.285 Anderson-Fabry centre between August 2017 and August 2018. 61 patients were established on migalastat, 41 switched from enzyme replacement therapy and 20 were treatment naïve. At baseline, 270 average age of the patients was 53 years old, the youngest was 19 Preliminary results demonstrate engraftment with minimal and the oldest was 73. 41 were male and 20 female. 28 had GLA p. neutropenia with MGTA-456, a CD34+expanded cord blood (CB) N215S mutations, 5 had GLA p.P259R mutations, the rest had other product in patients transplanted for inherited metabolic disor- amenable mutations. Mean lyso Gb3 levels were 7.9, minimum 0.7 N ders (IMD) and maximum 55.4. 23 patients had an eGFR 90mL/min, 31 between 60-90mL/min, 2 between 45 - 59mL/min and 2 between 30 - 44mL/min. No patient had an eGFR less than 34 at initiation. 6 Paul J. Orcharda, Glen D. Raffelb, Carolyn E.H. Condonb, Catherine A. had neurological symptoms at initiation of treatment. The baseline Monaghanb, Jennifer A. Brauna, Ryan Shanleya, Troy C. Lunda, Ashish echocardiogram demonstrated a mean interventricular septal diam- Guptaa, Anthony E. Boitanob, Michael Cookeb, Davis C. Davisb, John E. eter of 1.4mm, minimum 0.7mm and maximum 2.4cm. 8 had severe, Wagnera, aUniversity of Minnesota, Minneapolis, MN, United States, 10 had moderate, 23 had mild LVH and 17 had no LVH. Mean LV bMagenta Therapeutics, Boston, MA, United States mass was 239g, maximum 554g, minimum 104g. 15 patients had severely elevated LV mass index 5 moderate, 3 mild and 33 normal. Mucopolysaccharidosis type IH (Hurler Syndrome), metachro- One patient stopped migalastat indefinitely following an unrelated matic leukodystrophy (MLD), globoid cell leukodystrophy (GLD) and diagnosis, three had a hiatus in treatment secondary to side-effects cerebral adrenoleukodystrophy (cALD) are progressive diseases all of which were re-established on treatment successfully. Side treatable with allogeneic hematopoietic stem cell transplantation effects reported included headaches, tiredness, nausea, vomiting, (HSCT). While cord blood is a common stem cell source, it may be bloating and muscle aches. The follow-up period ranged from 29 to associated with prolonged neutropenia and graft failure. MGTA-456 is 431 days, mean 277, median 292 days. Migalastat has been a cell therapy produced from a single CB unit using an aryl successfully used in this cohort, comparing baseline RCT data with hydrocarbon receptor antagonist in a 15-day expansion culture of real-world baseline is useful when observing a drugs application in CD34+cells. In previous studies, patients with hematologic malignan- clinical practice. cies demonstrated a median 324-fold expansion of CD34+cells, and the time to neutrophil recovery was significantly reduced by a median + of 9 days compared to historical controls. As higher CD34 doses have doi:10.1016/j.ymgme.2018.12.287 been correlated with improved engraftment and outcomes in inherited metabolic disease patients, we initiated a Phase 2, open- label trial to enroll up to 12 patients with a diagnosis of MPS1, cALD, MLD, and GLD utilizing CB units matched at ≥6 of 8 HLA loci using 272 allele-based typing. The HSCT regimen consists of anti-thymocyte Functional evaluation of an AAV9 based vector expressing alpha- globulin (days -9 to -6), fludarabine (days -5 to -2) and busulfan (days galactosidase A for potential gene therapy of Fabry disease -5 to -2), with cyclosporin and methylprednisolone as graft vs. host disease prophylaxis. In four patients treated thus far, a marked Saida Ortolanoa, Maria G. Biferib, Beatriz San Millána, Olga Soutoa, expansion of CD34+cells (median 482-fold) was observed. Two Irene Vieiteza, Jose R. Fernández-Lorenzoa, Julian J. Fernandez- patients had no days of neutropenia and 2 patients had 1 and 4 days Martina, Martine Barkatsb, aInstituto de Investigacion Sanitaria Galicia respectively, in contrast to a mean of 7.8 days in a historical Minnesota Sur, Vigo, Pontevedra, Spain, bInstitut de Myologie (UMRS974/INSERM/ cohort (N=27) with identical conditioning. Donor myeloid chimerism UPMC/AIM), Paris, France (≥98%) was achieved by day +14 in all patients. One patient developed autoimmune cytopenia, a known complication unrelated to MSGT 456, Fabry disease (FD) is treated by enzyme replacement therapy resulting in death at day +143. Based on these promising data, MGTA- (ERT) or pharmacological chaperons (in UE). However, ERT has a 456 has potential to improve hematopoietic cell recovery and short half-life and cannot cross the blood brain barrier (BBB), while transplant-related outcomes in patients undergoing HSCT for patients pharmacological chaperons are not indicated for all diagnosed with Inherited metabolic disorders. patients. We aim to test the efficacy of a gene therapy vector for FD, based on Adeno-Associated-Virus9 (scAAV9). The vector (pAAV9_PGK_GLA) expresses human α-GalactosidaseA (rh-α-GalA) doi:10.1016/j.ymgme.2018.12.286 under the promoter phosphoglycerate-kinase1. This vector can potentially achieve systemic expression of rh-α-GalA and cross the BBB. Similar therapies that are being developed are excluded from 271 the CNS, therefore our vector could complement their effects. α Real-world baseline data in patients established on migalastat for Although FD is not prevalently a neurological disorder, -GalA is Fabry disease expressed in neurons, astrocytes and microglia and its defect maybe related to stroke in FD patients. scAAV9_PGK_GLA was injected in hemizygous B6;129-Glatm1Kul/J newborns (P1, Nº12) and the ani- Chris Orsborne, Lorraine Thompson, Ana Jovanovic, Salford Royal mals were sacrificed 3 or 5 months post-injection. rh-α-GalA activity Foundation Trust, Manchester, United Kingdom was measured in brain, spinal cord, liver, heart and kidney and S114 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 glycosphynsolipid deposits were detected by immunohistochemis- 274 try. rh-α-GalA is functional in all the analyzed tissues including the Dysfunctional autophagy impairs muscle regeneration in lyso- CNS and remains stable after 5 months. Gb3 deposit are less somal diseases abundant in treated animals compared with not injected knock outs. Currently we are analyzing tissues from adult injected animals of Ron Padilla, Suleiman A. Igdoura, McMaster University, Hamilton, ON, 10 both sexes, which were treated at two doses (15-50x10 vg/Kg) and Canada ages (1 month or 3 months). Preliminary results indicate that rh-α- GalA is expressed and functional in all the analyzed tissues (brain, Lysosomal diseases are metabolic diseases which occur because of heart, kidney, liver) of the adult mice. These results confirmed that adeficiency of one of the essential lysosomal enzymes. A mutation in the vector lead the expression of rh-α-GalA when it is injected at the gene encoding one of these enzymes leads to an accumulation of early stages (P1) and possibly when injected in presymtomatic (1 unwanted substrates, resulting in a variety of clinical manifestations. month) or symptomatic (3 months) adults. We will analyze Lyso- A common symptom found in lysosomal diseases is skeletal muscle Gb3 (Mass_Spec), viral genome distribution and antibody production dysfunction, which includes muscle weakness, atrophy and loss of (by Feb19). This project is sponsored by the French patients muscle mass. One metabolic pathway which has been shown to be association Vaincre les Maladie Lisosomales. essential for maintenance of skeletal muscles is autophagy. This study sought to investigate the impact of lysosomal diseases on doi:10.1016/j.ymgme.2018.12.288 autophagy and how this may potentiate muscle dysfunction. We utilized in-vivo and in-vitro models of sialidosis, Sandhoff disease, and GM1-gangliosidosis to assess autophagy and its impact on myogenic differentiation in skeletal muscles. Our results demon- 273 strated that in lysosomal diseases, autophagy is induced upstream, Qualitative interviews with patients with mucopolysaccharidosis i.e., ULK1 phosphorylation but is hindered at the autophagosome to type I (MPS I) and caregivers to evaluate the Muco- lysosome fusion step, i.e., p62 accumulation which led to impairment polysaccharidosis Health Assessment Questionnaire (MPS-HAQ) of myoblast fusion and myogenic differentiation. We conclude that blocking autophagy impairs myogenic differentiation, which poten- a b c Anureet K. Pabla , Leighann Litcher-Kelly , Laurence Pollissard , tiates the muscle dysfunction observed in lysosomal diseases. This b b b b Andrew Yaworsky , Sarah Ollis , Masami Kelly , Brad Padilla , Alaa work highlights autophagy as a new pathway of interest and possible a a b Hamed , Sanofi Genzyme, Cambridge, MA, United States, Adelphi therapeutic target to alleviate muscle dysfunction in lysosomal c Values, Boston, MA, United States, Sanofi, Chilly-Mazarin, France diseases.

Mucopolysaccharidosis I (MPS-I) is a rare autosomal recessive genetic disorder that can have physical, social, and emotional impacts. doi:10.1016/j.ymgme.2018.12.290 The MPS Health Assessment Questionnaire (MPS-HAQ) was developed to assess these impacts and includes 52 items across 10 domains of activities of daily living (i.e., eating/drinking, dressing, bathing, 275 grooming, tooth brushing, toileting, mobility, walking, stairs, and Platform technology for treatment of the brain in lysosomal caregiver assistance). It is intended for patient or caregiver comple- diseases: Application to NCL1 Batten disease tion. Cognitive debriefing interviews were conducted with patients and/or caregivers to evaluate the readability, comprehensibility, relevance, and comprehensiveness of the MPS-HAQ. Interviews were William M. Pardridge, Eric K.-W. Hui, Jeff Z. Lu, Huilan Lin, Ruben J. semi-structured. Participants were asked to “think aloud” about how Boado, ArmaGen Inc., Calabasas, CA, United States they chose an answer for each MPS-HAQ item to identify any words or concepts they did not understand and to provide feedback on whether Batten disease has primary brain manifestations. Progress in the the items assess impacts experienced by individuals with MPS-I. All treatment has primarily been limited to the fact that recombinant interviews were audio-recorded, transcribed verbatim, and transcripts enzymes are large molecules that do not cross the blood-brain were coded for qualitative analysis. Interviews were conducted in the barrier (BBB). BBB-penetration of enzyme therapeutics is enabled by US via telephone representing the experiences of 19 individuals with re-engineering the recombinant enzyme as a bi-functional IgG- MPS-I. Among these individuals with MPS-I, ages ranged from 2-37 enzyme fusion protein, wherein the IgG domain is a monoclonal fi years (median age of 15 years) and over half were female (n=13, antibody (MAb) that targets a speci c endogenous receptor- 68.4%). Patients had a diagnosis of Hurler Syndrome (n=14, 73.7%) or mediated transporter within the human BBB, such as the human Hurler-Scheie (n=5, 26.3%). Preliminary analysis of the interview data insulin receptor (HIR). A proof of concept phase II clinical trial in indicates that patients and caregivers are able to interpret the content pediatric subjects with severe Hurler MPSI with a BBB penetrating of the MPS-HAQ as intended and that the MPS-HAQ evaluates concepts HIRMAb-iduronidase fusion protein (AGT-181, valanafusp alpha) relevant to the MPS-I experience. Formal qualitative data analyses produced stabilization of cognition, and reduced were conducted on the coded transcripts to evaluate readability, hepatosplenomegaly. The aim of the present investigation was to comprehensibility, relevance, and comprehensiveness of the MPS- determine if a similar BBB penetrating IgG-fusion protein can be HAQ with the addition of relevant participant quotations. The results engineered for the treatment of neuronal ceroid lipofuscinosis type 1 from the qualitative interviews support the content validity of the (NCL1), which is caused by mutations in the CLN1 gene encoding for MPS-HAQ for assessing the impacts experienced by patients with MPS- palmitoyl-protein thioesterase type 1 (PPT1). Genes encoding the I. Sponsor: Sanofi Genzyme heavy chain and the light chain of a HIRMAb-PPT1 fusion protein were engineered followed by stable transfection of Chinese hamster ovary (CHO) cells. The identity of the CHO derived HIRMAb-PPT doi:10.1016/j.ymgme.2018.12.289 fusion protein (AGT-194) was confirmed by hIgG1 and PPT1 Western blot analyses and the purity by SDS-PAGE and SEC-HPLC. Fusion Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S115 protein binding to the HIR was saturable with an ED50 in the low nM 277 range and comparable to the binding of the parental HIRMAb. The Rare GBA1 genotype in two siblings with a severe bone phenotype PPT1 activity in the HIRMAb-PPT1 fusion protein was comparable to of type 1 Gaucher disease recombinant PPT1. The brain uptake of HIRMAb-lysosomal enzymes in non-human primates approximates 1% of injected dose per brain. Livia D. Paskulin, Rodrigo T. Starosta, Vitoria S. Zizemer, Suelen This level of brain uptake is able to produce therapeutic levels of Basgalupp, Ida Vanessa D. Schwartz, Universidade Federal do Rio PPT1 enzyme activity in the brain. Successful development of a BBB Grande do Sul, Porto Alegre, Brazil penetrating HIRMAb-PPT1 fusion protein may be transformational for the treatment of the brain in NCL1 disease. Introduction: Gaucher disease type 1 (GD1) is a lysosomal storage disorder characterised by a variable phenotype ranging from oligosymptomatic patients to severe visceral and doi:10.1016/j.ymgme.2018.12.291 bone manifestations. Herein we present two cases of siblings diagnosed with GD1 with a rare genotype and a particular phenotype. 276 Case report: Patient 1 is a 47 years-old female diagnosed with fi Capturing the MPS IIIA patient and family voice in orphan drug GD1 when 42 after evaluation for hyperferritinaemia. At rst development to appreciate what is important in managing the evaluation she had chronic lumbar pain with Bone Marrow disease and improving quality of life Burdeen score = 14 (severe), mild splenomegaly, ferritin = 588.2ng/mL, normal platelets and Hb values, chitotriosidase activity = 9,609nmol/mL/h. Patient started therapy with miglustat Samantha Parkera, Sophie Oliviera, Anupam Chakrapanib, Roberto (since bone pain was a main feature of her phenotype) but Giuglianic, Bénédicte Herond, Nicole Muschole, Fofana Fofanaf, Frits changed to alfa-taliglucerase due to gastrointestinal adverse effects Wijburgg, aLysogene, Neuilly sur Seine, France, bGreat Ormond Street and worsening of hyperferritinaemia due to allergic reaction to Hospital, London, United Kingdom, cUFRGS and Medical Genetics alfa-taliglucerase infusion, therapy was changed to imiglucerase Service, HCPA, Porto Alegre, Brazil, dHôpital Armand Trousseau La (current treatment). Patient 2, the older sister of patient 1, is 50 Roche Guyon, Paris, France, eUniversity Medical Center, Hamburg, years old and was diagnosed with GD1 when 45 after an Germany, fMAPI, an ICON PLC Company, Amsterdam, Netherlands, osteonecrosis of the left femur and a total hysterectomy for gAcademic Medical Center, Amsterdam, Netherlands uncontrollable bleeding. At first evaluation, she had bone pain Mucopolysaccharidosis type IIIA (MPS IIIA) (OMIM 252900) is a with a Bone Marrow Burden score = 14 (severe), mild rare, neurodegenerative, pediatric and lethal disease. Understanding splenomegaly, anaemia (Hb = 11.5 g/dL), normal platelets count, the impact of the disease and the needs of patients and families chitotriosidase activity of 2970nmol/mL/h. Patient started treat- related to treatment benefits, risks and disease burden is an ment with miglustat, and after 48 months changed to alfa- important addition to assess treatment effect in clinical trials. taliglucerase due to bone pain and platelets decrease. Upon Lysogene launched an observational study for MPS IIIA in five genotyping with next-generation sequencing, both patients were countries (NCT02746341), to function as a non-concurrent control discovered to be compound GBA1 heterozygotes for E349K and for their phase 2-3 pivotal gene therapy trial (NCT03612869). The S366N mutations. fi observational study includes mixed methods research using face-to- Conclusion: To our knowledge, this is the rst case presented face interviews, questionnaires, and digital health technology. of the GBA1 genotype E349K/S366N in patients with GD. The case Twenty-three children were included in the study and twenty-two is remarkable in that the bone disease is the most prominent parents were interviewed at baseline. Data from nineteen patients feature in the phenotype, with both patients presenting BMB and sixteen parent interviews are available at 12 months. Patient age scores as high as 14, and in that visceral and haematological ranged from 2 to 10 years mean age at baseline was 61 months manifestations are only mild. (range 28 – 105). Four children had been recently diagnosed and aged 2, 4, 4, and 6 years. The age range was important in providing doi:10.1016/j.ymgme.2018.12.293 an accurate development story covering the journey from diagnosis to the age of ten, highlighting issues that each group may face. At baseline, 43% of patients reported sleep disorder. Eight children were taking melatonin. Actigraphy showed overall stability over the 278 course, with high inter- and intra-patient variability. The Child NEO1 and NEO-EXT studies: Long-term safety of repeat Behavior Checklist scores showed 30%, 50% and 60% of children had avalglucosidase alfa dosing for 4.5 years in late-onset Pompe behavior scores for internalizing, externalizing and total problems disease patients above the 90e percentile, respectively. Externalizing scores increased over the study period. Data shows well-preserved Health Utility Loren Penaa,1, Richard J. Barohnb, Mazen M. Dimachkieb, Priya S. Index (HUI) scores in patients with high cognitive DQ and lower Kishnanic, Shafeeq Ladhad, Karl-Eugen Mengele, Sabrina Sacconif, scores in patients with more advanced cognitive impairment. The Philip Van Dammeg, John Vissingh, Kristina An Haacki, Christopher parent interviews confirmed two of the main challenges of this Hugj, Judith Johnsonj, Charlotte Sensingerj, Benedikt Schoserk on disease: disturbed sleep and behavior. The presentation will include behalf of the NEO-EXT investigators, triangulating the qualitative data with data from the QOL, sleep, aDuring study participation: Duke University Medical Center, Durham, cognitive and behavior questionnaires. NC, United States, bUniversity of Kansas Medical Center, Kansas City, KS, United States, cDuke University Medical Center, Durham, NC, United States, dBarrow Neurological Institute, Phoenix, AZ, United States, eVilla doi:10.1016/j.ymgme.2018.12.292 Metabolica, Centre for Pediatric and Adolescent Medicine, University S116 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Medical Center, Mainz, Germany, fNeuromuscular Diseases Centre, Mutations in GBA1, which encodes the lysosomal acid beta- Department of Clinical Neurosciences, University Hospital of Nice glucosidase (GCase), cause GCase deficiency in Gaucher disease (CHU), Nice, France, gKU Leuven (Catholic University of Leuven) - (GD) and is the most common genetic risk factor for Parkinson Department of Neurosciences, VIB - Center for Brain & Disease Research, disease (PD). There are no effective treatments available for University Hospitals Leuven - Department of Neurology, Leuven, neuronopathic GD and PD that can stop or slow neurodegeneration. Belgium, hCopenhagen Neuromuscular Center, Rigshospitalet, University In this study, we evaluated a novel non-invasive cell transplantation of Copenhagen, Copenhagen, Denmark, iSanofi Genzyme, Chilly-Maza- therapy in a GD mouse model (9H/PS-NA) exhibiting alpha- rin, France, jSanofi Genzyme, Cambridge, MA, United States, kFriedrich- synuclein aggregation, a key PD-relevant phenotype. A subclass of Baur-Institut, Department of Neurology Klinikum München, München, iPSC-derived neural progenitor cells (NPCs) expresses VLA4 which Germany; 1Current affiliation: Cincinnati Children's Hospital Medical allows systemically-delivered NPCs to cross the blood-brain-barrier. Center and University of Cincinnati College of Medicine, , Cincinnati, OH, We established VLA4+ mouse iPSC-derived NPCs harboring United States. lentiviral-mediated overexpression of wild-type human GCase (hGCase). In-vitro, VLA4+ hGCase+ NPCs were expandable without Safety, tolerability, pharmacokinetics, pharmacodynamics, and loss of multipotency and secreted GCase which was taken up by exploratory efficacy of repeat avalglucosidase alfa dosing (5, 10, or lysosomes of adjacent Gba1-null cells. In-vivo efficacy was evaluated 20 mg/kg qow) for 6 months were evaluated in NEO1 in 9H/PS-NA mice. VLA4+ hGCase+ NPCs were intravenously (NCT01898364) in late-onset Pompe disease patients either treat- administered by weekly tail-vein injection and mice were analyzed ment-naïve (Naïve) or having received alglucosidase alfa for ≥9 at 14 weeks of age for effects on neuropathic and PD phenotypes. months (Switch). In NEO-EXT (NCT02032524), an ongoing NEO1 Injected cells engrafted throughout the brain, including thalamus, extension, long-term safety and pharmacokinetics of repeat cortex, brainstem and midbrain, and differentiated into neural avalglucosidase alfa dosing will be monitored over 6 years. Interim lineages. Human GCase protein was detected in the transplanted safety results after 4.5 years of NEO-EXT are reported here. Mean mouse brains with specific anti-human GCase antibody. Histological ages at NEO1 enrollment were: Naïve: 44.8 (SD:20.3, range:20-78) analyses of brain sections showed reduced neurodegeneration by years and Switch: 46.7 (SD:14.1, range:21-68) years. Of the 24 NEO1 Fluoro-Jade C staining in the regions of NPC migration. CNS participants, 19 entered NEO-EXT (8/10 Naïve 11/14 Switch) 17 are inflammation, detected by anti-CD68 and anti-GFAP antibodies, was currently participating (7/8 Naïve 10/11 Switch) 2 participants significantly decreased in the brain of transplanted mice. Compared discontinued NEO-EXT (personal reasons). During 2016, all NEO- to vehicle treated mice, VLA4+ hGCase+ NPC-transplanted mice EXT participants switched to avalglucosidase alfa 20 mg/kg qow. By showed ~50% reduction of α-synuclein aggregates in the substantia May 2018, total infusions reached 731 (Naïve) and 1174 (Switch) nigra. Together, these results demonstrate the efficacy of non- mean infusions were Naïve: 73 (SD:43, range:9-113) and Switch: 84 invasive delivery of iPSC-derived NPCs overexpressing hGCase in (SD:40, range:8-118). Mean avalglucosidase alfa exposure duration GD mouse model and establish the feasibility of combined cell and reached 1025 (SD:611, range:109-1572) days for Naïve and 1179 gene therapy for GBA1-associated PD. This approach provides a (SD:570, range:102-1658) days for Switch participants. Treatment- potential treatment for both rare and common neurological diseases. emergent adverse events (AEs) were generally mild across doses. Most frequently reported treatment-related AEs were nausea, fatigue, and headache (3 participants each) and dizziness, dyspnea, doi:10.1016/j.ymgme.2018.12.295 erythema, myalgia, muscle spasms, and rash (2 participants each) and are unchanged since the last safety update (WORLDSymposium™ 2018). One Switch participant discontinued 280 NEO1 for a study drug-related serious AE (respiratory distress/chest Case control study to identify the prevalence of menstrual and discomfort) no deaths/life-threatening serious AEs have occurred. pregnancy complications in women with mucopolysaccharidosis Anti-avalglucosidase IgG/IgM antibodies developed in 9/10 Switch (median titer:1600, range:200-51,200) and 9/14 Naïve (median Merlene Peter, Stephanie Cagle, Emory University, Atlanta, GA, United titer:400, range:50-12,800) participants. No patient developed IgE States antibodies or tested positive for enzyme activity inhibition. In NEO- EXT, the avalglucosidase alfa safety profile is consistent with the Mucopolysaccharidosis (MPS) are a group of inherited lysosomal initial 6 months’ treatment during NEO1, with no additional storage disorders which are caused due to deficiencies in enzymes participants developing anti-avalglucosidase alfa IgG/IgM antibodies leading to accumulation of glycosaminoglycans. There is extensive or new allergic reactions. Funding: Sanofi Genzyme. literature available on the systemic impact of MPS. However, there is adeficit in the number of publications about the prevalence of doi:10.1016/j.ymgme.2018.12.294 gynecological problems in women with MPS. In this study, we aim to identify if there is a difference in the menstrual patterns, gynecological problems, and pregnancy complications in women with MPS in comparison to the control population. The study is designed as a case 279 control study in which the cases are women diagnosed with MPS Intravenous infusion of iPSC-derived neural progenitors express- over the age of 12 who have begun menstruating, and the controls ing GCase ameliorates alpha-synuclein aggregates in a mouse are women over the age of 12 without a diagnosis of MPS. Women model of Gaucher disease who underwent a hematopoietic stem cell transplant were excluded from the study as the drugs used for this procedure could influence Yanyan Penga, Benjamin Lioua, Venette Inskeepa, Rachel Blackwooda, menstruation and fertility. The study was conducted in the form of a Sheila Flemingb, Christopher N. Mayhewa, Ying Suna, aCincinnati 34-question survey. The survey is divided into questions regarding Children's Hospital Medical Center, Cincinnati, OH, United States, demographics, menstruation and pregnancy outcomes. The men- bNortheast Ohio Medical University, Rootstown, OH, United States strual questionnaire was scored, and chi square analysis was Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S117 performed to analyze the differences between the case and control 282 group. Data regarding pregnancy complications, age at menarche and Safety, tolerability and pharmacokinetics of oral venglustat in use of enzyme replacement therapy were portrayed using descrip- Parkinson disease patients with a GBA mutation tive statistics. The results of the study will be presented at the conference. Through this study we shed light on the menstrual and M. Judith Peterschmitta, Thomas Gasserb, Stuart Isaacsonc, Jaime pregnancy complications found in women with MPS. We hope that Kulisevskyd, Pablo Mire, Tanya Simunif, Anne-Marie Willsg, Leonor this study will be the first step towards identifying any effect MPS Correia Guedesh, Per Svenningssoni, Cheryl Watersj, Allena J. Jik, Jian disease may have on the reproductive system. Lia, Pascal Mininil, Blandine Nembom, Stéphane Saubadul, Jyoti Sharmam, Tanya Fischera, aSanofi Genzyme, Cambridge, MA, United States, bUniversitätsklinikum Tübingen, Tübingen, Germany, doi:10.1016/j.ymgme.2018.12.296 cParkinson’s Disease and Movement Disorders Center, Boca Raton, FL, United States, dHospital de la Santa Creu i Sant Pau, Barcelona, Spain, eInstituto de Biomedicina de Sevilla (IBiS), Hospital Universitario 281 Virgen del Rocío/CSIC/Universidad de Sevilla, Sevilla, Spain, f g Correlations between glucosylsphingosine (lyso-GL-1) and base- Northwestern University, Chicago, IL, United States, Massachusetts h line disease severity as well as response to treatment in two General Hospital, Boston, MA, United States, Hospital de Santa Maria, i clinical trials of eliglustat in treatment-naïve adults with type 1 Lisbon, Portugal, Karolinska Universitetssjukhuset Huddinge, Stock- j Gaucher disease holm, Sweden, Neurological Institute, Columbia University Medical Center, New York, NY, United States, kSanofi Genzyme, Framingham, l fi m fi M. Judith Peterschmitta, Meredith Fostera, Kate Zhangb, Allena Jia, MA, United States, Sano , Chilly-Mazarin, France, Sano , Bridgewa- Gerald Coxb, aSanofi Genzyme, Cambridge, MA, United States, bEditas ter, NJ, United States Medicine, Cambridge, MA, United States Mutations in GBA are associated with increased risk of developing Patients with Gaucher disease type 1 have deficient acid β- Parkinson disease (PD), characterized by younger onset, higher glucosidase activity resulting in accumulation of the major substrate prevalence of cognitive impairment, and more rapid disease glucosylceramide (GL-1) and the minor substrate progression. Part 1 of the phase 2 MOVES-PD study (NCT02906020) glucosylsphingosine (lyso-GL-1), the deacylated form of GL‑1. Lyso- was an up to 36-week randomized, placebo-controlled, double-blind, GL-1 is increasingly recognized as a key biomarker of Gaucher sequential cohort study of once-daily venglustat at 3 escalating ≥ disease, as elevations are highly specific to the disease and in the doses. PD patients age 18-80 years with symptoms 2 years and ≤ causal pathway. We report correlations between plasma lyso-GL-1 Hoehn & Yahr stage 2 at baseline who were heterozygous carriers of and baseline disease burden and treatment response to oral a GBA mutation were eligible. The primary endpoint was the safety eliglustat in two Sanofi-Genzyme-sponsored trials (with extensions) and tolerability of venglustat. Secondary endpoints included plasma fl of treatment-naïve adults with Gaucher disease type 1 (GD1): an and cerebrospinal uid (CSF) pharmacokinetics (PK). Exploratory open-label Phase 2 trial in patients with moderate to severe disease endpoints included pharmacodynamics in plasma and CSF. Seven- with 8-year follow-up (NCT00358150) and a placebo-controlled teen patients (13M, 4F) were randomized to placebo (n=4) or Phase 3 trial in patients with mild to moderate disease with up to venglustat (n=13). Mean age at enrollment was 58.4 years. Mean 4.5-year follow-up (ENGAGE, NCT00891202). At baseline, all patients years since symptom onset was 6.7, and since diagnosis was 5.2. in both trials had substantial elevations of lyso‑GL-1, with median Twelve patients on venglustat and 4 on placebo reported at least 1 values 119 times(x) the upper limit of normal (range: 29x-314x) and treatment-emergent adverse event (TEAE) most were mild or 61x the upper limit of normal (range: 12x-208x) in Phase 2 (N=24) moderate and resolved without corrective treatment during the and ENGAGE (N=38), respectively. Moderate-to-strong baseline study. The most common TEAEs were psychiatric, neurological, and correlations were observed for lyso-GL-1 with spleen volume, liver gastrointestinal events consistent with common motor/non-motor volume, and hemoglobin levels (spleen: r=0.49 and r=0.76 liver: PD symptoms or known side effects of concurrent PD medications. r=0.43 and r=0.72 hemoglobin: r= ‑0.53 and r= ‑0.49 in Phase 2 No serious AEs or deaths occurred. Two patients on venglustat and ENGAGE, respectively all pb0.05) but not with platelet count discontinued due to TEAEs after the primary analysis period (Week (r= ‑0.18, p=0.40 r= ‑0.29, p=0.08). After eliglustat treatment, 4). Venglustat exposure in plasma and CSF increased in a close to median lyso-GL-1 decreased by 92% in Phase 2 and 84% in ENGAGE. dose-proportional manner. Plasma and CSF glucosylceramide (GL-1) Change in lyso-GL-1 correlated with change in clinical parameters levels decreased from baseline in a dose-dependent manner over 4 after treatment (spleen: r=0.72 and r=0.63 liver: r=0.28 and weeks. CSF GL-1 decreased 74.3% (higher dose). The data demon- fi r=0.57 hemoglobin: r= ‑0.65 and r= ‑0.34 and platelets: r= ‑0.26 strate a favorable safety and tolerability pro le of venglustat at all and r= ‑0.18, in Phase 2 and ENGAGE, respectively all pb0.05). Our doses investigated for up to 36 weeks of treatment. Dose-dependent data, showing marked elevations in lyso-GL-1 in all patients before plasma and CSF exposure and reduction of plasma and CSF GL-1 treatment, correlations with baseline disease, and correlations with were observed. Part 2 of MOVES-PD, a 52-week randomized, double- fi improvements after treatment, underscore the utility of lyso-GL-1 as blind, placebo-controlled study, is ongoing. Funding: Sano Gen- a clinically useful biomarker for GD1. zyme. doi:10.1016/j.ymgme.2018.12.297 doi:10.1016/j.ymgme.2018.12.298 S118 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

283 hypertrophic cardiomyopathy, stroke and/or proteinuria developed Modeling CLN6 with IPSC-derived neural cells in all males after the age of 30. In females the severity of Fabry disease did not vary from classic multiorgan disease (60% of females) Tyler Mark Pierson, Maria Gabriella Otero, David Fabian Nonis, to a mild disease as expected but from a classic disease to a disease Jaemin Kim, Cedars-Sinai Medical Center, Los Angeles, CA, United States that mimiced late onset cardiac variant (40% of females). No one was totally asymptomatic and without any sings of Fabry disease. Cardiac Neuronal ceroid lipofuscionosis type 6 (CLN6) is a neurodegen- MRI was the most sensible method to detect early cardiac erative disease associated with dementia, seizures, and retinopathy. involvement. It was performed in eight patients of 14. Of those, four The disorder is due to mutations in the CLN6 gene encoding a (50%) had late gadolinium enhancement (LGE) in the typical location resident ER transmembrane protein of unknown function. Similar to in posterolateral wall. Cardiac symptoms and wall hypertrophy in other NCLs, the cellular pathology associated with CLN6 includes the ultrasound and in electrocardiography became evident between 30- abnormal accumulation of autofluorescent storage material, with 60 years of age. Cardiac involvement should effectively be detected subunit C of the mitochondrial ATP synthase (SUBC) being the in females before judging them as asymptomatic mutation carriers or predominant component. Four CLN6 patient-derived IPSC lines were Fabry patients with so mild disease that they do not need regular generated and differentiated into neuroprogenitor cells (NPCs), follow-up. In males the mutation R227X caused a classic Fabry cortical neurons and oligodendroglia in order to model human disease as expected but in females the disease course was more CLN6. CLN6-IPSCs and CLN6-NPCs did not possess any overt storage variable. material and were similar to control cells with regards to subcellular organelle morphology and organization. IPSC-derived CLN6-neurons were generated with two neuronal differentiation protocols, a more doi:10.1016/j.ymgme.2018.12.300 conventional dual-SMAD protocol and an inducible neurogenin-2 (I3N) protocol. CLN6-neuronal cultures differentiated with the dual- SMAD protocol consisted of a mixed population of neural cells 285 (mitotic NPCs ad post-mitotic neurons) and had significantly Intranasal delivery of the CRISPR-Cas9 system for gene editing in increased SUBC+ puncta after ~30-40 days in culture compared to MPS II mice control lines. Alternatively, the I3N-CLN6 cultures generated post- mitotic neurons in only 3-5 days (consisting of N95% TUJ1+ cells) that had accumulated SUBC+ puncta during the same period. Luisa Pimentel, Esteban Gonzalez, Angela Tavares, Roselena Schuh, Preliminary studies with IPSC-derived oligodendroglia from CLN6 Edina Poletto, Graziella Rodrigues, Paola Barcellos Carneiro, Helder lines also had increased SUBC+ puncta. These findings indicate that Teixeira, Roberto Giugliani, Guilherme Baldo, Hospital de Clínicas de molecular markers for this disorder can be monitored in IPSC- Porto Alegre, Porto Alegre, Brazil derived neural cells. Differentiated neurons and oligodendroglia are undergoing further phenotypic characterization and RNA evaluation The CRISPR-Cas9 system allows precise gene editing, being a to identify other cellular markers of CLN6 disease. These molecular potential new gene therapy strategy to treat lysosomal disorders. In markers can then be used to measure efficacy of therapeutic this present study, we used a lipossomal vector applied nasally fi interventions including small molecule compounds, anti-sense aiming to correct enzyme de ciency in MPS II mice (caused by fi oligonucleotides, and gene therapy. de ciency of iduronate-sulfatase or IDS). Lipossomes were instilled in the nostril of young adult MPS II mice for 30 days. Mice were sacrificed at 6 months of age to evaluate IDS activity in the tissues. As doi:10.1016/j.ymgme.2018.12.299 control groups, we compared the results with wild-type (normal) and untreated MPS II mice. We found a significant increase in IDS activity in the heart and the lungs compared to untreated mice, although enzyme levels were still lower than those from normal 284 mice. We also evaluated different brain areas, and IDS activity was The natural course of Fabry disease in a Finnish R227X cohort elevated in the olfactory bulb, but not in the cortex, cerebellum or hippocampus. Based on these results, we conclude that the nasal a a b Päivi Pietilä-Effati , Jukka T. Saarinen , Eliisa Löyttyniemi , Reijo route allows the lipossomes with the CRISPR-Cas9 system to reach a c c a Autio , Maria Saarenhovi , Ilkka Kantola , Vaasa Central Hospital, the circulation and also the brain, although the distribution of the b c Vaasa, Finland, University of Turku, Turku, Finland, Turku University vector in the central nervous system is still limited. Current studies Hospital, Turku, Finland are aiming to identify if the improvements observed here lead to reduction of neuroinflammation or alterations in behavior. Fabry disease caused by a genetic deficiency of alfagalactosidase activity results in accumulation of glycosphingolipids especially in fi vascular endothelium. Progressive organ dysfunction signi cantly doi:10.1016/j.ymgme.2018.12.301 shortens life span in males and in less extent in females. In general, classical mutations cause more severe disease than late onset mutations. Here we describe the natural course of a common, classic Fabry disease causing mutation R227X in Finland. The cohort 286 consisted of four males and ten females in one extended family Exploring surrogate biomarkers of skeletal and joint disease making this cohort very homogenous. The mean age was 46 years progression in mucopolysaccharidosis type I (SD 18.4). Six patients (43% of all patients) had severe cardiomyopathy, 3 patients (21%) had ischemic stroke but no one had severe Lynda E. Polgreena, Jeremiah Menkb, Kyle Rudserb, Ellen B. Fungc, kidney dysfunction. Three patients (21 %) had atrial fibrillation and Bradley S. Millerb, Klane Whited, Paul J. Orchardb, Troy C. Lundb, aLos two of those (14% of all patients) had also pacemaker. Severe Angeles Biomedical Research Institute at Harbor-UCLA, Torrance, CA, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S119

United States, bUniversity of Minnesota, Minneapolis, MN, United enzyme reactions to be combined in a single reaction, increasing States, cChildren's Hospital Oakland Research Institute, Oakland, efficiency. We report validation results for a new 6-plex assay for the CA, United States, dSeattle Children's Hospital, Seattle, WA, diagnosis of five Mucopolysaccharidosis (MPS) disorders (MPS II, United States IIIB, IVA, VI and VII) and neuronal ceroid lipofuscinosis type 2 (CLN2) in DBS using UPLC-MS/MS. Normal reference ranges were developed Background: Skeletal and joint disease persists and slowly using a minimum of 236 DBS samples from unaffected controls or worsens in individuals with mucopolysaccharidosis type I (MPS I) patients with an alternate diagnosis, and 100% clinical sensitivity was despite treatment with hematopoietic cell transplantation (HCT) established using 99 samples from patients affected with one of the and/or enzyme replacement therapy (ERT). Surrogate biomarkers are six disorders. Both intra-day and inter-day precision were acceptable crucial for efficient testing of the effects of new and adjunctive (b 20% CV) and 6 month long term sample stability has been verified. therapies on bones and joints in MPS I. Our laboratory can now analyze 18 enzymes in DBS (12 utilizing MS/ Methods: As part of a 9-year observational study of individuals MS substrates and 6 utilizing 4-MU substrates) to diagnose 20 with MPS I, hip, and spine radiographs, and range-of-motion different LSDs (including Multiple Sulfatase Deficiency and (ROM) were measured annually in years 7-9 and height in years Mucolipidosis II/III). Furthermore, we can now offer a 7 enzyme 1-9. Fasting morning plasma and urine samples of inflammatory MPS panel in DBS, which will significantly enhance the worldwide and bone biomarkers were measured by ELISAs in years 1 and 2. A diagnosis of MPS patients. Finally, the development of this 6-plex has novel summary score for skeletal disease (i.e. hip and spine significant implications for newborn screening, as FDA approved radiographs and height Z-score standardized by age and sex) and therapies are available for five of the six included conditions. joint disease (i.e. ROM in shoulders, elbows, and knees) were calculated. Linear regression, adjusting for age, was used to test for doi:10.1016/j.ymgme.2018.12.303 associations of change in biomarkers over the first year with the annualized change in summary bone and ROM score, and height Z-score over years 7-9 and 1-9 respectively. Sensitivity analyses removed biomarkers greater than 3 standard deviations from the 288 mean. Utility of multiple myeloma screening in Gaucher disease Results: The cohort included eighteen participants (MPS IH N=14, MPS IA N=4), age 5-16 years, who were 6.7±3.0 years Carlos Prada, Katherine Abell, Sarah Chadwell, Laurie Bailey, since HCT (MPS IH) or 7.1±2.8 years since ERT initiation (MPS Cincinnati Children's Hospital, Cincinnati, OH, United States IA). Sensitivity analysis showed an association between increasing interleukin-6 and more rapid worsening of ROM (-31 Gaucher disease (GD) is a lysosomal storage disease characterized pg/ml: 95%CI -45 to -18 pg/ml pb0.001) and between increasing by deficiency of glucocerebrosidase within lysosomes, altering urine pyridinolines (PYD) and more rapid decrease in height Z- degradation of glycosphingolipids, which causes accumulation of score (-3.9e-4 nmol/mmol creatinine: 95%CI -6.8e-4 to -0.9e-4 glycosylceramide. Patients with GD type 1 (GD1) have an increased p=0.013). incidence of gammopathy (i.e., hypergammaglobulinemia, monoclo- Conclusions: We have identified two potential candidate nal gammopathy of undetermined significance [MGUS]), and multi- surrogate biomarkers of change in skeletal or joint disease over ple myeloma (MM) when compared to the general population. No time. Once validated, these biomarkers may prove useful in formal evaluation of gammopathy screening has been reported in the determining early efficacy of skeletal or joint directed therapies literature. We conducted an IRB approved retrospective analysis of in MPS I. clinical and laboratory data, including screening for MM, in adults with GD at Cincinnati Children’s Hospital Medical Center between 2010 and 2018. During the 8-year study period, 20 adults with GD doi:10.1016/j.ymgme.2018.12.302 (11 females/9 males) ages 19 to 71 (mean = 51.95 years) were screened for MM using serum protein electrophoresis (SPE) and immunofixation (IFE). Of those, 10 were treated with substrate reduction therapy (SRT) and 9 with enzyme therapy (ET). We 287 identified no patients with MM, 2 with MGUS, and 1 with persistent Multiplex DBS enzyme assay for MPS II, IIIB, IVA, VI, VII and CLN2 polyclonal gammopathy. The 2 MGUS patients were 48 and 61 years- via LC-MS/MS expands clinical utility of DBS enzyme testing old, female and male respectively, with intact spleens. SPE and IFE follow-up has been stable without progression. The polyclonal Laura Pollard, Tim Wood, Greenwood Genetic Center, Greenwood, SC, gammopathy patient was a 54-year-old splenectomized male. All United States patients with abnormal SPE or IFE were on therapy (2 ET/ 1 SRT). Only one abnormal patient’s GBA genotype was known (p. Enzyme analysis is the gold standard for the diagnosis of Asn409Ser, p.Leu483Pro). Other SPE abnormalities included slight lysosomal storage disorders (LSDs). However, enzyme analysis in increases/decreases in albumin, protein, and beta globulin, or slight leukocytes requires that whole blood samples arrive to the testing decreases in alpha 2 or gamma globulin without clinical significance. laboratory within 24-48 hours of collection, and requires a relatively Family history of cancer was documented in 10 individuals, with no large volume of blood. This is problematic for the international MM or blood-related malignancies reported. Our data show that as in shipment of specimens and for testing infants. The adaptation of the general population and previous GD studies, gammopathies are these enzyme assays for use in dried blood spots (DBS) has more common in patients greater than 50 years. These data support ameliorated these issues for a large number of LSDs. Initially enzyme screening in GD1 for gammopathy greater than age 40 years. analysis in DBS exclusively utilized 4-methylumbelliferone (4-MU) fluorogenic substrates. The limitation of this methodology is that each enzyme must be measured individually. The development of doi:10.1016/j.ymgme.2018.12.304 tandem mass spectrometry (MS/MS) substrates has allowed multiple S120 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

289 proteins (including multiple Batten disease proteins) in 5 years. We Evaluation of lentiviral vectors in Morquio syndrome type A will make enough protein to test the most promising candidates in patients' fibroblasts commercially available patient derived cells to determine if they can return enzyme activity in vitro and enable in vivo testing in the Alejandra Puentes-Telleza, Felipe Rojas-Rodrígueza, Diego Suarezb, appropriate knock out mouse models with a collaborator network. Oscar A. Hidalgoa, Carlos J. Almeciga-Diaza, aPontificia Universidad The ultimate goal is to create a sustainable pipeline that leverages Javeriana, Bogota, Colombia, bUniversidad Nacional de Colombia, the decades of insight and research into developing ERTs, and Bogota, Colombia proteins in general. We anticipate that the development of new ERTs might open new avenues for a future lysosomal storage disease Mucopolysaccharidosis IVA (Morquio syndrome type A) is a patient cell line bank that would be useful for testing ERTs that have lysosomal storage disease produced by deficiency in the N- been developed and for screening to identify small molecule acetylgalactosamine-6-sulfate sulfatase (GALNS) enzyme, leading to chaperones. Besides these benefits, scaling ERT development for lysosomal accumulation of keratan sulfate and chondroitin-6-sulfate. more diseases might also accelerate the development of diagnostics Although enzyme replacement therapy has shown impact in some and earlier routine screening of these diseases once more treatment disease manifestations, it has limited impact in bone and cardiac options are available. Our approach to creating a pipeline for protein problems and requires a weekly infusion of the recombinant enzyme. development could accelerate treatment development and reinvig- The use of gene therapy represents an alternative that can help to orate the pipelines of biotechs with ERTs. overcome some of these current therapies limitations. In this work, we evaluated the use of lentiviral vectors to mediate the transduction of GALNS (lenti-GALNS) and SUMF1 (lenti-SUMF1) cDNAs to fibroblasts derived from three Morquio syndrome type A patients. doi:10.1016/j.ymgme.2018.12.306 Vector transduction was monitored by intra and extracellular GALNS activity, lysosomal mass by LysoTracker staining, and autophagy evaluation by monitoring p62 and LC3B. The results showed an 291 increase in GALNS activity after lenti-GALNS transduction and, in Developing an enzyme replacement therapy and small molecule some cases, a higher increase after lenti-SUMF1 co-transduction. We chaperone combination as a potential treatment for multiple observed a wide cell-to-cell variation in GALNS activity, with sulfatase deficiency fibroblasts with severe mutations showing a higher increase in enzyme activity. In addition, we observed that high vector titers and Ana C. Puhl, Ekins, Collaborations Pharmaceuticals, Inc., Raleigh, NC, co-transduction with lenti-SUMF1 showed a deleterious effect in United States GALNS activity in some treated fibroblasts. Noteworthy, it was observed that the treatment with the lentiviral vectors allowed a Multiple sulfatase deficiency (MSD) is a fatal, inherited lysosomal reduction in lysosomal mass, as observed by changes in the disease characterized by reduced activities of all 17 sulfatases in LysoTracker staining, and a change in the autophagy biomarkers patients. It combines features of individual sulfatase deficiencies such p62 and LC3B. Overall, this study offers valuable data towards the as metachromatic leukodystrophy, mucopolysaccharidosis, cho- use of lentiviral vectors as tools for the design of a gene therapy ndrodysplasia punctata, hydrocephalus, ichthyosis, neurologic dete- fi strategy for Morquio syndrome type A patients, and shows the rst rioration and developmental delay. Sulfatases are activated by post- evidence of the autophagy impairment in Morquio syndrome type A translational modification of a conserved cysteine in their active site fi broblasts. to the catalytic Ca-formylglycine (FGly) residue in the endoplasmic reticulum (ER) by the formylglycine-generating enzyme (FGE), codified by the SUMF1 gene. To date, about 40 different MSD- doi:10.1016/j.ymgme.2018.12.305 causing SUMF1mutations have been reported, and the majority of these were predicted to affect the intracellular stability of FGE. FGE variants carrying MSD causing missense mutations showed clearly 290 detectable activity in vitro. In the cell, however, these variants were Industrializing enzyme replacement therapy development rapidly degraded, which established that reduced intracellular stability is the major determinant for severity in MSD manifestation. Ana C. Puhl, Sean Ekins, Collaborations Pharmaceuticals, Inc., Raleigh, An emerging strategy for the treatment of lysosomal diseases is NC, United States pharmacological chaperone therapy (PCT), based on the use of molecules that assist the folding of mutated enzymes and improve Today there are 10 FDA approved enzyme replacement therapy their stability and lysosomal trafficking. We are evaluating the (ERTs), yet there are thousands of rare diseases where ERTs could possibility of combining enzyme replacement therapy (ERT) with extend patient lives. The pathway to develop ERTs is well pharmacological chaperones to rescue mutated protein activity. understood. Besides the high costs of GMP manufacturing, we think Initially, we tested the enzyme FGE expression in bacteria and the development of ERTs is being inhibited at the preclinical stage by mammalian cell systems. The protein was purified by ion-exchange the current research paradigm focusing on individual groups chromatography and gel filtration. FGE activity assays was performed working on a ‘single protein-single lab’ approach until a point where as described in the literature, using the synthetic ASA-derived they have proof of concept data. We could learn from other areas peptide P23 as substrate. Patient’s fibroblasts carrying the mutation using higher throughput approaches, such as the Structural Geno- A279V were treated with the recombinant FGE (as an ERT) and mics Consortium where over 1500 proteins have been produced in sulfatase activity measured. Additionally, we screened small mole- an efficient collaborative manner and shared. We now propose to cules as potential chaperones in patient’s fibroblasts carrying the integrate several commercially available technologies together to mutation A279V to verify if they could rescue protein activity. The create a pipeline that will allow us to develop at tens to hundreds of discovery of such molecules that could stabilize FGE, may lead to Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S121 future studies to evaluate the combination of ERT+PCT in vitro and monthly at the nationally designated lysosomal storage disorders in vivo. unit at Royal Free London NHS Foundation Trust, UK. 207/375 patients are on DMTs. Migalastat became available for use in the UK in July 2017. As of August 2018, we report data for 75 patients on doi:10.1016/j.ymgme.2018.12.307 migalastat. Median age in males 59.1 years (21.5 to 79.2 years) and in females 55.7 years (19.0 to 75.2 years), treatment naïve (n=16) and switched from ERT (n=59). Prior to switch, median duration on ERT was 6.5 years (range 0.7 - 15.4 years). Baseline data on 292 demographics, genetics, renal, cardiac, biomarkers, treatment, reason fi Molecular description of the rst case of Hunter syndrome for switch and side effects were systematically recorded. Migalastat burdened with Turner syndrome in Russian girl was prescribed at 123mg every other day as per the SmPC. 42/75 had the GLA cardiac variant missense mutation N215S (c.644ANG). At Alexander Pushkov, Kirill Savostyanov, Sergey Trufanov, Dmitry baseline, cardiac ECHO: left ventricular hypertrophy in 16/26 Sladkov, Natalya Zhurkova, Leyla Namazova-Baranova, FSAI, females and 24/28 males and where available, Lysogb3 was National Medical Research Center for Children's Health, Moscow, abnormal (N1.8 ng/ml) in 22/48. The median duration on migalastat Russian Federation was 0.8 years (range 0.1 to1.2 years). The reasons for switch included convenience, difficult venous access, infusion associated Hunter syndrome is the X-linked recessive hereditary disorder reactions to ERT. Common side effects included headaches and caused by IDS gene mutations. Clinical implications of Hunter gastrointestinal symptoms, in the first few weeks. 4 patients stopped syndrome develop in women in very rare cases and can be caused due to side effects. One-year follow up data will be presented on by structural anomalies of X-chromosome, homozygous mutations of patients receiving migalastat. the IDS gene or X-inactivation. We represent a case of molecular genetic and molecular cytogenetic diagnostics of Hunter syndrome doi:10.1016/j.ymgme.2018.12.309 burdened with Turner syndrome in Russian girl. The whole blood sample from the girl at the age of 4 years 5 months with clinical implications of Turner syndrome and Hunter syndrome was brought 294 for molecular analysis in our department. At the first stage of Fabry disease mobile phone application - A new service improve- diagnostics the array CGH analysis was carried out, as a result the ment tool molecular karyotype arr(1-22)x2,(XX)x1 was defined. The extended deletion of chrX:100,015,846-155,199,833 of 55,183,987 bp size a a a including the IDS gene was revealed in the chromosomal region Uma Ramaswami , Mark Mckie , Giuseppina Grillo , Derralynn a a b a Xq22.1q24-Xq28. At the second stage we found the hemizygous Hughes , Atul Mehta , Michael Hughes , Lysosomal Disorders Unit, b deletion c.1438_1442del leading to a frameshift of p.P480Ffs*17 in London, United Kingdom, Health Touch Ltd, Chislehurst, BR7 6NX, the exon 9 of IDS gene used the Sanger sequencing. The deletion United Kingdom wasn’t described in the international base HGMD professional earlier. Symptoms of Anderson Fabry disease (FD) (OMIM 301500) Existence of other mutations and recombinations between the IDS include neuropathic pain, gastrointestinal (GI) symptoms, and gene and the IDSP pseudogene wasn’t revealed. The absence of a fatigue, which influence health-related quality of life (QoL). Cur- mosaicism in data of the array CGH allowed us to assume that the rently we record FD symptoms 6-12 monthly at routine clinical partial monosomy was inherited from gametes of one of the parents visits. Frequent monitoring of day to day variations of these whose biomaterial wasn't available. The case that we have revealed symptoms is required to assess its true impact on QoL. To enable represents the first-ever molecular description of Hunter syndrome this, using existing technology from Health Touch Ltd, a novel Fabry burdened with Turner syndrome in girl with the partial monosomy specific mobile phone application (app) was developed. We designed on one X-chromosome and the novel deletion c.1438_1442del in IDS a template with FD -relevant questions on activity, pain (a visually gene on the other X-chromosome. interactive scoring tool), GI and sweating. A patient passport allows voluntary entry of personal information. Monthly QoL and disease specific questionnaires are completed by the patient. Medications doi:10.1016/j.ymgme.2018.12.308 can be recorded by patients themselves. Fabry specific therapies such as enzyme replacement therapies are entered by the clinician in a non-editable format. In addition a messaging facility allows commu- 293 nication with patients. Patient data entry takes less than 5 minutes. Migalastat: Single centre experience of adult patients with Fabry Consent forms and information governance, including General Data disease from the Royal Free London NHS Foundation Trust, UK Protection Regulation (GDPR) compliance, were approved by the Hospital Trust. Patients control their data and consent to sharing data Uma Ramaswami, Mark Mckie, Giuseppina Grillo, Atul Mehta, with their clinician. The data is secured in accordance with UK Derralynn Hughes, Lysosomal Disorders Unit, London, United Kingdom privacy policies. The clinician accesses data via a secure password protected website. Patient participation commenced in June 2018. 41 Anderson Fabry Disease (FD), OMIM 301500, is a lysosomal patients with a confirmed diagnosis of FD consented. In the first storage disorder due to a deficiency of the enzyme alpha galactosi- three months, 25% are actively using the app (more than once a dase A. Disease modifying therapies (DMTs) include intravenous week). We will present the data from patients on both long-term enzyme replacement (ERT) and oral pharmacological chaperone treatment and newly diagnosed patients on diseases modifying therapy (PCT). We describe a single centre experience of patients therapies. We anticipate our novel Fabry phone app will improve with FD treated with a novel PCT, migalastat (GalafoldTM). 375 care and will encourage patients to take ownership of their own patients with a confirmed diagnosis of FD are reviewed 6-12 health. It would allow the clinician to monitor both, day-to-day S122 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 variations in Fabry symptoms and treatment responses. This may Chapel Hill, Chapel Hill, NC, United States, cClinical Genome Resource, enable personalised therapy goals. Chapel Hill, NC, United States doi:10.1016/j.ymgme.2018.12.310 Although publication of the ACMG-AMP criteria was an important step in standardizing variant interpretation, discrepancies in patho- genicity classification between laboratories can occur due to the fi fi 295 general nature of these criteria. Gene and disease-speci c modi ca- Real-world outcomes in pregnant imiglucerase-treated patients tions are required to minimize these discrepancies. The Clinical with Gaucher disease: Data from the global safety database and Genome Resource (ClinGen) Lysosomal Storage Diseases Variant International Collaborative Gaucher Group (ICGG) Gaucher regis- Curation Expert Panel, composed of individuals with expertise in try pregnancy sub-registry maintained by Sanofi Genzyme clinical genetics, clinical laboratory diagnostics, clinical trials, research, and newborn screening, has focused initial efforts on modification of the ACMG-AMP criteria for interpretation of variants within the GAA Lewis Raynor, Julie Batista, Laura Carloni, Badari Gudivada, Grace gene associated with Pompe disease. Our expert panel has proposed Lewis, Davorka Sekulic, So-Fai Tsang, Sanofi Genzyme, Cambridge, MA, modification or specification for 14 of the 28 originally published United States criteria, based on broader ClinGen guidance or addressing gene or disease specific characteristics. Adjustments include the addition of Women with Gaucher disease are at risk for pregnancy complica- specifications, such as allele frequencies and functional assays, and tions. The EU Summary of Product Characteristics advises that women changes to criteria weighting based on the available evidence. These with Gaucher disease begin or continue imiglucerase before conception criteria will be piloted on ~50 variants. Two curators will examine each and continue treatment throughout pregnancy. The US Package Insert variant independently, and discrepant interpretations will be review recommends that imiglucerase not be administered during pregnancy by the expert panel. We will also assess the impact of using case-level except when the indication and need are clear and potential benefit data from laboratories with extensive internal variant databases on substantially justifies the risk. We analyzed cumulative data up to May variant classification, as well as the application of the PM2 criterion 2018 reported on women with Gaucher disease exposed to imiglucerase which, according to the ACMG-AMP criteria, can be utilized when a (Cerezyme, Sanofi Genzyme) before/during pregnancy from two variant is absent from control databases or at extremely low frequency sources: the Sanofi Genzyme Global Safety Database and the Interna- in the case of recessive conditions. Our long-term goal is to use our tional Collaborative Gaucher Group Gaucher Registry (NCT00358943) validated criteria for interpretation of a larger number of GAA variants Pregnancy Sub-Registry. The safety database, which tracks all reported and to submit those interpretations to ClinVar for use by the scientific pregnancy and lactation cases in imiglucerase-treated patients regard- and medical communities. Once the GAA variant curation process is less of association with an adverse event, reported data on 760 established, we will turn our attention to additional lysosomal storage pregnancies with 399 known pregnancy outcomes. There were 320 disorders, with priority assigned to those disorders currently being (80.2%) live births (including twins), 67 (16.8%) spontaneous abortions examined as part of the newborn screening, such as MPS I. (some also classified as fetal deaths), 15 (3.8%) stillborn/fetal deaths, and 6 (1.5%) therapeutic/elective terminations. Neonatal outcomes were normal in 240/395 (60.8%) pregnancies and unknown in 130/395 (32.9%) pregnancies. Neonatal complications were reported in 25/395 doi:10.1016/j.ymgme.2018.12.312 (6.3%) pregnancies, of which 10 (2.5%) were congenital anomalies. Of 64 outcomes reported in the Pregnancy Sub-Registry, there were 52 (81.3%) live births, 2 (3.1%) spontaneous abortions, 1 (1.6%) stillborn 297 N ( 20 weeks), 2 (3.1%) elective/therapeutic terminations, and 7 (10.9%) Longitudinal follow-up (4 years) of lyso-Gb1 in children with unknown outcomes. Among 61 pregnancies with reported complication Gaucher disease in a single center cohort status, 36 (59%) were uncomplicated, 15 (24.6%) had complications, and 10 (16.4%) had incomplete information. In 37/61 (60.7%) pregnancies, Shoshana Revel-Vilka, Tama Dinura, Michal Becker Cohena, Noa there were no neonatal complications, 19/61 (31.1%) had incomplete Horvitza, Claudia Cozmab, Marina Hovakimyanb, Sebastian neonatal information, and 5/61 (8.2%) had complications and/or Oppermannb, Laura Dermuthb, Arndt Rolfsb, Aya Abramova, Ari congenital anomaly. Spontaneous abortion frequency in both datasets Zimrana, aShaare Zedek Medical Center, Jerusalem, Israel, bCentogene, is within published general population rates (11.4-22%). The birth Rostock, Germany defect rate in the safety database is within published general population rates (3-6%). Glucosylsphingosine (Lyso-Gb1) a downstream metabolic product of doi:10.1016/j.ymgme.2018.12.311 glucosylceramide, which is also degraded by the glucocerebrosidase, has been identified as a sensitive and specific biomarker for the diagnosis and monitoring of patients with Gaucher disease (GD). This study aims to describe the lyso-Gb1 levels in a longitudinal cohort of treated and 296 untreated children (b18 years) with type I (GD1) and type III GD (GD3). Improving Pompe diagnostics through modification of the ACMG- Methods: All children followed in SZMC with GD were eligible for this AMP variant classification criteria by the Clinical Genome study. Since 2014, lyso-Gb1 levels (ng/ml) are routinely measured during Resources (ClinGen) Lysosomal Storage Diseases Variant Curation the semi-annual/annual visit. Results: A total of 270 tests were done for Expert Panel 82 children (median age 10 (1-18) years) GD1, N370S homozygous (n=31), GD1, N370S compound heterozygous (n=39) and GD3 Catherine Rehdera, Jennifer Goldsteinb, Deeksha Balia, ClinGen (n=12). A non-linear correlation was found between the first lyso-Gb1 Lysosomal Storage Diseases Variant Curation Expert Panelc, aDuke measurement and platelet count (pb0.001), but not with age at the time University, Durham, NC, United States, bUniversity of North Carolina at of sampling. Although lyso-Gb1 was higher for GD3 (median 107, 11- Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S123

1270) and GD1 compound heterozygous (median 95, 6.1-667) compared 299 to GD1 N370S homozygous (median 80, 9.5-200), these differences did Clinical program to evaluate safety, preliminary efficacy, and dose not reach statistical significance. During a median follow-up of 2.3 (0.5- selection of AAV8 gene therapy in patients with infantile and late 4.1) years, the levels of lyso-Gb1 increased in 30 children (14 treated) and onset Pompe disease (IOPD and LOPD) decreased in 36 children (27 treated) (treated vs. untreated, p= 0.03). The increase in lyso-Gb1 was associated with the decrease in platelet Salvador Ricoa, Benedikt Schoserb, David Dimmockc, Michael W. count (p=0.01). Clinically significant decrease in platelet count Lawlord, Emma Jamesa, Magda Mortona, Suyash Prasada, aAudentes 3 (N50x10 /ml) was preceded with an increase in lyso-Gb1. Change in Therapeutics, San Francisco, CA, United States, bLudwig-Maximilians- lyso-Gb1 was not associated with age, follow-up time and change in University, Munich, Germany, cRady Children’s Institute for Genomic hemoglobin level and spleen and liver volume. In conclusion, this is the Medicine, San Diego, CA, United States, dMedical College of Wisconsin, first report of a longitudinal follow-up of lyso-Gb1 in children with GD. Milwaukee, WI, United States Also in children, lyso-Gb1 seems to be a sensitive marker for Gaucher burden and could be used for monitoring the need for therapy in Pompe disease is a rare neuromuscular disease in which acid α- untreated children and compliance to therapy in treated children. glucosidase (GAA) deficiency leads to glycogen accumulation in all tissues. It presents as a spectrum ranging from the severe, rapidly progressive IOPD to the more slowly progressive LOPD. Currently, doi:10.1016/j.ymgme.2018.12.313 the only treatment is enzyme replacement therapy (ERT) with recombinant GAA. While ERT improves prognosis, limitations are associated with clinical progression, functional decline, and burden on the lives of patients/caregivers. Gene therapy with AT982 - a 298 recombinant adeno-associated viral vector serotype 8 (rAAV8) Screening for serum free light chains in patients with type 1 expressing human GAA - offers potential for a single administration Gaucher disease type 1 with targeted GAA expression in the most affected tissues, including skeletal and cardiac muscle and the nervous system. Here, we Shoshana Revel-Vilk, Majdolen Istaiti, Dafna Frydman, Tama Dinur, describe the study design of INQUIRO, a non-interventional, clinical- Michal Becker Cohen, Yotam Kolben, Heftziba Ivgy, Ari Zimran, Rosa assessment study of subjects with IOPD or LOPD to evaluate baseline Ruchlemer, Shaare Zedek Medical Center, Jerusalem, Israel disease characteristics of the disease, and INTEGRO and FORTIS, the Phase 1/2, open-label, ascending dose studies to assess safety, Gaucher disease type 1 (GD1) is associated with a high prevalence preliminary efficacy, and dose selection of AT982 in IOPD and LOPD of polyclonal and monoclonal gammopathies. However, risk factors patients. Both interventional trials are designed to assess three dose associated with gammopathies in patients with GD1 are still unclear. levels to determine the optimal dose in each population. The IOPD ® In this study, serum free light chains (FLC) (Freelite , Birmingham, U. trial is planned to only include ERT-experienced patients (n=12), K.), a useful screening tool for identifying patients with monoclonal while the LOPD trial will include ERT-experienced (n=12) and gammopathies, were measured in cryopreserved serum samples from -naïve patients (n=5). Both IOPD and LOPD ERT-experienced 98 patients (42 male) with GD1. All patients had N370S mutations (69 subjects will receive ERT until confirmation of GAA activity in muscle homozygous and 29 compound heterozygous). Serum immunoglob- biopsies and subsequent weaning. INQUIRO will explore neuromus- ulin and glucosylsphingosine (Lyso-Gb1) levels from the same date of cular function, quality of life assessments, biomarkers and the status sampling were retrieved from the hospital records. At the time of of AAV and GAA antibody titers. INTEGRO and FORTIS will include sampling, the median (range) age of patients was 63 (50-92) years. assessment of adverse events, laboratory tests (including immuno- Sixty-four were receiving enzyme replacement therapy and two logical parameters), 12-lead ECGs, echocardiograms, vital signs and substrate replacement therapy for a median (range) of 5 (1-28) years. physical examinations. Primary efficacy endpoints include GAA The median free-kappa and free-lambda levels were 23.7 (range, 8.9- activity in muscle, serum, lymphocytes, and whole blood change in 222) ng/ml and 18.6 (range, 6.5-82.6) ng/ml, respectively. An GAA protein, mRNA, glycogen content, morphology, and vector copy N N increased free kappa ( 19.6 ng/ml) and FLC ratio ( 1.65) was detected number. Additional functional, imaging and quality of life endpoints in 19 patients (19%, 95% CI 12%-28%). Patients with an increased FLC will be discussed. ratio were significantly older, median (range) age 72 (52-92) years, as compared to those with a normal FLC ratio, median (range) age 61 (50- 82) years (p=0.001, odd ratio (OR) 1.12, 95% 1.05-1.19). Increased FLC doi:10.1016/j.ymgme.2018.12.315 ratio was not associated with gender, underlying mutation (homozygous vs. compound heterozygous), treatment and previous splenectomy. Interestingly, increased FLC ratio was associated with increased lyso-Gb1 levels (N140 ng/ml), 8/19 vs. 14/79 (p=0.03, OR 3.37, 95% CI 300 1.15-9.93), adjusted to age and GD specific treatment. The older age Assessment of bone mineral density and FRAX tool analysis in a association with increased FLC ratio in our cohort is similar to reports cohort of male Fabry disease patients and GLA gene GVUS subjects from the non-GD1 population. The association with lyso-Gb1 levels suggests that “Gaucher burden” may be a risk factor for the Nilton S. Rosa Neto, Judith C.B. Bento, Liliam Takayama, Valeria F. development of monoclonal gammopathy, however larger prospec- Caparbo, Rosa Maria R. Pereira, University of Sao Paulo, Sao Paulo, tive studies with confirmatory tests including serum protein electro- Brazil phoresis and immunofixation are needed. Osteoporosis/osteopenia are described in Fabry disease (FD) patients and attributed to multifactorial etiology. We evaluated the doi:10.1016/j.ymgme.2018.12.314 prevalence of osteoporosis/osteopenia in Brazilian male FD patients and GLA gene genetic variants of unknown significance (GVUS) S124 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 subjects compared to controls. To estimate 10-year fracture risks by disease is associated with elevated levels of IL-6 and TNF-α despite ERT FRAX© tool adapted to Brazil. Sixteen FD patients, 8 GLA GVUS treatment however IL-1β levels are not different from controls in this subjects and 39 sex/age-matched healthy controls were evaluated by cohort. It is not possible to exclude effect of ERT on this result. Higher Dual-energy X-Ray Absorptiometry (DXA). Scans were performed at MSSI scores were correlated with higher levels of IL-6 and TNF-α. the lumbar spine, right total hip and femoral neck and non-dominant Patients on ERT had higher levels of IL-6 and TNF-α than those not on distal 1/3 radius. FRAX© risks were estimated for patients age ≥40 treatment, reflecting greater disease burden. years (n=10). High risk was defined as risk of hip fracture N3% or of major osteoporotic fracture N20%. Mean age (years) for patients was 44.4±11.6, GVUS 48.5±12.3 and controls 42.9±10.6, (pN0.05). Body doi:10.1016/j.ymgme.2018.12.317 Mass Index (kg/m2) was lower in patients (22.8±3.8) than GVUS (28.3±2.6, p=0.001) and controls (26.9±2.6, pb0.001). Four patients had previous major fractures (26.7%). Lumbar bone mineral density (BMD) (g/cm2) was lower in patients (0.891±0.121) than 302 controls (1.019±0.108, pb0.001) but not GVUS (0.997±0.110, Depression, sleep disturbances, disability and quality of life in a p=0.06). Hip BMD was lower in patients (0.824±0.134) than cohort of Brazilian Fabry disease patients and GLA gene GVUS controls (1.002±0.111, pb0.001) and GVUS (0.956±0.109, subjects p=0.04). Neck BMD was lower in patients (0.694±0.103) than controls (0.859±0.123, pb0.001) and GVUS (0.804±0.090, p=0.03). Nilton S. Rosa Neto, Judith C.B. Bento, Rosa Maria R. Pereira, Radius BMD was lower in patients (0.667±0.116) than controls University of Sao Paulo, Sao Paulo, Brazil (0.764±0.063, pb0.001) but not GVUS (0.741±0.086, p=0.15). There were 4 Osteopenia (25%), 9 Osteoporosis (56.3%) in patients Fabry disease (FD) is an X-linked sphingolipidosis whose 3 Osteopenia (37.5%), 2 Osteoporosis (25%) in GVUS 21 Osteopenia mutations on the GLA gene affect alpha-galactosidase A activity. As (53.8%), 5 Osteoporosis (12.8%) in controls. FRAX© estimates for hip/ a progressive storage disease, FD patients are known to have more major osteoporotic fractures were lower than 3%/20% for patients depression and worse sleep quality, functionality and quality of life. and GVUS. Osteoporosis/osteopenia are prevalent in male FD We evaluated prevalence of depression, sleep disturbances, disability patients and GLA GVUS subjects. These results reinforce the and impact on quality of life in a cohort of Brazilian FD patients and fi incorporation of DXA analysis in routine follow-up of patients compare between groups strati ed by the Mainz Symptom Severity fi following local guidelines. FRAX© may not be an adequate tool for Index (MSSI) and to GLA gene genetic variants of unknown signi - fi fracture risk estimation in this population. cance (GVUS) subjects. Thirty-seven FD genotype con rmed patients (16:21 M:F) and 19 GLAgene GVUS subjects (8:11 M:F) were assessed by the Hamilton Depression Rating Scale (HAM-D), doi:10.1016/j.ymgme.2018.12.316 Pittsburgh Sleep Quality Index (PSQI), Health Assessment Question- naire Disability Index (HAQ-DI), and Short-Form Health Survey 36 (SF36) questionnaires. Depression was referred by 11 patients [29.7%] and 7 GVUS subjects [36.8%]. Nevertheless, HDS indicated 301 depression in 21 patients and 9 GVUS subjects. Insomnia and/or Inflammation in Fabry disease: Correlation with Mainz Severity unrefreshing sleep were reported by 17 patients [45.9%] and 9 GVUS Score Index (MSSI) subjects [47.3%]. However, PSQI depicted sleep disturbances in 22 patients and 10 GVUS subjects. Mean HAQ-DI according to MSSI Nilton S. Rosa Neto, Judith C.B. Bento, Valeria F. Caparbo, Rosa Maria status was 0.490 (severe), 0.274 (moderate) and 0.157 (mild) in R. Pereira, University of Sao Paulo, Sao Paulo, Brazil patients and 0,458 (moderate) and 0,127 (mild) in GVUS subjects, (pN0.05). SF-36 analysis showed differences in raw score between FD Fabry disease (FD) is an X-linked lysosomal storage condition where patients and GVUS subjects in functional capacity (56.08±24.75 vs. defective enzyme alpha-galactosidase A contributes to accumulation of 73.68±20.19, p=0,01) and general health status (48.89±21.03 vs. substrate in several organs, with varying degrees of severity, and 70.68±17.06, pb0.001). Depression, sleep disturbances and disability subsequent loss of function. Chronic inflammation in thought to were under-recognized in FD patients as well as in GLA gene GVUS contribute to organ damage in these patients. We evaluated levels of subjects in this cohort. HAQ-DI revealed worse disability according to inflammatory cytokines in FD patients compared to healthy controls MSSI severity status in both groups but not between groups. General and correlation with Mainz Severity Score Index (MSSI). Thirty-six FD health status is poorer for FD patients than GVUS subjects. genotype confirmed FD patients (M:F 16:21) and 25 age-matched healthy controls (M:F 20:8) had an array of cytokines measured using Milliplex© technology. Mean age (years) for FD patients was 43.1±15.4 doi:10.1016/j.ymgme.2018.12.318 and controls 47.4±12.2, (pN0.05). Enzyme replacement therapy (ERT) was given to 22 patients (59.5%). FD patients had higher Interleukin-6 [IL-6] (pg/ml) levels than controls (3.04±2.56 vs. 1.47±0.55, p=0.01), higher levels of tumoral necrosis factor alpha [TNF-α] (pg/ml) than 303 controls 1.67±0.87 vs. 0.88±0.39, pb0.001) but not levels of interleu- Dual-energy X-ray absorptiometry body composition assessment kin-β1[IL-1β] (pg/ml) (0.44±0.18 vs. 0.38±0.14, p=0.14). Patients on in male Fabry disease patients and GLA gene GVUS subjects ERT (n=22) had higher levels of IL-6 (pg/ml) (3.97±2.86 vs. 1.74±1.17, p=0.01) and higher levels of TNF-α (pg/ml) (1.92±0.84 vs. 1.32±0.79, Nilton S. Rosa Neto, Judith C.B. Bento, Liliam Takayama, Valeria F. p=0.04) than those not on treatment. MSSI scores correlated with Caparbo, Rosa Maria R. Pereira, University of Sao Paulo, Sao Paulo, levels of IL-6 (r=0.60, pb0.001) and TNF-α (r=0.45, p=0.005). Fabry Brazil Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S125

Fabry disease (FD) is an X-linked sphingolipidosis whose for age was present in 24 patients (64.8%) fibromyalgia was mutations on the GLA gene affect alpha-galactosidase A activity. diagnosed in 16 patients (43.2%)lymphedema was present in 8 Dual-energy X-ray absorptiometry (DXA) is mostly dedicated to the patients (21.6%) Charcot foot was present in 4 patients, 2 of whom evaluation of bone mineral status, but is an available technique to required finger amputation. Other diagnosis included chronic low assess body composition in terms of fat mass and lean mass. We back pain (5), hip/knee osteoarthritis (5) out (1) clinodactyly (1) evaluated body composition by DXA in Brazilian male FD patients yperCKemia (1) carpal tunnel syndrome (2). Past or present Fabry and GLA gene genetic variants of unknown significance (GVUS) crises were referred by 16 patients (43.2%). Six autoimmune/ subjects compared between them and to controls and correlation of autoinflammatory diseases were diagnosed in 4 patients (10.8%), all body composition parameters with Lyso-Gb3 levels. Sixteen FD female, including celiac disease, autoantibody positive hyper- and genotype confirmed patients, 8 GLA gene GVUS subjects and 39 sex/ hypothyroidism, IgA nephropathy, psoriasis and vitiligo. In conclu- age-matched healthy controls had body composition evaluated by sion, rheumatic manifestations are very frequent in FD, related or not DXA and lyso-Gb3 levels assessed. Mean age (years) for FD patients to the physiopathology of the disease, and some patients may was 44.4±11.6, GVUS 48.5±12.3 and controls 42.9±10.6, (pN0.05). present with concomitant autoimmune/autoinflammatory diseases. Body Mass Index (kg/m2) was lower in FD patients (22.8±3.8) than These results reinforce the need to raise awareness and increase GVUS (28.3±2.6, p=0.001) and controls (26.9±2.6, pb0.001). Bone knowledge about Fabry disease among non-specialist physicians, mineral content (g) was lower in FD patients (2175.8±332.5) than notably rheumatologists. controls (2563.3±364.7, p=0.001) but not GVUS (2406.3±262.8, pN0.05). Lean mass (g) was lower in FD patients (47681.9±5201.8) than controls (58912.3±6439.2, pb0.001) and GVUS (61243.2 doi:10.1016/j.ymgme.2018.12.320 ±4880.8, pb0.001). There were no differences between patients and controls regarding fat mass (g) (14357.2±5668.1 vs. 16958.1 ±5374.6, pN0.05) or fat percentage (%) (21.76±5.9 vs. 21.3±5.1, pN0.05). GVUS subjects had higher fat mass (g) (21653.7±4511.9, 305 fl pb0.001) but not fat percentage (%) (25.2±3.9, pN0.05) or bone In ammation in GLA gene GVUS subjects mineral content (2406.3±262.8, pN0.05). Lyso-Gb3 levels were inversely correlated with lean mass (r=-0.63, p=0.007), fat mass Nilton S. Rosa Neto, Judith C.B. Bento, Rosa Maria R. Pereira, (r=-0,68, p=0.003) and fat percentage (r=-0.64, p=0.006). In University of Sao Paulo, Sao Paulo, Brazil conclusion Brazilian FD patients in this cohort have lower body mass index, bone mineral content and lean mass than controls and lower Fabry disease (FD) is a X-linked lysosomal storage disorder due to body mass index, lean mass and fat mass than GVUS subjects. Lyso- mutations on the GLA gene, located on the X chromosome, affecting Gb3 accumulation is associated is lower lean mass, lower fat mass alpha-galactosidase A activity and is associated with chronic fl and lower fat percentage in FD patients. in ammation. Some mutations have little interference on enzyme activity and mild or absent characteristic FD signs and symptoms being considered genetic variants of unknown significance (GVUS). fl doi:10.1016/j.ymgme.2018.12.319 In ammatory markers were not evaluated in this population. We evaluated levels of inflammatory cytokines in GLA gene GVUS subjects compared to healthy controls. 12 GLA gene GVUS subjects (M:F 4:8) and 25 age-matched healthy controls (M:F 20:8) had an 304 array of cytokines measured using Milliplex© technology. Mean age Rheumatic and autoimmune/autoinflammatory manifestations (years) for GVUS subjects was 40.±15.1 and controls 47.4±12.2, in a cohort of Brazilian Fabry disease patients (pN0.05). Enzyme replacement therapy (ERT) was given to 2 patients (16.7%). Levels of Interleukin-6 [IL-6] (pg/ml) 1.61±1.09 vs. 1.47 Nilton S. Rosa Neto, Judith C.B. Bento, Rosa Maria R. Pereira, ±0.55, pN0.05 and levels of interleukin-β1 [IL-1β] (pg/ml) 0.41 University of Sao Paulo, Sao Paulo, Brazil ±0.14 vs. 0.38±0.14, pN0.05 were not different from controls. Levels of tumoral necrosis factor alpha [TNF-α] (pg/ml) had a tendency of Fabry disease (FD) is an X-linked lysosomal storage condition being higher than controls 1.24±0.60 vs. 0.88±0.39, p=0.051). where defective enzyme alpha-galactosidase A contributes to When separated by gender, male GVUS subjects had similar values of accumulation of substrate in several organs, with varying degrees IL-6 and TNF-α (2.32±1.64 vs. 1.49±0.62, pN0.05 / 1.47±0.87 vs. of severity, and subsequent loss of function. Early diagnosis is the 0.99±0.39, pN0.05) but female subjects did had more elevated TNFα clue to better treatment outcomes. Patients may present with values (1.14±0.36 vs. 0.63±0.20, p=0.01) than controls. This is the different rheumatic and autoimmune/autoinflammatory manifesta- first study to evaluate inflammation in carriers of GLA gene mutations and few studies addressed this topic. We reviewed the tions considered to be nonpathogenic. GLA gene GVUS subjects have rheumatic and autoimmune/autoinflammatory manifestations in a a tendency of elevated TNF-α values in this cohort and female cohort of Brazilian FD patients. FD genotype confirmed patients were subjects are prone to having more elevated TNF-α than controls. The interviewed and examined by a rheumatologist. Disease character- low number of subjects and the effect of ERT in some of them may istics are described. Thirty-seven consecutive patients were evalu- have influenced the results. Whether these results derive from the ated (16:21 M:F), with a mean age of 43,1 years (range, 12-72). gene mutation itself is yet to be determined but attention should be Rheumatic manifestations were present in 36 patients (97.3%). The paid to chronic inflammation on those patients. only patient who did not present any rheumatic symptoms at the time of study was the youngest female at age 12. Pain in the extremities (nociceptive and/or neuropathic) was present in 26 doi:10.1016/j.ymgme.2018.12.321 patients (70.3%) osteopenia/osteoporosis/low bone mineral density S126 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

306 initiation were reviewed. Thirty-seven consecutive patients were Bone microarchitecture as measured using HR-pQCT of male interviewed (16:21 M:F) and 33 were considered symptomatic. Fabry disease patients and GLA gene GVUS subjects Misdiagnosis occurred in 11 males (62.5%) and 13 females (61.9%) of which 10 males and 9 females being previously diagnosed with one Nilton S. Rosa Neto, Judith C.B. Bento, Liliam Takayama, Valeria F. or more rheumatological conditions, most frequently rheumatic Caparbo, Rosa Maria R. Pereira, University of Sao Paulo, Sao Paulo, fever or “rheumatism” (unspecified rheumatic disorder). Median Brazil age at first symptoms 12.0 years (5-55) for males and 16.5 years (9- 66) for females, p=NS. Mean age at diagnosis (including asymp- Osteoporosis/osteopenia are described in Fabry disease (FD) tomatic patients) was 35.5 years for males (8-55) and 35.9 years for patients and attributed to multifactorial etiology. High-resolution females (4-71) p=NS. Median time for diagnosis was 16.0 years (0- peripheral Quantitative Computed Tomography (HR-pQCT) assesses 52) overall 20.5 years (1-42) for males and 14.0 years (0-52) for trabecular and cortical bone structural parameters and total bone females, p=NS. 22 patients were considered for ERT initiation (13:9 volume and density in high-definition mode, allowing digital M:F) and mean time to ERT initiation after FD diagnosis was 3.7 constructs of bone microarchitecture. Bone microarchitecture was years (0-15) for males and 0.4 year (0-1) for females, p=NS. never evaluated in this setting. We evaluated bone micro- Misdiagnosis occurred in 64.8% FD patients, commonly for rheuma- architecture using HR-pQCT of male FD patients and GLA gene tological conditions. Median time for correct diagnosis was 16 years. genetic variants of unknown significance (GVUS) subjects compared These results reinforce the need to raise awareness and increase between them and to controls. Sixteen FD genotype confirmed knowledge about Fabry disease among non-specialists, particularly patients, 8 GLA gene GVUS subjects and 39 sex/age-matched rheumatologists. healthy controls were evaluated using HR-pQCT (distal tibia and distal radius). Mean age (years) for FD patients was 44.4±11.6, GVUS 48.5±12.3 and controls 42.9±10.6, (pN0.05). Body Mass doi:10.1016/j.ymgme.2018.12.323 Index (kg/m2) was lower in FD patients (22.8±3.8) than GVUS (28.3±2.6, p=0.001) and controls (26.9±2.6, pb0.001). Radius analysis revealed lower cortical thickness (mm) in FD patients 308 (0.63±0.21) compared to controls (0.91±0.22, pb0.001) but not to Rheumatic manifestations and misdiagnosis in Brazilian GLA gene GVUS (0.81±0.19, pN0.05). Tibia analysis revealed significant lower GVUS subjects values for FD patients compared to controls in volumetric densities (mg HA/cm3): total bone mineral density (263.4±71.3 vs. 328.4 ±61.4, p=0.002) trabecular bone mineral density (149.2±53.3 vs. Nilton S. Rosa Neto, Judith C.B. Bento, Rosa Maria R. Pereira, 182.3±42.1, p=0.02) and structural parameters (mm): bone University of Sao Paulo, Sao Paulo, Brazil volume ratio (0.12±0.04 vs. 0.15±0.04, p=0.02) trabecular separation (0.53±0.15 vs. 0.45±0.09, p=0.03) and inhomogeneity of Fabry disease (FD) is a X-linked lysosomal storage disorder due to network (0.27±0.14 vs. 0.20±0.06, p=0.02). At tibia level, the only mutations on the GLA gene, located on the X chromosome, affecting significant difference between patients and GVUS subjects was alpha-galactosidase A activity. Some mutations have little interfer- cortical thickness (0.99±0.21 vs. 1.22±0.26, pb0.001). This is the ence on enzyme activity and, alongside mild or absent characteristic first study that evaluated bone microarchitecture using HR-pQCT in FD signs and symptoms, are considered genetic variants of unknown fi FD patients and GLA gene GVUS subjects. The results show bone signi cance (GVUS). We reviewed rheumatic manifestations and deterioration at trabecular and cortical densities and structural misdiagnosis in a cohort of Brazilian GLA gene GVUS subjects. GLA fi parameters in FD patients that may predispose to increased fracture gene GVUS patients were identi ed during hemodialysis units risk. screening procedures. Subjects were interviewed and examined by a rheumatologist. Clinical characteristics and previous rheumatic diagnosis are described. Nineteen consecutive subjects were evalu- doi:10.1016/j.ymgme.2018.12.322 ated (8:11 M:F), with a mean age of 39.6 years (range, 9-66). R118C was detected in 15 patients and A143T in 4. Lyso-Gb3 levels were normal. Two patients were on Enzyme Replacement Therapy. Application of the Mainz Severity Score Index for FD depicted those 307 2 patients as “Moderate” severity, which was accounted for renal Misdiagnosis and delay in treatment initiation in a cohort of (subnephrotic proteinuria/kidney transplantation) and neuropsychi- Brazilian Fabry disease patients atric symptoms. Thirteen patients (68.4%) presented rheumatic manifestations. Pain in the extremities in 10 patients (52.6%) Nilton S. Rosa Neto, Judith C.B. Bento, Rosa Maria R. Pereira, osteopenia/osteoporosis in 11 patients (57.9%) fibromyalgia in 7 University of Sao Paulo, Sao Paulo, Brazil patients (36.8%). Other diagnosis included chronic low back pain (3), osteoarthritis (3) rigger finger (1) leg cramps (1). Four patients [3 Fabry disease (FD) is commonly misdiagnosed, most frequently A143T/ 1 R118C] referred past/present pain crises comparable to FD. for rheumatological conditions. Incorrect diagnosis may lead to Furthermore, 4 patients were previously misdiagnosed with a incorrect therapies and delay in FD recognition and proper treatment rheumatic disease: rheumatoid arthritis, rheumatic fever, and initiation. We reviewed diagnostic errors in a cohort of Brazilian FD “rheumatism”, 3 of those affected by pain crises. In conclusion patients, time to correct FD diagnosis and time to treatment rheumatic manifestations are frequent in GLA gene GVUS subjects. initiation in those whose enzyme replacement therapy (ERT) were FD-like pain crises might account for misdiagnosis, and whether the indicated. FD confirmed patients were interviewed and previous symptoms derive from the gene mutation is yet to be determined. diagnosis, age at first symptoms, age at FD diagnosis and age at ERT Renal and neuropsychiatric manifestations may warrant further Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S127 investigation to exclude other diagnosis. These results reinforce the leads to severe symptoms, including short stature, coarse facial role of rheumatologists in determining disease burden. features, skeletal dysplasia, cardiorespiratory disease, declining quality of life, and shortened lifespan. Enzyme replacement therapy (ERT) with galsulfase has emerged as a treatment option, with doi:10.1016/j.ymgme.2018.12.324 current literature suggesting that earlier ERT initiation improves patient outcomes. However, there is considerable variability in the phenotypic spectrum and it is unclear if the effectiveness of therapy is dependent on age at initiation or severity of the disease. Sibling 309 studies minimize genotype-phenotype differences and provide ideal Newborn screen for MPS1 (Hurler syndrome) - The Washington, case control data but only limited reports exist in the literature. Here, DC experience we report on a sibship of two patients. The younger sibling started ERT at 6 years of age compared to 10 years of age for the older Tamanna R. Roshan Lal, Christina Grant, Kathleen Crosby, Rebecca sibling. The younger sibling has maintained height at 75th percentile Hicks, Sarah Viall, Pranoot Tanpaiboon, Rare Disease Institute - while the older sibling had decline in growth velocity to 3rd Childrens National Medical Center, Washington, DC, United States percentile and has tracked at 3rd percentile since therapy initiation. Facial features and skeletal gestalt are milder in the younger sibling Introduction: Newborn screening for MPS1 (Hurler syndrome) and unlike the older sibling, there is no respiratory disease. Extent of was recommended by the RUSP in 2016. Screening was started in the cardiac disease, corneal clouding and skeletal disease is similar in District of Columbia (DC) in January, 2018 whereby all patients both. Compared to baseline, both siblings demonstrate improvement fi identi ed by NBS are referred to CNRDI for follow-up. Method: Eight in 6 minute walk test and decrease in urine GAGs. Both report fi patients have been identi ed by NBS through September 2018. All improvements in energy, joint mobility and quality of life with patients were evaluated by genetics and investigations including galsufase. In this sibling case control study, we demonstrate repeat dried blood spot (DBS) IDUA activity, DBS and urine improved outcomes especially in growth, facial features and glycosaminoglycans (GAGs) analysis (LC-MS), and molecular analy- respiratory disease with earlier initiation of ERT. Together, these sis were performed. Results: Urine samples collected in 6 of 8 two patients demonstrate both the benefit of early ERT initiation, as patients, all revealed normal qualitative and quantitative GAGs. DBS well as the need for further study in this patient population. GAGs collected in 7 of 8 patients all revealed normal dermatan and heparan sulfate levels. Five of 8 patients had low DBS-IDUA activity 2 were found to be carriers of pathogenic variants in IDUA, and 3 were doi:10.1016/j.ymgme.2018.12.326 found to carry pseudodeficiency alleles. Other 3 patients had normal DBS-IDUA enzyme activity 1 out of 3 had pseudodeficiency allele, 1 had negative molecular test and 1 did not have molecular testing. 311 Discussion: Multiple pseudodeficiency alleles commonly seen in Real-world/health economic publication patterns before and individuals of African descent have been identified, the data is after enzyme replacement therapy approval across lysosomal consistent with population distribution of Washington, DC, making diseases these results likely in our catchment population. In our cohort, majority of positive NBS patients were found to have either a a b pseudodeficiency alleles or be carriers of pathogenic variants. All Tom Rouwette , Rosalie Gadiot , Michael L. Friedman , Tessa E. a a c c patients had normal urine and DBS GAGs which helped clinicians to Hartog , Yunyu Huang , Liina Nevalaita , Stefaan Sansen , Pascale c a b rule out the diagnosis of MPS1 prior to completion of molecular Vincendon , Excerpta Medica, Amsterdam, Netherlands, Excerpta c fi testing. The utility of repeat DBS enzyme analysis is unclear. Based on Medica, Los Angeles, CA, United States, Sano Genzyme, Naarden, our experience and practicality of sample collection, we feel Netherlands performing DBS GAGs measurement would reduce time and expense Real-world and health economic (RW/HE) evidence is becoming to resolving diagnosis with a positive newborn screen for MPS1. In addition, number of false positive newborns may be reduced if DBS increasingly important to clinical decision-making in many therapeutic areas. The proportion of RW/HE publications has increased GAGs was implemented in the second tier analysis. over the past 10-15 years in major therapeutic areas, though little is known about rare diseases like lysosomal storage disorders (LSDs). This study quantifies the RW/HE publications on enzyme replace- doi:10.1016/j.ymgme.2018.12.325 ment therapy (ERT) relative to drug approval in selected LSDs. EMBASE was searched to identify original research articles about ERT in Fabry disease, Gaucher disease, Pompe disease, and muco- 310 polysaccharidosis type I (MPS I). Each eligible article was coded as Galsulfase treatment in two siblings with mucopolysaccharidosis clinical publication or RW/HE publication (defined as observational, type VI: A case report registry, claims data, electronic medical records and modeling studies). A 12-year time frame was established from the first ERT Alexandra Rostona, Clara Hungb, Alicia Chanb, Shailly Jain-Ghaib, approval date for each LSD. Descriptive statistics and Chi-square tests aFaculty of Medicine and Dentistry, Edmonton, AB, Canada, bUniversity were used to compare changes. Before approval, a low number of of Alberta, Edmonton, AB, Canada ERT publications was identified all described data from clinical studies. After approval, overall, a strong increase in RW/HE Mucopolysaccharidosis VI (MPS VI, Maroteaux-Lamy Syndrome) publications in the first 4 years (from 0.0% to 49.2%, p=0.041) was is a lysosomal storage disorder that results from a deficiency in the observed and this continued to increase up to 62.8% 12 years post- enzyme arylsulfatase B, which in turn causes glycosaminoglycans approval (p=0.023). Similar patterns were observed across included (GAGs) to accumulate in body tissues. This progressive accumulation LSDs. The strongest increase over time was observed in Fabry disease S128 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

(78.9% 12 years post-approval), whereas for Gaucher disease, Pompe 313 disease and MPS I the proportion of RW/HE publications ranged from The pro-inflammatory immunophenotype of a Farber disease 42.9% to 50.0% 12 years post-approval. Most RW/HE publications mouse model is ameliorated by repeated dosing with RVT-801, a were observational studies (75.8%) followed by registry studies developmental enzyme replacement therapy for Farber disease (14.3%). In this study, a strong increase in RW/HE publications was observed for all LSDs, confirming RW/HE publication trends previ- Brante P. Sampeya, Katie S. Dukea, Gavin C. Sampeya, Eric J. Gaukela, ously observed in other therapeutic areas. These results could Alexander Solyomb, Stephen A. Wringc, Xingxuan Hed, Changzhi suggest increasing interest in publication of RW/HE evidence for Zhud, Victor DeAngelisd, Edward H. Schuchmand, Christine M. health-care decision-making purposes, and to overcome inherent Coquerye, aEnzyvant, Durham, NC, United States, bEnzyvant, Basal, challenges in rare disease studies including the paucity of Switzerland, cAltavant, Durham, NC, United States, dIchan School of randomised, placebo-controlled clinical trials, small patient popula- Medicine, New York, NY, United States, eImmunovant, Durham, NC, tions, and disease heterogeneity. United States doi:10.1016/j.ymgme.2018.12.327 Farber disease (FD) is an ultra-rare disease where ASAH1 gene mutations result in non- or minimally functional acid ceramidase (AC), leading to accumulation of pro-inflammatory ceramide and 312 severe inflammation. RVT-801 is a novel recombinant human AC in Mucopolysaccharidosis type VI: Identification of novel mutations development as an enzyme replacement therapy (ERT) to reduce on the arylsulphatase B gene (ARSB) in Moroccan patient ceramide in FD patients. Treatment of FD mice with RVT-801 has demonstrated reduced ceramide content in multiple tissues and Es-Said Sabira, Naima Marzoukib, Karima Lafhalc, Rabiy Elqadiryd, lowered monocyte chemoattractant protein (MCP)-1. Although Nourrdin Radaad, Mohammed Bouskraouid, Naima Fdila,b, aLaboratory upregulation of MCP-1 and histiocytic infiltration of tissues has been of Biochemistry Faculty of Medicine of Marrakech, Cadi Ayad University, described in FD, the inflammatory cells contributing to disease Marrakech, Morocco, bClinical Analysis Laboratory, Hospital Med VI, pathology have not been well characterized. Using a FD mouse Marrakech, Morocco, cFaculty of Medicine and Pharmacy, Marrakech, model, flow cytometry was employed to characterize the immune Morocco, dPediatric Service, Hospital Med VI, Marrakech, Morocco cell repertoire in blood and key tissues (lung, liver, spleen). Compared to wild-type mice, multiple pro-inflammatory immune Mucopolysaccharidosis type VI (MPS VI, Maroteaux-Lamy syn- cells were broadly elevated in FD mice, including neutrophils (2-7- drome), is inherited as an autosomal-recessive disorder which occurs fold increase), activated monocytes (3-6-fold increase) and activated due to the deficiency of N-acetyl galactosamine-4- sulfatase B cells (2-fold increase). In contrast, total T cells were decreased in (Arylsulfatase B or ARSB) involved in catabolism of Glycosaminogly- the blood and all FD tissues examined. Changes in immune cell cans dermatan sulfate resulting from disease-causing variations in composition were observed in Farber mice as young as 4 weeks and the ARSB gene [1]. This study was carried out to identify the remained comparable or slightly exacerbated with increasing age. To mutational spectrum in MPS VI patients in Morocco and to describe assess whether RVT-801 directly impacted the immune cell com- the genotype, phenotype association and functional outcomes of partment, FD mice were treated with four once-weekly intraperito- these mutations. The patient was a 12-yr-old boy presenting with neal doses at 10 mg/kg/dose - the maximally effective dose in the skeletal deformity, short stature, facial dysmorphy, kyphosis, heart model. RVT-801 decreased pro-inflammatory immune cells, with key defects and delayed motor development. The first diagnostic was reductions in neutrophils and activated monocytes to wild-type based on the quantitative and qualitative biochemical analysis by control levels in the lung, liver, blood, and to a lesser extent the using dried blood filter paper (DBFP) samples and confirmed by spleen. These findings, together with the previously reported data genetic tests using polymerase chain reaction (PCR) sequencing on showing MCP-1 and ceramide reduction, demonstrate amelioration the entire coding region of the ARSB gene. Analysis of the molecular of the pro-inflammatory status of the FD mouse model following structure of the ARSB gene in Moroccan MPS VI patients revealed a repeat dose treatment with RVT-801 and further support its nonsense deletion mutation of two thymines at position 770 and 771 potential as an ERT for Farber disease. Moreover, the study identifies (c. [770_771delTT] [770_771delTT]). Analyses of ARSB gene muta- potential biomarkers that may be relevant to Farber patients. tions have been reported for many world populations. However, so doi:10.1016/j.ymgme.2018.12.329 far, no scientific study has been carried out regarding the Mucopolysaccharidoses syndrome in Moroccan population. Data from the Moroccan Demographic and Health Survey indicates that Morocco is a country with a high level of consanguinity, as other 314 Muslim populations of North Africa, Middle East, South and Central Gaucher disease and associated plasma cell neoplasia: A diagnos- Asia [2]. Moroccan population represents a challenge for genetic tic dilemma studies as its high rate of consanguineous marriages could be a contributing factor to the high incidence of some rare autosomal Angela Sanchez, Robert A. Ali, Neal J. Weinreb, Nicole K. Osorio, recessive diseases. Until now, the best of our knowledge, this is the Daniel P. Cassidy, Kyle White, Jennifer R. Chapman, James E. first MPS VI case with novel frame shift ARSB gene mutation to be Hoffman, Francisco Vega, Offiong F. Ikpatt, Offiong F. Ikpatt, confirmed in Morocco. University of Miami Leonard M Miller School of Medicine, Miami, FL, United States doi:10.1016/j.ymgme.2018.12.328 Gaucher disease (GD) patients have an increased risk of cancer, including plasma cell neoplasm. The clinical manifestations of GD and plasma cell myeloma overlap. Both may present with anemia, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S129 lytic bone lesions, bone pain and osteopenia/osteoporosis. Here, we pneumoniae [control 5.90 (5.42-5.90), lyso-Gb3 5.92 (5.88-5.95), p describe the clinical features, laboratory parameters and molecular ns], or C. perfringens [control 4.64 (4.42-4.64), lyso-Gb3 4.75 (4.73- features of 9 patients with GD who developed an M-spike. We reviewed 4.87), p ns], but promoted the growth of B. fragilis [control 5.92 records of 67 adult GD patients. Nine patients had an M-spike (5.45-6.02), lyso-Gb3 6.15 (6.13-6.30), p b0.05]. Conclusions: Lyso- demonstrated by SPEP and confirmed with immunofixation and plasma Gb3 favors the growth of B. fragilis in a multispecies biofilm. Since free light chain measurements. Two patients had overt myeloma 7 had changes in the intestinal microbiota have been associated with both monoclonal gammopathy of undetermined significance (MGUS). M- local symptoms and systemic cardiovascular and renal effects, spike patients were older on average than non-affected patients (72 ± 5 further studies should address the contribution of glycolipids to gut v 59 ± 19 y), more likely male (M/F 7:2 v 1:2) and less likely Jewish (91% dysbiosis and of dysbiosis to Fabry disease gastrointestinal symp- v 56%). Anemia (Hb LT10g/dl) [44% v 16%] and lytic bone lesions [44% v toms and systemic complications. 29%) were more common in M-spike patients. Renal dysfunction was more common in M-spike patients (44% v 26%) although an age effect could not be excluded. Renal dysfunction is not usually caused by GD but doi:10.1016/j.ymgme.2018.12.331 rather by co-morbidities. GBA1 genotype including N409S homozygosity was a non-discriminant. In M-spike patients, abnormal free light chain ratio was seen in 4 patients. M-spike was LT 3mg/dl in all. All had IgG isotype. Clonal plasma cells over 10% were seen in 3 patients with 316 bone marrow biopsies. Karyotype and myeloma specific FISH were Study to determine predictive potential of an algorithm for normal in 2. One showed del13. One myeloma patient was diagnosed earlier diagnosis of Gaucher disease: Retrospective biobank study with GD only after a myeloma diagnosis. The other developed myeloma utilizing real-world data available in Finland despite years of treatment with ERT and more than 4 years of eliglustat a b c d treatment. The overlap of symptoms between GD and plasma cell Markku Savolainen , Antti Karlsson , Iiro Toppila , Kaisa Elomaa , e c a neoplasia complicate efforts at differential diagnosis. Patients with GD Zoya Panahloo , Maija Wolf , Oulu University Hospital, Oulu, Finland, b c d should be monitored for MGUS and plasma cell neoplasm particularly Auria Biobank, Turku, Finland, Medaffcon Oy, Espoo, Finland, Shire, e but not exclusively after age 50. Espoo, Finland, Shire, Zug, Switzerland

Diagnosis of Gaucher disease (GD), caused by deficiency of β- fi doi:10.1016/j.ymgme.2018.12.330 glucocerebrosidase, can be signi cantly delayed due to lack of disease awareness or misdiagnoses owing to the spectrum of symptoms that overlap with other conditions. Diagnostic delay can lead to disease complications, increased morbidity and reduced 315 quality of life. The experts of the Gaucher Earlier Diagnosis Lyso-Gb3 modulation of gut microbiota biofilms: Potential Consensus (GED-C) initiative have recently reached consensus contribution to Fabry disease gastrointestinal symptoms and regarding signs and co-variables that can be classified as major or systemic complications minor early indicators of GD types 1 and 3. These 32 indicators can be used to generate guidance to facilitate early diagnosis and forms Maria Dolores Sanchez-Niño, Patricia Madrazo, Alberto Ortiz, Jaime the basis of a prototype point-scoring system (PSS) for GD. Finnish Estegan, John Jairo Aguilera Correa, IIS-Fundacion Jimenez Diaz, biobanks collect and provide biological human material for high- Madrid, Spain quality research. Here, a retrospective biobank study, supported by Shire, was conducted to locally adjust and apply the prototype PSS to Introduction/Aim: Fabry disease is characterized by the accumu- electronic patient record data in Southwest Finland. The aim of the lation of globotriaosylceramide (Gb3) and globotriaosylsphingosine study was to determine the feasibility of using hospital patient (Lyso-Gb3). Glycolipid accumulation leads to gastrointestinal man- records for assessing the clinical indicators of GD, and to use the ifestations (diarrhea, pain). Lyso-Gb3 is biologically active and obtained data to prioritize suspected patients for further sample excreted in bile, where it may reach higher concentrations than in analytics to confirm or exclude GD diagnosis. Electronic patient plasma (500 nM). We hypothesized that lyso-Gb3 secretion into the record data available from ~170,000 individuals, previously treated gut may modulate the gut microbiota. Since the gut microbiota at the Hospital District of Southwest Finland, were available for data grows as a biofilm on the enterocyte surface, we now describe the mining. Accessed data included diagnoses, laboratory measurements, effect of Lyso-Gb3 on the growth of a mixed biofilm composed of operations, imaging results, demographics and unstructured patient different bacterial species commonly found in the intestinal micro- record texts. Adjustment of the GED-C criteria was performed to biota. Methods: The formation of biofilm was evaluated by the match the local data format, Finnish language and data availability. method of Stepanovic (APMIS 2007), on TSB+1% glucose with a Using this approach, all 170,000 individuals were successfully point- mixture of 106 CFU/mL E. coli ATCC 25922 and K. pneumoniae ATCC scored for 28 parameters and a sub-cohort ‘GD suspects’, with high 23357 and 108 CFU/mL B. fragilis ATCC 25285 and C. perfringens scores, was identified. Testing of the corresponding biobanked ATCC 13124 with/without 500 nM Lyso-Gb3 for 24h in anaerobiosis. samples will be performed in the next phase. To summarize, this Enterobacteria were quantified on chromID® CPS® Elite agar, study shows the potential of using hospital patient record data in Schaedler neomycin-vancomycin agar and colistin-nalidixic acid identifying suspected patients with a rare disease for further blood agar by the drop plate method. The experiment was repeated diagnostic testing. five times. The data were analyzed in using the non-parametric Wilcoxon test. Results: The results are expressed as Log10[CFU/mL]), median (Q1-Q3). Lyso-Gb3 did not modulate the growth of E. coli doi:10.1016/j.ymgme.2018.12.332 [control 5.60 (5.16-5.64), lyso-Gb3 5.30 (5.08-5.34), p ns], K. S130 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

317 biomarker for the first stage screening of GD among Russian patients. Lyso-Gb3 is as a primary biomarker for Fabry disease screening Lyso-GL1 level was measured by UPLC-MS/MS in DBS in more than among high-risk contingents 3,500 patients with clinical manifestations of GD from 69 different subjects of the Russian Federation, who were admitted to the Kirill Savostyanova, Alexander Pushkova, Natalya Mazanovaa, Lolita diagnosis during the last 30 months. All samples with Lyso-GL1 Paka, Lyudmila Kuzenkovaa, Tatyana Podkletnovaa, Alexander values above the cut-off were subjected to molecular genetic analysis Pakhomova, Alexey Sukhozhenkoa, Sergey Moiseevb, aFSAI, National by the Sanger method. As a result molecular genetic testing was Medical Research Center for Children's Health, Moscow, Russian performed in 23 patients with elevated Lyso-GL1 levels and all of Federation, bFirst Moscow State Medical University of the Ministry of them had mutations in the GBA gene and demonstrated a reduced Health (Sechenov University), Moscow, Russian Federation activity of β-glucocerebrosidase. Among the revealed mutations five have not been described previously in the HGMD database. They Fabry disease (FD) is a rare X-linked inherited disorder that were two novel missense mutations: c.487GNC (p.A163P) and affects mainly males with mutation in the GLA gene. It causes c.863TNA (p.L288Q), two novel nonsense mutations: c.849CNA (p. accumulating of lysosomal globotriaosylceramide (Lyso-Gb3) in the Y283*) and c.544CNT (p.Q182*) and one novel deletion c.958del (p. different body's cells and reducing of α-galactosidase A activity. We L320Wfs*16) At the next stage, the Lyso-GL1 concentration was investigated more than 17500 samples from patients of high-risk measured in patients with GD confirmed earlier with molecular contingents for the period of 30 months from 31 different subjects of genetic analysis and measurement of enzyme activity, which the Russian Federation. In all patients measurement of Lyso-Gb3 by biomaterial was available before the beginning of ERT. In total, we UPLC-MS/MS method were carried out and 36 patients (0,21%) with analyzed 153 samples of patients with GD. All of them showed a Lyso-Gb3 values above the cut-off were detected, after that a family significant increase levels of Lyso-GL1 359.6 ng/ml (173.8-545.4 95% analysis was performed. The analysis of all coding and adjacent CI) compared to healthy controls 1.8 ng/ml (1.4-2.2 95% CI). Lyso- intron regions of GLA gene by the Sanger method revealed mutations GL1 concentration correlated with other biomarkers: chitotriosidase, in all 62 patients. Among the 31 different variant of GLA gene eight hepatomegaly and splenomegaly. However, associations between the mutations have not been described previously in the HGMD level of Lyso-GL1 biomarker and the type of mutations in the GBA database. They were five novel missense mutations: c.203TNC (p. gene were not found. In our study, Lyso-GL1 showed high sensitivity Leu68Pro), с.804АNС (p.Leu268Phe), c.821GNA (p.Gly274Asp) and specificity and can be used as a key biomarker for screening of c.895GNC (p.Asp299His), c.920CNG (p.Ala307Gly), one novel non- Gaucher's disease. sense mutation c.1134TNA (p.Cys378X) one novel deletion: с.949del (p.Ile317Leufs*31) and one insdel: c.539_547+9del (p. L180_G183delinsC). The next step was a retrospective analysis and doi:10.1016/j.ymgme.2018.12.334 measured the Lyso-Gb3 concentration in previously proven patients with FD. In total for 154 patients with 63 different mutations of the GLA gene we received values for the Lyso-Gb3 concentration, which 319 allowed us to correlate with the genotype. The high level of Lyso-Gb3 The European Reference Network Program for Hereditary Meta- was indicated predominantly in patients with nonsense mutation bolic Diseases (MetabERN) and deletions. In addition, it is interesting that two patients from one related family with a severe classical course of FD and missense mutation с.782GNT (p.Gly261Val) demonstrated the high level of Maurizio Scarpa, Helios Dr. Horst Schmidt Klin, Wisbaden, Germany Lyso-Gb3, and two brothers with atypical cardiac form of FD and mutation c.644ANG (p.Asn215Ser) opposite demonstrated the low The European Reference Network for Hereditary Metabolic level Lyso-Gb3. Based on our data, we can conclude, that Lyso-Gb3 Diseases (MetabERN) has been set up in response to the European can be used as a primary biomarker for FD screening. Commission call to establish European Reference Networks (ERNs) following the directive on patients’ rights in cross-border healthcare. MetabERN represents the first most comprehensive, pan-metabolic, pan-European, patient-orientated platform aimed to transform how doi:10.1016/j.ymgme.2018.12.333 care is provided to patients with inherited metabolic diseases (IMDs) in Europe. MetabERN represents 69 certified Healthcare Providers (HCP) from 18 European countries, 44 Patients Organizations and is 318 endorsed by the SSIEM. MetabERN involves 1681 experts, 52% of Glucosylfingosine (Lyso-GL1) may be the primary biomarker for which are specialized medical doctors. It manages 42.427 patients, screening Gaucher disease in Russian patients 68%of which are pediatric. More than a half of the patients are affected by lysosomal disorders or amino acid and organic acids Kirill Savostyanova, Alexander Pushkova, Lubov’ Mura’vovaa, Goar related diseases (23% and 39% respectively). The group of more than Movsisyana, Anastasia Rykunovaa, Rodion Ponomarevb, Kira Lukinab, 700 ultra rare metabolic diseases is structured in 7 subnetworks for Elena Lukinab, Leyla Namazova-Baranovaa, aFSAI, National Medical different metabolic diseases and divided in 8 workpackages (Coor- Research Center for Children's Health, Moscow, Russian Federation, dination and management, Dissemination, Evaluation, Guidelines bFSBI, National Research Center for Hematology, Moscow, Russian and Care Pathways, Virtual Counseling Framework, Research, Federation. transitional activities and clinical trials, Capacity building and training and Continuity of Care). There is also a patient involvement Gaucher disease (GD) is the autosomal recessive lysosomal working group recently created. MetabERN aims to accommodate storage disease caused by mutations in GBA gene which leads to and interconnect expertise harmonise data collection optimise accumulation of glucosylsphingosine (Lyso-GL1) and significant prevention, diagnostics, management and treatment develop and damage in various organs and tissues. We selected Lyso-GL1 as the implement guidelines stimulate cross-border research and Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S131 innovative treatments develop training and education opportunities activity against peripheral manifestations and potential activity and interact with patients. Patient organizations will work in close against progression of neurological manifestations of GD3. collaboration with HCP to map and understand patients’ needs and identify solutions. MetabERN will is facilitating and harmonizinge doi:10.1016/j.ymgme.2018.12.336 patients’ access to diagnosis and best treatment across the EU. By implementing collaboration with academia, politics, insurance companies and industry it aims to capture the most innovative 321 medical advances and tailors them to patient needs. It is also Improved description of clinical features of multiple sulfatase fostering collaboration with other ERNs in areas where overlapping deficiency: A meta-analysis of published cases exists.

Lars Schlotawaa, Joana Preiskorna, Rebecca Ahrens-Nicklasb, Laura A. Adangb, Jutta Gärtnera, Tim Friedea, aUniversity Medical Center doi:10.1016/j.ymgme.2018.12.335 Goettingen, Goettingen, Germany, bChildren's Hospital of Philadelphia, Philadelphia, PA, United States

320 Multiple sulfatase deficiency (MSD, MIM #272200) is an ultra- Venglustat in adult Gaucher disease type 3: Preliminary safety, rare lysosomal storage disorder that results from mutations in the pharmacology, and exploratory efficacy from a phase 2 trial in SUMF1 gene, which encodes formylglycine-generating enzyme combination with imiglucerase (LEAP) (FGE). Pathogenic variants in SUMF1 lead to misfolding and degradation of FGE a necessary protein for the post-translational activation Raphael Schiffmanna, Eugen Mengelb, Nigel Crawfordc, Sebastiaan of all sulfatases. Because of reduced sulfatase activities, individuals Gaemersd, Derralynn Hughese, Ana Jovanovicf, Pascal Mininig, Jyoti affected by MSD present with a severe multisystemic disorder with fi Sharmac, Tanya Fischerh, aBaylor Research Institute, Dallas, TX, United overlapping features of single sulfatase de ciencies, including States, bUniversitätsmedizin Mainz, Mainz, Germany, cSanofi Genzyme, metachromatic leukodystrophy (MLD) and several Bridgewater, NJ, United States, dSanofi Genzyme, Naarden, Netherlands, mucopolysaccharidoses (MPS) subtypes. Due to the extreme rarity eBaylor Research Institute, London, United Kingdom, fSalford Royal NHS of this disorder, clinical studies and natural disease history data on Foundation Trust, Salford, United Kingdom, gSanofi Genzyme, Chilly- larger cohorts of patients is limited. Without a comprehensive Mazarin, France, hSanofi Genzyme, Cambridge, MA, United States understanding of the onset, symptoms, and natural course of MSD, the design of future clinical trials may be difficult. In this work, we Venglustat is a selective, CNS-penetrating glucosylceramide (GL- did a systematic search for case reports, case series and other clinical fi 1) synthase inhibitor under evaluation for treatment of Gaucher studies reporting on MSD in published literature since its rst disease type 3 (GD3) manifestations that do not respond to description. As available from these published reports, we extracted enzyme-replacement therapy (ERT). The ongoing LEAP trial the systemic symptoms, age of onset, and disease trajectory and (NCT02843035/Sanofi Genzyme) assesses the safety and tolerability completed a meta-analysis of data where appropriate. From the fi of venglustat in combination with imiglucerase and disease-specific literature, we identi ed 66 relevant publications, which described 80 biomarker changes (glucosylsphingosine [lyso-GL-1] and GL-1) in individuals with 54 unique SUMF1 mutations. We found a broad cerebrospinal fluid (CSF) and plasma. Secondary objectives include phenotype spectrum, with variable severity, organ involvement, and pharmacokinetics and exploratory efficacy in neurologic and age of presentation. In this retrospective study, our primary aim was systemic disease. Eligible GD3 patients are age ≥18 with horizontal to establish a foundation for future natural history studies, to identify saccade eye movement (HSEM) impairment, received ERT for ≥3 key variables that may distinguish unique clinical subtypes, and to years with a stable imiglucerase dose for ≥6 months before understand the breadth of the clinical and mutation spectrum. enrollment, and achieved therapeutic goals. This interim analysis Additional investigations are needed to better characterize the reports 26-week data from 5 patients (mean age 24 years). At week complicated genotype-phenotype relationship of MSD. Additionally, 26, patients had measurable venglustat concentrations in CSF our results will guide an ongoing international MSD natural disease (mean 4.71 ng/ml [SD=±2.50]). At week 26, GL-1 was reduced history study and further our clinical trial readiness. from baseline by a mean 59.1% (SD=36.8%) in CSF and 60.1% doi:10.1016/j.ymgme.2018.12.337 (41.2%) in plasma, and lyso-GL1 by a mean 19.4% (63.7%) and 44.01% (35.5%), respectively. Neurological manifestations measured by the Modified Severity Scoring Tool and HSEM velocity showed no deterioration from baseline. Although no patient was thrombo- 322 cytopenic at baseline, mean platelet count increased by 16.7% Protein disulfide isomerase is a possible target for disease (SD=24.3%). At baseline, all patients had evidence of interstitial modification in multiple sulfatase deficiency lung disease via high-resolution computed tomography, which improved at week 26 in all patients except one who displayed Lars Schlotawaa, Michaela Wachsb, Olaf Bernharda, Franz J. Mayerc, reduced venglustat exposure over weeks 12-26. There were no Thomas Dierksb, Bernhard Schmidta, Karthikeyan Radhakrishnanb, clinically meaningful changes in pulmonary function tests, hemo- aUniversity Medical Center Goettingen, Goettingen, Germany, bBielefeld globin levels or liver volumes, and no serious adverse events or University, Bielefeld, Germany, cBruker Daltronics, Bremen, Germany adverse events of special interest. All 5 patients continue in the study. In conclusion, combination treatment with this oral substrate Multiple sulfatase deficiency (MSD) is an untreatable, ultra-rare reduction therapy and imiglucerase over 26 weeks is well tolerated, inherited metabolic disease, which results from mutations in the improves biomarkers in CSF and plasma, and shows encouraging SUMF1 gene encoding the formylglycine-generating enzyme (FGE). S132 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

FGE posttranslationally activates all known cellular sulfatases and sample size and lack of control data (for example, administration of impaired FGE function, due to mutation-induced misfolding, results PROMIS to other MPS types) and merit further inquiry. in catalytically impaired sulfatases. MSD patients show variable symptoms due to reduced or absent sulfatase activities and the progress of the disease is determined by residual activity and doi:10.1016/j.ymgme.2018.12.339 stability of misfolded FGE. Clinical symptoms in MSD overlap as features of single sulfatase deficiencies like metachromatic leukodystrophy and mucopolysaccharidosis in a severe multisystem disorder. In this current study, we report that protein disulfide 324 isomerase (PDI) serves as a quality control mechanism for misfolded Preliminary patient-reported outcomes and safety of advanced α FGE proteins. PDI recognizes a designated molecular signature in and targeted acid -glucosidase (AT-GAA (ATB200/AT2221) in misfolded FGE and facilitates an early and fatal protein degradation patients with Pompe disease from the ATB200-02 trial that determines disease severity in MSD. Overexpression of PDI a b c reduces the residual activity of misfolded FGE, whereas inhibition of Benedikt Schoser , Drago Bratkovic , Barry J. Byrne , Paula R. d e f g PDI function increases sulfatase activities in MSD patient fibroblasts. Clemens , Ozlem Goker-Alpan , Priya Kishnani , Xue Ming , Mark h i j These results highlight a role for PDI as a disease modifier in MSD. Roberts , Peter Schwenkreis , Kumaraswamy Sivakumar , Jacquelyn k k k k Our findings serve as a first step in the understanding of molecular Wright , Sheela Sitaraman , Jay A. Barth , Hjalmar Lagast , Tahseen l m a quality control and degradation mechanisms in MSD and the Mozaffar , Ans T. van der Ploeg , Friedrich-Baur-Institut, Neurologi- discovery of potential targets for therapeutic intervention. sche Klinik, Ludwig-Maximilians-Universität München, Munich, Ger- many, bPARC Research Clinic, Royal Adelaide Hospital, Adelaide, Australia, cUniversity of Florida, Gainesville, FL, United States, d doi:10.1016/j.ymgme.2018.12.338 University of Pittsburgh and Department of Veterans Affairs Medical Center, Pittsburgh, PA, United States, eO&O Alpan LLC, Fairfax, VA, United States, fDuke University Medical Center, Durham, NC, United States, gRutgers New Jersey Medical School, Newark, NJ, United States, 323 hSalford Royal NHS Foundation Trust, Salford, United Kingdom, Emotional, social, behavioral, and pain self-report measures and iNeurologische Klinik und Poliklinik des Berufsgenossenschaftlichen, outcomes in Morquio syndrome Universitätklinikum Bergmannsheil, Bochum, Germany, jNeuromuscular Research Center, Phoenix, AZ, United States, kAmicus Therapeutics, Inc., Ashley Schneider, Sarah Lewis, Kelly King, Chester Whitley, Brenda Cranbury, NJ, United States, lUniversity of California, Irvine, CA, United Diethelm-Okita, University of Minnesota, Minneapolis, MN, United States, mErasmus Medical Center, Rotterdam, Netherlands States Late-onset Pompe disease (LOPD) is a rare, inherited metabolic Mucopolysaccharidosis IV, Morquio syndrome (MPS IV), is an disorder that poses significant burden to patients. In LOPD, acid α- autosomal recessive disorder caused by the deficiency of N- glucosidase (GAA) deficiency results in progressive muscle weakness acetylgalactosamine-6-sulfate sulfatase, resulting in wide clinical and respiratory impairment. Recombinant human GAA enzyme heterogeneity characterized by significant skeletal manifestations replacement therapy (rhGAA ERT alglucosidase alfa) is the current (Peracha, 2018) and severe short stature. In the literature, there is standard treatment. ATB200-02 (NCT02675465) is a first-in-human, minimal information regarding behavior, emotional, and social open-label, phase 1/2 trial evaluating AT-GAA (ATB200/AT2221), an outcomes in MPS IV however, one previous study has demonstrated ERT/chaperone regimen, in adults with LOPD. The study enrolled 20 score elevation on scales assessing anxiety, depression, and with- patients: ERT-switch ambulatory (Cohort 1, n=11), ERT-switch drawal (Davison, 2013). The objective of this cross-sectional study is nonambulatory (Cohort 2, n=4), and ERT-naive ambulatory (Cohort to explore the behavioral, emotional, social, and pain outcomes of 3, n=5). This analysis evaluates the 12-month interim data for MPS IV individuals utilizing the NIH-developed PROMIS question- patient-reported outcomes, including activities of daily living naires. MPS IV participants who were enrolled in a multicenter study (Rasch-built Pompe-specific Activity [R-PAct] Scale), Rotterdam of brain structure and function in MPS disorders (U54NS065768) Handicap Scale, Fatigue Severity Scale, and Subject Global Impression were administered the questionnaires to assess anger, anxiety, of Change (SGIC). Improvements in R-PAct scores (scale: 0-36, relationships, support, fatigue, and pain. Eight subjects completed higher scores indicate less difficulty performing daily activities) were the questionnaires (6 females, 2 males). Half were adult (N N 18yo = demonstrated for Cohort 1 (mean [SD], n=10: month 6, 1.50 [3.03] 4) and half were children (N b 18yo = 4) with an age range of 11-53. month 9, 0.60 [3.60] month 12, 1.70 [3.74]) Cohort 2 (n=4: month 6, A standardized T-score of 50 (±1 SD=10) is the mean score for the 1.50 [2.38] month 9, 1.00 [2.00] month 12, 1.00 [2.00]) and Cohort 3 general population. Children and adults were given similar assess- (n=5: month 6, -0.20 [0.84] month 9, 1.00 [1.23] month 12, 2.60 ments respective of age. In both groups, no mean T-scores were ± 1 [3.51]). Cohorts 2 and 3 improved on the Rotterdam Handicap Scale SD or greater for any domains. Mean T-scores N ±0.5 SDs were found and all cohorts demonstrated improvements in the Fatigue Severity only on the fatigue (T-score: 55.525, range: 45.4-63.6) scale for Scale. Most patients reported improved overall physical well-being children and the anxiety (T-score: 56.0, range: 50.8-60.7), pain on the SGIC (Cohort 1, n=6/10 Cohort 2, n=4/4 Cohort 3, n=5/5). interference (T-score: 56.6, range: 40.7-63.5), emotional support (T- The most common treatment-emergent adverse events were ab- score: 55.75, range: 52.1-63.5), and informational support (T-score: dominal pain (n=8), diarrhea (n=8), and nasopharyngitis (n=6). 57.875, range: 48.9-69.1) scales for adults. Children and adults Three events of infusion-associated reactions (skin discoloration, scored similarly on questionnaires relating to pain quality and peer erythema, pruritus) occurred in ≥550 infusions. These data demon- relationships/isolation. This pilot data found no significant elevations strate benefits in activities of daily living with AT-GAA (ATB200/ for MPS IV in areas described above. However, caveats include small AT2221) in ERT-naive patients and both ambulatory and Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S133 nonambulatory ERT-experienced patients, suggesting that AT-GAA CLN2 disease, a rare, inherited, pediatric, neurodegenerative (ATB200/AT2221) has the potential to be a well-tolerated treatment lysosomal storage disorder caused by TPP1 deficiency, is character- for LOPD that reduces the disease burden. Updated 24-month ized by seizures, language and motor function loss, blindness, and efficacy/safety data will be included in the presentation. early death. An open-label study demonstrated that intracerebroventricular (ICV) infusion of 300 mg cerliponase alfa, a recombinant doi:10.1016/j.ymgme.2018.12.340 human TPP1 enzyme, every other week for 48 weeks slowed deterioration in motor and language function. This extension study (NCT02485899) assesses the long-term safety and efficacy of 325 cerliponase alfa for up to 240 weeks. Subjects who completed the Newborn genome editing improves phenotype, cardiovascular, initial study continued receiving 300 mg cerliponase alfa q.o.w. in respiratory, and bone disease in mucopolysaccharidosis type I this open-label extension study. Cumulative data from both studies mice were used to evaluate long-term safety (assessed by adverse events (AEs) frequency) and efficacy (assessed by changes in the CLN2 clinical rating scale motor and language (ML) domains). 24 subjects Roselena S. Schuh, Esteban A. Gonzalez, Angela M.V. Tavares, Roberto were treated with cerliponase alfa in the open-label study (9 male, Giugliani, Ursula Matte, Helder F. Teixeira, Guilherme Baldo, 15 female, mean (SD) age 4.3 years (1.24)) 23 subjects enrolled in Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil the extension study (162 to 239 weeks treatment, median 193.7 Mucopolysaccharidosis type I (MPS I) is a lysosomal disorder weeks) 22 remained on study for at least 168 weeks. Common AEs caused by deficiency of alpha-L-iduronidase (IDUA), which promotes included pyrexia, vomiting, and convulsion. Fewer subjects reported fi multisystemic storage of glycosaminoglycans (GAG). Accumulation convulsions over time (21 (88%) during the rst 24 weeks of therapy of dermatan and heparan sulfate in bone, cartilage, heart valves, vs 6 (26%) during the last 24 weeks). Twenty (83%) subjects lungs, and widespread tissues likely contributes to reduced mobility experienced serious AEs including hypersensitivity and device- b and increased morbidity in untreated MPS I individuals. We related infections. The responder ( 2 point loss per 48 weeks) rate previously reported that neonatal hydrodynamic injection of liposo- at 160 weeks was 83% (p=0.0013). The rate of decline in ML score b mal CRISPR/Cas9 complexes expressing murine Idua resulted in long- (mean (95% CI): 0.20 (0.10, 0.30) points/48 weeks, p 0.0001) was fi lasting IDUA activity in MPS I mice, and secreted IDUA could be taken signi cantly attenuated compared with a rate of decline of 2.0 up from blood by multiple organs, except for the brain. In the present points/48 weeks in untreated patients, and has improved since prior study, we performed additional analysis and report the effect of this analysis at 48 weeks (0.40 points/48 weeks). These data suggest that therapy on the alterations found in the cardiovascular, respiratory, enzyme replacement therapy with ICV-administered cerliponase alfa fi bone, and brain tissues. There were improvements in facial has an acceptable safety pro le and a sustained treatment effect. morphology and large bones, specifically in zygomatic and femoral width. The heart had reduced systolic and diastolic diameters, and doi:10.1016/j.ymgme.2018.12.342 cardiac mass, although heart valves presented cell hyperplasia and lysosomal storage. In addition, MPS I treated mice had improved aortic diameter and reduced elastin breaks. The respiratory function was evaluated and lung resistance was normalized. However, as 327 expected, the brain had no improvements, as neuroinflammation Genetic approaches for diagnosis of two Brazilian patients with was still present. Furthermore, these mice had just partial improve- mucolipidosis II/III alpha/beta ments in behavioral tests, suggesting deterioration in brain function. We conclude that neonatal gene therapy can improve some aspects Ida Vanessa Doederlein Schwartz, Nataniel Floriano Ludwig, Devora of bone, heart, and lung disease in MPS I mice. However, better Random, Fernanda Sperb Ludwig, Universidade Federal do Rio Grande delivery of our gene product to these and other “difficult-to-treat” do Sul, Porto Alegre, Brazil tissues will be necessary to achieve more profound effects in bone, heart, and especially in brain. (Support: FIPE-HCPA, CNPq Brazil) Mucolipidosis II and III (ML II/III) alpha/beta are autosomal recessive lysosomal disorders characterized by the abnormal trafficking of lysosomal enzymes due to pathogenic variants in GNPTAB and GNPTB doi:10.1016/j.ymgme.2018.12.341 genes. These genes encode the alpha and beta subunits of N- acetylglucosamine-1-phosphotransferase, a key enzyme for mannose- 6-phosphate marker. Genetic analysis by Sanger sequencing is able to identify 90% of pathogenic variants in ML II/III patients. The objective of 326 present work is to describe the genetic investigation of two Brazilian Persistent treatment effect of cerliponase alfa in children with patients with biochemical and clinical diagnosis of ML II and III. DNA CLN2 disease: A 3 year update from an ongoing multicenter from both patients (1 and 2) was extracted from peripheral blood. RNA extension study of patient 2 was extracted and cDNA was synthetized. GNPTAB gene was analyzed in both patients by Sanger sequencing. The patient 2 had Angela Schulza, Nicola Specchiob, Paul Gissenc, Emily de los Reyesd, additional analyzes in DNA by next generation sequencing (NGS) and in Heather Cahane, Peter Slasore, David Jacobye, Temitayo Ajayie, cDNA by Sanger sequencing. The DNA sequencing of patient 1 was aUniversity Medical Center Hamburg-Eppendorf, Hamburg, Germany, concluded easily by Sanger, identifying the pathogenic variant bBambino Gesù Children’s Hospital, Rome, Italy, cGreat Ormond Street c.2249insA in homozygosis. In patient 2 the same analysis identified Hospital for Children, London, United Kingdom, dNationwide Children’s the heterozygous allele c.242GNT in the patient and in his father. NGS Hospital, The Ohio State University, Columbus, OH, United States, presented the same results and was negative for alterations in GNPTG. eBioMarin Pharmaceutical Inc., Novato, CA, United States The sequencing of cDNA concluded the diagnosis identifying the S134 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 r.86_116delins86+19_49 and c.242GNT. Some cases are easily solved Tabrizi, Raphael Schiffmann, Ajay Goel, Baylor Research Institute, and others are extremely challenging in molecular diagnostics. The Dallas, TX, United States variant r.86_116delins86+19_49 is novel. Sanger and NGS present limitations in detecting large insertions/deletions and sometimes only Epigenetic modifications are implicated in the development of RNA analysis can conclude the case. Further analyses will confirm which human diseases however, the role of epigenetics in the pathogenesis DNA change is causing r.86_116delins86+19_49. RNA analysis may be of lysosomal storage disorders remains unclear. The aim of present an important tool to conclude the genetic diagnosis and to perform study was to unravel the epigenetic basis of Fabry disease by genetic counseling. Support: FIPE/HCPA, CNPq, CAPES, FAPERGS. analyzing DNA methylation in patient’s cells. To test whether α-gal A deficiency can affect DNA methylation, we generated two isogenic models using Fabry patient-derived endothelial cells (IMFE1): 1) doi:10.1016/j.ymgme.2018.12.343 enzyme deficiency in IMFE1 was corrected by stable expression of normal α-gal A gene, and this corrected IMFE1 was compared with mock-treated IMFE1 (the genetic model) 2) IMFE1 cells were treated with deoxygalactonojirimycin, a potent inhibitor of α-gal A, for 2 328 weeks, and these cells were compared with mock-treated IMFE1 (the The utility of the swallow study in the diagnosis and management chemical model). Deoxygalactonojirimycin inhibited residual activity of type 2 Gaucher disease of IMFE1 cells, leading to significantly elevated globotriaosylceramide level compared to mock-treated cells. First, Gurpreet K. Seehra, Beth Solomon, Emory Ryan, Tamanna Roshan Lal, by using bisulfite pyrosequencing, we examined methylation of Grisel Lopez, Ellen Sidransky, National Institutes of Health, Bethesda, specific CpG regions in androgen receptor (AR) gene, whose MD, United States expression is upregulated in Fabry disease. We found that, in both the ‘genetic’ and the ‘chemical’ models, decreased α-gal A activity Gaucher disease (GD) is an inherited deficiency of the enzyme was associated with hypomethylation of one of the CpG regions in glucocerebrosidase, resulting in diverse phenotypic manifestations AR promoter. We further performed genome-wide DNA methylation including non-neuronopathic (Type 1), acute neuronopathic (Type 2), array that covers more than 865,000 CpG sites. There were N165,000 and chronic neuronopathic (Type 3) forms. Type 2 (GD2) is the most differentially methylated probes (DMPs) and N65,000 differentially severe form of GD, presenting as early as prenatally, and resulting in methylated regions (DMRs) between α-gal A-deficient cells and the death within the first years of life. There is not yet a treatment that cells with normal activity (the genetic model). We also identified 42 effectively targets the neurological manifestations of GD, although DMPs and 4,699 DMRs in the ‘chemical model’. Collectively, these new therapeutic strategies are under development. While discerning results suggest that α-gal A deficiency causes dysregulated DNA GD2 from other types of GD in the first months of life can be methylation that may play important roles in the pathogenesis of challenging, we have constantly noted neurological involvement Fabry disease. Further analysis of methylome and transcriptome of resulting in deteriorating swallow functions in patients with GD2, these samples will likely lead to identification of new target genes when compared to patients with Type 1 (GD1) and 3 (GD3) Gaucher and pathways that may be useful to better understand Fabry disease disease. A dysfunctional swallow can lead to respiratory aspiration and and to develop novel therapies. subsequent pneumonia or failure to thrive. Early diagnostic measures, like videofluoroscopic swallow studies, are extremely useful in ’ directing management of these patients and establishing the infant s doi:10.1016/j.ymgme.2018.12.345 neurological status. In this study, we reviewed charts of patients with GD2, focusing on documented results of swallow studies in the context of other neurological and visceral manifestations. We scored the swallow study results into four carefully selected scales and describe 330 the results in a series of 10 patients with complete neurological Phase I/II gene transfer clinical trial of scAAV9.U1a.hSGSH for MPS profiles. The evaluations demonstrate that impaired swallow is an IIIA vector shedding results over 6 months post-gene transfer early and universal finding in patients with GD2, which provides insight into the rapid non-linear disease progression. This illustrates Pavel Shiyanov, Scott Kerns, Linas Padegimas, Timothy J. Miller, Juan that early impaired swallow function can be used to confirm the Ruiz, Abeona Therapeutics, Cleveland, OH, United States diagnosis of GD2, anticipate neurological decline, and help direct management of patients with aspiration. Furthermore, this test may Sanfilippo syndrome type A (mucopolysaccharidosis type IIIA), a have utility as an efficacy parameter when considering therapeutic lysosomal storage disorder resulting from mutations in the SGSH (N- trials for acute neuronopathic GD. sulphoglucosamine sulphohydrolase) gene, gives rise to intracellular glycosaminoglycan accumulation and results in cellular dysfunction and death. A global, open-label, dose-escalation Phase I/II gene doi:10.1016/j.ymgme.2018.12.344 transfer trial using intravenous administration of a recombinant scAAV9.U1A.hSGSH vector with tropism for both CNS and relevant somatic tissues has demonstrated significant transduction in multiple tissues. We evaluated pharmacokinetic metabolism throughout 329 the study in the high dose cohort by measuring vector shedding in Dysregulated DNA methylation in the pathogenesis of Fabry subjects treated at the high dose of 3E13 vg/kg. Tissue samples were disease analyzed by qPCR to quantitate AAV9.hSGSH virus shedding in plasma, feces, saliva and urine samples. We optimized DNA isolation Jin-Song Shen, Kunitoshi Shigeyasu, Yoshinaga Okugawa, Uthra methods and qualified the absolute qPCR quantitation assay with a Balaji, Jinyan Chan, Jinghua Gu, Taniqua Day, Siamak Jabbarzadeh- LLOQ = 10 v.g. per reaction mix or 1 v.g. per 2 μL of specimen. Vector Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S135 shedding analysis revealed the highest viral DNA copy numbers in drug approved as enzyme replace therapy (ERT). This report plasma specimens 4 hours after the treatment. After 14 days virus describes the case of a 38-year-old male Fabry patient who started titers in plasma dropped in magnitude, but were still detectable 180 to have hypersensitivity reactions to Fabrazyme® after 14 years of days after dosing. The vector shedding profile varied by subject, and his first drug infusion and after a 6-month- shortage of Fabrazyme® in most cases no more than 109 v.g./mL (for feces and saliva) and no in Brasília, Brazil, due to local government purchase problems. The more than 106 v.g./mL (for urine) were detected. For the majority of AF disease male patient received Fabrazyme® home infusion for patients, shedding vector titers in saliva and urine dropped to more than 13 years and at the day after the first infusion following undetectable levels in 60-90 days after dosing. These results can help shortage, he experienced fever, chills, abnormal edema in hands and prospectively guide future clinical trial designs for gene transfer feet, diffuse myalgia and headache. He needed several antihista- studies and patient sample handling. minics and analgesics intravenous drugs before symptoms relief. During the following 15 days before next infusion he experienced the recurrence of above symptoms in lower severity. After the doi:10.1016/j.ymgme.2018.12.346 second infusion he had upper airways symptoms such as sore throat, hoarseness and dyspnea. He was diagnosed with anaphylaxis and since that, he had a change in the Fabrazyme® infusion place to hospital. Moreover, the ERT was made slower, fractioned and the 331 patient would be previously treated with a desensitization scheme The genetics of Mendelian disorders is not always simple: Lessons which included morphine, ranitidine, dipyrone, sodium from Gaucher disease montelucaste and cetirizyn. In spite of this disensitization scheme, he experienced low severity hypersensitivity reactions that de- Ellen Sidransky, Yushiro Tanima, Nahid Tayebi, Nahid Tayebi, Nahid creased his quality of life and kidney function. At that time, he was Tayebi, NHGRI, Bethesda, MD, United States investigated for other diseases and a rare gene mutation in Factor 12 was discovered. We decided to treat him with danazol and he had a fi Gaucher disease, the inherited de ciency of lysosomal complete relief of all hypersensitivity reactions. glucocerebrosidase is a rare lysosomal storage disorder with a recessive pattern of inheritance. However careful genotypic analyses doi:10.1016/j.ymgme.2018.12.348 of patients with this disorder reveal that the genetics is not always as straightforward as would be anticipated, as reflected in examples of patients with atypical inheritance. In our cohort we have observed 333 two patients who inherited two copies of a mutated paternal allele, Attenuated multiple sulfatase deficiency: Description of two occurring as a result of uniparental disomy. We also have three Brazilian patients showing interesting clinical and genetic patients where no mutation could be found in the mother’s DNA, findings suggesting that the second mutant allele arose either by germline mosaicism or as a result of a new mutations in the proband. The Thiago O. Silvaa, Carolina F.M. Souzaa, Joshua W.B. Rochaa, Ana C. glucocerebrosidase (GBA1) locus on chromosome 1q21 is a gene rich Brusius-Facchina, Kristiane Michellin-Tirellia, Maira G. Burina, region with multiple genes and pseudogenes located in close Roberto Giugliania, Karthikeyan Radhakrishnanb, Lars Schlotawac, proximity. Some mutant alleles encountered in patients result from Thomas Dierksb, Ida V.D. Schwartza, aHospital de Clínicas de Porto recombination events occurring between the GBA1 gene and Alegre, Porto Alegre, Brazil, bUniversity of Bielefeld, Bielefeld, Germany, pseudogene, including duplications and deletions. Furthermore, cUniversity Medical Center Göttingen, Göttingen, Germany some patients have been identified who carry more than two discrete mutations in GBA1. The complexity observed is not unique Multiple sulfatase deficiency (MSD) is a rare lysosomal disease to Gaucher disease, as next generation sequencing strategies are caused by biallelic loss of function mutations in the SUMF1 gene, increasingly revealing such events more frequently. Thus, clinicians leading to a functional defect in the post-translational activation of must be aware that exceptions to traditional genetics exist, and that all sulfatases. The clinical presentation comprises symptoms from the such events can impact diagnostic testing and genetic counseling for individual sulfatase deficiency disorders, with variable severity. Here Mendelian disorders. we describe two cases of milder presentations of MSD, both female. Patient 1 was first referred to medical geneticist at 17 months to investigate mild developmental delay and minimal dysmorphic doi:10.1016/j.ymgme.2018.12.347 features (flat face, bifid uvula, long fingers), with history of parental consanguinity and one male sibling prematurely deceased. Initial workup, including neuroimaging and basic metabolic screening 332 (lysosomal tests not included), had normal results. At subsequent Description of a patient with Anderson Fabry disease and a evaluations, dry skin was observed. She lost follow-up at our clinics mutation in factor XII: A case report and returned at 8 years old, presenting neurological regression, aggressive behavior, ichthyosis and short stature. Neuroimaging Sandra M. Silvaa, Camila Veronezb, Carolina Sanches Arandab, Jose showed dysgenesis of the corpus callosum, white matter hyper- Sobral Netoc, Joao B. Pesquerob, aInstituto Hospital de Base do Distrito intensities and cerebral and cerebellar atrophy. Urinary glycosami- Federal, Brasilia, Brazil, bUniversidade Federal de São Paulo, Sao Paulo, noglycans chromatography was requested and presented abnormal Brazil, cCENTROCARD, Brasilia, Brazil results, leading to a new extensive metabolic workup which demonstrated MSD. Next generation sequencing (NGS) of SUMF1 Anderson Fabry (AF) disease is a rare X-linked lysosomal disorder showed no abnormalities, but Sanger sequencing revealed a N caused by the alfa galactosidase A deficiency which leads to Gb3 previously reported variant c.1A G mutation (p.Met1Val) in homo- fi deposition in different organs. In Brazil, Fabrazyme® is an orphan zygous state. Patient 2 was rst referred to medical geneticist at 10 S136 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 years of age with previous investigation but without conclusive Cahani, Anita Groveri, Andrew Meltoni, Lynn Smithi, Stephen M. diagnosis, presenting mild developmental delay, hepatomegaly and Maricichi, Fatih Ezgub, aUniversidad del Rosario, Bogota, Colombia, hypercholesterolemia since infancy. We found the activity of bGazi University Hospital, Ankara, Turkey, cGreat Ormond Street arylsulfatase A and other sulfatases to be deficient. A novel Hospital, London, United Kingdom, dHospital Clínico Universitario de pathogenic variant p.Arg224Gln (c.671GNA) in SUMF1 was detected Santiago, A Coruña, Spain, eUCSF Benioff Children’s Hospital Oakland, by NGS, in heterozygosis. The second variant has not been detected Oakland, CA, United States, fRoyal Children’s Hospital, Melbourne, yet. In-silico analysis and 3-D modelling confirm the mutation is Australia, gMackay Memorial Hospital, Taipei, Taiwan, hUniversity pathogenic. These two cases demonstrate how MSD diagnosis may Medical Center Hamburg-Eppendorf, Hamburg, Germany, iBioMarin be a real challenge, mainly with the milder presentations. Pharmaceutical Inc., Novato, CA, United States

Sanfilippo syndrome type B (MPS IIIB) is caused by deficiency of doi:10.1016/j.ymgme.2018.12.349 the α-N-acetylglucosaminidase (NAGLU) enzyme and subsequent heparan sulfate (HS) accumulation in the brain. Sanfilippo syndrome type B patients experience progressive neurocognitive, behavioral and motor decline, typically succumbing to the disease in the second 334 or third decades of life. To date no studies have comprehensively or fi Clinical pro le of mucopolysaccharidosis type I patients from a quantitatively characterized Sanfilippo syndrome type B progression Brazilian reference center in young patients. This information is essential for defining measures of therapeutic efficacy in treatment studies. We initiated an ongoing Joselito Sobreira, Marco Antônio Curiati, Carmen Silvia Curiati observational study of Sanfilippo syndrome type B patients aged 1- Mendes, Maret Holanda Rand, Ana Maria Martins, Universidade 10 years (BMN 250-901 NCT02493998) in an effort to better Federal de São Paulo, São Paulo, Brazil characterize multiple facets of disease progression in this population. All enrolled subjects had deficient NAGLU activity and cognitive Mucopolyssacharidosis type I (MPS I) is an autosomal recessive developmental quotient (DQ) ≥50 at time of enrollment. Cognitive inborn error of metabolism caused by deficiency in the lysosomal function and behavior is assessed every 12 weeks measurement of enzyme alpha-L-iduronidase (IDUA), necessary for the degradation heparan sulfate levels, abdominal and brain MRI imaging, assessment of heparan sulfate and dermatan sulfate. The aim of this study is to of hearing function and quality of life measures are conducted every describe the clinical profile of 35 MPS I patients followed at a 24 weeks. Preliminary data demonstrate that CSF heparan sulfate Brazilian reference center. In our cohort of 35 patients, 20 (57,1%) levels are universally elevated in Sanfilippo syndrome type B were male. The patients had a median age of diagnosis of 5y, ranging patients, but to widely varying degrees. Cognitive decline occurs in from 9m to 27y. The median age for starting ERT was 5y, ranging either a progressive linear or stepwise fashion as patients’ age and is from 3m to 25y. Consanguinity was observed in 8/35 (22%). Age of similar to but slower than that seen in young Sanfilippo syndrome symptom onset ranged from 1m to 8y (mean of 2y). Age of symptom type A patients. Baseline measures of cognitive function are strongly onset ranged from 1m to 8y (median of 12m). The main symptoms correlated with caregiver assessments of adaptive function, demon- reported by parents at diagnosis were developmental delay, chronic strating good construct validity of the utilized cognitive tests. diarrhea, upper airway infections (UAI), and osteoarticular deformi- Abdominal MRI imaging demonstrates baseline enlargement of liver ties. At the examination, the patients presented coarse facies in and spleen that tends to worsen over time. Whole brain volume 81.8%, joint limitation in 80%, recurrent UAI in 78.8%, tends to decrease and ventricular volume tends to increase over time hepatosplenomegaly in 71.4%, umbilical hernia in 68.5%, macrocrania these parameters are outside normal range in a subset of patients. was observed in 42.8%, corneal clouding in 40%, hearing impairment These data represent a systematic, quantitative evaluation of in 31.4% and cardiac involvement in 25%. From the total of 35 Sanfilippo syndrome type B disease progression in young patients. patients, 5 lost regular follow-up, 2 opted to be followed in their This rich dataset will serve as an important comparison dataset for hometown, and 13 died. To date, we have 15 patients in regular an ongoing treatment trial (BMN 250-201 NCT02754076). follow-up and ERT. We conclude from this cohort that coarse facies, joint limitation, recurrent UAI, hepatosplenomegaly and umbilical fi hernia were the most frequent ndings. The prevalence of our doi:10.1016/j.ymgme.2018.12.351 findings is in line with the findings found in the literature. The recognition of phenotypes by the pediatrician is fundamental for the diagnosis and early onset of ERT, which is related to a better prognosis. 336 Farber disease (acid ceramidase deficiency): Data from an ongoing natural history study doi:10.1016/j.ymgme.2018.12.350 Alexander Solyoma, Carlos Ferreirab, John Mitchellc, Norberto Guelbertd, Neslihan Mungane, Fatma Bulute, Bo Magnussonf, Erik Sundbergf, Christina Lampeg, Nur Arslanh, Balahan Makayh, Ratna 335 Purii, Sunita Bijarnia-Mahayi, Laila Selimj, Iman Gamal el Dinj, Seema fi Natural history data for young subjects with San lippo syndrome Kapoork, Maja Di Roccol, Seza Ozenm, Ezgi Deniz Batum, Gulden type B (MPS IIIB) Gokcayn, Marta Torcolettio, Alan Kimurap, Paul Harmatzq, aEnzyvant, Basel, Switzerland, bChildren's National Medical Center, Washington, a b c Martha Solano Villarreal , Ilyas Okur , Maureen Cleary , Maria Jose de DC, United States, cMontreal Children's Hospital, Montreal, QC, Canada, d e f g Castro Lopez , Paul Harmatz , Joy Lee , Shuan-Pei Lin , Maria Luz dChildren's Hospital of Cordoba, Cordoba, Argentina, eCukurova Univer- d h f i Couce , Nicole Muschol , Heidi Peters , Adam J. Shaywitz , Heather sity Hospital, Adana, Turkey, fKarolinska University Hospital, Stockholm, Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S137

Sweden, gHelios Dr. Horst Schmidt Kliniken, Wiesbaden, Germany, attention and behavioral control in the MPS IVA population (J Inherit hDokuz Eylul University Hospital, Izmir, Turkey, iSir Ganga Ram Metab Dis (2013) 36:323–328). The objective of this cross-sectional Hospital, New Delhi, India, jCairo University Children's Hospital, Cairo, study was to investigate potential neurocognitive abnormalities in Egypt, kLok Nayak Hospital and Maulana Azad Medical College, New MPS IVA via direct measurements of neuropsychological functioning. Delhi, India, lIstituto Giannina Gaslini, Genoa, Italy, mHacettepe MPS IVA participants from a multicenter study of brain structure and University Hospital, Ankara, Turkey, nInstanbul University Hospital, function in MPS disorders were administered a comprehensive Istanbul, Turkey, oUniversity of Milan, Milan, Italy, pEnzyvant, Boston, neuropsychological assessment of intellectual functioning, attention, MA, United States, qUCSF Benioff Children's Hospital Oakland, Oakland, verbal and visual memory, visual-spatial skills, and adaptive CA, United States functioning. Test performance yields standardized scores with μ=100 and SD=15, with qualitative ranges of average (85-115), Farber disease is a rare lysosomal storage disorder caused by below average (70-84), and impaired (b70). The study cohort mutations in both alleles of the ASAH1 gene. The resulting deficiency (N=11) consisted of approximately equal pediatric and adult of the lysosomal enzyme acid ceramidase causes accumulation of the participants (N b 18yo = 6 N ≥ 18yo = 5) with a median age of 17 pro-inflammatory and pro-apoptotic sphingolipid ceramide. The (range 11-53). Results indicated average functioning across all resulting broad spectrum of symptoms and severity may delay domains except attention. All ages were below average on a scale diagnosis or lead to misdiagnosis. The ongoing study described here related to consistency of focus, with mean standard scores of 71 for is the first comprehensive and systematic clinical study of the natural pediatrics (SD 27 median 73 range 40-106) and 78 for adults (SD 12 history of Farber disease. The Observational and Cross-Sectional median 76 range 64-92). In addition, pediatric participants were Cohort Study of the Natural History and Phenotypic Spectrum of below average for sustained attentional vigilance, with a mean Farber Disease (NCT03233841) is designed to collect retrospective standard score of 72 (SD 36 median 68 range 40-110). These and prospective data including demographics, clinical presentation, observations offer quantifiable evidence of attention problems in phenotype, and diagnostic history of patients diagnosed with Farber patients with Morquio syndrome. Findings should be considered in disease who have or have not undergone hematopoietic stem cell the context of mounting evidence that neurocognitive problems exist transplantation (HSCT), along with specific prospective clinical even in MPS types with normal IQs, such as MPS I and MPS II. evaluations (including measures of symptom impact through National MPS Society, Lysosomal Disease Network U54NS065768, standardized physical assessments and patient reported outcomes) UMN Center for Neurobehavioral Development in living patients. From November 2017 to September 2018, 34 patients (19 living, 15 deceased) have been enrolled in the study. The average age of the living patients is 6.9 years, with ages ranging doi:10.1016/j.ymgme.2018.12.353 from 1 to 28 years. 16 living patients (84%) are below 18 years of age. 80% (12/15) of deceased patients were born in the last decade. Patient countries of birth include Afghanistan, Argentina, Canada, Egypt, Germany, India, Iraq, Italy, Mexico, Sweden, Syria, Turkey, and 338 fi the USA. Patients representing the full breadth of the known Liver biopsy ndings in patients with Gaucher disease: Experience phenotypic spectrum of Farber disease, from rapidly progressive of the reference center of Rio Grande do Sul, Brazil (severe), to slowly progressive (attenuated) have been enrolled from 15 centers in 9 countries. Demographic data and numbers of patients Rodrigo T. Starostaa, Filippo P. Vairob, Suelen P. Basgaluppa, Carlos enrolled in less than 12 months indicate that Farber disease is likely Thadeu S. Cerskia, Mário R. Álvares-da-Silvaa, Ida Vanessa D. not as rare as previously thought. This unique dataset will hopefully Schwartza, aUniversidade Federal do Rio Grande do Sul (UFRGS), Porto expand and deepen the understanding of Farber disease and Alegre, Brazil, bCenter for Individualized Medicine, Mayo Clinic, contribute to the information available for the evaluation of future Rochester, MN, United States potential therapies. The extent of liver disease in Gaucher disease (GD) ranges from hepatomegaly to cirrhosis. Non-invasive methods may not accurately doi:10.1016/j.ymgme.2018.12.352 detect or diagnose liver abnormalities in GD patients, so a biopsy should be considered a suitable approach. The objective was to review the liver findings of a cohort of GD patients. Data from 41 patients followed at the Reference Center for GD of the HCPA, Brazil 337 were retrieved. Nine treated patients with GD type 1 (pts A-H, 23 to Evidence of attention problems in Morquio syndrome 67 years-old) underwent liver biopsy, yielding 10 biopsies. Biopsy indications were: (i) staging of fibrosis (ii) consistently elevated liver Katherine Spurlocka, Brenda Diethelm-Okitaa, Ashley Schneidera, enzymes and (iii) high ferritin level despite treatment. One patient Kelly Kinga, Leigh Charvetb, Sarah Lewisa, Chester B. Whitleya, Julie (pt G) without symptoms of liver disease or laboratory abnormalities Eisengarta, aUniversity of Minnesota, Minneapolis, MN, United States, was biopsied during cholecystectomy, and steatohepatitis was bNew York University, New York City, NY, United States detected. No patient had serious complications from the procedure. On liver ultrasounds, four patients (pts A-D) had steatosis one had Morquio syndrome, mucopolysaccharidosis type IVA (MPS IVA), cirrhosis (pt E), and four (pts F-I) had normal findings. Biopsy is caused by deficiency of the degrative lysosomal enzyme N- confirmed bridging fibrosis and Gaucher cells (GC) in pt A mild acetylgalactosamine-6-sulfatase, leading to cellular glycosaminogly- steatosis and haemosiderosis in pt B teatosis, mild inflammation, and can deposition and progressive, severe skeletal dysplasia, dental cholestasis in pt C steatohepatitis in pts D and G cirrhosis and small abnormalities, corneal clouding, and severe short stature. Morquio nodules of atypical GC and, in a second biopsy, a hepatocellular syndrome has not typically been associated with neurocognitive carcinoma (HCC) in pt E perisinusoidal fibrosis and GC in pt F mild deficits but a recent publication suggests evidence of problems with inflammation, haemosiderosis, nuclear glycogenosis, and GC in pt H S138 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 active hepatitis B in pt I. All patients had findings that were not 340 accurately determined non-invasively. Two cases of steatohepatitis Critical care situations in adult patients with muco- and one of HCC were detected, prompting specific treatment. polysaccharidosis (MPS) Significant non-cirrhotic fibrosis was diagnosed in one patient, leading to a regular specific follow-up. Hemosiderosis was detected Karolina Stepiena, Anahit Gevorkyanb, Christopher Hendrikszc, in several patients, reinforcing the need of following iron status Tinatin Lobhanidzeb, Jordi Pérez-Lópezd, Mireia del Torod, Nato closely. We propose that liver biopsy should be indicated during the Vashakmadzeb, Christina Lampee, aSalford Royal NHS Foundation regular follow up of patients with GD who are at risk of liver Trust, Salford, United Kingdom, bResearch Center for Children's Health, compromise. Moscow, Russian Federation, cUniversity of Pretoria, Pretoria, South Africa, dVall D'Hebron University Hospital, Barcelona, Spain, eHelios Dr. Horst Schmidt Kliniken Weisbaden, Weisbaden, Germany doi:10.1016/j.ymgme.2018.12.354 Mucopolysaccharidoses (MPS) are rare, inherited disorders associated with enzyme deficiencies that result in glycosaminoglycan (GAG) accumulation in multiple organ systems. Because of advances in 339 diagnostic, medical and surgical procedures, more patients with MPS Parent report of school functioning and behavior in children with are surviving to adulthood. However, MPS can result in a range of Pompe disease critical clinical situations in adult patients, who often require multiple surgeries and complex care planning throughout their lives. We Mihaela C. Stefanescu, Gail A. Spiridigliozzi, Stephanie L. Austin, Priya illustrate challenges and practical solutions to optimise care of adult S. Kishnani, Duke University Medical Center, Durham, NC, United States patients with MPS through case studies. Eight cases (MPS I, II, IVA and VI ages 21-38 years) were provided from four leading European Due to the effectiveness of enzyme replacement therapy, centres for inherited metabolic diseases. Critical care situations children with Pompe disease are living longer. However, the impact included surgical procedures (spinal decompression, cardiac valve or of this condition on their school functioning and developmental corneal replacement, and tracheostomy), pregnancy, and a thrombus abilities, such as attention, executive function, and social compe- in a port-a-cath. Major surgical challenges included managing fi tence remain unclear. We describe our ndings from a cross- complex cardiac and respiratory dysfunction, and adapting procedures sectional analysis of behavior checklists completed by parents of to patients’ short stature, particularly in the context of limited children with Pompe. Twenty participants (5-18 years, median age expertise in MPS. These were resolved by involving paediatric- and 9.5 years) with infantile (17/20) and late-onset (3/20) Pompe adult-care specialists with MPS expertise, including from external disease enrolled in a longitudinal study at Duke University were centres when needed. Medical challenges included disrupting enzyme assessed yearly using specially designed and standardized parent- replacement therapy during pregnancy and breastfeeding, resulting in administered questionnaires. These included a survey of school increased risk of infection. This was addressed by regular follow-ups interventions the Child Behavior Checklist For Ages 6-18 (CBCL) as and antibiotics as required. The thrombus in the port-a-cath was a measure of behavior problems, mood and social competence the resolved by Hickman line insertion, using sterile methods, and disease Connors-3 Parent (Connors-3) targeting behaviors associated with management support for the patient. These cases highlight the fi attention-de cit/hyperactivity disorder and the Behavior Rating heterogeneity of MPS and the diverse factors that should be considered Inventory of Executive Function-2 (BRIEF-2) to assess aspects of when coordinating MPS care plans. Where adult-care clinicians have executive function. Data from Year 1 were analyzed. Seventeen limited practical skills in MPS management, expertise can be sought children were based in general education classrooms, and three from more experienced paediatric teams. As well as standardised were based in special education classrooms. The most commonly approaches, with multidisciplinary input from paediatric- and adult- reported classroom accommodations were extra time to complete care experts who are familiar with the clinical needs of patients with assignments, seating closer to the teacher, frequent breaks as MPS, individualised support is required for all adult patients to manage fi needed, and use of a device for written work. CBCL ndings their specific symptoms and possible clinical complications. comparing the children’s social competence and participation in activities to same-aged peers will be presented. Our clinical observations of behaviors suggestive of ADHD and/or weakness in doi:10.1016/j.ymgme.2018.12.356 executive function in some children will be systematically explored using the Conners-3 and BRIEF-2. Our findings suggest that most children with Pompe benefit from accommodations within a general education classroom. Although standardized parent rating 341 scales appear to be good screening tools for problem behaviors and Hormonal dysfunction in adult patients with muco- executive functioning deficits in children with Pompe, their motor polysaccharidosis type I post haematopoietic stem cell weaknesses, general fatigue and other physical challenges need to transplantation considered as contributing factors. A portion of this study was funded by Genzyme Corporation (Sanofi Aventis). Karolina M. Stepiena, Simon Jonesb, Robert Wynnc, Christian J. Hendrikszd, aSalford Royal NHS Foundation Trust, Salford, United Kingdom, bWillink Genetics Unit, Manchester, United Kingdom, doi:10.1016/j.ymgme.2018.12.355 cManchester Foundation Trust, Manchester, United Kingdom, dSteve Biko Academic Hospital, Pretoria, South Africa Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S139

Haematopoietic stem cell transplantation (HSCT) is used to treat administered between 1 and 18 times over 20 years. Mean verbal, children with Hurler syndrome, mucopolysaccharidosis IH (MPS IH), performance, and full-scale intelligence quotients (IQs) were 86.2, a lysosomal storage disease caused by deficiency of alpha- 79.3, an 81.0, respectively. Variation in all IQ domains was not linear iduronidase. Gonadal dysfunction is the most frequent endocrine with time and was best characterized using the coefficient of variation complication in long-term survivors of HSCT (Orio et al. 2014). Not (CV, standard deviation/mean) for each individual. Mean (sd) verbal, much is known about endocrine function in adult patients with performance, and full-scale CVs were 0.08 (0.05), 0.09 (0.06), and 0.07 MPSIH after HSCT. Our clinical protocol outlines endocrine tests as (0.05), respectively. Mixed effects regressions were used to determine part of a long-term follow-up of patients post HSCT. Hormonal if IQ was associated with clinical features. Models were optimized profile includes thyroid function tests, gonadotrophins, estradiol using backward step reduced fit with Satterthwaite degrees of (NR: 72-529 pmol/L) for females and testosterone (NR: 8.4-28.7 freedom. Whether a patient underwent splenectomy was strongly nmol/L) for males. Results from 12 adult patients who underwent associated with IQ and behavioral issues, ataxia, myoclonus, attention HSCT in childhood were available (5F/7M). TFTs were all normal. deficit disorders, and academic challenges were specifically associated Among females (median age 30 years, range, 18-37), 3 had with performance IQ. In enzyme-treated GD type 3, IQ varied roughly amenorrhea and 1/3 was on hormonal replacement therapy (HRT). 2-13% non-linearly over time, with no clear trajectory. It is likely that In 2/5 cases BMI was b18kg/m2 and resulted in suppressed IQ varies similarly in other neuronopathic diseases. Therefore, to gonadotrophins (secondary hypogonadism): LH b2U/L , FSH was reliably use IQ as an efficacy measure in any neurotherapeutic clinical low/normal and serum estradiol was 84 and 120 pmol/L respectively. trial, multiple baseline evaluations are necessary both prior to and post 2 females had regular menstruation and ovulation. Ovarian reserve intervention. was not assessed. Among males (median age 23 years, range, 18-29), median LH was 8.6 U/L (range, 2.9-18.3), median FSH was 20.7 U/L (range, 4.7-31.4) and median male testosterone was 14.4 nmol/L doi:10.1016/j.ymgme.2018.12.358 (range, 9.3-21.4). 5/7 males had raised gonadotrophins despite serum testosterone concentration being in normal reference ranges. Median Sex-Hormone-Binding Globulin (SHBG) was 37 (range, 22- 51) and was used to assess testosterone deficiency (1 male). 343 fi Endocrine abnormalities are common in MPS IH patients post HSCT, Quanti cation of glucosylsphingosine in plasma/serum by UPLC- in particular those who were exposed to chemotherapy (busulfan) MS/MS and/or total body irradiation. Patients with MPSIH live into their adult years and therefore determining long-term outcomes and Ashlee Stiles, Deeksha Bali, Haoyue Zhang, James Beasley, Patricia hormonal complications has become clinically important. We McCaw, Sarah Young, Duke University, Durham, NC, United States recommend annual screening for endocrine dysfunction in this group. Early diagnosis and replacement of hormonal deficiencies Background: Gaucher disease is an autosomal recessive lysosomal are critical for general well-being and bone health in adulthood. storage disorder that results in accumulation of glucocerebroside and its deacylated-derivative, glucosylsphingosine (lyso-Gb1), in lysosomal macrophages. We developed a stable isotope dilution ultra- doi:10.1016/j.ymgme.2018.12.357 performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to quantify lyso-Gb1 in plasma as recent data suggests lyso-Gb1 may be useful for monitoring the disease burden in patients on treatment. Method: Plasma was combined 13 342 with C6-glucosylsphingosine internal standard (IS) and extracted The natural history of cognition in Gaucher disease type 3 using dichloromethane:methanol (1:2, v:v). The precipitate was removed by centrifugation and the supernatant dried under a a a Alta M. Steward , Edythe Wiggs , Taylor Lindstrom , Somto nitrogen. The sample was further extracted using H2O:butanol (1:2, Ukwuania, Emory Ryana, Nahid Tayebia, Tamanna Roshan Lala, Grisel v:v) and the organic layer was dried under nitrogen and rec- Lopeza, Raphael Schiffmannb, Ellen Sidranskya, aNational Institutes of onstituted in a solvent matrix. Samples were analyzed on an Acquity Health, Bethesda, MD, United States, bBaylor Scott & White Research UPLC-Xevo TQ MS system. Lyso-Gb1 was separated from gal- Institute at Dallas, Dallas, TX, United States actosylsphingosine (GalSph) on an Acquity UPLC BEH HILIC column (2.1mm x 100mm) by gradient elution and detected by selected In Gaucher disease (GD), homozygous mutations in GBA1 lead to reaction monitoring in positive ion mode. The peak area ratio of lyso- deficiency of the lysosomal enzyme glucocerebrosidase. GD type 3 is Gb1 to IS was converted to a concentration using a 1/x weighted characterized by varying degrees of chronic neurological involvement. linear fit calibration curve. Results: The interday imprecision of The most common neurological feature is horizontal supranuclear quality control samples was ≤5%. Mean inaccuracy of calibrators gaze palsy, though manifestations range from intellectual delay to ranged from -2.3% to 1.8% over the calibration range of 1.25 to 2000 progressive myoclonic epilepsy. Current therapies for GD have not nmol/L. The lower limit of quantification is 1.06 nmol/L where been proven to affect neurological manifestations. Because identifica- accuracy and precision were acceptable (RSD ≤5%). Lyso-Gb1 in tion of relevant efficacy parameters is essential in designing clinical plasma is stable when stored at ambient temperature, 4oC and -20oC, trials, we focused on acquiring an accurate description of the natural over 5 freeze-thaw cycles from and showed no degradation over history of cognitive function in GD type 3. Our cohort, the largest to time. Preliminary studies showed mild to marked elevations of lyso- date, includes 36 patients who completed neuropsychological evalu- Gb1 in patients with Gaucher disease. Conclusion: We developed an ations at the National Institutes of Health Clinical Center between 1982 accurate and precise method for plasma lyso-Gb1 determination and 2017. Most received enzyme replacement therapy. Individuals using isotope dilution-UPLC-MS/MS. We will analyze plasma from were tested using age-appropriate Wechsler Intelligence Scales, Gaucher patients attending Duke’s Metabolic Clinic and correlate S140 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 lyso-Gb1 levels with clinical data to determine if lyso-Gb1 can be long-term ERT in a real-world setting. Additionally, early initiation of used as a reliable marker for organ involvement. alglucosidase alfa may have a greater impact in preserving FVC in patients with higher FVC before treatment onset doi:10.1016/j.ymgme.2018.12.359 doi:10.1016/j.ymgme.2018.12.360

344 Impact of time from diagnosis to treatment on lung function 345 among patients with late-onset Pompe disease: Data from the Reconstructive aortic valve surgery in a type 3 Gaucher patient Pompe registry homozygous for the p.D409H mutation

David W. Stocktona, Matthias Boentertb, Barry Byrnec, Juan Llerena, Katharina M. Stumpfe, Luise Ammer, Anja F. Köhn, Benjamin Jrd, Priya Kishnanie, Mark Robertsf, Ans van der Ploegg, Roberto Lohmöller, Peer Hauck, Arlindo Riso, Evaldas Girdauskas, Nicole M. Araujoh, Sonia S. Marutih, Nathan Thibaulth, Kenneth I. Bergeri, Muschol, UKE, Hamburg, Hamburg, Germany aDivision of Genetic, Genomic and Metabolic Disorders, Departments of Pediatrics and Internal Medicine, Wayne State University and Children’s Patients homozygous for p.D409H mutation display a particular Hospital of Michigan, Detroit, MI, United States, bDepartment of Sleep type of Gaucher’s disease, namely Gaucher type IIIc. The major Medicine and Neuromuscular Disorders, University Hospital of Münster, feature of Gaucher type IIIc patients is a progressive calcification of Münster, Germany, cDepartment of Pediatrics, University of Florida, cardiac valves and the aorta, which is presumed to cause early death. Gainesville, FL, United States, dInstituto Fernandes Figueira (FIOCRUZ), A 15 year old boy with consanguineous parents of Palestinian origin Departamento de Genética Médica, Rio de Janeiro, Brazil, eDivision of presented with gaze palsy, typical corneal opacities as well as mild Medical Genetics, Department of Pediatrics, Duke University Medical splenomegaly and hematopoietic changes. Anamnesis revealed a Center, Durham, NC, United States, fSalford Royal NHS Foundation Trust, history of speech delay and poor school performances. The cognitive Salford, United Kingdom, gCenter for Lysosomal and Metabolic Diseases, testing showed an IQ of 80. MRI displayed T1 hypointense lesions in Erasmus MC University Medical Center, Rotterdam, Netherlands, hSanofi vertebrae, femora and tibiae (Düsseldorf Score of 4). The diagnosis Genzyme, Cambridge, MA, United States, iDivision of Pulmonary, Critical was confirmed by enzyme assay and molecular genetic analysis, Care and Sleep Medicine, New York University School of Medicine, and which proved Glucocerebrosidase deficiency and a homozygous p. the André Cournand Pulmonary Physiology Laboratory, Bellevue D409H mutation in the GBA-gene, respectively. The echocardiogra- Hospital, New York, NY, United States phy detected hyperdense structures on the mitral and aortic valves, mitral regurgitation and aortic stenosis. The patient was started on Chronic respiratory insufficiency resulting from progressive enzyme replacement therapy (ERT), under which cardiac function respiratory muscle weakness causes significant morbidity and was monitored closely. Unfortunately, aortic valve stenosis was premature death in Pompe disease patients. Skeletal and respiratory progressive despite ERT. Due to a severe increase of the aortic muscle function has improved and stabilized during enzyme pressure gradient, the patient was scheduled for reconstructive replacement therapy (ERT) with alglucosidase alfa in clinical trials aortic valve surgery. Intraoperative findings (functional bicuspid and follow-up studies. Few studies examined the association aortic valve, aortic valve calcification, hypoplastic aorta ascendens between long-term ERT with alglucosidase alfa and outcomes in and small aortic root) and postoperative outcome as well as present late-onset Pompe disease (LOPD). We examined respiratory muscle challenges (re-increase of gradient despite successful valve replace- strength (measured by upright forced vital capacity [FVC]) during ment, re-surgery required) will be discussed. alglucosidase alfa therapy (≤5 years) in LOPD patients enrolled in the Pompe Registry (NCT00231400) sponsored by Sanofi Genzyme. Longitudinal data from Registry patients with LOPD (symptom onset doi:10.1016/j.ymgme.2018.12.361 N12 months or ≤12 months without cardiomyopathy) were analyzed. Participation required that patients had FVC assessments at ERT initiation and ≥2 valid post-baseline assessments. Among 379 eligible patients, median baseline FVC was 66% predicted (range: 9.3-126.0). 346 Longitudinal analyses were conducted using a linear mixed-model Design of a prospective, multicenter, multinational, observational regression. FVC remained stable during the 5-year follow-up safety and outcomes registry in Fabry disease patients treated (slope=‑0.15, P=0.27). We also compared impact of a shorter versus with migalastat and untreated patients longer delay from diagnosis to start of alglucosidase alfa. Groups a b c were defined by median time (1.8 years) from diagnosis to Gere Sunder-Plassmann , Daniel G. Bichet , Martynas Davidonis , d e f treatment initiation: Shorter-Time (0-1.8 years, n=190) versus Dominique P. Germain , Roberto Giugliani , Robert Hopkin , Ana g h i j Longer-Time (N1.8 years, n=189). Baseline FVC was significantly Jovanovic , Ales Linhart , Kathleen Nicholls , Peter Nordbeck , Ulla k l l l higher in the Shorter-Time versus Longer-Time group (69.2 vs. 63.6% Feldt-Rasmussen , Jasmine Rutecki , Joe Giuliano , Nina Skuban , Juan m a b predicted). The between-group difference (4.34%, Pb0.01) persisted Politei , Medical University of Vienna, Vienna, Austria, Hôpital du c throughout the 5-year follow-up. FVC remained stable and did not Sacré-Coeur, University of Montréal, Montreal, QC, Canada, Vilnius d decline in both groups (slopes: -0.14, P=0.48 [Shorter-Time] and University, Vilnius, Lithuania, University of Versailles, Paris-Saclay e ‑0.17, P=0.38 [Longer-Time]). Improvement of respiratory function University, Montigny, France, Medical Genetics Service, HCPA, and f was not observed, demonstrating that an unmet need remains for Department of Genetics, UFRGS, Porto Alegre, Brazil, Cincinnati LOPD patients with respiratory insufficiency. The minimal decline in Children's Hospital Medical Center, Cincinnati, OH, United States, g h FVC over time suggests that respiratory function is preserved during Salford Royal NHS Foundation Trust, Salford, United Kingdom, Charles Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S141

University, Prague, Czech Republic, iRoyal Melbourne Hospital, Univer- globotriaosylceramide (GL-3) and globotriaosylsphingosine (lyso- j sity of Melbourne, Melbourne, Australia, Medizinische Klinik I des Gb3) in many key organs. Until recently, treatment options were Klinikum Vest, Recklinghausen, Germany, kRigshospitalet, National limited to enzyme replacement therapy (ERT) with agalsidase alfa or University Hospital, Copehhagen University, Copenhagen, Denmark, agalsidase beta infusions. Migalastat is a first-in-class, oral pharma- lAmicus Therapeutics, Inc., Cranbury, NJ, United States, mDr Chamoles cological chaperone approved for treating adults with Fabry disease Neurochemistry Laboratory, Buenos Aires, Argentina and migalastat-amenable GLA mutations. Migalastat has demonstrated efficacy across multiple organs in both ERT-naive and Fabry disease is a rare, X-linked disorder caused by α-galactosidase -experienced patients with Fabry disease and amenable mutations. A(α-Gal A) deficiency due to mutations in the GLA gene. Reduction in This post hoc analysis of the phase 3, randomized, active-controlled α-Gal A activity results in an accumulation of glycosphingolipids, ATTRACT study (NCT01218659) evaluates clinical outcomes for 4 which leads to the clinical manifestations of Fabry disease, including groups of patients with amenable GLA mutations who were previ- neuropathic pain, skin disorders, ophthalmological disturbances, ously treated with ERT and completed the 18-month randomized gastrointestinal signs and symptoms, pulmonary problems, renal treatment period: patients who switched to migalastat from failure, cardiomyopathy, cerebrovascular events, and early mortality. agalsidase beta (n=11) or agalsidase alfa (n=20) and patients who Migalastat is a first-in-class, oral pharmacological chaperone that stayed on agalsidase beta (n=5) or agalsidase alfa (n=11). Renal binds to amenable mutant forms of α-Gal A, thus stabilizing the function was stable over 18 months in patients who switched to enzyme and restoring lysosomal trafficking. Migalastat is approved for migalastat from agalsidase beta or agalsidase alfa. The mean (SD 95% the treatment of patients with Fabry disease and amenable GLA CI) annualized rates of change in eGFRCKD-EPI of the 4 groups were variants. A prospective, multicenter, multinational, observational -1.0 (3.6-3.4, 1.4), -0.9 (3.0-2.3, 0.5), 2.2 (3.2-1.8, 6.2), and -4.4 (6.7- registry was designed to evaluate effects of treatment on long-term 8.9, 0.1) mL/min/1.73 m2, respectively. Switching to migalastat from safety, effectiveness, and quality of life (QoL) in patients with Fabry either ERT led to numerically larger reduction in left ventricular mass disease in real-world settings. This unique, patient-focused registry index (LVMi) from baseline compared with patients who stayed on will include ~250 patients receiving migalastat, ~100 patients the ERT (mean [SD 95% CI]: -14.1 [13.4-23.1, -5.1], -4.4 [10.5-9.3, receiving enzyme replacement therapy, and ~100 patients not 0.6], -3.5 [11.0-21.0, 14.0], -2.4 [17.7-17.2, 12.4] g/m2, respectively). receiving any Fabry disease therapy but who are eligible for migalastat Plasma lyso-Gb3 levels remained stable in all 4 groups. White blood treatment from 80 centers worldwide. Safety endpoints include cell α-Gal A activity (assessed in males only) increased after occurrence of serious adverse events, serum creatinine, urine protein, switching to migalastat. Taken together, these data indicate that urine albumin, plasma globotriaosylsphingosine (lyso-Gb3) levels, patients with Fabry disease and amenable GLA mutations can benefit eGFR, urine albumin-creatinine ratio, echocardiograms, and cardiac from switching to migalastat from either agalsidase alfa or agalsidase MRI for up to 5 years of follow-up. Effectiveness endpoints include beta. occurrence of cardiac, cerebrovascular, and renal clinical events and overall survival. Patient-reported outcomes include overall QoL and key aspects of physical and mental health (12-item short form health doi:10.1016/j.ymgme.2018.12.363 survey, EuroQol-5D, Treatment Satisfaction Questionnaire for Medi- cation-9, Brief Pain Inventory, and Fabry Disease Patient-Reported Outcome–Gastrointestinal Signs and Symptoms Questionnaire). Data from this registry will be made available to patients and researchers. 348 This patient registry, which began August 2018, will collect valuable Ketogenic therapy as an adjunct to ERT for Pompe disease safety, effectiveness, and patient-reported outcome data to help to evaluate the effect of a novel treatment in Fabry disease in the real Mark Tarnopolskya, Michael Croziera, Alessia Di Carloa, Mats Nilssonb, world. aMcMaster University, Hamilton, ON, Canada, bExerkine Corporation, Hamilton, ON, Canada doi:10.1016/j.ymgme.2018.12.362 Pompe disease (PD) is a rare autosomal recessive disorder characterized by the deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA), leading to buildup of intra-lysosomal glycogen and autophagic debris, mitochondrial dysfunction, acceler- 347 ated organ deterioration, and premature death. Currently, enzyme Clinical outcomes after switching to migalastat from agalsidase replacement therapy (ERT) is available for PD however, the efficacy alfa or agalsidase beta in patients with Fabry disease: Post hoc of the exogenously administered enzyme is limited partially be poor analysis from ATTRACT uptake in type II skeletal muscle fibers. Adjunctive therapies, such as dietary modification and supplementation, may improve both Gere Sunder-Plassmanna, Ana Jovanovicb, Ulla Feldt-Rasmussenc, primary (lysosomal) and secondary (mitochondrial) pathologies in Vipul Jaind, Markus Pecenyd, Nina Skuband, Roberto Giuglianie, PD. A ketogenic diet (KD) or administration of exogenous ketones, aMedical University of Vienna, Vienna, Austria, bSalford Royal NHS such as 1,3-butanediol (1,3-BD), may improve autophagic function, Foundation Trust, Salford, United Kingdom, cRigshospitalet, National while mitochondrial function enhancers (‘mito-enhancers’) may University Hospital, Copenhagen University, Copenhagen, Denmark, alleviate oxidative damage. In this study, we investigated the effect dAmicus Therapeutics, Inc., Cranbury, NJ, United States, eMedical of combining ERT with a standard KD, 1,3-BD, or a mito-enhancer Genetics Service, HCPA, and Department of Genetics, UFRGS, Porto diet (MD) on PD pathology in GAA-KO mice (6neo/6neo) following 3 Alegre, Brazil months of therapy. We found that exogenous 1,3-BD administration raised circulatory ketone levels to ≥2mM (similar to KD), attenuated Fabry disease results from GLA gene mutations that lead to autophagic buildup in type II muscle fibers, and maintained α-Gal A deficiency and the subsequent accumulation of functional capacity in 3-mo-old, ERT-treated GAA-KO mice (P b S142 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

0.05) however, it did not augment glycogen clearance. Surprisingly, 350 the KD did not provide any benefit beyond ERT alone and the Mortality in Fabry disease cohort: One centre experience combination of ERT + 1,3-BD + MD was superior to ERT and ERT + KD, but it did not provide significant benefits above ERT + 1,3-BD Lorraine A. Thompson, Karolina M. Stepien, Reena Sharma, Salford alone. Collectively, our results suggest that combining exogenous Royal NHS Foundation Trust, Manchester, United Kingdom ketones (1,3-BD) and mitochondrial enhancers with ERT may slow cellular pathology and functional decline in PD, and may be an useful Fabry disease is an, X-linked metabolic disorder caused by adjunctive therapy in other lysosomal storage disorders and in aging reduced lysosomal α-galactosidase activity. Accumulation of glyco- conditions. Funding: Canadian Institute for Health Research and lipids in various tissues leads to variable clinical manifestations. The Exerkine Corporation. signs and symptoms of Fabry disease are most severe among classical males however, heterozygous females often exhibit disease. Lifespan is considerably shortened in males. Retrospective data was collected doi:10.1016/j.ymgme.2018.12.364 and analysed from deceased Fabry patients including cause of death, genetics and monitoring data who attended this centre over last 10 years. The cohort included classical as well as variant disease patients. Among 320 patients, 22 (6.9%) died between 2008-2018 349 (14M/8F). The median age at death was 58.8 years (range, 50-69) How do Fabry disease patients find their way to a metabolic and 60.5 years (range, 38-78) for males and females respectively. 13/ physician? Referral pathway to a tertiary metabolic centre over 14 males were treated with ERT up to the time of death and the ten years average duration of treatment was 5.9 years (0-10years). 6/8 females had received ERT for 9.7 years (0-18years) at the time of their Lorraine A. Thompson, Ana Jovanovic, Karolina M. Stepien, Salford deaths. Among deceased males, non Fabry disease-related cause Royal NHS Foundation Trust, Manchester, United Kingdom accounted for 14% (2 patients). The remaining died of Fabry disease- related complications: 72% cardiac, 14% renal and cerebrovascular. Anderson-Fabry disease (AFD) is a rare X-linked metabolic Among deceased females, non- Fabry disease-related cause of disorder caused by the partial or complete absence of the lysosomal mortality was identified in 6 patients (75%), one patient had α enzyme -galactosidase A. Clinical manifestation is multisystem cerebrovascular disease, one another had Fabry disease-related with more severe phenotype among males. Disease awareness and cardiac disease. 5/14 (36%) males and no females had renal targeted information on the disease is essential to reduce both replacement therapy (3 renal transplant). 9 out of 14 males and 1 diagnostic and therapeutic delays. Further complications to a out of 8 females had received implantable cardioverter defibrillators. misdiagnosis can result in mental disorders, depression, social Cardiac complications appear to be the most prevalent cause of death isolation, poorly managed pain. This project aimed to review the among our cohort of male patients with Fabry disease. In majority of impact of educational intervention on the number and source of male patients the cause of death was relayed to Fabry disease as referrals of patients with Fabry disease. A retrospective audit of the compared to deceased females. The longevity data in males not source of referral among our patients with Fabry disease was changed as compared to the published data (Waldek et al. Genet conducted. Educational intervention included direct patient discus- Med 2009). sion with index cases and subsequent at risk family members, pedigree drawing, leaflets, MPS Society website, Fabry focus days, conference proceedings and local presentations. The cohort of 324 doi:10.1016/j.ymgme.2018.12.366 patients (155m/169F) was compared with a previous audit in 2007 of 120 patients (67M/53F). Among females, the mean age at diagnosis was 50 years (range: 18-81) and among males, the mean age at diagnosis was 41 years (range: 19-78). Sources of referral 351 included: clinical genetics (35%), cardiology (8%), renal (5%), stroke Long-term efficacy of olipudase alfa in adults with acid neurology (5%), transition (11%), cardiac genetic nurse counsellor sphingomyelinase deficiency (ASMD): Further clearance of he- (13%) and General Practitioner in partnership with specialist centre patic sphingomyelin is associated with additional improvements (23%). These results show a significant increase of referrals of in pro- and anti-atherogenic lipid profiles after 3.5 years of patients with AFD by 62% to Adult Metabolic Medicine Department treatment over 10 years. The number of referrals as a direct result of family screening has increased significantly and reflects the effect of disease Beth L. Thurberga, George A. Diazb, Robin H. Lachmannc, Thomas awareness, education and access to biochemical and genetic testing. Schianob, Melissa P. Wassersteind, Allena J. Jie, Atef Zahere, Ana In conclusion, we demonstrated that a targeted approach on Fabry Cristina Pugaa, aSanofi Genzyme, Cambridge, MA, United States, bIcahn disease, provided over a period of ten years among general School of Medicine at Mount Sinai, New York, NY, United States, cardiologist, nephrologists, geneticists and General Practitioners cNational Hospital for Neurology and Neurosurgery, London, United had a direct clinical impact by allowing the diagnosis of Fabry Kingdom, dAlbert Einstein College of Medicine, Bronx, NY, United States, disease and early therapy. eSanofi Genzyme, Framingham, MA, United States

The liver is a major site of lipoprotein synthesis and metabolism. doi:10.1016/j.ymgme.2018.12.365 Liver manifestations of chronic visceral ASMD include hepatomegaly, fibrosis, elevated liver function tests (LFTs) and a pro-atherogenic Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S143 lipid profile. As plasma sphingomyelin (SM) levels in patients are be operational for at least 10 years. The first patient was enrolled in normal, measurements of SM levels in liver biopsies, along with lyso- September 2013. As of September 2018, TALIAS had enrolled ~104 SM in plasma, were used as pharmacodynamic biomarkers. Five patients and is active in the US, Israel, and Albania, the countries adult patients with chronic visceral ASMD were enrolled in a 26- with the greatest numbers of patients using taliglucerase alfa as part week phase 1b trial of olipudase alfa (NCT01722526) followed by an of their standard of care. As part of this PMR, a pregnancy and ongoing long-term extension study (NCT02004704). We present the lactation exposurerelated substudy is also being conducted with the changes in hepatic SM levels, plasma lyso-SM, and lipoprotein intent to follow all female patients who are/become pregnant and/or profiles for the cumulative 42 months (3.5 years) of treatment. At breastfeed while in the Drug Registry and their infants up to the baseline, histologic SM storage ranged from 9.8% to 53.8% of the child’s first birthday. Baseline clinical and demographic characteris- microscopic field. After 6 months, SM storage in post-treatment tics, including median age at entry and proportions of patients who biopsies ranged from 1.2% to 9.5% of the microscopic field, are female, have Type 1 GD, are of Ashkenazi Jewish descent, and corresponding to an 84% to 92% reduction from baseline. Similar who report a family history of GD, are consistent with the reductions in plasma lyso-SM were observed. Improvements in LFTs characteristics monitored by two other global GD registries. (ALT, AST), and pro-atherogenic lipid profiles (total cholesterol, LDL and VLDL subfractions, and triglycerides) were observed at month 6. Conversely, there were increases in anti-atherogenic markers, HDL doi:10.1016/j.ymgme.2018.12.368 and apolipoprotein A-I. After 3.5 years, SM storage was further reduced from baseline by N99% in all five patients. LFTs remained normal lyso-SM levels were stable. This was accompanied by further reductions from month 6 levels in pro-atherogenic markers: total 353 cholesterol (2/5 patients), LDL (3/5 patients), VLDL (2/5 patients) Newborn screening of mucopolysaccharidoses: Past, present, and and triglycerides (2/5 patients). Remarkably, HDL increases ranged future from 60% to 200% over baseline levels after 3.5 years of treatment. These data demonstrate that hepatic clearance of SM during Shunji Tomatsu, Nivethitha Arunkumar, Stapleton Molly, Nemours/ olipudase alfa treatment over 3.5 years is associated with overall Alfred I. duPont Hospital for Children, Wilmington, DE, United States improvements in liver function and the cardiac risk-related lipid profiles of ASMD patients. Mucopolysaccharidoses (MPS) is a rare inherited metabolic disease that occurs from a deficiency of the enzymes involved in the glycosaminoglycan (GAG) breakdown pathway. MPS is catego- This study was sponsored by Sanofi Genzyme. rized as a lysosomal storage disorder, caused by the accumulation of doi:10.1016/j.ymgme.2018.12.367 GAGs. Unless there is a history of MPS already within the family, MPS is diagnosed when clinical symptoms have already arisen during childhood. The progression of MPS disorder provides a negative impact in various developments of the patients, leading to CNS 352 involvement, severe bone dysplasia, hepatosplenomegaly, and car- Baseline characteristics of patients with Gaucher disease enrolled diac and respiratory issues. MPS significantly decreases the quality of in the taliglucerase alfa surveillance (TALIAS) registry life within patients and require appropriate treatment and management as soon as possible. A way to diagnose MPS patients as early as Lina Titievskya, Tilman Schustera, Rong Wangb, Pamela Beckerc, Neal possible is by newborn screening. Newborn screening for MPS I and Weinrebd, Paige Kaplane, Christina Chambersf, Kabir Quazia, Betina II has been implemented in several countries. In this review paper, Hernandeza, Michael P. Wajnrajcha, aPfizer Inc, New York, NY, United we look at the newborn screening implemented in several countries, States, bPfizer Inc, Groton, CT, United States, cUniversity of Washington especially the United States. We also discuss the different tiers of School of Medicine, Seattle, WA, United States, dUniversity Research screening that could take place. Newborns are not born with Foundation for Lysosomal Storage Disease, Boca Raton, FL, United States, symptoms at birth so that it is crucial to focus on the biomarkers eThe Children’s Hospital of Philadelphia, Philadelphia, PA, United States, that would be present at birth. The tiered methods and instrumen- fUniversity of California San Diego, La Jolla, CA, United States tation used should be quick, cost-effective, sensitive, and specificto the biomarkers that are screened for. The screening of biomarkers in Taliglucerase alfa for injection is a recombinant active form of the patients with MPS before/shortly after birth may not only be human lysosomal enzyme β-glucocerebrosidase that is expressed in important for a quick diagnosis but also will lead to forms of genetically modified carrot root cells and produced in a disposable treatments that take advantage of possible immune tolerance at the bioreactor system. Taliglucerase alfa is approved in many countries neonatal period and maximize therapeutic efficacy. around the world for long-term enzyme replacement therapy in adult and pediatric patients with a confirmed diagnosis of Type 1 (non-neuronopathic) Gaucher disease (GD) and may also be used for doi:10.1016/j.ymgme.2018.12.369 the hematologic manifestations in pediatric patients with Type 3 GD. Taliglucerase Alfa Surveillance (TALIAS) is the third global GD registry and is being conducted as PostMarketing Requirement (PMR) to the US Food and Drug Administration. The registry was 354 designed to further characterize the long-term safety and effective- Enzyme replacement therapy for mucopolysaccharidoses: Past, ness of taliglucerase alfa in the real-world post-marketing setting. present, and future TALIAS is a single product registry that collects data under routine local clinical practice and is open to all pediatric and adult GD Shunji Tomatsu, Hui-hsuan Chen, Kazuki Sawamoto, Nemours/Alfred patients receiving taliglucerase alfa treatment. TALIAS is intended to I. duPont Hospital for Children, Wilmington, DE, United States S144 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Mucopolysaccharidoses (MPS) are a group of lysosomal storage exhibit poor growth as untreated patients even when ERT is disorders that lack an enzyme corresponding to the specific type of administered under 5 years of age, indicating that conventional ERT MPS. The standard therapeutic option for some types of MPS has cannot fix bone growth. been enzyme replacement therapy (ERT) because of the ability to start immediate treatment and provide palliative results. Currently, MPS I, II, IVA, VI, and VII have approved conventional intravenous doi:10.1016/j.ymgme.2018.12.371 ERT. While the feasibility is the most advantage for ERT, there are several disadvantages for conventional ERT 1) limited impact to the brain and avascular bone, 2) the weekly or biweekly infusions for 4-5 hours, 3) immune response against infused enzyme, and 4) the high 356 cost. Clinical studies of ERT have shown limited efficacy in Biomarkers in patients with mucopolysaccharidosis types II and preventing or resolving progression in neurological, cardiovascular, IV and skeletal diseases, especially in the severe phenotypes. The main focus is crossing the blood-brain barrier (BBB) to deliver enough Shunji Tomatsu, Honoka Fujitsuka, Kazuki Sawamoto, Nemours/Alfred enzyme to the brain since conventional ERT does not cross the BBB. I. duPont Hospital for Children, Wilmington, DE, United States Novel ERT strategies are under development to reach the brain 1) utilizing a fusion protein with monoclonal antibody to target a Glycosaminoglycans (GAGs), dermatan sulfate (DS), heparan receptor on the BBB, 2) using a protein complex from plant lectin, sulfate (HS), and keratan sulfate (KS), are the primary biomarkers glycan, or insulin-like growth factor 2, and 3) direct infusion across in patients with mucopolysaccharidoses (MPS) however, little is the BBB. Tremendous research on whether the modified enzyme- known about other biomarkers. To explore potential biomarkers and protein complex can penetrate BBB is required to consider novel ERT their correlation with GAGs, blood samples were collected from 46 as a standard treatment. The use of alternate administrations, such as MPS II patients, 34 MPS IVA patients, and 5 MPS IVB patients. We fl intrathecal (IT) or intracerebroventricular (ICV), are also under evaluated the levels of 8 pro-in ammatory factors, collagen type II, investigation. This review article summarizes the current progress and DS, HS (HS0S, HSNS), and KS (mono-sulfated, di-sulfated) in fi and limitations on ERT for MPS and describes the new technology blood. Eight biomarkers measured were signi cantly elevated in investigated to overcome the obstacles of conventional ERT. untreated and ERT-treated MPS II patients, compared with those in normal controls: EGF, IL-1β, IL-6, HS0S, HSNS, DS, mono-sulfated KS, and di-sulfated KS while three biomarkers remained elevated in doi:10.1016/j.ymgme.2018.12.370 post-HSCT MPS II patients: EGF, mono-sulfated KS, and di-sulfated KS. Ten biomarkers were significantly elevated in untreated MPS IVA patients: EGF, IL-1β, IL-6, MIP-1α, MMP-1, MMP-2, MMP-9, HSNS, mono-sulfated KS, and di-sulfated KS while four biomarkers 355 remained elevated in MPS IVA patients under ERT: IL-6, TNF-α, Effect of enzyme replacement therapy on the growth of patients mono-sulfated KS, and di-sulfated KS. Two biomarkers were with Morquio syndrome type A significantly elevated in untreated MPS IVB patients: IL-6 and TNF- α. Collagen type II level was significantly decreased in untreated and Shunji Tomatsu, Caitlin Doherty, Molly Stapleton, Nemours/Alfred I. ERT-treated MPS II patients and untreated MPS IVA patients. We also duPont Hospital for Children, Wilmington, DE, United States found that in untreated MPS IVA patients, MMP-2 levels correlated positively with the level of di-sulfated KS and that in all patients with Mucopolysaccharidosis IVA (MPS IVA) is a degenerative systemic MPS II, IVA, and IVB, EGF moderately correlated with DiHS0S, skeletal dysplasia, in which children exhibit marked short stature DiHSNS, and DS. Post-HSCT patients with MPS II showed that 4 and become physically handicapped. The purpose of this study was biomarkers were normalized as HS and DS levels decreased. to evaluate the growth patterns of patients treated with ERT, Meanwhile, five biomarkers were significantly different between compared with those of untreated patients. In this study, cross- untreated MPS IVA patients and ERT-treated MPS IVA patients sectional and longitudinal data of heights and weights were collected although there was no reduction of KS in the ERT-treated patient. from 128 MPS IVA patients and compared with the growth charts of In conclusion, selected pro-inflammatory factors can be potential MPS IVA [Tomatsu et al., 2011]. Eleven patients (5 males, 6 females) biomarkers in patients with MPS, correlated with GAGs levels. starting ERT before 5 years old were treated for at least 2 years. Six out of 11 patients with ERT over 2 years stopped growing between 94 and 98 cm with a mean height of 95.1 ± 2.2 cm from 5.0 years to doi:10.1016/j.ymgme.2018.12.372 9.0 years of age (mean age of 6.2 ± 1.6 years). Other 5 patients exhibited a marked slow growth velocity from 3.6 years to 7.7 years. Thus, MPS IVA males and females starting ERT even before 5 years of age failed to show significant growth improvement. Treated and 357 untreated patients reached their final heights by approximately 10 Development of AAV gene therapy for Morquio syndrome type A years of age. Patients with ERT still showed a reduced pubertal growth spurt, which contributes to marked short stature as seen in Shunji Tomatsu, Kazuki Sawamoto, Shaukat Kahn, Molly Stapleton, untreated patients. Compared with the growth charts for untreated Subha Melethil, Olivier Danos, Nemours/Alfred I. duPont Hospital for patients, patients with ERT did not show any significant increase in Children, Wilmington, DE, United States growth in any age group. The proportion of obese and overweight males and females was higher in treated and untreated patients, Mucopolysaccharidosis IVA (MPS IVA) is characterized by severe compared with the normal population, and thus ERT itself did not bone pathology, not improved by conventional enzyme replacement prevent obesity in MPS IVA patients in this study. Overall, ERT- therapy (ERT). There is an important unmet need that can be treated patients do not experience growth improvement and still addressed by AAV gene therapy. Current study used recombinant Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S145

AAV8 vectors to express deficient GALNS enzyme, with and without levels of two or more different GAGs improves separation of MPS a bone-targeting signal, into two mouse models of the disease, MPS patients from unaffected controls, indicating that GAG measurement IVA KO and Mtol immunotolerant mice. We evaluated AAV8 vectors in DBS is a potentially valuable tool for first-tier newborn screening expressing native human hGALNS under the liver-specific promoter of MPS, especially by the fact that a single test can screen for several TBG (TBG-hGALNS) and hGALNS with an aspartic acid octapeptide distinct MPS. (D8) (TBG-D8-hGALNS). The vectors were delivered intravenously into 4-week-old MPS IVA KO and Mtol mice at a dose of 5x1013 GC/ kg body weight. These mice were monitored for 12-14 weeks post- doi:10.1016/j.ymgme.2018.12.374 injection. We carried out a biweekly collection of blood and assayed for GALNS activity and keratan sulfate (KS). At necropsy, bone pathology was evaluated by histopathological analysis. GALNS enzyme activity was elevated significantly in plasma of all treated 359 mice two weeks post-injection. The increase in the activity observed Hematopoietic stem cell transplantation for was 5-100-fold of the endogenous activity in WT mice and was kept mucopolysaccharidoses past, present, and future throughout the monitoring period. Plasma enzyme activity levels in Mtol mice treated with AAV8-D8-hGALNS vector were significantly Shunji Tomatsu, Madeleine Taylor, Shaukat Khan, Nemours/Alfred I. higher than those in mice treated with AAV8-hGALNS vector. In MPS duPont Hospital for Children, Wilmington, DE, United States IVA KO mice, both vectors resulted in a reduction of mono-sulfated KS levels to WT levels two weeks post-injection and these normal Allogenic hematopoietic stem cell transplantation (HSCT) has levels were kept until necropsy at 12 weeks post-injection. been shown to be a treatment option for a select group of patients Histopathological evaluation showed that both vectors reduced the with mucopolysaccharidoses (MPS) (MPS I, II, IVA, VI, and VII). storage in articular cartilage, ligaments, meniscus, and growth plate Early diagnosis and timely referral to an expert in MPS are in tibia and femur. Current data outline a strategy for developing a necessary, followed by a complete examination and evaluation novel treatment to address the bone disease in MPS IVA using AAV with a multidisciplinary team, including a transplant physician. gene therapy. Results from this study conducted in relevant animal Treatment recommendations for MPS are based on multiple factors fi models of the disease provide critical information to design a human such as biological, sociological, and nancial effects which include clinical trial. type of MPS, clinical severity and prognosis, present clinical signs and symptoms (disease stage), age at onset, the rate of progression, family significances and expectations, financial bur- fi doi:10.1016/j.ymgme.2018.12.373 dens, feasibility and availability, and risks and bene ts with available therapies: HSCT, enzyme replacement therapy (ERT), surgical interventions, and other supportive care. To evaluate therapeutic efficacy and adverse effects of utilizing HSCT for MPS, 358 international collaboration and accumulation of outcome data are Newborn screening for mucopolysaccharidoses by GAG assay with critical. Since the first attempt of HSCT in a patient with MPS in tandem mass spectrometry 1981, collaborative efforts on assessment of HSCT for MPS have been made continuously. Accumulation of data has made it Shunji Tomatsu, Molly Stapleton, Robert Mason, Nemours/Alfred I. possible to identify early outcomes (transplant outcomes) and duPont Hospital for Children, Wilmington, DE, United States long-term disease-specific outcomes resulting from HSCT. Recent identification of predictive factors and innovative regimens has Mucopolysaccharidoses are a group of inborn errors of metabo- significantly improved the outcomes of both engraftment failure lism that are progressive and usually result in severe skeletal, and transplant mortality. Assessment of long-term outcomes has visceral and/or CNS manifestations, highlighting a need for early also been described under consideration of a variety of factors: diagnosis, especially as many of these conditions are now treatable. type of MPS, graft-type, age at transplant which affects disease This pilot study analyzed 17,467 dried blood spots (DBS) from progression, etc. Formidable studies on the long-term outcomes newborns and 14 DBS from newborn patients with MPS 7MPS I, 2 are considered a key achievement for the HSCT in MPS commu- MPS II, 5 MPS III. Disaccharides were produced from polymer GAGs nities. These studies have shown the effects and limitations of by digestion with chondroitinase B, heparitinase, and keratanase II. HSCT to improve disease manifestations and quality of life. In this Heparan sulfate (0S, NS), dermatan sulfate (DS) and mono- and di- review, we summarize the efficacy, side effects, risks, and cost of sulfated KS were measured by liquid chromatography tandem mass HSCT for each type of MPS. spectrometry LC-MS/MS. Median absolute deviation (MAD) was used to determine cutoffs to distinguish patients from controls. Cutoffs were defined as median + 7x MAD from general newborns. Results: doi:10.1016/j.ymgme.2018.12.375 The cutoffs were as follows: HS-0SN 90ng/mL HS-NSN 23 ng/mL, DSN 88 ng/mL mono-sulfated KSN 445 ng/mL di-sulfated KSN 89 ng/mL and ratio di-KS in total KSN 32%. 2.3% of the samples were above the cut off values in HS-0S. 2.4% of the samples were above the cut off 360 values in HS-NS. 0.9% of the samples were above the cut off values in Study of telomeres in monocytes from a cohort of Spanish DS. The rate of false positives for MPS I and II was 0.07% based on a patients with Gaucher disease combination of HS-0S, HS-NS, and DS, and for MPS III was 1.4% based upon a combination of HS-0S and HS-NS. All MPS I and II samples Miguel-Angel Torralba-Cabeza, José Luis Sierra Monzón, Susana were above the cutoffs for HS-0S, HS-NS and DS, and all MPS III Olivera González, Ana María Camón Pueyo, Sebastián Menao Guillén, samples were above cutoffs for HS-0S and HS-NS. Combination of Lozano Blesa Zaragoza University Hospital, Zaragoza, Spain S146 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

Background: The "telomeres" are the extremes of the chromo- Patients: 0,86-7,18) Maltose with Acarbose: (Controls: 1,15-9,92/ somes of eukaryotic cells that are necessary for the control of cell Patients: 0,09-1,19) Glycogen no Acarbose: (Controls: 0,41-4,71/ division and for maintaining the integrity and stability of chromo- Patients: 0,21-1,54) Glycogen with Acarbose: (Controls: 0,41- 2,93/ somes. At present, 4 physiopathological models of telomeric disease Patients: 0,0-0,24). Activities expressed in nmol/mg protein/hour). This (TD) are known depending on the size of the telomeres (decreased comparison shows the increased selectivity of pure glycogen in the or increased) and the activity of telomerases (low or high). TD are discrimination between patients and controls which is full in acarbose characterized by affecting the bone marrow, fibrosis, liver disease, presence. In the other hand we show the poor selectivity of 4-MUG for elevation of TGF-B and increased incidence of cancer, events that this discrimination being the less recommended substrate for assessing occur in Gaucher disease (GD). this enzyme activity. Hypothesis: GD could be an entity due in part to telomere dysfunction. doi:10.1016/j.ymgme.2018.12.377 Material and methods: 9 GD stable cases and 9 healthy controls matched by age and gender were subjected to a real-time PCR. As an internal control, the 36B4 gene was used, which never modifies its expression. The measurement was carried out by determining the Ct 362 = Pending Increase. The samples were studied in duplicate, High-risk population screening for Sly syndrome: Application of a obtaining the average of the readings. The determination of the Ct micro-method in collected dried blood spots of the cases and of the controls was modified according to the Ct of the control gene: Ct obtained/Ct gene 36B4. Subsequently, the results Alfredo Uribe, Patricia Moreno, Jaqueline Benavides, Universidad de obtained were analyzed by applying the SPSS statistical program. los Andes, Bogotá, Colombia Results: The telomere size was significantly higher in patients with GD than in their respective controls (p = 0.019), and it must be Mucopolysaccharidosis type VII (MPS VII) or Sly syndrome is a interpreted as a monocyte activation. The size of telomeres lysosomal glycosaminoglycans metabolic alteration caused by an correlated positively with the GAUSS-I score (p = 0.001), GD-DS3 autosomic recessive inherited deficit of Beta-glucuronidase (EC score (p = 0.001), plasmatic Chitotriosidase (p = 0.002), CCL-18 (p 3.2.1.31). This deficit leads to progressive and multi-systemic accumu- = 0.002), and Ferritin (p= 0.02), but not with bone disease (p = lation of these glycosaminoglycans, affecting tissue functionality 0.27) and the gender (p = 0.8). including central nervous system tissues. Clinical manifestations share Conclusions: 1. Telomere dysfunction is present in GD. 2. The similar findings to those in other glycosaminoglycan disorders including study of telomeres size in monocytes of GD patients provides facial and skeletic dysmorphism, gross skin, hepatosplenomegaly, and important data in the knowledge the physiopathology of this entity. retinal degeneration among others. MPS VII has been considered an ultra-orphan disease (reported frequency 1:300,000 to 2,000,000). doi:10.1016/j.ymgme.2018.12.376 Nevertheless, registries imprecision has been considered and therefore, entity’s sub-diagnosis. These results express the importance of a pilot test realized in DBS simplifying referral of samples for suspected patients. Our purpose is to report the results of beta-glucuronidase 361 activity values on DBS (1,2 mm cut) in patients with clinical suspect of Leucocitary alpha-glucosidase performance in Pompe patients mucopolysaccharidosis (n=895) and the statistic comparison with and normal controls: A comparison of different substrates Colombian population controls. Methods/Results: By using artificial substrate 4-methylumbelliferyl-β-D-glucoronide, we standardized the Alfredo Uribe, Patricia Moreno, John Gamba, Universidad de los Andes, micro-method, stablishing a reference range for control population: Bogotá, Colombia 31,2 - 242,6 nmol/hour/ml (median: 110,3). High-risk population measured include ages between 1 month and 32 years-old (90% b20 Metabolic myopathies conform an important group of disorders years). Activity values were distributed between 20,05 and 1209,1 involving muscle cell metabolism. Among them, Pompe disease has nmol/hour/ml. One case was reevaluated for Beta-glucuronidase been widely studied and its metabolic pathway has started been deficiency presenting values 0,18 nmol/hour/ml (0,16% residual dilucidated. The pathologic cause of the disease is glycogen accumula- activity). Is essential to maintain study continuity to establishing real tion in myocytes produced by a deficiency of lysosomal alpha- frequencies of this disease in Colombian population. glucosidase inherited in an autosomic recessive way. Diagnostic of Pompe disease is made by quantification of low enzyme levels in at least doi:10.1016/j.ymgme.2018.12.378 two different tissues. In diagnostic process of this disease, assessment of catalytic reaction of alpha-glucosidase in dried blood spot samples has proved been useful as screening method, and subsequent analysis of the enzyme in leucocyte isolates, as confirmation method. This confirma- 363 tion must be accomplished carefully exploring performance of the Alglucosidase alfa inhibitory antibodies and clinical correlates in reaction when enzyme is faced to natural and artificial substrates. The treatment-naive late-onset Pompe disease patients in the late- presence of non-disease-related enzymatic isoforms which also operate onset treatment study (LOTS) over 78 weeks: A new post hoc degrading substrate, can lead to confusing results. In this work, we analysis presented a comparison using alternate analytic pathways, including a b a different selective substrates. We use 4-methylumbelliferyl-Alpha-D- Vassili Valayannopoulos , Crystal Sung , Quanhong Lei , Meredith a c a a glucopyranoside (4-MUG), maltose, and pure glycogen, with and Foster , Kristina An Haack , Roberto Araujo , Christopher Hug , a a a b without acarbose. These were the activity ranges for alpha-glucosidase Edward Neilan , Nathan Thibault , Susan Sparks , Susan Richards , a fi b fi with different substrates. 4-MUG No Acarbose: (Controls: 4,8 - 252,4/ Sano Genzyme, Cambridge, MA, United States, Sano Genzyme, c fi Patients: 16,5 - 65,9) 4-MUG with Acarbose: (Controls: 1,2 - 99,0/ Framingham, MA, United States, Sano Genzyme, Chilly-Mazarin, Patients: 0,9 - 14,7) Maltose no Acarbose: (Controls: 3,38-14,15/ France Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S147

In late-onset Pompe disease (LOPD), inhibitory anti-alglucosidase rapid diagnosis of LSDs in the EU as often a significant diagnostic delay alfa antibodies (IAb) are less thoroughly studied than anti-drug is still present. • improve and standardize management of LSDs in the antibodies (ADA). This retrospective analysis examined IAb associ- EU. • improve prospects of patients with LSDs in the EU by initiating ations, trajectories, and effects on alglucosidase alfa efficacy and and contributing to research and implementation of innovative safety in Late Onset Treatment Study data (LOTS NCT00158600). therapies. The Lysosomal Subnetwork consists of 45 Health Care Treatment-naïve LOPD patients, aged ≥8 years, received Providers (HCPs) with expertise in the field of LSDs from 13 EU- alglucosidase alfa infusions 20 mg/kg (n=60) or placebo (n=30) countries representing 6588 patients: 2987 pediatric and 3601 adults. qow for 78 weeks. Co-primary efficacy endpoints were 6-minute Working groups are being formed to address different important walk test (6MWT) and % predicted forced vital capacity (FVC). topics within the field of lysosomal diseases. These include: • Uptake and catalytic inhibition were assayed in ADA-positive sera Prevention & Screening for lysosomal diseases. • Diagnosis of (new) positive titers only occurred for uptake inhibition. Post-hoc analyses lysosomal diseases • Management of lysosomal diseases • Epidemiol- assessed relationships of IAb status with baseline characteristics (χ2 ogy & Outcome • Education and Training • Virtual Counselling • Patient test), GAA genotypes and GAA protein expression, ADA titers at Empowerment • Research These activities will be performed in close Weeks 10-20 and 40-52, change from baseline in efficacy outcomes collaboration with LSD patient organizations. Activities in the first year (Wilcoxon rank-sum test), and safety data. 59/60 patients had post- of the LSD subnetwork have focused on setting up an organizational treatment data all 59 had ADA 18/59 (31%) were IAb-positive. 14/35 structure, making an inventory of patients with different LSDs, starting (40%) ambulatory and/or respiratory support users at baseline and 4/ development of clinical guidelines and mapping teaching and training 24 (17%) device-independent patients became IAb-positive capabilities. We now aim at initiating collaborative research projects (P=0.0559). 13/31 (42%) patients with baseline FVC b55% predicted and mapping diagnostic capabilities. versus 5/28 (18%) with baseline FVC ≥55% predicted became IAb- positive (P=0.0449). IAb status was unaffected by genotype or residual GAA protein level. Most patients whose maximal ADA titers doi:10.1016/j.ymgme.2018.12.380 during Weeks 10-20 were ≥6400 (12/14 [86%]) or mean ADA titers ≥6400 during Weeks 40-52 (13/15 [87%]) were IAb-positive. IAb- positive patients had lower median changes from baseline in 6MWT (13 vs 15 m, P=0.9021), and lower median changes from baseline in 365 FVC % predicted (−0.5 vs 1.0, P=0.3180) differences by IAb status Clinical criteria for the early detection of mucopolysaccharidosis were significant only for % predicted leg quantitative muscle tests type I in pediatric practice (−1 vs 2, P=0.0296). Patients with greater baseline LOPD severity a a a and higher early and sustained ADA titers were likely to be IAb- Nato Vashakmadze , Leyla Namazova-Baranova , Anahit Gevorkyan , a a a positive. Rates of treatment-emergent/serious adverse events and Ludmila Kuzenkova , Tatyana Podkletnova , Mariia Fadeeva , Sofya b a a infusion-associated reactions appeared unaffected by IAb status. IAb- Demura , Nataliya Zhurkova , National Medical Research Center for b efficacy relationships require further study. Funding: Sanofi Children's Health, Moscow, Russian Federation, First Moscow State Genzyme. Medical University of the Ministry of Health of the Russian Federation, Moscow, Russian Federation doi:10.1016/j.ymgme.2018.12.379 Early diagnosis of mucopolysaccharidosis type I (MPS I) is important for the prediction of life. MPS excludes the possibility of large cohort and randomized controlled studies to create protocols of diagnosis and therapy. The study included 16 patients (3m-15y.o.) 364 with MPS I, 59 patients (3m-16.7y.o.) with suspected on MPS, but the Lysosomal subnetwork of MetabERN: Objectives and organiza- diagnosis not confirmed. We have also included the results of tional structure screening for MPS I, 738 patients from different regions of the Russia. In 7 patients in this group was confirmed by low alpha-L-iduronidase Ans Van der Ploega, Giancarlo Parentib, Dominique Germainc, Hidde activity. Questionnaire compiled according the literature data and Huidekopera, Mireia del Torod, Giovanni Cianae, Dominique Rolandf, included clinical synopsis, typical for patients with MPS I. The Christina Lampeg, Maurizio Scarpag, aErasmus Medical Center, Rotter- statistical analysis was included: Pearson's χ2 Criterion, odds ratio dam, Netherlands, bFederico University, Naples, Italy, cUniversity of criterion, cluster analysis. Main clinical criteria of MPS I was divided by Versailles, Montigny, France, dHospital Vall d'Hebron, Barcelona, Spain, odds ratio criterion. The most significant criterion (5 points) was an eAMC Hospital of Udine, Udine, Italy, fCentre de Genetique Humaine, increase in the level of glycosaminoglycans. The first group of Gosselies, Belgium, gHelios Dr. Horst Schmidt Klin, Wiesbaden, Germany symptoms (3 points) was assigned the following criteria - coarse face, joint contractures, cloudy corneas, to group II symptoms - heart valves The European Reference Network for Hereditary Metabolic Dis- pathology, hepato-splenomegaly, macroglossia, bullet-shaped pha- eases (MetabERN) has been set up in response to the European langes, claw-hand deformity, umbilical/inguinal hernia, frequent Commission (EC) call to establish European Reference Networks respiratory infections. Group III symptoms (1 point) - noisy breathing, (ERNs) following the adoption of the Directive on patients’ rights in macrocephaly, short stature, kyphosis/scoliosis, progressive mental cross-border healthcare in 2011. MetabERN is structured in 7 deterioration. The analysis of signs was carried out in 59 patients with Subnetworks (SNWs) of homogeneous diseases and has organized an unconfirmed diagnosis of MPS I for the validation of this criteria activities and expected outcomes into a set of defined work packages system. It showed that patients in this group have 15 points very rarely (WPs). We here report the aims and organizational structure of the and only 8% had a result of at least 15 points. The system of criteria was lysosomal subnetwork (LSD SNW) of MetabERN. The Lysosomal tested on 23 patients with a confirmed diagnosis of MPS I: all patients Subnetwork aims to: • gain an overview of patients with LSDs in the of this group more than 15 points. The frequency of detection of EU. • promote awareness towards LSDs in the EU. • facilitate the more patients MPS among patients with suspected MPS was 1:105 S148 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 accordingly, the frequency of detection MPSI among such patients is London, United Kingdom, cUniversity of Sydney, Sydney, Australia, 10:1050.We grouped more important signs and symptoms for clinical dRoyal London NHS Foundation Trust, London, United Kingdom diagnostic of MPS I. This system of clinical criteria is the probably elevate of diagnosing MPS I in patients at an early age Fabry disease (FD) is characterized by progressive left ventricular hypertrophy (LVH), fibrosis and heart failure. Cardiovascular magnetic resonance (CMR) imaging is the gold standard for evaluation of doi:10.1016/j.ymgme.2018.12.381 LV mass, while tissue characterization using T1 mapping detects early myocardial lipid deposition. The aim of this study was to define changes in cardiac structure and function in FD on serial CMR. 71 patients with genetically-confirmed FD were included in a longitu- 366 dinal, multicenter study. All underwent CMR imaging (1.5T Avanto) The survival of patients with mucopolysaccharidosis: Analysis of using a standardized clinical protocol, including T1 mapping 117 cases of Russian patients performed on mid LV short-axis slices using a Modified Look-Locker Inversion recovery sequence and late enhancement imaging follow- Nato Vashakmadzea, Leyla Namazova-Baranovaa, Nataliya Zhurkovab, ing injection of gadolinium. A gradient estimating equation regres- Anahit Gevorkyanb, Ludmila Kuzenkovab, Tatyana Podkletnovab, sion model was used to evaluate serial change, accounting for Anastasia Rykunovac, Tatiana Privalovaa, Elena Komarovab, aPirogov variable follow-up. 182 patient visits were assessed over a median Russian National Research Medical University (RNRMU), Moscow, follow-up period of 32-months (range 9-90). The mean age at Russian Federation, bNational Medical Research Center for Children's baseline was 41±14 years, with 43% ♂ (n=30). At baseline, males Health, Moscow, Moscow, Russian Federation, cFirst Moscow State had a higher LV mass and lower T1 time compared to females (♂119 Medical University of the Ministry of Health of the Russian Federation, vs. ♀ 68g/m2 ♂ 891 vs. ♀ 939ms (pb0.05)). A progressive increase in Moscow, Russian Federation LV mass was measured in males (↑ 1.7% per year, p=0.007) and was detectable despite the use of enzyme replacement therapy (ERT) (↑ Mucopolysaccharidoses (MPS) is group of inherited metabolic 2.1% per year, pb0.001). In females, there was no change in LV mass disease, resulting accumulation glycosaminoglycans in cell, caused by over time. T1 time however, fell in those not on therapy (-9.4% per mutations in the gene encoding lysosomal enzymes. Patients with year, p=0.007) but increased in those on ERT (5.1% per year, severe phenotypes of all types MPS have progressive cardiovascular, p=0.112). In males, increasing LV mass is a marker of advancing respiratory failure, progressive deterioration, skeletal anomaly and die Fabry cardiomyopathy and using CMR, we have demonstrated that at an early age. Enzyme replacement therapy used in Russia since 2008 progression occurs irrespective of therapy. Over the time frame in for MPS II, and more later - for other form MPS. Most of the children this study, LV mass did not change in females but there was evidence began to receive ERT at the age of 10 years. All of them had severe of ongoing myocardial sphingolipid deposition when not receiving clinical manifestations of the disease. We analyzed 117 patients in our ERT, consistent with a sex-dependent response to therapy. clinic with differential types of MPS: MPSI - 23, MPSII - 55, MPSIII - 24, MPSIV - 7, MPSVI - 8. Eleven patients was died since 2008 to 2017: MPSI -1, MPSII - 6, MPS3 -2, MPSVI -2. All patient have severe phenotype of doi:10.1016/j.ymgme.2018.12.383 disease. Kaplan-Meier survival analysis to assesses survival performed using Stats Direct and STATA(version 8) software for all patients. Kaplan-Meier survival analysis, median survival estimates and log-rank tests performed for all our patients. The time of death oldest patient was 368 20 years old, the youngest - 10 months. We used time intervals, A multicentre study of cardiac device implantation, arrhythmic multiples of 5 years - 5, 10, 15 and more than 15 years. 57% patients burden and risk factors in Fabry cardiomyopathy death registered to 15 years. Median survival estimated from ERT patients was 16,27±1,08 years, 95% CI:14,16-18,39, from non-ERT Ravi Vijapurapua, Ana Jovanovicb, Nigel Wheeldonc, Abbasin Zegarda, patients 12,11±0,55 years, 95%CI-11,03-13,2. Two case of patient death William Bradlowa, Francisco Leyvad, Patrick Deegane, Rosemary was at 9 years and as result - sharp decline in the survival curves from Ruske, James C. Moonf, Derralynn A. Hughesg, Peter Woolfsonb, 100% to 78%. Survival curves of patients with ERT treatment was Richard P. Steedsa, Tarekegn Geberhiwota, aUniversity Hospital gradual. There was no statistically significant effect of ERT on patient Birmingham NHS Foundation Trust, Birmingham, United Kingdom, survival (pN0.05). Short time that ERT has been available and small bSalford Royal Hospital, Salford, United Kingdom, cSouth Yorkshire number of patients, which get ERT-treatment from fist years of life can Cardiothoracic Centre, Sheffield, United Kingdom, dAston Medical not possible to model survival of these patients post-treatment. Research Institute, Birmingham, United Kingdom, eAddenbrookes Hos- pital, Cambridge, United Kingdom, fBarts Heart Centre, London, United Kingdom, gRoyal London NHS Foundation Trust, London, United doi:10.1016/j.ymgme.2018.12.382 Kingdom

Fabry disease (FD) is an X-linked lysosomal storage disorder, in which accumulation of myocardial sphingolipid leads to progressive 367 cardiac disease. Atrial and ventricular arrhythmia is variably reported The natural progression of cardiac involvement in Fabry disease with multiple arrhythmic risk predictors described. No guidance exists however, to risk stratify patients and determine when to Ravi Vijapurapua, Shanat Baiga, Sabrina Nordinb, Joao Augustob, implant cardiac devices. We aimed to evaluate current implantation Rebecca Kozorc, James C. Moonb, Derralynn A. Hughesd, Tarekegn practice, quantify arrhythmia burden and assess potential risk Geberhiwota, Richard P. Steedsa, aUniversity Hospital Birmingham NHS factors. 109 patients from four UK Fabry centres with genetically Foundation Trust, Birmingham, United Kingdom, bBarts Heart Centre, confirmed FD who have undergone cardiac device implantation were Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S149 included in an observational, retrospective study. Demographic and 370 baseline clinical data were collected (ECGs, echocardiography and Evaluation of a potential of chaperone therapy for muco- cardiovascular magnetic resonance imaging). Information regarding polysaccharidosis type IIID arrhythmia burden and treatment modification was captured from device follow-up and clinical notes. Arrhythmia rate and ICD Feng Wang, LA BioMed, Torrance, CA, United States utilisation was compared with age-matched hypertrophic cardiomyopathy (HCM) patients who had an implantable cardiac defibrillator Mucopolysaccharidosis type IIID (MPS IIID Sanfilippo syndrome (ICD). Of the 880 patients under follow-up in these centres, 12.4% type D) is a devastating pediatric neurodegenerative disorder with had a device implanted. Permanent pacemakers were inserted no cure or effective treatment available. The fundamental cause of following clinically significant bradycardia events in 92.1%, whereas MPS IIID is an inherited mutation in glucosamine (N-acetyl)-6- ICD insertion was more variable with 32.6% implanted for primary Sulfatase (GNS) required to catabolize heparan sulfate (HS). We will prevention outside current guidance, but based on suspected report our progress on evaluation of a potential chemical chaperone arrhythmic risk. The incidence of any arrhythmic event was higher (JSF-3358) with purified rhGNS, with patient fibroblast cells and in FD (61% vs. 44% in HCM, pb0.05). The rate of arrhythmia requiring iPSC-derived neural stem cells. Our preliminary data indicated that medical treatment however, was similar in both cohorts (FD vs. JSF-3358 can activate rhGNS activity (40%) and slightly enhance cell HCM: atrial fibrillation – 30% vs. 25%, non-sustained ventricular survival of MPS IIID fibroblasts versus control fibroblasts. We are tachycardia (NSVT) – 23% vs 25%. The incidence of VT with currently evaluating the ability of JSF-3358 to restore GNS activity in haemodynamic compromise was greater in FD compared to the three cell types: MPS IIID fibroblastsa, iPS cells, and neural stem cells. HCM cohort (28% over 5-years vs. 12% over 6-years, respectively In addition, we are evaluating its potential to enhance GNS stability (pb0.05)). The prevalence of risk factors was greater in FD patients during production of GNS and cellular uptake. who developed arrhythmia. Indications for device implantation are variable in FD. The burden of life-threatening arrhythmia and prevalence of risk predictors in FD are similar to other hypertrophic doi:10.1016/j.ymgme.2018.12.386 cardiomyopathies, supporting the need for Fabry-specific guidance. doi:10.1016/j.ymgme.2018.12.384 371 Intra-articular AAV9 α-iduronidase gene therapy in the canine model of mucopolysaccharidosis type I results in rapid synovial 369 and cartilage iduronidase expression, clearance of heparan The Fabry PRO online tool for secure and continuous patient sulfate, and high serum α-iduronidase levels follow-up and communication Raymond Y. Wanga,b, Shih-Hsin Kanc, Haoyue Zhangd, Afshin Susanne Walls, Ilkka Kantola, Miia Rainto, Turku University Hospital, Aminianc, Elizabeth M. Snellae, Jackie K. Jense, Patricia I. Dicksonf, Turku, Finland Sarah Youngg, N. Matthew Ellinwoode, aCHOC Children's Specialists, Orange, CA, United States, bUniversity of California Irvine School of Finland has 106 Fabry patients of whom 55 are on treatment. Most Medicine, Orange, CA, United States, cCHOC Children's Hospital, Orange, of the patients are in Fabry registries. Up to now paper documents and CA, United States, dDuke University Health System, Durham, NC, United outpatient visits have been used in the follow-up of the patients. States, eIowa State University, Ames, IA, United States, fWashington However, technology is rapidly changing healthcare and through University, St Louis, MO, United States, gDuke University School of technology we can improve both follow-up and communication Medicine, Durham, NC, United States between patients and healthcare providers in rare diseases. The Finnish Fabry Disease Centre of excellence at Turku University hospital Despite intravenous enzyme replacement and stem cell therapies, uses a web-based application for follow-up and communication treated MPS I patients still struggle with significant skeletal and joint between the Fabry Centre of excellence and Finnish Fabry patients. disease. This is thought to be in large part due to limited delivery of The application Fabry PRO tool uses Kaiku Health technology platform enzyme to joint cartilage and tissues. Previously, we showed that for secure communication. The platform supports all the major Fabry intra-articular IDUA injection of MPS I dog joints reduced joint tissue related questionnaires and they are accessible via web browser on any inflammation and cleared synoviocyte and chondrocyte lysosomal device. The goal is to improve and personalize patient management. storage. Since repeated intra-articular enzyme injections is not The application supports and encourages patient involvement. clinically feasible, we are studying the safety, efficacy, durability, Patients have access to their own saved data on the application. The and dose response of intra-articular AAV9-IDUA gene therapy in application supports patient-provider communication and it simplifies canine MPS I and report our early preliminary findings. Baseline and improves patient monitoring through automatic alerts and synovial and cartilage biopsies were obtained at 6 weeks from personalized instructions for patients. It enables a secure and bilateral shoulder and stifle joints. Four weeks later, the left shoulder continuous communication between the patient and the health care and stifle were administered 5e12 vector genomes (vg)/joint of providers. In the future we want to involve all Finnish Fabry patients, AAV9-eGFP (control) he right shoulder and stifle were administered able to use a computer or smartphone, to be part of this follow-up and 5e12 vg/joint of AAV9-canine IDUA (cIDUA). Serum, synovium, and communication channel. Using the web instead of traditional papers cartilage were obtained two weeks’ post-treatment IDUA and tissue and mail supports sustainable development. glycosaminoglycan concentrations were quantified. Treatment was well-tolerated without pain or joint effusions. Baseline serum cIDUA level was below limit of detection (b0.3 units/mL) post-treatment doi:10.1016/j.ymgme.2018.12.385 serum cIDUA was 3.6 units/mL (mean heterozygote cIDUA 3.35 units/mL). Post-treatment control synovial cIDUA levels were S150 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 undetectable, versus 43.5 and 153 units/mL in the AAV9-cIDUA- 0.13 mL/min/1.73m2/year P-difference=0.80). Different eGFR change treated joints. The difference between baseline and post-treatment patterns were found between groups of HRI/LRI (P-interac- synovial heparan sulfate in control joints was +6.7%, versus -96% in tion=0.0002) or ever/never RAS-blocker use (P-interaction=0.066). AAV9-cIDUA-treated joints. Heparan sulfate in cIDUA-treated joint The median first-AB age for 42 unique patients (50% classic) with cartilage was reduced 48%, compared with a 52% increase in control cardiac wall thickness measures was 50 years (follow-up: pre-AB 3.5, joints. One injection of intra-articular AAV9-cIDUA results in post-AB 3.6 years). Compared to the significant increase during the appreciable cIDUA enzyme levels in joint tissues and serum, effecting pre-AB period, both LVPWT and IVST did not progress pre- vs. post-AB large reductions in synovial and cartilage heparan sulfate substrate slopes were significantly different for LVPWT (slope difference=-0.41 storage. Tissue histopathology and gene expression analyses are mm/year, P-difference=0.0026) and approached significance for IVST ongoing vector biodistribution, dose response, durability of cIDUA (slope difference=-0.32 mm/year, P-difference=0.068). In conclu- expression, and long-term effects upon substrate concentration and sion, compared to the treatment-naïve period, overall eGFR decline joint morphology in additional animals are being evaluated. remained unchanged after treatment with more prominent decline among females with more renal involvement. Cardiac hypertrophy did not progress. Funding (Fabry Registry, abstract): Sanofi Genzyme. doi:10.1016/j.ymgme.2018.12.387

doi:10.1016/j.ymgme.2018.12.388

372 Renal and cardiac outcomes in female patients with Fabry disease treated with agalsidase beta: A Fabry registry analysis of pre- 373 versus post-treatment comparison Analysis of the baseline characteristics of Fabry disease patients screened for the pegunigalsidase alfa phase III BALANCE study Christoph Wannera, Ulla Feldt-Rasmussenb, Ana Jovanovicc, Aleš Linhartd, Meng Yange, Eva Brandf, Dominique P. Germaing, Derralynn David Warnocka, Wallace Erica, Raphael Schiffmannb, Myrl Holidac, A. Hughesh, John L. Jefferiesi, Ana M. Martinsj, Albina Nowakk, Bojan Ozlem Goker-Alpand,RobertHopkine, Pilar Giraldof, Nedd Khang,Bojan Vujkovacl, Frank Weidemannm, Michael L. Westn, Alberto Ortizo, Vujkovach, William Wilcoxi,AlesLinhartj, Camilla Tøndelk, Bruce A. aUniversity Hospital Würzburg, Würzburg, Germany, bRigshospitalet, Barshopl,AmelKaraam, Maria Judit Molnarn, Patrick Deegano,Jerry Copenhagen University Hospital, Copenhagen, Denmark, cSalford Royal Vockleyp,WilliamRheadq, Virginia Kimonisr,RaulChertkoffs,Nicola NHS Foundation Trust, Salford, United Kingdom, dCharles University and Longot, aUAB, Birmingham, AL, United States, bBaylor University Medical General University Hospital, Prague, Czech Republic, eSanofi Genzyme, Center, Dallas, TX, United States, cUniversity of Iowa, Iowa City, IA, United Cambridge, MA, United States, fUniversity Hospital Münster, Münster, States, dO&O Alpan LLC, Fairfax, VA, United States, eCincinnati Children's Germany, gUniversity of Versailles, Paris-Saclay University, Montigny, Hospital Medical Center, Cincinnati, OH, United States, fHospital de Dia France, hRoyal Free London NHS Foundation Trust and University Quiron, Zaragoza, Spain, gInfusion Associates, Grand Rapids, MI, United College London, London, United Kingdom, iUniversity of Tennessee States, hGeneral Hospital Slovenj, Gradec, Slovenia, iEmory University Health Science Center and Methodist University of Tennessee, Memphis, School of Medicine, Atlanta, GA, United States, jCharles University, Praha, TN, United States, jFederal University of Sao Paulo, Sao Paulo, Brazil, Czech Republic, kHaukeland University Hospital, Bergen, Norway, kUniversity Hospital of Zürich and University of Zürich, Zürich, lUniversity of California San Diego, San Diego, CA, United States, Switzerland, lGeneral Hospital Slovenj Gradec, Slovenj Gradec, Slovenia, mMassachusetts General Hospital, Boston, MA, United States, nSemmelweis mKlinikum West, Knappschaftskrankenhaus, Recklinghausen, Germany, University, Budapest, Hungary, oCambridge University, Cambridge, United nDalhousie University, Halifax, NS, Canada, oIIS-Fundación Jiménez Díaz/ Kingdom, pUniversity of Pittsburgh, Pittsburgh, PA, United States, qMedical UAM, IRSIN and REDINREN, Madrid, Spain College of Wisconsin, Milwaukee, WI, United States, rUniversity of California Irvine Center, Irvine, CA, United States, sProtalix Biotherapeutics, Carmiel, Fabry disease is an X-linked disorder caused by GLA variants and Israel, tUniversity of Utah, Salt Lake City, UT, United States cellular accumulation of glycolipids. The phenotypic spectrum in female patients is wide but few progress to end-stage renal disease Enzyme replacement therapy (ERT) may lead to formation of anti- compared with classic males. This Fabry Registry analysis drug antibodies (ADA) in “classical” patients with Fabry disease (FD). (NCT00196742) compared renal and cardiac outcomes pre- and Recent studies provided evidence that neutralizing antibodies (nAb) post-treatment with agalsidase beta (AB, 1 mg/kg EOW) in adult may interfere with ERT efficacy. Higher doses of ERT have been female patients. Analyses included estimated glomerular filtration rate recently shown to overcome the activity inhibition caused by ADAs. (eGFR, CKD-EPI equation), interventricular septum thickness (IVST) BALANCE (PB-102-F20 NCT02795676) is a double blind head-to-head and left ventricular posterior wall thickness (LVPWT). We included study comparing the effect of pegunigalsidase-alfa to agalsidase-beta patients receiving AB ≥2 years, having ≥2 pre- and ≥2 post-AB records (1mg/kg every 2 weeks) on renal function. The screening strategy within -5/+5 years of AB initiation, excluding patients with dialysis or selected FD patients with progressive loss of kidney function despite kidney transplant. GLA variants of patients were classic, unclassified long-term ERT (1-16 years). The current analysis describes the (fabry-database.org), or had not been reported. Piece-wise mixed baseline characteristics of FD patients screened for this study. Among model was used to compare pre- vs. post-AB slopes. Effect modifiers male patients currently screened for the study, 55% were positive for included renal involvement (low [LRI] vs. high [HRI]) and renin- both binding and neutralizing ADA to agalsidase-beta, with in-vitro angiotensin system (RAS)-blocker use. The median first-AB age for 86 mean enzyme activity inhibition of ~80% and IgG titers of 60 to patients (53.5% classic) with eGFR assessments was 46.3 years (follow- 127,933. One female was ADA-positive (binding ADA only). In-vitro up: pre-AB 3.6, post-AB 4.1 years). eGFR slopes declined consistently evaluation of the cross-reactivity of the nAb-positive samples toward within a normal range (b1 mL/min/1.73m2/year) pre- and post-AB pegunigalsidase-alfa showed a lower mean enzyme activity inhibition without significant differences between slopes (slope difference=- of ~64%, resulting in a greater amount of effective (non-inhibited) Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S151 enzyme of ~40% vs ~20%. The lower enzyme activity inhibition Since 1991, type 1 Gaucher disease (GD1) has been treated primarily combined with ~40-fold longer plasma half-life of pegunigalsidase- with pharmacological enzyme replacement therapy (ERT). In the pre- alfa compared to agalsidase-beta (~80 h vs ~2h) are expected to result ERT era, sequelae of fibrotic liver disease were a common cause of early in a greater amount of effective enzyme available in the circulation of death. Ultrasonic and MR elastography suggest that increased liver patients switching from a long-standing treatment with agalsidase- stiffness attributed to fibrosis correlates with GD1 disease severity beta to pegunigalsidase-alfa. Further evaluation of ADA status and scores. Here, we evaluate the impact of initial and post-ERT GD1 DS3 renal function of this cohort indicates that 64% (14/22) of the ADA- severity scores on probability for liver fibrosis (LF) in 30 patients using 4 positive FD male patients also have significant proteinuria (UPCR≥ 500 well-established LF prediction scores (AST/Platelet Ratio Index, Fibrosis- mg/gr). The unique characteristics of pegunigalsidase-alfa, 4 Index, Lok Score and NAFLD Fibrosis Score. We also studied 9 never- encompassing an improved pharmacokinetic profile, lower immuno- treated pts and 7 pts treated for less than 18 m and examined the genicity, and lower cross reactivity to pre-existing anti-agalsidase nAb, relationship between LF, disease severity scores and D-dimer, ferritin, may result in higher levels of effective enzyme reaching target organs, chitotriosidase and GBA1 genotype. 26/30 pts (86.7%) treated for at least with the potential to improve long-term clinical outcomes and 3 y had low probability of LF at last evaluation. 4/9 (44.4%) ERT-naïve pts attenuation of renal function deterioration in FD patients. and 3/7 (42.9%) pts treated for ≤ 18 m had high probability for LF. 5/7 (71.4%) pts with marked DS3 severity had high LF probability compared to 2/14 (14.3%) with moderate DS3 severity and 3/25 (12.0%) with mild doi:10.1016/j.ymgme.2018.12.389 DS3 severity. There was no correlation between plasma chitotriosidase, serum ferritin or D-dimer levels and a high probability for LF. 7/10 patients with high probability for LF were N370S homozygous. Per elastography studies reported by others, we find that LF probability 374 based on non-invasive laboratory risk stratification is often increased Promise of AAV9 gene therapy in the treatment of Batten disease: parallel to GD1-DS3 score. However, in contrast to the pre-ERT era, most Systematic approach in therapy design reduces pathological and adult GD1 patients treated with ERT for a median 20 y, including those fi behavioral de cits and prolongs survival in mouse models of with initial marked disease severity, appear unlikely to develop LF. CLN3-, CLN6-, and CLN8-Batten disease Exceptions may be attributed to confounding concurrent illnesses or to poor treatment compliance. We emphasize that N370S homozygosity a a a a Jill Weimer , Jacob Cain , Tyler Johnson , Katherine White , Shibi can be associated with severe GD1 manifestations including LF and such b b a Likhite , Kathrin Meyer , Sanford Research, Sioux Falls, SD, United pts should be treated accordingly. States, bThe Research Institute at Nationwide Children’s Hospital, Columbus, OH, United States doi:10.1016/j.ymgme.2018.12.391 Batten disease, also referred to as neuronal ceroid lipofuscinosis, is an autosomal recessive, lysosomal storage disorder resulting from mutations in one of 13 genes. While rare, affecting 1:100,000 live births, these disorders collectively represent the most prevalent 376 pediatric neurodegenerative disease. The disease leads to cognitive, Valvular heart disease complicating advanced Fabry disease: perceptual, and motor coordination impairment ultimately ending in Association with chronic kidney disease premature death. Here we explore gene therapy as a potential treatment for multiple subtypes of this disease. We use a self- Michael L. Westa, Mathieu C. Castonguaya, Emily Chisholma, Kaye complementary adeno-associated virus serotype 9 (scAAV9) to target LeMoineb, aDalhousie University, Halifax, NS, Canada, bQE II Health the nervous system and introduce a healthy copy of the human CLN3, Sciences Centre, Halifax, NS, Canada CLN6, and CLN8 genes, separately. A single intracerebroventricular (ICV) injection to a postnatal day one mouse resulted in widespread Cardiac involvement is common in Fabry disease, which is transduction of scAAV9 payload throughout the brain and spinal cord, characterized by a deficiency of alpha-galactosidase A activity with and prevented or drastically reduced many of the common pathologies accumulation of neutral glycosphingolipids in lysosomes of all cells. of the disease, such as autofluorescent storage material, astrocytosis, Patients develop cardiac hypertrophy with dysrhythmias and heart and microglial activation. Moreover, there were significant improve- failure. Valvular heart disease has rarely been reported in patients with ments in motor performance, learning/memory deficits, and survival Fabry disease. We report a retrospective review of valvular heart in treated mutant mice. These studies highlight the promising future of disease among 107 adults with Fabry disease in Nova Scotia enrolled in gene therapy for treating patients with Batten disease. the Canadian Fabry Disease Registry since 2007. Eight FD patients (M7, F1) mean age 51.5 years all of the Nova Scotia kindred with A143P mutation, severe classical phenotype, mean MSSI 50/76 developed doi:10.1016/j.ymgme.2018.12.390 severe symptomatic valvular heart disease with cardiac hypertrophy. Seven were receiving enzyme replacement therapy (ERT) with agalsidase infusions for a mean of 108 months. Mean LVMI was 218.2 g/m2. Most (87.5%) had a chronic dysrhythmia pre-op. Six has stage 5 375 CKD with 4 on dialysis and 2 had been on dialysis prior to a kidney fi Gaucher disease type 1 severity and risk for liver brosis in transplant. Valve surgery was done in 7 patients: AVR 5, TAVI 1, MVR 1, untreated patients and in patients treated primarily with enzyme MV annuloplasty 1, concurrent CABG 1. A female patient with severe replacement therapy for a median of 20 years heart failure and TR did not have surgery and died. Removed valves showed glycosphingolipid in 4/4, myxomatous change in 3/6 and Neal J. Weinreb, Robert A. Ali, University of Miami Leonard M Miller calcification in 6/6 of whom 5 had stage 5 CKD. Major post-operative School of Medicine, Miami, FL, United States complications included a watershed stroke, heart block requiring S152 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 pacemaker insertion and a paravalvular leak. During median 26 Lexington, MA, United States, bChildren's Hospital of Philadelphia, months follow up 2 patients died, 12 and 21 months after surgery Philadelphia, PA, United States, cUniversity Children’s Hospital Tübingen, from cardiac arrest and aortic dissection with perforation 5 remain Tübingen, Germany stable. Valvular heart disease in Fabry disease has a high prevalence (16.8%) in adults in the Nova Scotia Fabry disease kindred despite ERT. Metachromatic leukodystrophy (MLD) is an autosomal recessive Most patients had stage 5 CKD. The valve damage appears to be of disease caused by arylsulfatase A (ASA) deficiency, leading to mixed origin including Fabry disease, uremic calcification, myxoma- sulfatide accumulation and neuronal demyelination. Children with tous degeneration and heart failure. late-infantile MLD experience severe, rapidly progressing neurodegenerative symptoms. There are no approved MLD therapies. Phase 1/2 data (NCT01510028) suggest that intrathecal administration of doi:10.1016/j.ymgme.2018.12.392 100 mg of recombinant human ASA (SHP611 rhASA) every other week may slow motor function decline in some children with late- infantile MLD. A global, phase 2b, open-label trial is planned to assess the efficacy of once-weekly intrathecal dosing with 150 mg of rhASA 377 in patients with late-infantile MLD (target enrollment: N=35). The Radiographic progression of hip disease in Morquio syndrome safety and tolerability of rhASA and its delivery via the SOPH-A-PORT type A: A natural history study Mini S intrathecal drug delivery device will also be assessed. Four treatment groups will be included based on age and degree of motor Klane Whitea, Yu-Ji Lib, Eveline Langereisc, I. Jung Fengb, aSeattle dysfunction: Group A (18-48 months Gross Motor Function Classi- CHildren's Hospital, Seattle, WA, United States, bChi-Mei Medical Center, fication in MLD [GMFC-MLD] level 1-2 n=16), Group B (18-72 Tainan, Taiwan, cAcademisch Medisch Centrum, Amsterdam, months GMFC-MLD level 3 n≤8), Group C (18-72 months GMFC‑MLD Netherlands level 4 n≤8) and Group D (b18 months pre-symptomatic siblings of participants, with the same ASA allelic constitution n=3). Symptom Morquio syndrome type A (MPS IVA) is associated with progressive onset must be before 30 months in Groups A-C. The primary hip disease. Hip dysplasia and proximal femoral epiphyseal dysplasia endpoint will be the proportion of children in Group A with an result in subluxation and arthrosis with associated pain and disability. increase in their GMFC-MLD classification level (indicating motor There are no studies assessing the natural history of untreated hip function decline) of no more than 2 levels from baseline to 2 years. disease in children with MPS IVA. This purpose of this study is to The key secondary endpoint will be the proportion of children in describe the radiographic natural history of hip disease of children with Group A with a Gross Motor Function Measure-88 total score of N40 MPS IVA. Images were obtained from the elosulfase alfa phase III clinical at 2 years. Other secondary endpoints will include changes from trial program in MPS IVA patients. Radiographs were prospectively baseline at 2 years in sulfatide levels in cerebrospinal fluid, brain N- collected for all subjects and performed at 72-week intervals. Patients acetylaspartate/creatine ratios and Expressive Language Function were aged 5-20 years. Of the 176 patients enrolled, 156 had hip Classification-MLD levels. Outcomes will be compared with a radiograph images available and included in the study, for a total of 444 propensity-score-matched, historical MLD control group. Shire images. Images were excluded for incomplete clinical information, age funded this study and medical writing support. exceeding 20 years, previous hip surgery, and poor image quality. 135 patients (389 hips) were included. Hip radiographs were evaluated for ’ Tonnis angle, Reimer smigrationindex,andTonnisgradeforarthrosisof doi:10.1016/j.ymgme.2018.12.394 the hips. The mean age of the 135 patients was 10.32 yrs. There are 43 patients younger than 7 years old, 29 patients are between the age 7 and 10, 36 patients are between the age 10 and 13, and 27 patients are older than 13 years old. An increase in Tonnis angle was observed with age. 379 (pb0.01) Tonnis angle progressed 0.5°/year. (pb0.01) RMI trended Chitotriosidase as a biomarker for central nervous system upward with age. Tonnis grade significantly worsened over time, with inflammation in the gangliosidosis diseases 96.3% Tonnis grade III hips seen in patients older than 13 years. (Pb0.0001) Hip disease is progressive in MPS IVA, as demonstrated by Chester B. Whitley, Donald Bystrom, Jeanine R. Jarnes-Utz, University worsening acetabular dysplasia and joint degeneration. Significant joint of Minnesota, Minneapolis, MN, United States disease is found in N95% of adolescents over 13 years of age. A better understanding of the natural history of hip disease in MPS IVA is gained Chitotriosidase has been a useful biomarker in evaluating the through this large, natural history study. response to treatment of Gaucher disease, and elevations have been thought to be indicative of inflammation in some other conditions. However, it has not been useful in evaluating the pathology or doi:10.1016/j.ymgme.2018.12.393 response to treatment in other lysosomal diseases. Toward clinical trial readiness for treatment of gangliosidosis diseases, this study evaluated chitotriosidase in plasma and cerebrospinal fluid in a natural history study of these conditions. Patients affected Tay-Sachs 378 disease, Sandhoff disease, and GM1-gangliosidosis have been en- Intrathecally administered recombinant human arylsulfatase A in rolled in the Natural History of Gangliosidosis Diseases and patients with late-infantile metachromatic leukodystrophy: undergoing longitudinal assessments including behavioral assess- Phase 2b clinical trial design ments, MRI imaging, and biochemical testing. To gain an understanding of the progression and severity of disease, patients David Whitemana, Adeline Vanderverb, Ingeborg Krägeloh-Mannc, Jill undergoing lumbar spinal taps to assess for hydrocephalus were Graya, James Wua, Margaret Wasilewskia, aShire International GmbH, included, covering a range from infantile, juvenile and late-onset Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S153 phenotypes over a range of ages. Results were compared to values of 381 patients with other lysosomal diseases. Results showed remarkable Evaluation of the frequency of pre-motor symptoms of Parkinson patterns that illustrate the inflammatory or “macrophage activation” disease in adult patients with type 1 Gaucher disease of liver disease, but also central nervous system disease, and provides a new marker for future potential therapies. (Supported Matheus V.M.B. Wilkea, Alicia D. Dornelesb, Artur S. Schuhc, Filippo P. by NIH Lysosomal Disease Network U54 NS065768.) Vairob, Tatiele Nalind, Tassia Tononb, Ida V.D. Schwartze,f,g, aPostgraduate Program in Genetics Applied to Medicine, Department of Genetics, Faculdade de Medicina, Universidade Federal do Rio Grande do doi:10.1016/j.ymgme.2018.12.395 Sul, Porto Alegre, Brazil, bMedical Genetics Service, Hospital de Clínicas de Porto Alegre, Porto Alegre, Brazil, cDepartment of Pharmacology, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil, dHospital Moinhos de Vento, Porto Alegre, Brazil, eMedical Genetics 380 Service, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS, Brazil, Design, baseline characteristics, and 18-24 months follow-up fDepartment of Genetics, Universidade Federal do Rio Grande do Sul, from the MPS IIIA natural history study Porto Alegre, Brazil, gBRAIN Laboratory, Hospital de Clínicas de Porto Alegre, Porto Alegre, Brazil Frits Wijburga, Samantha Parkerb, Philippe Drevotb, Alice Le Meurb, c d e Anupam Chakrapani , Virginie Coutinho , Thilo Diehl , Roberto Parkinson's disease (PD) is the second most common neurodegen- f g h e Giugliani , Benedicte Heron , Jan Pieter Marchal , Nicole Muschol , erative disease. The classic motor symptoms of PD may be preceded by c e f Imogen Newsom-Davies , Anna Pereze , Solanger Perrone , Sophie many non-motor symptoms (NMS), such as hyposmia, REM sleep b a Olivier , Academic Medical Center, Amsterdam, Netherlands, behavior disorder, constipation, cognitive impairment, and depres- b c Lysogene, Neuilly sur Seine, France, Great Ormond Street Hospital, sion. Population studies have identified mutations in GBA1 as the main d e London, United Kingdom, Hopital Trousseau, Paris, France, University risk factor for idiopathic PD. This is an observational, cross-sectional f Medical Center, Hamburg, Germany, UFRGS and Medical Genetics study aiming at the evaluation of the prevalence of NMS of PD in a g Service, HCPA, Porto Alegre, Brazil, Hôpital Armand Trousseau La Roche cohort of South Brazilian adult patients with Gaucher Disease (GD) h Guyon, Paris, France, University Medical Center, Amsterdam, type 1. Cognition was evaluated by the Montreal Cognitive assessment Netherlands (MoCa), daytime sleepiness by the Epworth Scale, depression by Beck´ s Inventory, constipation by the Unified Multiple System Atrophy Mucopolysaccharidosis type IIIA (MPSIIIA) (OMIM252900) leads Rating Scale and REM sleep behavior disorder by the single-question to early-onset neurodegeneration and premature death. To reinforce screen. Hyposmia was assessed with Sniffin’ Sticks. The sample published data on the natural history of MPS IIIA and expand the included 22 patients (mean age: 51.25 ±15.3, range: 22-67 years). geographic outreach, Lysogene launched an observational study We identified REM sleep behavior disorder in 4 patients, constipation fi ("NHS") in ve countries (NCT02746341), which will function as a in 3, daytime sleepiness in 9, depression in 4 patients. A low MoCa non-concurrent control for the Phase 2-3 gene therapy trial score was found in 7 patients and hyposmia was identified in only one (NCT03612869). Lysogene coordinated discussions with experts patient. PD affects 2% of the population over 60 years and 4% of the ensuring that the selection of parameters measured in the NHS and population over 80 years. Since the mean age of our sample is b60 the gene therapy trial are robust and clinically meaningful. This NHS years, a longitudinal follow-up of our patients is necessary, especially is completely compliant to the International MPS Consensus for for those ones who present NMS. Cognitive Endpoints recommendations (van der Lee, 2017). The NHS enrolled twenty-three patients, less than 9 years of age, with the classical form of MPS IIIA. Subjects are from Brazil, France, Germany, doi:10.1016/j.ymgme.2018.12.397 Netherlands and UK. Median age at enrollment was 61 months (range, 28 to 105). Median age at diagnosis was 37 months. Baseline data showed a mean BSID-III development age (DA) of 15.2 months (range, 4.67 to 29) with mean developmental quotient (DQ) score of 382 31.52 (n=23). At six months the mean DQ score was 30.52 (n=20) Immune modulation for a female Hunter syndrome patient prior at twelve months the mean score was 25.71 (n=19). While DA to starting idursulfase increased slightly at 6 and 12 months (8.58% and 3.35%, respectively), DQ declined by 2.33% and 17.72% at 6 and 12 months, Kara Woolgara, Daniel Juliena, Holly Millera, Laura Pollardb, Zoheb respectively. At baseline, the mean DA as measured by the VABS-II Kazic, Kristin Lindstroma, Priya Kishnanic, aPhoenix Children's Hospital, was 19.32 months. Most patients showed decline in VABS-II DA and Phoenix, AZ, United States, bGreenwood Genetic Center, Greenwood, SC, DQ scores over 12-months. Mean VABS-II DQ were slightly higher United States, cDuke University, Durham, NC, United States than mean BSID DQ scores but correlate in terms of decline -0.84% and -14.07% at 6 and 12 months, respectively. Lysogene’s NHS has A 3.5-year-old female presented with coarse facial features, formed the largest international cohort in MPS IIIA. The use of macrocephaly, macroglossia, hepatomegaly, and no family history of a consensus agreed outcome measures and instruments for patient lysosomal storage disorder. She was diagnosed with Hunter syndrome evaluation allows data pooling with other studies using the same (MPS II) by enzyme and molecular testing. Genetic evaluation found an assessment tools and quality standards. undetectable plasma iduronate-2-sulfatase enzyme activity and an inversion between intron 7 of the IDS gene and a region near exon 3 of IDS-2. Patients with Hunter syndrome who have large insertions, doi:10.1016/j.ymgme.2018.12.396 deletions, or inversions in IDS-2 are at an increased risk of developing enzyme-neutralizing antibodies as they are CRIM negative when S154 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 compared to those who have missense mutations. Because of the 384 inversion of the IDS gene, there was concern that she was at high risk of Intravenous 2-hydroxypropyl-beta-cyclodextrin for a Niemann- developing high and sustained antibody titers to idursulfase which Pick disease type C1 infant with liver cirrhosis would limit her benefit from enzyme replacement therapy (ERT). Neutralizing antibodies to idursulfase are associated with poor clinical Kara Woolgara, Daniel Oryb, Sylvie Lebela, Kyrieckos Alecka, aPhoenix outcomes such as decreased pulmonary function due to antibody- Children's Hospital, Phoenix, AZ, United States, bWashington University mediated inhibition of idursulfase’s therapeutic activity, elevated urine Metabolomics Facility, St. Louise, MO, United States glycosaminoglycans, and also an increased risk for hypersensitivity reactions. Therefore, she underwent immune modulation to make it less Niemann-Pick C (NPC) is an autosomal recessive, life threatening likely that she would develop high sustained antibody titers. The neurodegenerative disease. Currently there is no FDA approved protocol included rituximab weekly for four weeks (4 doses), treatment. In preclinical studies, treatment with 2-Hydroxypropyl-β- methotrexate three times a week for three weeks (9 doses) and Clyclodextrin (HP-β-CD) has been shown to reduce both cholesterol monthly IVIG through B-cell recovery. Immune modulation using this and sphingolipid storage and prolong survival, and has been same protocol has been successful for patients with CRIM negative advanced to clinical trials for intrathecal delivery (IT). While IT Pompe disease. One year later, she is doing well. Her IgG level is within delivery in the clinical trials directly addresses the neurodegenera- normal range, she no longer requires IVIG, anti-drug antibodies tive component of NPC disease, visceral manifestations of the disease continue to be negative, and urine total glycosaminoglycans are in the are left untreated. In infantile and juvenile forms of NPC disease, normal range. This immune modulation protocol could be considered patients typically present with neonatal cholestasis or for patients with other lysosomal storage disorders with available hepatosplenomegaly, and in severe cases may progress to liver enzyme replacement therapy who may be at high risk of developing failure. Limited data is available regarding the effect of HP-β-CD anti-drug antibodies. upon human NPC liver disease. A 3 month old boy presented with hepatomegaly, conjugated hyperbilrubinemia, transaminitis, elevated alkaline phosphatase, and elevated GGT, without jaundice doi:10.1016/j.ymgme.2018.12.398 and with a normal neurological exam. A liver biopsy indicated the presence of cirrhosis. Two pathogenic NPC1 mutations were identified. At 6 months of age, he was started on intravenous HP-β- CD every two weeks under an FDA-approved expanded access 383 program. HP-β-CD was administered at an initial dose of 500mg/kg, Superior vena cava syndrome in an infantile onset Pompe patient increased to 1000 mg/kg for the next five infusions, and then increased to 1500mg/kg/dose due to ongoing concerns of abnormal Kara Woolgar, Molly Maenchen, Kyrieckos Aleck, Phoenix Children's liver function studies and only slightly improved hepatomegaly. Hospital, Phoenix, AZ, United States After fourteen months of HP-β-CD treatment, his laboratory liver function tests and liver size are improved from initial presentation A 9 year old with infantile onset pompe disease (IOPD) post spinal He has had no infusion associated reactions and clinically is doing fusion surgery had complications including superior vena cava very well. At 20 months of age he continues on HP-β-CD intravenous syndrome (SVC). He was diagnosed with IOPD at 1 month of age. His every two weeks at 1500mg/kg and shows no signs of progressive CRIM status was positive and he was started on alglucosidase alfa at liver disease. His growth and development are normal for his age. 20mg/kg every 2 weeks. He tolerated infusions well with no infusion This demonstrates that early treatment with intravenous HP-β-CD related reactions. Due to lack of meeting motor milestones, severe may change the clinical course for NPC patients who present within a hypotonia and increased Hex4 levels his dose was increased over a 6- couple months of life with liver cirrhosis. month period to 40mg/kg weekly. His anti-drug antibody titers have fluctuated between +400 and +1600. At 7 years old, his left subclavian port malfunctioned. A venogram showed an occluded left subclavian doi:10.1016/j.ymgme.2018.12.400 and multiple collateral vessels. A port was placed in his right internal jugular (IJ) vein that seemed to be slightly deeper than the more superficial collateral veins. At 9 years old, he underwent a posterior spinal fusion. Post op day two he developed superior vena cava 385 syndrome (SVC) which resulted in altered mental status and respiratory Fabry disease has been found by using of the tumor mutational distress. An echo, ultrasound and venogram showed complete bilateral burden analysis of 3000 Japanese cancer genomes using whole occlusion of major venous vessels and reliance on multiple collaterals, exome and targeted gene panel sequencing: Project Hightech suggesting long-term hypercoagulability and venous occlusion. A Omics-based Patient Evaluation (Project HOPE) Heparin infusion did not resolve the SVC syndrome. Post op day 7, tPA was started and after 24 hours, a venogram demonstrated complete Hiroyuki Yamakawaa, Hiroyuki Matsubayashb, Seiichiro Nishimurab, resolution of acute thrombus within multiple collateral veins and Yasue Horiuchib, Yoshimi Kiyozumib, Satomi Higashigawab, Kei Iidab, resolution of SVC syndrome. His clinical course was complicated by two Takahiro Tsukimurac, Tadayashu Togawac, Kenjiro Kosakia, Hitoshi pulmonary embolisms, pleural effusions, two episodes of SVT, and Sakurabac, Keiichi Fukudaa, Masatoshi Kusuharab, aKeio University, difficulty with extubation. A previous study by Nayak et al., suggested Tokyo, Japan, bShizuoka Cancer Center, Shizuoka, Japan, cMeiji Pharma- hypercoagulability with high doses of rhGAA and high anti-rhGAA ceutical University, Tokyo, Japan. antibodies in the mouse model for Pompe disease. Background: Project HOPE (High-Tech Omics-Based Patient Evaluation) began in January 2014 at the Shizuoka Cancer Centre. doi:10.1016/j.ymgme.2018.12.399 The project goal is the integrated analysis of multiomic specimen data and clinical information, to achieve a clear understanding of Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156 S155 mechanisms of cellular oncogenesis and microenvironment changes, Methods: In our nationwide NBS program, more than one million to better elucidate cancer states. Collation and comparison of newborns were screened for Pompe disease. We have diagnosed and patient-specific clinical and omics information facilitate treated around forty percent of Pompe patients in Taiwan. individualised medicine, enabling holistic treatment of cancer Results: After 2010, the mean age at first enzyme-replacement patients. Using blood cells, we conduct an all-exon analysis of the therapy (ERT) for our IOPD patients was 9-day-old. Our patients had germ line, obtaining insight into the genetic makeup of the patient. better biological, physical, and developmental outcomes and lower We can then predict the risk of developing inherited cancers and anti-rh acid α-glucosidase antibodies. The mean age for independent inherited non-cancer diseases and can provide preventative pre- walking was 11.6 ± 1.3 months, the same age as normal children.our crisis care, recognizing that appropriate medical intervention lowers IOPD patients who had more severe form of this disease have better morbidity and mortality rates. clinical outcomes and comparing to other study groups of IOPD. Our Methods: In this example, a 48-year-old female patient under- patients have better outcomes in all aspects. went surgery for ascending colon cancer at the Shizuoka Cancer Conclusions: ERT for patients with Pompe disease should be Centre. At admission, we obtained the patient’s approval and initiated as early as possible before irreversible damage occurs. included her in the HOPE project. An α-galactosidase A (GLA) gene Starting ERT even a few days earlier might be the critical factor of anomaly was discovered, which is associated with Fabry disease, better outcomes. already reported. Mulberry bodies were observed in the patient’s urine. The patient’s eldest son (28-year-old) had been diagnosed doi:10.1016/j.ymgme.2018.12.402 with ulcerative colitis and had undergone surgery in 2014 he had suffered from limb pain and chronic diarrhoea since his teens. Blood fi tests revealed signi cantly lowered GLA activity. Mulberry bodies 387 were also observed in his urine. Analysis of cognitive ability and adaptive behavior assessment Conclusions: Project HOPE enabled the successful diagnosis of tools used in an observational study of patients with Hunter Fabry disease, a non-cancerous inherited illness, as a secondary syndrome finding. Diagnosis of Fabry disease in the patient’s son was possible and we could begin enzyme replacement therapy. Developments in Karen Yeea, Yanyu Wua, Magdalena Harringtonb, Susan E. Waisbrenc, genome therapy can lead to further secondary diagnoses of Fabry aShire, Cambridge, MA, United States, bPfizer, Cambridge, MA, United disease. It is vital that symptoms are holistically evaluated and States, cBoston Children's Hospital, Boston, MA, United States treatment plans are carefully selected. Additionally, we must investigate the relatives of diagnosed patients and ensure that we Cognitive and adaptive behavior tools used in an observational diagnose possible cases of Fabry disease. study (NCT01822184) of patients with mucopolysaccharidosis type II (MPS II Hunter syndrome), a rare lysosomal disease characterized by doi:10.1016/j.ymgme.2018.12.401 iduronate-2-sulfatase deficiency, were assessed in a post-hoc analysis. Patients aged ≥2–b18 years with a score of 55–85 on the Differential Abilities Scales-II (DAS-II), General Conceptual Ability 386 (GCA) composite scale were included. DAS-II (early years [EY] or Very early treatment for infantile-onset Pompe disease contrib- school age [SA] battery), Vineland Adaptive Behavior Scales, Second utes to better outcomes: 10-year experience of nationwide NBS in Edition (VABS-II) and the Hunter Syndrome-Functional Outcomes for Taiwan Clinical Understanding Scale (HS-FOCUS) tests were performed at baseline and 3-month intervals for 2 years. A mixed-effect model for Chia-Feng Yanga, Chen Chang Yanga, Hsuan-Chieh Liaob, Ling-Yi repeated measures was used to assess item-domain score associa- Huangc, Chuan-Chi Chiangd, Hui-Chen Hoe, Chih-Jou Laia, Tzu-Hung tions of each tool. Patients (n=38) had a mean ± SD age of 5.84 ± Chua, Tsui-Feng Yanga, Ting-Rong Hsua, Wen-Jue Soonga, Dau-Ming 3.44 years. Mean ± SD DAS-II GCA and VABS-II Adaptive Behavior Niua, aTaipei Veterans General Hospital, Taipei City, Taiwan, bNational Composite (ABC) scores at baseline were 73.1 ± 14.4 and 81.8 ± Yang-Ming University, Taipei City, Taiwan, cTaipei City HospitalHeping 11.8, respectively, which were relatively stable over time (mean − − Fuyou Branch, Taipei City, Taiwan, dThe Chinese Foundation of Health change 5.54 ± 13.1 and 0.31 ± 6.32, respectively at month 24). Neonatal Screening Center, Taipei City, Taiwan, eTaipei Institute of Change from baseline over time in EY verbal and SA non-verbal Pathology, Taipei City, Taiwan reasoning ability cluster scores, and EY pattern construction, naming vocabulary, and picture similarities subtest scores was similar to that Objective: Newborn screening is the way to initiate the very early for DAS-II GCA scores. DAS-II GCA and VABS-II ABC scores were diagnosis and treatment. Our team, the Taipei Veterans General positively associated (β=0.1104, p=0.0316) over time. DAS-II GCA Hospital (TVGH) began Pompe newborn screening from 2008, score was negatively associated with 2/5 domains (grip/reach and testing approximately two-thirds of the newborn population in walking/standing) and 6/21 items of the HS-FOCUS. VABS-II ABC Taiwan. Until now, more than one million newborns were enrolled in score was negatively correlated with 2 domains (activities and grip/ our series. From 2010, we had established an effective newborn reach) and 2 items but positively associated with 1 item (sleep screening program with rapid diagnostic strategies, and almost all of without snoring) of the HS-FOCUS. In conclusion, DAS-II was a the suspected IOPD infants could be diagnosed correctly within 2 suitable measure of change in cognitive ability in pediatric patients hours and receive ERT within 4 hours of admission. With such an with MPS-II over 2 years, whereas VABS-II, a parent-report measure, effective system, the average age of our IOPD patients started their and HS-FOCUS did not reflect this change. Shire funded this study ERT is less than 10-day-old, the earliest group all over the world. To and medical writing support. evaluate whether very early treatment for our IOPD patients would result in better outcomes, we analyzed our IOPD patients who were diagnosed by NBS program by multiple clinical and biological doi:10.1016/j.ymgme.2018.12.403 parameters. This is a ten-year cohort study. S156 Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156

388 Evaluations were conducted at one site and followed a standardized Airway stenting in MPS IVA (Morquio syndrome type A) protocol designed for long-term neurodevelopmental outcomes. Nineteen late-infantile patients underwent HSCT between March Mildrid Yeo, Sergei Korenev, Richard Hewitt, Derek Roebuck, 1997 and January 2018 (15 boys, 4 girls). Five were asymptomatic and Maureen Cleary, Anupam Chakrapani, Great Ormond Street Hospital, 14 were symptomatic when transplanted. Sixteen were evaluated London, United Kingdom longitudinally from December 2001 to June 2018 (median follow-up 4.75 years, range 0.42-8.83) as two died and one was lost to follow-up. Patients with MPS IVA syndrome often develop progressive airway All asymptomatic children have normal cognitive function, 3 symp- obstructions which are challenging to manage. These extend from the tomatic have near-normal, 1 symptomatic is severely delayed but nasopharynx to the intrathoracic trachea. Tracheal narrowing and improving, and the remaining plateaued below 2 years of age. Gross tortuosity has been attributed to shortened spinal height, laxity of motor is most affected with 3 asymptomatic children developing tracheal tissue, glycosaminoglycan (GAG) deposits, and extrinsic normally and 2 plateauing at 3 and 1.5 years of age. All symptomatic vascular compression. We present the first case of tracheal stenting in children plateaued or regressed around 6 months of age. Fine motor MPS IVA syndrome in the United Kingdom. Tracheal problems followed a similar trend. Language was least affected with 7 children encountered and surgical interventions are discussed. A 15.5-year-old developing normally for receptive language and 8 for expressive. For boy with MPS IVA (diagnosed age 1 year) presented with respiratory both, the remaining symptomatic children plateaued or regressed. failure following an admission for exacerbation of asthma. Lung HSCT improves the survival and functional abilities of late-infantile function tests and sleep studies were normal leading up to this. Krabbe patients. Children who underwent HSCT while asymptomatic Microlaryngoscopy and bronchoscopy revealed a 90% reduction in the experienced normal development in all domains except 2 patients tracheal cross-sectional area (6cm distal to vocal cords 2cm proximal to who plateaued in motor skills at 1.5 and 3 years respectively. carina) with jagged narrowing from GAG deposits. Multiple failed extubation attempts and unsuitability for tracheostomy insertion led to doi:10.1016/j.ymgme.2018.12.405 the decision for tracheal stenting. Two 16mm x 30mm 6F self- expanding nitinol tracheal stent were inserted. However failure to extubate required a third stent to be placed in the upper trachea. These 390 procedures were undertaken after in-depth discussions with patient, Deletion of chromosomal region Xq28 cause of Hunter syndrome parents and medical teams. Patient developed increased respiratory in a patient with mild clinical phenotype secretions post-stent insertion. After the last stent insertion, he suffered from respiratory failure secondary to pneumothorax and left lung collapse. Bronchoscopy confirmed patency of the stents. At nine Natalia V. Zhurkova, Kirill Victorovich Savostyanov, Alexandr months’ post procedure, he had a chest infection necessitating a brief Andreevich Pushkov, Tatiana Vladimirovna Podkletnova, Nato period of ventilation. At one-year follow-up his respiratory secretions Dzhumberovna Vashakmadze, Evgeny Leonidovich Penkov, Ludmila and symptoms remain stable. Airway compromise is a leading cause of Michaylovna Kuzenkova, National Medical Research Center of ’ mortality in MPS IVA. Current recommended schedule of respiratory Children s Health, Moscow, Russian Federation assessments may not provide early warning of airway obstruction. Tracheal stenting should be considered after exhausting other available Background: Mucopolysaccharidosis II (MPS II) is a rare lyso- options. Patients may require multiple stents. Increased secretions and somal storage disorder caused by mutation in IDS gene, leading to fi infection risk post-stenting occur due to mucocilliary inhibition. de ciency of iduronate-2-sulfatase and accumulation of glycosaminoglycans in different tissues and organs. doi:10.1016/j.ymgme.2018.12.404 Case report: The 10 year old boy presented with history of recurrent respiratory infections progressive joint stiffness since 4 years. Patient had normal intelligence, mild growth retardation, overweight, joint contractures, asymmetry of face, upper and lower limbs SND, valgus deformation 389 of the lower limbs, mild dysostosis multiplex, minimal coarsening of Long-term neurodevelopmental outcomes of hematopoietic stem facial features, pebbly skin lesions, carpal tunnel syndrome, sensorineural cell transplantation for late-infantile Krabbe disease hearing loss left-side, hypermetropia. Нe had no cognitive impairment, seizures, respiratory problem. GAG urine excretion was increased. Isabel C. Yoona, Nicholas Bascoub, Michele Poeb, Maria L. Escolarb, Iduronate-2-sulfatase activity in blood cells was low: 3,039 nM 4MU aChildren's Hospital of Pittsburgh, Pittsburgh, PA, United States, \mg protein/4h (NN17 nM 4MU\mg protein/4h). bUniversity of Pittsburgh, Pittsburgh, PA, United States Results: DNA was isolated from blood samples using a standard, semi-automated method. We did not find IDS gene on Sanger Krabbe disease is a rare neurodegenerative disorder caused by a sequencing. Array CGH was proved in Affymetrix CytoScan 750K. deficiency in the lysosomal enzyme galactocerebrosidase, resulting in Analysis showed a 363 kb deletion of genomic material at Xq28 demyelination. It is divided into four subtypes (early-infantile, late- (148,294,415-148,657,098), which contains IDS gene. infantile, juvenile, and adult) based on age of symptom onset. The only Discussion: Despite the considerable length of the identified 363 effective treatment for Krabbe disease is hematopoietic stem cell kb deletion of the Xq28 chromosomal region, our patient demon- transplantation (HSCT). Studies have shown the efficacy of HSCT for strates the development of a soft MPS II phenotype with normal improving lifespan and functional abilities in early-infantile patients intelligence. This boy get enzyme replacement therapy Idursulfase who underwent transplantation before the onset of overt symptoms. with a positive effect. However, there is a lack of studies that evaluate the efficacy of HSCT for late-infantile patients. In this prospective longitudinal study, patients doi:10.1016/j.ymgme.2018.12.406 were evaluated by neurodevelopmental pediatricians, speech pathol- ogists, audiologists, physical therapists, and psychometricians. Molecular Genetics and Metabolism 126 (2019) S157–S168

Contents lists available at ScienceDirect

Molecular Genetics and Metabolism

journal homepage: www.elsevier.com/locate/ymgme

Author Index

Abaoui, Mona 212 Amschl, David 258, 260 Abasolo, Ibane 1 Andersen, Henning 159 Abdel-Hamid, Hoda 41 Anderson, Nicole 140 Abdelwahab, Magy 2, 233 Andersson, Gudrun 177 Abel, Florian 30 Andrade, Helida M. 253 Abell, Katherine 288 Andrade-Campos, Marcio M. 16, 17, 123 Abily-Donval, Lenaig 39 Angel, Kim 73 Abramov, Aya 297 Anguela, Xavier 21 Abu Ali, Qais 229 An Haack, Kristina 278, 363 Aburachis, Alyssa T. 3, 4 Anjum, Muhammad N. 61 Acosta, Angelina X. 200 Annala, Antti-Pekka 184 Adam, Jacqueline 5, 6, 7 Anstett, Kara 18, 19 Adang, Laura A. 8, 14, 321 Antonini, Mathilda 20 Adedipe, Dee 126 Appleby, Matt 195 Aerts, Hans 52 Aranda, Carolina S. 332 Aﬂaki, Elma 9 Arash-Kaps, Laila 157 Afonso, Carlos 39 Araujo, Roberto 344, 363 Aggarwal, Vipin 87 Arenaz, Izaskun 16 Aguiar, Mario 83 Arepalli, Sampath 115 Aguiar, Patricio 10, 11 Arguello, Annie 158 Aguilera Correa, John Jairo 315 Arif Hossain, Mohammad 173 Ahern-Rindell, Amelia 12 Arkadir, David 37, 92 Ahmed, Seemin S. 113 Armour, Sean M. 21 Ahn, Sujin 13 Armstrong, Dustin 22 Ahrens-Nicklas, Rebecca 14, 321 Arnold, M. 186 Ai, Lingbao 149 Arribas, A.I. 17 Ait Sab, Imane 88 Arslan, Nur 336 Ajayi, Temitayo 326 Arunkumar, Nivethitha 353 Akbari, Ayub 212 Asai, Katsuhito 174 Akiyama, Keiko 166, 173 Asher, Stephanie B. 95 Alaez, Carmen 116 Ashmead, Rhea E. 23, 65 Alcalay, Roy N. 108 Ashton, Helen 24 Aldámiz-Echevar, Luis J. 88 Astarita, Giuseppe 158 Aldámiz-Echevarria, L. 57 Astolﬁ, Annalisa 25 Aldrich, A. 186 Atanacio, Nora G. 26, 97 Aleck, Kyrieckos 383, 384 Auer, Ewald 258, 259, 260 Alexanderian, David 244 Augusto, Joao 367 Alfonso, Pilar 220 Auranen, Mari 159 Ali, Nafeeza 84 Auray-Blais, Christiane 27, 176, 212, 232 Ali, Robert A. 314, 375 Austin, Stephanie L. 28, 198, 339 Almeciga-Diaz, Carlos J. 15, 289 Autio, Reijo 284 Almon, Einat 160, 218 Avanti, Mahima 191 Alon, Sari 160, 218 Ayers, Dieter 83 Alonso-Jimenez, Alicia 91 Azevedo, Leila M. 127 Alonso-Pérez, Jorge 91 Azevedo, Olga 10, 11 Altmann, Vivian 33 Aziz, Nadine 22 Álvares-da-Silva, Mário R. 338 Alyward, S. 106 Babu, Deepti 221 Amaral, Fernanda G. 79 Baek, Rena 120 Amaral, Olga 239 Baig, Shanat 367 Amartino, Hernan 244 Bailey, Laurie 226, 288 Amato, Dominick 261 Bailey, Rachel M. 29, 225 Amici-García, Alba V. 56, 58 Baker, Nancy C. 197 Aminian, Afshin 371 Balaji, Uthra 329 Amit-Cohen, Bat-chen 160, 218 Baldo, Guilherme 285, 325 Ammer, Luise 345 Bali, Deeksha S. 90, 296, 343 Amraoui, Yasmina 157 Balistreri, William 30 https://doi.org/10.1016/j.ymgme.2018.12.407

Available online 09 January 2019 1096-7192 Molecular Genetics and Metabolism 126 (2019) S157–S168

Balmaseda-Serrano, E. 57 Brand, Stefan-Martin 214, 215 Balwani, Manisha 30, 31, 236 Bratkovic, Drago 68, 196, 324 Barba, Miguel Angel 128 Braun, Jennifer A. 270 Barber, Dwayne 232 Braunlin, Elizabeth 47, 48, 49, 140, 178 Barbey, Frederic 264 Bravo-Hidalgo, Alberto 116 Barch, Joshua 153 Brea-Hernando, A. 57 Barcos-Martínez, Montserrat 228 Brenchley, Paul 121 Bareiro, Rodolfo 200, 201, 240 Breyer, Sandra 203 Barez, Abelardo 16, 17 Briones, Eduardo 228 Barisoni, Laura M. 32 Bröijersén, Anders 50 Barkats, Martine 272 Brooks, Melissa L. 129 Barohn, Richard J. 278 Brown, T.M. 199 Barrabes, Jose A. 89 Brudvig, Jon 54 Barros Mendes, Pedro H. 163 Bruno, Irene 137 Barshop, Bruce A. 373 Brusius-Facchin, Ana C. 200, 202, 333 Barth, Anneliese L. 163, 164, 165 Bueno, Dolores 1 Barth, Jay A. 42, 68, 104, 181, 196, 324 Bueno Delgado, Maríadel Amor 131 Bascou, Nicholas 389 Bulut, Fatma 336 Basgalupp, Suelen P. 33, 55, 277, 338 Burgess, Rob 225 Basheeruddin, Khaja 51, 51 Burin, Maira G. 74, 200, 202, 333 Baskfield, Amanda 15 Burke, Derek G. 148, 150, 151 Batista, Julie 87, 295 Burton, Barbara K. 51, 244, 245, 246, 247 Batra, Gauri 121 Byrne, Barry J. 68, 196, 324, 344 Bay, Luisa 34 Bystrom, Donald 379 Beasley, James 343 Beaton, Brendan 35, 98 Cabasso, Or 52 Bebout, Cassie 36 Cabrera, Gustavo H. 161, 162 Becker, Pamela 352 Cador, Bérengère 255 Becker Cohen, Michal 37, 92, 289, 297 Cagin, Umut 21, 53, 70 Bedard, Catherine 158 Cagle, Stephanie 280 Bedreddine, Najya 262 Cahan, Heather 67, 326, 335 Bedwell, David 38 Cain, Jacob T. 54, 374 Bekri, Soumeya 39, 40, 262 Caiozzo, Vince 192 Bellcross, Cecelia 221 Caldeira, Raíssa P. 55 Belmatoug, Nadia 31, 209, 255 Calderón-Sandubete, Enrique 228 Belmonte, Izaskun 91 Camara, Daniely P. 45 Beltran-Quintero, Maria L. 41, 183 Camelo, José S. 87 Benavides, Jaqueline 362 Camón Pueyo, Ana María 360 Bento, Judith C. 300, 301, 302, 303, 304, 305, 306, 307, 308 Campbell, Samantha 233 Bentz Pino, Gessi 136 Canaan-Kühl, Sima 214 Berger, Kenneth I. 344 Canal, Maria 122 Berger, Marc 255 Canavese, Miriam 235 Bergquist, Lars 177 Cano, Horacio 16 Bernat, John 160, 227 Canossa, Sueli G. 164 Bernhard, Olaf 322 Cao, Liching 171 Berry, Lisa 204 Capablo, Jose L. 16 Bertuccio, Salvatore N. 25 Caparbo, Valeria F. 300, 301, 303, 306 Bhalla, Akhil 158 Carabajal, Ricardo 112 Bhatnagar, Caleb 192 Carbone, Marco 203 Bhattarai, Sajag 182 Carloni, Laura 295 Bichet, Daniel G. 42, 346 Carneiro, Paola B. 285 Biferi, Maria Grazia 70, 272 Carnicer, Clara 89 Bigger, Brian W. 43, 122, 266 Carozza, Patricia 112 Bijarnia-Mahay, Sunita 336 Carrasco-Rozas, Ana 91 Bisinoto, Joyce R. 127 Carrer, Michele 193 Bissoli, Larissa F. 45 Carrillo-Sanchez, Karol 116 Bittencourt, Rosane I. 55 Carson, Vince 41 Blackwood, Rachel 279 Casas, Josefina 1 Blumenfeld, Jessica 158 Case, Laura 28, 195 Boado, Ruben J. 44, 170, 222, 275 Cassidy, Daniel P. 314 Boentert, Matthias 344 Castelein, Rene M. 203 Boitano, Anthony E. 129, 270 Castelli, Jeffery P. 32 Bolton, Shaun 172 Castillo-García, Daniela 56, 58 Bonanni, Maria 95 Castonguay, Mathieu C. 376 Bondar, Constanza M. 76, 243 Castro Moreira, Maria L. 200, 201, 202, Bonner, Nicola 101 Cathey, Sara 62 Booth, Claire 86 Cattaneo, Federica 137, 156 Borges, Frederico M. 74 Cebolla, Jorge J. 16, 17, 57 Borgwardt, Line 137, 156 Cerón-Rodríguez, Magdalena 56, 58 Bostwick, J.Robert 38 Cerski, Carlos Thadeu S. 338 Botha, Jaco 209, 210, 245, 246 Cervantes-Barragan, David 116 Bouraghouat, Aicha 88, 206 Chabaa, Laila 206 Bouskraoui, Mohammed 88, 312 Chadwell, Sarah 288 Boutin, Michel 176, 212 Chaivorapol, Christina 152 Bozzo, Pedro Paulo F. 45 Chakrapani, Anupam 24, 267, 276, 380, 388 Braddock, Stephen R. 51 Chambers, Christina 352 Bradley, Colm 46 Chan, Alicia 310 Bradlow, William 368 Chan, Jinyan 329 Bragge, Timo 54 Chan, Yunghang 167 Brand, Eva 161, 162, 214, 215, 216, 372 Chandler, Randy 84

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Chandrasekharappa, Settara 99, 115 Curiati, Marco Antônio 164, 334 Changsila, Erk 217 Curran, Christopher 82 Chapman, Jennifer R. 314 Cybulla, Markus 214 Charles, Severine 69 Charrow, Joel 75, 179, 236 Dala-Ali, Benan 85 Charvet, Leigh 337 Dalén, Per 50 Chauhan, Anuj 59 D’Almeida, Vania 77, 78, 79 Chavez-Cintora, Edwin 60 Dalton, Nancy 167 Cheema, Huma A. 61 Daniel, Amanda V. 80 Cheggour, Mouna 206 Daniele, Nathalie 53 Chen, Hui-hsuan 354 Danos, Olivier 357 Chen, Steven 198 D'Antiga, Lorenzo 30 Chen, Wu 102 Dao, Julia 81, 175 Chen, Xin 62 da Silva, Adalberto S. 253 Chen, Yanjun 191 Das Mahapatra, Pronabesh 82, 83, 143 Chertkoff, Raul 160, 218, 373 Davidonis, Martynas 346 Chiang, Chuan-Chi 386 Davidson, Cristin 84 Chirino, Andrea 97 Davis, Crystal 225, 270 Chisholm, Emily 376 Davis, Samantha 54 Chisté, Yuri L. 163 Davison, James E. 85, 86, 151 Choi, Chulhee 63 Day, Taniqua 176, 329 Choi, Hojun 63 De Angelis, Victor 313 Choi, Kyungsun 63 De Armey, Stephanie M. 108 Choi, Soongyu 13 de Castro Lopez, Maria J. 67, 335 Chou, Tsui-Fen 64 De Cosmo, Joseph A. 149 Choy, Francis Y. 23, 65 Deegan, Patrick 87, 160, 209, 368, 373 Christensen, Chloe L. 23, 65 de Ferrán, Ceila P. 200 Christensen, Katherine M. 51 delas Heras, J. 57 Chu, Dorna 142 del Castillo, Francisco J. 88, 238 Chu, Samuel S. 65 delos Reyes, Emily 326 Chu, Tzu-Hung 386 Delpino, Victoria 76 Chuang, Wei-Lien 66 del Toro, Mireia 89, 207, 340, 364 Chung, Sooyong 13 de Medeiros, Paula F. 200 Chung, Wonhui 13 Demir, Hulya 138, 139 Ciana, Giovanni 364 Demura, Sofya 365 Cimms, Tricia 229 Deniz Batu, Ezgi 336 Ciubotariu, Crina 88 Dennis, Mark S. 158 Clark, Alex M. 197 Dermuth, Laura 297 Clarke, Lorne A. 34 Derrien, Valérie 262 Clatterbuck, Cinde L. 125 Desai, Ankit K. 90 Clausen, Dana 213 De Silva, De Silva 169 Cleary, Maureen A. 6, 20, 24, 67, 85, 86, 151, 335, 388 Desnick, Robert J. 171, 263 Clemens, Paula R. 68, 196, 324 Diaz, Dolo 158 Cloyd, James C. 185 Diaz, George A. 351 Colella, Pasqualina 21, 53, 69, 70 Díaz-Manera, Jordi 91 Collaud, Fanny 21, 69 Díaz-Riascos, Zamira V. 1 Colvin, Robert B. 32 Di Carlo, Alessia 348 Condon, Carolyn E. 270 Dickson, Patricia I. 64, 224, 371 Conner, Therese 71, 72, 73 Diehl, Thilo 380 Consortium, Gaucherite 94 Dierks, Thomas 322, 333 Cook, Francesca 71, 72, 73 Diethelm-Okita, Brenda 323, 337 Cook, Jennifer 225 Dimachkie, Mazen M. 278 Cooke, Michael P. 129, 270 Dimmock, David 299 Cooney, Jeffrey W. 108 Dinur, Tama 37, 92, 289, 297 Cooper, Jonathan D. 182 Di Rocco, Maja 336 Cope, Heidi 28 Dixon, Phillip 59 Coquery, Christine M. 96, 313 Do, Jenny 93 Corbau, Romuald 180, 235 Doherty, Caitlin 355 Corchero, José L. 1 Dokmeci(Emre), Serap 138 Correia Guedes, Leonor 282 Dollive, Serena 113 Cossio, Gladys 200 Dominguez, Fernando 128 Costa, Maria Julia G. 74 Donald, Aimee 94 Costa, Marina C. 11 Donohue, Jeffrey 154 Costa Verdera, Helena 21, 69 Donohue, Yanique 154 Côté, Anne-Marie 212 Donovan, Frank 99, 115 Couce, Maria L. 67, 106, 335 Dorneles, Alicia D. 381 Coutinho, Virginie 380 Dorot, Orly 52 Cox, Gerald 281 Drack, Arlene V. 182 Cox, Timothy M. 75, 94 Drake, Elise 269 Cozma, Claudia 92, 297 Drevot, Philippe 380 Crawford, Nigel 320 Drivas, Theodore G. 95 Cristobal, Rafael 128 D'Souza, Precilla 126 Cristóbal-Lecina, Edgar 1 Du, Ming 38 Crivaro, Andrea N. 243, 76 Duarte, Ana Joana 239 Crosariol, Marco 21 Ducla Soares, José Luís 10, 11 Crosby, Kathleen 309 Duke, Katie S. 96, 313 Crozier, Michael 348 Dundar, Halil 38 Cruz-Villalba, Jordi 228 Dupuis-Siméon, Frédérique 262 Crystal, Ronald 267 Duque, Joseph 158

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Durand, Consuelo 26, 97, 112 Frabasil, Joaquin 26, 97, 112 Dusek, Petr 251 Fraile, Jesus J. 16 Dussol, Bertrand 262 Francone, Omar L. 113 Fraser, Graeme 232 Eastwood, Deborah M. 85, 203 Frauenheim, Jill 179 Ebrahim, Hatim Y. 98 Fried, Leah 141 Echols, Josh 38 Friede, Tim 321 Eghbali, Areian 99 Friedman, Michael L. 311 Eikani, Carlo 254 Frydman, Dafna 289 Eisengart, Julie B. 140, 194, 224, 337 Fujitsuka, Honoka 356 Ekins, Sean 64, 197, 290, 291 Fukao, Toshiyuki 114 Ekstrand, Agneta 184 Fukuda, Keiichi 385 Elkahloun, Abdel 99, 115 Fuller, M. 106 Ellinwood, N.Matthew 371 Fung, Ellen B. 286 Ellis, Hannah 134 Furtak, Vyalcheslav 217 Ellsworth, Jeﬀ L. 113 Furujo, Mahoko 114 Elomaa, Kaisa 316 Elqadiry, Rabiy 312 Gadiot, Rosalie 311 Elstein, Deborah 101 Gaeckle, Nathaniel 47 El Turk, Farah 100 Gaemers, Sebastiaan J. 31, 75, 320 Emrick, Lisa 267 Gaggioli, Daniela 112 Endo, Masahiro 173 Gallardo, Eduard 91 Enguita, Francisco 11 Gamalel Din, Iman 336 Eric, Wallace 373 Gamba, John 361 Eriksson, Maria 177 Gameleira, Flávia 200 Escolar, Maria L. 3, 4, 41, 183, 389 Gandía, Marta 88 Esmer-Sanchez, Carmen 116 Garber, Kathryn 221 Estegan, Jaime 315 Garbuio, Gabriela C. 130 Esteves, Ananda 78 Garcia, Eric Joshua 99, 115 Eto, Kaoru 102, 166 Garcia, Jose E. 116 Eto, Yoshikatsu 102, 166, 173, 237 Garcia, Pablo 128 Eyskens, Francois 103, 160 García-Aranda, Natalia 1 Ezgu, Fatih 50, 67, 160, 335 Garcia Fernandez, Jose M. 117, 118 Garcia-Frade, Javier 17 Fadeeva, Mariia 365 Garcia-Jimenez, I. 57 Falese, Lillian 171 García-Moreno, M.Isabel 117 Fallet, R. 186 García-Silva, María Teresa 88 Fang, Shona 30 Garin, Christophe 203 Fantin, Bruno 255 Gärtner, Jutta 321 Farmer, Cristan 126 Gary, Sam 93 Faulkner, Deiby 113 Gasser, Thomas 282 Fayyaz, Zafar 61 Gaukel, Eric J. 96, 313 Fdil, Naima 88, 206, 312 Gavrilov, Dimitar K. 136, 205, 257 Fehnel, Sheri 199 Geberhiwot, Tarekegn 6, 367, 368 Feldman, Ricardo A. 76 Gelb, Michael H. 119 Feldt-Rasmussen, Ulla 104, 161, 162, 346, 347, 372 Gelmetti, Elisabetta 137 Felipe, Ana 89 Genevaz, Delphine 262 Felizardo, Tania C. 248 George, Kelly 120 Feltrin, Letícia C. 130 Germain, Dominique P. 161, 162, 262, 346, 364, 372 Feng, I-Jung 377 Getz, Jennifer A. 158 Feriozzi, Sandro 168 Gevorkyan, Anahit 207, 340, 365, 366 Fernandez, Vivian 71, 72, 73 Ghosh, Arunabha 121, 122 Fernández-Lorenzo, Jose R. 272 Gibson, Alana 84 Fernandez-Martin, Julian J. 272 Giese, Tina 158 Fernández-Simon, Esther 91 Gil-Campos, Mercedes 156 Ferreira, Carlos 336 Gil Ortega, D. 57 Ficicioglu, Can 247 Gimeno-Blanes, Juan 228 Fidelia-Lambert, Marie N. 217 Giraldo, Pilar 16, 17, 57, 123, 124, 160, 209, 218, 220, 373 Figueroa Sauceda, Sergio R. 105 Giraldo-Castellano, Pilar 228 Finglas, Alan 14 Girdauskas, Evaldas 345 Fischer, Tanya 250, 282, 320 Gissen, Paul 24, 326 Fiúza, Manuela 11 Giugliani, Luciana 202 Flanigan, K.M. 106 Giugliani, Roberto M. 74, 125, 155, 200, 201, 202, 240, 276, 285, Fleischer, Nicole 268 325, 333, 346, 347, 380 Fleming, Sheila 279 Giuliano, Joe 346 Flores, Romina 97 Gjata, Bernard 53, 69, 70 Flores-Lagunes, Luis L. 116 Glase, Brianna 126 Flueckinger, Lauren B. 28, 107, 108 Gleitz, Helene 43 Fofana, Fofana 276 Goba, Heather 261 Foley, Allison 219 Godinho, Stella C. 127 Foley, Jonathan 235 Goel, Ajay 329 Foley, Ronan 232 Gokcay, Gulden 336 Folgering, Joost H.A. 177 Goker-Alpan, Ozlem 68, 81, 160, 175, 196, 217, 231, 324, 373 Fontain, Freja 159 Goldgrub, Rachel 83 Forshaw-Hulme, Stuart 109, 110, 111, 132 Goldstein, Jennifer 296 Forte, Gabriella 266 Gomez, Alejandra 141 Fortini, Sebastian 97 Gomez, Manuel J. 53 Foster, Meredith 31, 75, 281, 363 Gomez Cerezo, Jorge Francisco 128 Fouilhoux, Alain 262 Goncalves, Kevin A. 129 Fowler, Daniel H. 232, 248 Goncalves, Soﬁa A. 130

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Gondré-Lewis, Marjorie C. 217 Hicks, Rebecca 309 Gonzaga, Mariana Q. 163 Hidalgo, Oscar A. 289 Gonzalez, Esteban A. 285, 325 Hietaharju, Aki J. 159, 184 Gonzalez, Irene 128 Higaki, Katsumi 117, 118 Gonzalez-Jaimes, Ronald 105 Higashigawa, Satomi 385 González Meneses, Antonio 88, 131 Hitchins, Lauren 51 González-Mira, Elisabeth 1 Hlavata, Anna 251 Goodspeed, Kimberly 14 Ho, Hui-Chen 386 Gorton, Janet 111, 132 Hocquemiller, Michael 211 Grange, Dorothy K. 51 Hoenig, Megan 153 Grant, Christina 309 Hoffman, James E. 314 Grau, Josep M. 88 Hoganson, George E. 51 Gray, Jill 378 Holida, Myrl D. 160, 227, 373 Gray, John 152 Holley, Rebecca 43 Gray, Steven J. 29, 62, 133, 225 Hopkin, Robert J. 161, 162, 204, 346, 373 Green, Diane 109, 134 Horiuchi, Yasue 385 Green, Jim 172 Horovitz, Dafne D. 163, 164, 165 Grifoni, Daniela 25 Horowitz, Mia 52 Grillo, Giuseppina 135, 293, 294 Horvitz, Noa 297 Grossman, Tamar 193 Hossain, Mohammad A. 166, 237 Grover, Anita 67, 335 Hossein, Arif 102 Gu, Jinghua 329 Hosseini, Paniz 180, 235 Gu, Tansy 84 Hostutler, Robert 229 Gudivada, Badari 295 Hovakimyan, Marina 92, 297 Guelbert, Norberto 336 Howe, Steven 150 Guenzel, Adam 136 Hsu, Ting-Rong 386 Guerchet, Nicolas 53, 69, 70 Huang, Jeffrey Y. 167 Guffon, Nathalie 34, 125, 137, 156, 161, 162, 203 Huang, Ju 232, 248 Guilder, Laura 85, 151 Huang, Ling-Yi 386 Guillén-Navarro, Encarna 228 Huang, Yunyu 311 Gumus, Ersin 138, 139 Hug, Christopher 278, 363 Gunasekaran, Kannan 158 Hughes, Derralynn 6, 10, 11, 31, 35, 80, 98, 104, 160, 168, 169, Gupta, Ashish 49, 140, 270 209, 218, 293, 294, 320, 367, 368, 372 Gupta, Punita 141 Hughes, Michael 294 Gurnon, Stephanie 142 Hughes, Samuel 14 Gustavsson, Susanne 260 Huhtala, Tuulia 54 Gutiérrez-Solana, Luis González 88, 244 Hui, Eric K. 44, 170, 222, 275 Gwaltney, Chad 143 Huidekoper, Hidde 364 Huledal, Gunilla 50 Hagège, Albert 262 Humphrey, Paul 60 Haldal, Shilpa 41 Hung, Clara 310 Hamazaki, Takashi 104 Huston, Marshall W. 171 Hamed, Alaa 82, 83, 143, 250, 273 Hutter-Paier, Birgit 258, 259, 260 Hamroun, Dalil 255 Huynh-Do, Uyen 264 Han, Sang-Oh 144 Huysse, Steven 209 Han, Tae-Dong 13 Hyzy, Sharon L. 129 Hanby, Haley 21 Hanna, Rosenbaum 145 Iaconangelo, Charlie 143 Harmatz, Paul 50, 67, 146, 147, 155, 210, 244, 245, 247, Ibarretxe, Daiana 17, 57 335, 336 Igarashi, Junko 173 Harrington, Magdalena 387 Igdoura, Suleiman A. 274 Hartog, Tessa E. 311 Iida, Kei 385 Harvey, Katie 148, 151 Ikpatt, Offiong F. 314 Hassib, Nehal 2 Illa, Isabel 91 Hauck, Peer 345 Imrie, Jackie 172 Haurigot, Virginia 21 Inskeep, Venette 279 Hawkins, Kimberly E. 149 Irun, Pilar 16, 17, 57, 220 He, Xingxuan 96, 313 Isaacson, Stuart 282 Heales, Simon J. 148, 150, 151 Istaiti, Majdolen 289 Heffner, Garrett 152 Itagaki, Rina 166, 173 Heldermon, Coy D. 147, 149 Itoh, Kohji 174 Heliö, Tiina 184 Ivanova, Margarita 175, 217 Hemsley, Kim 211 Ivgy, Heftziba 289 Henckaerts, Els 266 Iwamoto, Takeo 166, 173, 237 Henderson, Nadene D. 153 Hendriksz, Christian J. 6, 154, 155, 207, 265, 340, 341 Jabbarzadeh-Tabrizi, Siamak 176, 329 Henne, Kirk R. 158 Jacoby, David 326 Hennermann, Julia B. 137, 156, 157 Jaensch, L. 106 Henry, Anastasia G. 158 Jain, Vipul 196, 256, 347 Hensman, Pauline 203 Jain-Ghai, Shailly 310 Hepburn, Michelle 122 James, Emma 299 Herbert, Mrudu 28 James, Lucy 152 Hermida, Alvaro 128 Jansen, Jeroen 83 Hermosin, N. 17 Janson, Juliette 177 Hernandez, Betina 352 Jaquez, Francisca 200 Hernandez, Dena 115 Jarnes-Utz, Jeanine R. 47, 178, 185, 379, 391 Hernandez Rivas, Jose Maria 17 Jayaraman, Susheela 179 Heron, Benedicte 267, 276, 380 Jefferies, John L. 161, 162, 372 Hewitt, Richard 388 Jennette, J.C. 32 Hickey, Rachel 51, 179 Jens, Jackie K. 371

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Jeyakumar, Jey 180, 235 Komarova, Elena 366 Ji, Allena J. 281, 282, 351 Kong, Younggyu 13 Jimenez Olivares, Marco 116 Konstantopoulou, Vassiliki 137 Jireckova, Jitka 251 Korenev, Sergei 388 Joers, James 185 Korlimarla, Aditi 198 Johnson, Chloe 101 Kosaki, Kenjiro 385 Johnson, Franklin K. 181 Koshimura, Yuri 189 Johnson, Jack 168 Kozor, Rebecca 367 Johnson, Judith 278 Krägeloh-Mann, Ingeborg 378 Johnson, Julie 146 Kramer-Golinkoﬀ, Julia 8 Johnson, Tyler B. 54, 182, 374 Krayenbuehl, Pierre-Alexandre 264 Johnston, Jean 126 Kreher, Nerissa 199 Jones, Mairead 110, 111, 132 Krijnse-Locker, Jacomine 53 Jones, Simon A. 6, 30, 94, 121, 122, 203, 244, 246, 341 Kruyt, Moyo C. 203 Jordan, Lauren 227 Kubaski, Francyne 200, 201, 202, 240 Jovanovic, Ana 6, 110, 132, 134, 161, 162, 218, 271, 320, Kuiper, Gé-Ann 203 346, 347, 349, 368, 372 Kulisevsky, Jaime 282 Julia, Koﬂer 183 Kunkler, K.L. 106 Julien, Daniel 382 Kusuhara, Masatoshi 385 Jurickova, Katarina 251 Kuter, David 233 Kuusisto, Johanna 184 Kahn, Shaukat 357 Kuzenkova, Ludmila M. 317, 365, 366, 390 Kaizer, Alexander 224 Kwon, Seulki 13 Kak, Manisha 195 Kyosen, Sandra O. 164 Kalcheva, Petya 180 Kallish, Staci 95 Labilloy, Anatalia 204, 253 Kamaly Asl, Ian 266 Labounek, Rene 252 Kampmann, Christoph 245, 246 Lacey, Jean M. 205, 257 Kan, Shih-Hsin 64, 371 Lachmann, Robin H. 104, 195, 351 Kananen, Kristiina 184 Lacombe, Didier 262 Kanters, Steve 83 Ladha, Shafeeq 278 Kantola, Ilkka 161, 162, 184, 284, 369 Lafhal, Karima 206, 312 Kantola, Taru 184 Lagast, Hjalmar 68, 196, 324 Kaplan, Paige 352 Lahoz-Gil, Carlos 57, 220 Kapoor, Seema 336 Lai, Chih-Jou 386 Karaa, Amel 161, 162, 373 Lambert, Jonathan 150 Karhan, Asuman N. 138, 139 Lampe, Christina 146, 207, 336, 340, 364 Karhu, Anne 184 Lamppu, Diana 113 Kariolis, Mihalis S. 158 Landy, Hal 195 Karlsson, Antti 316 Lane, Monica 120 Kärppä, Mikko 159 Laney, Dawn A. 208, 221 Kartha, Reena V. 185 Langereis, Eveline J. 203, 377 Kaur, Amrit 121 Laquerriere, Annie 40 Kazi, Zoheb B. 90, 382 Latre-Martínez, Paz 228 Keating, Armand 232 Lau, Heather A. 18, 19, 147, 209, 210, 247 Keeling, Kim 38 Laufer, Ralph 211 Keenan, Hillary 34 Lautredoux, Florent 255 Kehr, Jan 259 Lavalle, Lucia 98, 169 Kelly, Masami 273 Lavoie, Pamela 27, 212 Kenis, Vladimir 203 Lawlor, Michael W. 152, 299 Kerns, Scott 186, 187, 330 Le, Steven 64 Kevany, Brian M. 187, 188 Leão-Teles, Elisa 146 Khan, Aneal 87, 232 Leardini, Davide 25 Khan, Nedd 160, 373 Lebel, Sylvie 384 Khan, Shaukat 359 Leborgne, Christian 69 Kia, Azadeh 180, 235 Lechado, Anna 1 Kida, Sachiho 189 Lee, Chris W. 213 Kielian, Tammy 186 Lee, Doyoung 235 Kim, Do Jin 158 Lee, Joy 67, 335 Kim, Donghoon 13 Lehtimaki, Kimmo 54 Kim, Gee-Hee 190 Lei, Quanhong 363 Kim, Jae-Min 283 Leistner-Segal, Sandra 200, 202 Kim, Jung-Hwan 13 Le Meur, Alice 380 Kim, Yestle 31 Le Moine, Kaye 376 Kimonis, Virginia 191, 192, 193, 373 Lenders, Malte 214, 215, 216 Kimura, Alan 336 Leslie, Nancy 147 King, Kelly E. 194, 323, 337 Levin, Adeera 212 Kinoshita, Masafumi 189 Lewis, Grace 295 Kirsztajn, Gianna M. 253 Lewis, Sarah 323, 337 Kishnani, Priya S. 28, 68, 90, 107, 108, 195, 198, 236, 278, 324, Leyva, Francisco 368 339, 344, 382 Li, Jian 282 Kiyozumi, Yoshimi 385 Li, Rong 15 Klein, Jennifer J. 197 Li, Shuping 129 Klemen, Martin 101 Li, Songtao 144 Knight, Margaret 191 Li, Yu-Ji 377 Koeberl, Dwight D. 144 Liang, Nicholas 158 Kohlschütter, Alfried 223 Liao, Ai-Yin 43 Köhn, Anja F. 345 Liao, Hsuan-Chieh 386 Kokkonen, Jorma 184 Libri, Vincenzo 195 Kolben, Yotam 289 Lidove, Olivier 262

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Lieberman-Hernandez, Esther 116 Mathieson, Toni 172 Likhite, Shibi 374 Matlach, Juliane 157 Limgala, Renuka P. 81, 217, 231 Matsubayash, Hiroyuki 385 Lin, Huilan 44, 222, 275 Matte, Ursula 325 Lin, Shuan-Pei 67, 335 Mauer, Michael 161, 162, 249 Lindqvist, Daniel 50 Mayer, Franz J. 322 Lindstrom, Kristin 382 Mayhew, Christopher N. 279 Lindstrom, Taylor 342 Mazanova, Natalya 317 Linhart, Aleš 218, 346, 372, 373 Mc Bride, K.L. 106 Liou, Benjamin 279 Mc Caw, Patricia 343 Litcher-Kelly, Leighann 273 Mc Clain, Monica R. 236 Llauger, Jaume 91 Mc Intosh, Jenny 180, 235 Llerena, Juan Jr. 344 Mckie, Mark A. 80, 293, 294 Lobhanidze, Tinatin 207, 340 Mc Knight, Tracy 22 Loeffler, Tina 259 Mc Millan, Annabel 233 Lohmöller, Benjamin 345 Mc Nally, Alexander P. 149 Long, Valynne 219 Mc Nelly, Briony 207 Longo, Nicola 160, 373 Mc Sweeney, Mel 20 Loos, Mariana 26 Mc Vie-Wylie, Alison 120 Lopes, Felipe 66 Medeiros, Rommel B. 163 Lopes, Simone S. 200 Medin, Jeffrey A. 232, 248 Lopez, Grisel 93, 99, 115, 328, 342 Medrano-Engay, Blanca 123 Lópezde Frutos, Laura 16, 17, 220 Mehta, Atul 35, 98, 169, 233, 293, 294 Lopez-Royo, M.Pilar 123, 124 Mei, Miranda 152 Lopez Wolf, Daniel 128 Melethil, Subha 357 Loucopoulos, Georgia 221 Mellgren, Svein I. 159 Lourenco, Charles M. 45, 74, 127, 130 Melton, Andrew 67, 335 Löyttyniemi, Eliisa 284 Mena-Barragán, Teresa 117 Lozano Almela, ML 17 Menao Guillén, Sebastián 360 Lu, Jeff Z. 44, 170, 222, 275 Mendes, Carmen Silvia C. 334 Ludwig, Nataniel F. 327 Mendoza, Rosa 1 Lukacs, Zoltan 223 Mendoza-Ruvalcaba, Sandra C. 116 Lukas, Jan 214 Meng, Hui 152 Lukina, Elena 31, 75, 318 Mengel, Karl-Eugen 278, 320 Lukina, Kira 318 Menk, Jeremiah 286 Lund, Allan M. 137, 156 Messelodi, Daria 25 Lund, Troy C. 48, 49, 140, 224, 251, 252, 270, 286 Meyer, Kathleen 171 Luquette, Mark 48 Meyer, Kathrin 374 Lutz, Cathleen 29, 225 Meyerink, Brandon 54 Ly, Christine 142 Michaud, Langis 234 Lykken, Erik 133 Michelin-Tirelli, Kristiane 55, 200, 202, 333 Miettunen, Kelly 48, 49, 140 Mac Kenzie, Tippi 254 Miller, Bradley S. 286 Mackenzie, William G. 203 Miller, Holly 382 Mack-Thorley, Eileen 82 Miller, Timothy J. 133, 186, 187, 188, 330 Mac-Way, Fabrice 212 Miller, Weston 224 Madrazo, Patricia 315 Ming, Xue 68, 196, 324 Maenchen, Molly 383 Mingozzi, Federico 21, 53, 69, 70 Magner, Martin 251 Minini, Pascal 282, 320 Magnusson, Bo 336 Mir, Pablo 282 Mahan, Farrah R. 226 Miranda, Carlos J. 235 Mahon, Cathal 158 Mirzaian, Mina 52 Makay, Balahan 336 Mishra, Usha 185 Manavalan, Arulmani 213 Mistry, Pramod K. 75, 236 Mansbach, Hank 229 Mitchell, John J. 100, 336 Maor, Gali 52 Mitchell, Nadia 266 Maranda, Bruno 27 Miyajima, Takashi 102, 166, 173, 237 Marcellus, Samantha 227 Moen, Derek 64 Marchal, Jan Pieter 380 Moiseev, Sergey 317 Marguet, Florent 40 Mole, Sara E. 154 Maricich, Stephen M. 60, 67, 335 Molenkamp, Liz 250 Marinakis, Theodore 75 Molero, Maria T. 16, 17 Marin-Leon, Ignacio 228 Molina-Garay, Carolina 116 Marino, Jacira 10, 11 Molnar, Maria Judit 373 Marmier, Solenne 69 Moltó-Abad, Marc 1, 89, 128 Márquez Infante, Celedonio 228 Monaghan, Catherine A. 270 Márquez Rivas, Javier 131 Monia, Bengherbia 255 Marret, Stéphane 39, 40 Montano, Hector P. 200, 201, 202 Marsden, Deborah 229 Monteiro, Amanda C. 45 Martin, Nathaniel I. 117 Montiel, Elena 91 Martins, Ana M. 77, 78, 161, 162, 164, 334, 372 Moochhala, Shabbir 98 Maruti, Sonia S. 344 Moon, James C. 367, 368 Marzouki, Naima 312 Morado, Marta 17, 238 Mashima, Ryuichi 230 Morales, Monserrat 128 Maski, Manish 83 Moreira, Luciana 239 Masllorens, Francisca 26 Moreno, Patricia 241, 361, 362 Mason, Lauren M. 231 Moriarity, Branden 242 Mason, Robert 358 Morimoto, Hideto 189 Mateos, Georgina 97 Morrison, Alexandra 6, 7 Matern, Dietrich 136, 205, 257 Morton, Magda 299

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Mouad, Fatima Zahra 206 Olivier, Sophie 267, 276, 380 Movsisyan, Goar 318 Olivotto, Iacopo 104, 256 Moya-Nilges, Maryse 53 Ollis, Sarah 273 Mozaffar, Tahseen 68, 191, 195, 196, 324 Olsen, Amber 14 Mucci, Juan M. 76, 243 O'Neill, Cara 268, 269 Mueller, Bryon 252 Oppermann, Sebastian 92, 297 Muenzer, Joseph 34, 125, 194, 244, 245, 246, 247 Orchard, Paul J. 47, 48, 49, 140, 178, 203, 224, 251, 252, 270, Mungan, Neslihan 336 286 Muñoz, Gloria 88, 238 Ordoñez, Beatriz 124 Munoz-Rojas, Maria Veronica 125 Oreiro, M.T. 106 Muramatsu, Shin-Ichi 174 Orii, Kenji 114 Murav’ova, Lubov 318 Ormazabal, Maximiliano 76, 243 Murko, Simona 223 Orsborne, Chris 271 Murphy, Elaine 6 Ortega, David 1 Murphy, Tara 225 Ortega-Caballero, Fernando 118 Muschol, Nicole M. 67, 137, 156, 267, 276, 335, 345, 380 Ortiz, Alberto 228, 315, 372 Mussi, Gabriela D. 127 Ortiz, Damara 104, 153 Mygland, Åse 159 Ortiz Mellet, Carmen 117, 118 Ortolano, Saida 272 Nagata, Satoru 102 Ory, Daniel 384 Nagree, Murtaza S. 248 Osorio, Nicole K. 314 Najafian, Behzad 249 Otero, Maria G. 283 Nalin, Tatiele 55, 381 Oxborrow, Neil 203 Nalysnyk, Luba 250 Oz, Gulin 185 Namazova-Baranova, Leyla 292, 318, 365, 366 Ozen, Hasan 138, 139 Nanba, Eiji 117, 118 Ozen, Seza 336 Nascene, David 49, 251, 252 Nathwani, Amit C. 180, 235 Pabla, Anureet K. 82, 273 Neddens, Joerg 258, 259, 260 Pacheco, Joshua 66 Neilan, Edward 363 Padegimas, Linas 133, 186, 187, 188, 330 Nembo, Blandine 282 Padilla, Brad 273 Nestrasil, Igor 251, 252 Padilla, Ron 274 Neto, Jose S. 17, 332 Pagant, Silvere 171 Neto, José Tibúrcio M. 253, 332 Pak, Lolita 317 Neumann, Jon 167 Pakhomov, Alexander 317 Neußer, Leon P. 214 Palk, Katrin 88 Nevalaita, Liina 311 Palmer, David 266 Newsom-Davies, Imogen 380 Pan, Wayne 142 Nguyen, Quoc-Hung 254 Panahloo, Zoya 209, 316 Nguyen, Yann 255 Panter, Charlotte 101 Nicholls, Kathleen 104, 218, 256, 346 Pardridge, William M. 44, 170, 222, 275 Nickander, Kim K. 257 Parenti, Giancarlo 364 Nickel, Miriam 223 Parini, Rossella 146, 203 Niederau, Claus 236 Park, Jong-Suk 13 Niederkofler, Vera 258, 259, 260 Parker, Samantha 267, 276, 380 Niemietz, Christoph 215, 216 Paskulin, Livia D. 277 Nieves Cobos, Paulina 223 Pasqualli, Marzia 224 Nijmeijer, Stephanie C. 203 Passemard, Solène 1 Nilsson, Mats 348 Patel, Akshilkumar 8 Nimmo, Graeme A. 261 Patterson, Marc 172 Nishimura, Seiichiro 385 Patterson, Mary Anne 261 Niu, Dau-Ming 386 Paul, Sumit 52 Noël, Esther 262 Paulson, Amy 251 Nonis, David F. 283 Pavan, William 84 Nordbeck, Peter 214, 346 Pearce, David A. 182 Nordin, Jayme 21 Peay, Holly L. 269 Nordin, Sabrina 367 Peceny, Markus 347 Nowak, Albina 214, 263, 264, 372 Peck, Dawn 136 Noya, Maria Soledad 17 Pedrosa, Irene 91 Nuñez-Peralta, Claudia 91 Peltola, Niina 159 Nurmi, Antti 54 Pena, Loren 278 Peng, Yanyan 279 Oglesbee, Devin 136, 205, 257 Penkov, Evgeny L. 390 Oh, Se-Woong 13 Pereira, Catarina 240 Ohashi, Toya 104, 256 Pereira, Ester M. 253 Ohira, Mari 230 Pereira, Rosa Maria R. 300, 301, 302, 303, 304, 305, 306, 307, 308 Ohlin Sjöström, Elisabet 260 Perevalova, Yelena 55 Ohnishi, Yukiya 174 Pereze, Anna 380 Okazaki, Tetsuya 117 Pérez-López, Jordi 207, 340 Okugawa, Yoshinaga 329 Perez Saenz, M.A. 17 Okumura, Miho 189 Perrone, Solanger 380 Okur, Ilyas 67, 335 Pesquero, Joao B. 332 Okuyama, Torayuki 230 Pession, Andrea 25 Olcay, Lale 139 Pession, Annalisa 25 Oldham, Andrew 109, 265 Peter, Merlene 280 O'Leary, Claire 266 Peters, Heidi 67, 335 Oliva, Petra 120 Peterschmitt, M.Judith 31, 75, 281, 282 Oliveira, Ivana Helena R. 253 Phillips, Dawn 199 Oliveira, Maria L. 163, 164 Pierpont, Rene 224 Olivera González, Susana 360 Pierson, Tyler M. 283

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Pieters, Roland J. 117 Robinson, Amy 203 Pietila-Eﬀati, Päivi 184, 284 Roca, Mercedes 16 Pilipenko, Valentina 226 Roca-Espiau, Mercedes 123, 124 Pimentel, Luisa 285 Rocha, Joshua W. 333 Pintos-Morell, Guillem 1, 89 Rochmann, Camille 250 Piresde Mello, Paulo 165 Rodrigues, Graziella 285 Piresde Mello Valente, Mariana 165 Rodriguez Ciancio, Jose 24 Piris, Miguel 88, 238 Rodriguez-Lopez, Alexander 15 Pirondi, Stefania 156 Roebuck, Derek 388 Pitt, Matthew 85 Rogers, Chris 182 Pitz, Susanne 157 Rojas-Rodríguez, Felipe 289 Plana, N. 57 Roland, Dominique 364 Podkletnova, Tatyana V. 317, 365, 366, 390 Rolfs, Arndt 92, 297 Poe, Michele 3, 4, 41, 183, 389 Romero-Trejo, Juan M. 105 Poggio, Eugene 83 Ronco, Pierre 262 Poletto, Edina 285 Ronzitti, Giuseppe 21 Polgreen, Lynda E. 224, 286 Ros, Emilio 30 Politei, Juan 97, 112, 346 Rosa, Juan M. 88 Pollard, Laura 62, 142, 287, 382 Rosa Neto, Nilton S. 300, 301, 302, 303, 304, 305, 306, 307, 308 Pollissard, Laurence 273 Rosenberg, Amy S. 90 Ponce, Elvira 83 Roshan Lal, Tamanna R. 309, 328, 342 Ponomarev, Rodion 318 Roston, Alexandra 310 Porter, Katherine 269 Rouch, Lucie 267 Posadadela Paz, Manuel 228 Rouillon, Jeremy 53 Potnis, Kunal C. 108 Rouwette, Tom 311 Prada, Carlos E. 204, 226, 247, 288 Royo, Miriam 1 Prasad, Suyash 152, 299 Rozenberg, Alejandra 133 Preiskorn, Joana 321 Rozenfeld, Paula 76, 243 Presa, Maxmiliano 29, 225 Rubin, Hillard 113 Press, Rayomand 159 Rubio, Ana P. 197 Privalova, Tatiana 366 Ruchlemer, Rosa 289 Provenzale, James M. 198 Rudnicki, Michael 214 Prunet, Caroline 34 Rudser, Kyle 185, 224, 286 Prunty, Helen 151 Ruiz, Adriana 116 Puentes-Tellez, Alejandra 289 Ruiz, Juan 106, 330 Puga, Ana Cristina 351 Ruiz-Garcia, Matilde 244 Puhl, Ana C. 290, 291 Rupar, C.A. 232 Pulido, Daniel 1 Rusk, Rosemary 368 Puri, Ratna 336 Russell, Katharina 266 Pushkov, Alexander A. 292, 317, 318, 390 Russo, Daniel P. 197 Puzzo, Francesco 21, 53, 69 Rust, Stewart 122 Rutecki, Jasmine 346 Qin, Cathy 169 Ryan, Emory 93, 328, 342 Quazi, Kabir 352 Rykunova, Anastasia 318, 366 Quintero Bernabeu, J. 57 Rytkonen, Jussi 54

Raben, Nina 193 Saarenhovi, Maria 284 Rada, Noureddine 88, 312 Saarinen, Jukka T. 184, 284 Radhakrishnan, Karthikeyan 322, 333 Sabir, Es-Said 88, 206, 312 Radovic, Ana 159 Sacconi, Sabrina 278 Raﬀel, Glen D. 270 Sadhu, Chanchal 152 Rahim, Ahad 150 Saeed, Anjum 61 Rainto, Miia 184, 369 Saettini, Francesco 86 Rairikar, Mugdha 28 Sakuraba, Hitoshi 385 Ramani, Bhumi 192 Saleck, Sam 46 Ramaswami, Uma 35, 98, 169, 293, 294 Saltik Temizel, Inci N. 138, 139 Ramirez, Joany 200 Sampey, Brante P. 96, 313 Rand, Maret H. 334 Sampey, Gavin C. 313 Random, Devora 327 Samuelsson, Kristin 159 Rangel, Anthony D. 167 Sanchez, Sandra 314 Rangel-Miller, Vanessa 60, 71, 72 Sanchez-Niño, Maria Dolores 315 Rascati, Karen 71 Sandes, Kiyoko A. 200 Rasmussen, Lynn 38 Sandini, Lorenzo 184 Rawson, Kacey 250 Sandoval, Heidy M. 200, 201 Raymond, Kimiyo 205, 257 San Millán, Beatriz 272 Raynor, Lewis 295 Sansen, Stefaan 311 Reed, Matthew C. 98 Sansone, Ken 229 Rehder, Catherine W. 90, 296 Santamaría-Olmo, Rafael 228 Revel-Vilk, Shoshana 37, 92, 289, 297 Santer, René 223 Rhead, William 373 Sanz, Alejandro 238 Rhodes, Melissa 96 Saubadu, Stéphane 282 Ribeiro, Diogo 239 Sauni, Chelsee 64 Richards, Susan 90, 363 Savolainen, Markku 184, 316 Rico, Salvador 299 Savostyanov, Kirill V. 292, 317, 318, 390 Rinaldo, Piero 136, 205, 257 Sawamoto, Kazuki 354, 356, 357 Riso, Arlindo 345 Sbyea, Safya 206 Rísquez-Cuadro, Rocio 118 Scalco, Fernanda B. 163, 164 Rivero, Mónica 131 Scarpa, Maurizio 155, 319, 364 Rivero-Arias, Oliver 233 Schema, Lynn 391 Roberts, Mark 68, 196, 324, 344 Schenone, Andrea B. 26, 97, 112

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Schiano, Thomas 351 Sokolovskiy, Alexey 249 Schiffer, Victoria 259 Solanich, Xavier 128 Schiffmann, Raphael 2, 160, 176, 320, 329, 342, 373 Solano Villarreal, Martha 67, 335 Schlotawa, Lars 14, 321, 322, 333 Solomon, Beth 328 Schmidt, Bernhard 322 Solyom, Alexander 96, 313, 336 Schmidt, Hartmut 215, 216 Soong, Wen-Jue 386 Schmidt, Staffan 259 Souto, Olga 272 Schmitz, Boris 214, 215 Souza, Ariane 77, 78 Schneider, Ashley 323, 337 Souza, Carolina F. 164, 333 Schoser, Benedikt 68, 152, 196, 278, 299, 324 Sparks, Susan 363 Schubert Hjalmarsson, Elke 203 Specchio, Nicola 326 Schuchman, Edward H. 96, 243, 313 Sperb-Ludwig, Fernanda 55, 327 Schuh, Artur S. 381 Spiridigliozzi, Gail A. 339 Schuh, Roselena S. 285, 325 Spurlock, Katherine 337 Schulz, Angela 223, 326 Srivastava, Ankita 158 Schuster, Tilman 352 Stapleton, Molly 353, 355, 357, 358 Schwartz, Ida Vanessa Doederlein 33, 55, 277, 327, 333, 338, 381 Stappers, Franciska 215, 216 Schwartz, Simó 1 Starosta, Rodrigo T. 277, 338 Schwenkreis, Peter 68, 196, 324 Staton, Clarissa 54 Seehra, Gurpreet K. 328 Steeds, Richard P. 367, 368 Segady, Kayla 153 Stefanescu, Mihaela C. 198, 339 Segovia, Sonia 91 Stepien, Karolina M. 109, 110, 111, 132, 134, 265, 340, 341, 349, Segura, Antonio 89 350 Sekulic, Davorka 295 Steward, Alta M. 99, 115, 342 Selim, Laila 336 Stiles, Ashlee 343 Sellier, Pauline 53, 69, 70 Stinchcombe, Tim 152 Sengooba, Arnold 113 Stirnemann, Jérôme 255 Sensinger, Charlotte 278 St Martin, Susan 171 Serrano, Daniel 143 Stockton, David W. 344 Sevšek, Alev 117 Strocchi, Silvia 25 Seymour, Albert 113 Stubblefield, Barbara 93, 99, 115 Shah, Jeet 192 Studinski Jones, April 136 Shankar, Suma 104 Stumpfe, Katharina M. 345 Shanley, Ryan 270 Sturdevant, David A. 54, 182 Shao, Rong 51 Suarez, Diego 289 Shapiro, Elsa G. 194 Suarez-Calvet, Xavier 91 Sharief, Nida 175 Sudrié-Arnaud, Bénédicte 40 Sharma, Jyoti 282, 320 Sukhozhenko, Alexey 317 Sharma, Pankaj 99 Sun, Ying 279 Sharma, Reena 111, 134, 146, 350 Sundberg, Erik 336 Shaywitz, Adam J. 67, 335 Sunder-Plassmann, Gere 104, 346, 347 Shea, Jeremy 254 Sung, Crystal 66, 363 Shen, Jin-Song 176, 329 Suto, Mark 38 Sherbini, Omar 8, 14 Suzuki, Yasuyuki 114 Sheridan, Rose 180, 235 Suzuki, Yoshiyuki 117 Shigeyasu, Kunitoshi 329 Svatkova, Alena 252 Shil, Priya 149 Svenningsson, Per 282 Shintaku, Haruo 173 Sweeney, Zachary K. 158 Shiyanov, Pavel 330 Symeonidis, Argiris 46 Shults, Justine 8 Szlaifer, Mali 160, 218 Siddiqui, Amna 38 Sidhu, Alpa 227 Taddia, Alberto 25 Sidransky, Ellen 93, 99, 115, 328, 331, 342 Takayama, Liliam 300, 303, 306 Siebert, Marina 33, 55 Tallaksen, Chantal 159 Sierra Monzón, José Luis 360 Tambini-King, Lorien 141 Sileno, Anthony 181 Tanaka, Satowa 189 Silva, Marina S. 130 Tanima, Yuichiro 99, 331 Silva, Sandra M. 332 Tanniou, Guillaume 69, 70 Silva, Thiago O. 333 Tanpaiboon, Pranoot 309 Silverman, Adam P. 158 Tarnopolsky, Mark 348 Simmons, Morgan 221 Taroua, Mouna 176 Simmons, T.R. 106 Taurio, Jyrki 184 Simonaro, Calogera 243 Tavares, Angela M. 285, 325 Simon-Sola, Marcelo 69 Tayebi, Nahid 93, 99, 115, 331, 342 Simuni, Tanya 282 Taylor, Madeleine 359 Sisic, Zlatko 146 Tebani, Abdellah 39, 40 Sitaraman, Sheela 68, 196, 324 Teixeira, Helder F. 285, 325 Sivakumar, Kumaraswamy 68, 196, 324 Terkelsen, Astrid J. 159 Sivam, Debbie 146 Terluk, Marcia 185 Skuban, Nina 32, 42, 104, 181, 256, 346, 347 Thibault, Nathan 344, 363 Sladkov, Dmitry 292 Thompson, Lorraine A. 134, 271, 349, 350 Slasor, Peter 326 Thrasher, Adrian 86 Slitine, Nadia 88 Thurberg, Beth L. 351 Smith, Lucas 254 Thurm, Audrey 126 Smith, Lynn 67, 335 Tienari, Pentti 184 Smith, N.J.C. 106 Tifft, Cynthia 126 Snanoudj-Verber, Sarah 62 Titievsky, Lina 352 Snella, Elizabeth M. 371 Togawa, Tadayashu 385 Sobreira, Joselito 334 Tokel, Kursad 139 Sokn, Silvia 112 Tomatsu, Shunji 114, 353, 354, 355, 356, 357, 358, 359

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Ton, Jennifer 152 Wang, Rong 352 Tøndel, Camilla 160, 218, 373 Wanner, Christoph 214, 372 Tonon, Tassia 381 Warnock, David 373 Toppila, Iiro 316 Wasilewski, Margaret 378 Torcoletti, Marta 336 Wasserstein, Melissa P. 351 Torralba-Cabeza, Miguel-Angel 360 Watanabe, Ryo-suke 174 Torres, Danielle 165 Waters, Cheryl 282 Tortorelli, Silvia 136, 205, 257 Watman, Nora 31 Tran, Hai 158 Watts, Ryan J. 158 Trapp, Franciele B. 201, 202 Wechsler, Stephanie 219 Trufanov, Sergey 292 Wechsler, Thomas 171 Truxal, K.V. 106 Wehmeyer, Connie 204 Tsang, So-Fai 295 Weidemann, Frank 372 Tsuji, Daisuke 174 Weill, Yishay 37 Tsukimura, Takahiro 385 Weimer, Jill M. 54, 182, 374 Tuite, Paul 185 Weinreb, Neal J. 87, 185, 314, 352, 375 Tummolo, Albina 137 Weisberg, Daniel 122 Turgeon, Coleman 136 Weiss, Lan 193 Tylee, Karen 122 Wellby, Martin 266 Tylki-Szymanska, Anna 156 Wells, Robert C. 158 Tyynismaa, Henna 159 Wencel, Marie 195 West, Michael L. 160, 218, 232, 372, 376 Ukkola, Olavi 184 Wheeldon, Nigel 368 Ukwuani, Somto 342 White, Amy 136 Ullman, Julie C. 158 White, Katherine A. 54, 182, 374 Uribe, Alfredo 241, 361, 362 White, Klane K. 203, 286, 377 White, Kyle 314 Vaidya, Sagar A. 147, 247 Whiteman, David 378 Vairo, Filippo P. 33, 55, 338, 381 Whitley, Chester B. 47, 147, 178, 194, 247, 252, 267, 323, 337, Valadez-Juvera, Guillermo 105 379, 391 Valayannopoulos, Vassili 363 Widera, Shanna 179 Vallim, Julia R. 79 Wiggs, Edythe 342 Valtola, Kati 184 Wijburg, Frits A. 203, 267, 276, 380 Van Damme, Philip 278 Wikén, Margareta 50 Vander Lee, Johanna H. 203 Wilcox, Gisela 109, 110, 111, 134 vander Ploeg, Ans T. 68, 196, 324, 344, 364 Wilcox, William R. 160, 161, 162, 373 Vanderver, Adeline 8, 378 Wilke, Matheus V. 381 Vaneckova, Manuela 251 Wilks, Flyod 217 Van Hasselt, Peter M. 203 Williams, Hadis 42, 256 van Rijn, Rick R. 203 Wills, Anne-Marie 282 van Wittenberghe, Laetitia 69, 70 Wilson, Don P. 30 Vanyo, Todd 391 Wiseman, Samantha 7 Vashakmadze, Nato D. 207, 340, 365, 366, 390 Witt, Russell 254 Veciana, Jaume 1 Wittenberghe, Laetitia V. 53 Vega, Francisco 314 Wolf, Maija 316 Vellodi, Ashok 94 Wong Po Foo, Cheryl 147, 247 Venditti, Charles 84 Wood, Jill 60, 64, 268 Ventosa, Nora 1 Wood, Tim 142, 287 Verhoef, Daniel 180 Woolfson, Peter 368 Veronez, Camila 332 Woolgar, Kara 382, 383, 384 Veteläinen, Matilda 159 Wright, Jacquelyn 68, 324 Viall, Sarah 309 Wright, Teresa 113 Vidal, Jo Ann 60 Wring, Stephen A. 313 Vieitez, Irene 272 Wu, Chen 166, 173, 237 Vijapurapu, Ravi 367, 368 Wu, James 378 Vijay, Suresh 6 Wu, Yanyu 387 Villalon, Lucia 16, 17 Wu, Yuna 244 Villanueva, Mercedes 26 Wynn, Robert F. 121, 341 Villareal, Martha L. 200, 201 Wynne, David 12 Villarrubia, Jesús 16, 17, 88, 238 Villeda, Saul 254 Yamakawa, Hiroyuki 385 Vincendon, Pascale 311 Yamamoto, Joyce U. 77, 78 Viskochil, David H. 34, 125 Yanagisawa, Hiroko 102, 173, 237 Vissing, John 278 Yang, Chen Chang 386 Vockley, Jerry 373 Yang, Chia-Feng 386 Vujkovac, Bojan 218, 372, 373 Yang, Meng 161, 162, 372 Yang, Tsui-Feng 386 Wachs, Michaela 322 Yasuda, Makiko 171 Wagner, John E. 270 Yaworsky, Andrew 273 Waheed, Nadia 61 Yee, Karen 387 Waisbren, Susan E. 387 Yee, Richard 66 Wajnrajch, Michael P. 352 Yeo, Mildrid 388 Walkley, Steven U. 84 Yliaska, Iina 184 Wallace, Eric 160 Ylikallio, Emil 159 Walls, Susanne 184, 369 Yoo, Han-Wook 161, 162 Walters, Crista 195 Yoon, Isabel C. 389 Wang, Bowen 254 Young, Sarah 343, 371 Wang, Feng 64, 370 Yousﬁ, Karima 255 Wang, Junhua 158 Youshani, Amir Saam 266 Wang, Raymond Y. 167, 267, 371 Yu, Chen 60

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Yu, Howard 193 Zhang, Xiaoyi 64 Yu, Julie 104 Zheng, Wei 15 Yuce, Aysel 138, 139 Zhu, Changzhi 313 Zhurkova, Natalia V. 292, 390, 365, 366 Zaher, Atef 351 Zibert, Andree 215 Zambrano, Marina 202 Zimran, Ari 37, 92, 101, 209, 289, 297 Zegard, Abbasin 368 Zindel, Thea 157 Zhang, Haoyue 343, 371 Zizemer, Vitoria S. 277 Zhang, Kate 120, 281 Zorn, Kimberley M. 197

S168 Molecular Genetics and Metabolism 126 (2019) S169–S171

Contents lists available at ScienceDirect

Molecular Genetics and Metabolism

journal homepage: www.elsevier.com/locate/ymgme

Keyword Index

AAV 21, 29, 53, 62, 69, 84, 106, 113, chaperone 15, 32, 52, 68, 104, 110, 117, 133, 144, 147, 152, 171, 174, 118, 132, 166, 175, 181, 196, 180, 186, 187, 188, 211, 225, 197, 213, 215, 256, 271, 291, 235, 242, 247, 264, 266, 267, 293, 322, 324, 347 272, 299, 330, 357, 36, 371, cholesteryl ester storage disease 57 374 cognition 108, 210, 244, 380, 387 acid alpha-glucosidase 21, 22, 68, 90, 91, 144, 151, CRISPR 9, 12, 65, 167, 215, 239, 242, 152, 167, 191, 192, 193, 195, 253, 325 196, 278, 324, 348, 361, 386 cysteamine 59 acid beta-glucosidase 31, 235, 331 cystinosis 59 acid ceramidase deficiency 96, 100, 313, 336 cytokines 259, 301, 305 acid sphingomyelinase deficiency 222, 351 Danon disease 22, 303 activities of daily living (ADL) 45, 127, 199, 221 education 20, 111, 135, 141, 221, 241, agalsidase 1, 83, 143, 161, 162, 214, 234, 349 ffi 256, 347, 372 e cacy 1, 32, 62, 96, 104, 241, 313, alglucosidase 82, 344, 363 326, 363 alpha-galactosidase 79, 83, 105, 150, 159, 160, 166, eliglustat 17, 31, 75, 175, 236, 281, 314 169, 171, 180, 208, 218, 232, enzyme replacement therapy 7, 15, 18, 20, 21, 23, 27, 28, 44, 239, 272, 293, 317, 332, 373 (ERT) 47, 50, 57, 64, 66, 68, 72, 83, alpha-L-iduronidase deficiency 38, 127, 285, 325, 371 85, 87, 90, 92, 96, 104, 107, alpha-mannosidosis 118, 137, 156, 157, 251 110, 111, 114, 137, 138, 139, alpha-N-acetylglucosaminidase 23, 65, 116, 149 141, 146, 150, 153, 155, 156, antibodies 44, 121, 170, 214, 363 158, 161, 162, 164, 165, 166, aspartylglucosaminuria 62 170, 178, 189, 191, 195, 196, autophagy 53, 22, 175, 274, 289, 348 209, 214, 222, 223, 229, 254, Batten disease 54, 133, 154, 173, 182, 186, 256, 261, 275, 278, 290, 291, 275, 283, 290, 326, 374 295, 310, 311, 313, 314, 324, beta-galactosidase 12, 131 326, 332, 334, 344, 346, 347, biomarker 10, 11, 16, 27, 35, 36, 37, 54, 350, 354, 355, 363, 366, 367, 57, 78, 79, 81, 100, 119, 120, 372, 375, 376, 378, 386 123, 148, 151, 168, 185, 190, exosomes 63 fi 212, 224, 251, 252, 281, 286, eye ndings 59, 157, 182, 234 297, 313, 318, 342, 343, 351, Fabry disease 1, 5, 10, 11, 42, 58, 66, 79, 80, 353, 356, 358, 360, 375, 379, 83, 95, 98, 102, 103, 104, 105, 381 109, 110, 111, 124, 128, 132, blood brain barrier 13, 23, 43, 44, 158, 170, 177, 134, 135, 141, 143, 150, 153, 189, 222, 254, 272, 275 159, 160, 161, 162, 166, 168, bone 16, 18, 56, 75, 76, 87, 163, 203, 169, 171, 176, 179, 180, 181, 243, 277, 286, 300, 306, 355, 184, 187, 190, 199, 208, 212, 357, 365, 377 215, 216, 217, 218, 227, 228, bone marrow transplantation 47, 129, 178, 246, 270, 341, 231, 232, 234, 237, 239, 248, 359 249, 253, 256, 257, 262, 263, cardiomyopathy 161, 231, 271, 284, 350, 376 264, 271, 272, 284, 293, 294, cartilage 377 300, 301, 302, 303, 304, 305, central nervous system (CNS) 2, 62, 149, 158, 177, 183, 198, 306, 307, 308, 311, 317, 32, 211, 266, 283, 320 329, 332, 346, 347, 349, 350, 367, 368, 369, 372, 373, 376, 385

https://doi.org/10.1016/j.ymgme.2018.12.408

Available online 17 January 2019 1096-7192 Molecular Genetics and Metabolism 126 (2019) S169–S171

Farber disease 96, 336 longitudinal study 3, 41, 77, 126, 229, 297, 342, fucosidosis 140 389 galactosylceramidase 136, 213 lymphedema 124 gangliosidosis 379 lyso-GB3 10, 42, 66, 102, 190, 215, 293, Gaucher disease 2, 13, 16, 17, 18, 19, 25, 31, 33, 315, 317 35, 37, 46, 52, 55, 61, 63, 75, lysosomal acid lipase deficiency 30, 57, 121 76, 80, 81, 87, 89, 92, 93, 94, (LAL-D) 99, 101, 108, 115, 117, 123, lysosphingolipids 259 138, 139, 145, 175, 185, 209, magnetic resonance imaging 123, 124, 198, 251, 252 217, 220, 226, 233, 235, 236, (MRI) 238, 239, 243, 250, 255, 261, mass spectrometry 27, 39, 66, 100, 119, 205, 230, 277, 279, 281, 288, 289, 295, 259, 287, 318, 322, 343 297, 311, 314, 316, 318, 320, mesenchymal stem cells 76 328, 331, 338, 342, 343, 345, metachromatic leukodystrophy 8, 41, 74, 170, 257, 270, 378 352, 360, 375, 381 (MLD) gene editing 25, 65, 147, 193, 239, 242, 247, methylation 329 285, 325 microglia 9, 129, 254 gene therapy 21, 36, 43, 53, 62, 69, 70, 84, migalastat 32, 42, 104, 110, 132, 135, 169, 106, 113, 133, 147, 150, 152, 181, 215, 256, 293, 346, 347 171, 174, 180, 186, 187, 188, miglustat 68, 84, 196, 324 211, 225, 232, 235, 247, 248, miRNA 11, 93 267, 272, 279, 289, 29, 299, mitochondria 53, 150 330, 357, 371, 374 modifier gene 70, 93, 322 gene transfer 113, 180, 330 mortality 74, 165, 350, 368 genetic counseling 5, 95, 107, 179, 219, 221, 238, mouse 1, 38, 69, 70, 84, 152, 167, 176, 296, 331, 349, 350, 385 225, 235 genotype-phenotype 2, 25, 58, 61, 74, 93, 139, 141, mucolipidosis III; pseudo-Hurler 327 correlation 148, 179, 208, 238, 277, 284, polydystrophy (ML III) 312, 318, 328, 331 mucolipidosisI II; Leroy disease; 86, 257, 327 globoid cell leukodystrophy 183, 213, 270, 389 I-cell disease (MLII) globotriaosylceramide 66, 171, 176, 212, 232, 237, mucopolysaccharidosis (MPS) 4, 45, 6, 77, 78, 100, 127, 130, 249, 257, 329, 332 155, 163, 165, 200, 201, 202, globotriaosylsphingosine 176, 212, 231 205, 207, 240, 269, 273, 280, glucocerebrosidase 9, 13, 33, 52, 55, 63, 117, 258, 323, 340, 353, 354, 355, 358, 279, 282, 316, 345 359, 365, 366 glucosylceramide 13, 92, 282 mucopolysaccharidosis I (MPS I) 34, 71, 73, 114, 125, 147, 203, glucosylsphingosine; 81, 281, 297, 317, 318, 343 242, 273, 286, 309, 311, 325, lysoglucocerebroside 334, 340, 371 glycogen storage disease 28, 82, 195, 278, 361 mucopolysaccharidosis IH; Hurler 38, 48, 49, 88, 127, 189, 224, type II syndrome (MPS IH) 252, 270, 309, 341, 365 glycosaminoglycans 36, 38, 114, 122, 149, 158, 163, mucopolysaccharidosis II; Hunter 43, 51, 56, 72, 88, 109, 130, 178, 310, 312, 353, 354, 355, syndrome (MPS II) 158, 164, 194, 204, 210, 240, 356, 357, 359, 362 244, 245, 246, 247, 287, 292, glycosphingolipids 320 356, 387, 390 GM1-gangliosidosis 12, 36, 126 mucopolysaccharidosis IHS; 85 GM2-gangliosidosis 174, 250 Hurler-Scheie syndrome (MPS hematopoietic stem cell transplant 43, 129, 140, 155, 224, 252, IHS) (HSCT) 359, 389 mucopolysaccharidosis III; 3, 4, 39, 116, 122, 267, 268, Hunter Outcome Survey (HOS) 210, 245 Sanfilippo syndrome (MPS III) 269 hydrops fetalis 40, 97, 362 mucopolysaccharidosis IIIA; 3, 4, 50, 60, 88, 106, 142, 177, idursulfase 56, 244, 245, 246, 382, 390 Sanfilippo syndrome type A 211, 260, 267, 276, 330, 380 imiglucerase 87, 138, 295 (MPS IIIA) immune response 86, 313 mucopolysaccharidosis IIIB; 3, 23, 60, 65, 67, 116, 142, 149, immune tolerance 69, 90, 144, 170 Sanfilippo syndrome type B 200, 201, 202, 268, 335 immunology 86, 255, 259, 289 (MPS IIIB) immunomodulation 56, 90, 166, 382 mucopolysaccharidosis IIIC; 60, 116, 142, 200, 266 incidence 128, 226 Sanfilippo syndrome type C induced pluripotent stem cells 9, 15, 25, 65, 216, 237, 283 (MPS IIIC) (iPSC) mucopolysaccharidosis IIID; 60, 64, 142, 370 infantile onset 198, 299, 333, 383, 384, 389 Sanfilippo syndrome type D inflammation 185, 286, 301, 305 (MPS IIIC) infusion-related reaction 214, 332 mucopolysaccharidosis IV; 6, 15, 77, 287, 323, 337, 356 intracerebroventricular (ICV) 24, 26, 326 Morquio syndrome (MPS administration IV) intrathecal delivery 24, 29, 133, 225, 378 mucopolysaccharidosis IVA; 6, 7, 27, 45, 155, 200, 201, 265, kidney transplant 83 Morquio syndrome type A 289, 337, 340, 355, 357, 377, Krabbe disease 136, 183, 213, 389 (MPS IVA) 388 late-onset 26, 74, 98, 136, 195, 284, 299

S170 Molecular Genetics and Metabolism 126 (2019) S169–S171 mucopolysaccharidosis VI; 47, 114, 146, 155, 178, 200, Pompe disease; glycogen storage 21, 22, 28, 53, 68, 69, 70, 82, Maroteaux-Lamy syndrome 287, 310, 312, 340 disease type II 90, 91, 95, 107, 109, 120, 144, (MPS VI) 151, 152, 167, 188, 191, 192, mucopolysaccharidosis VII; Sly 78, 97, 112, 131, 229, 254, 362 193, 195, 196, 198, 217, 219, syndrome (MPS VII) 278, 296, 311, 324, 339, 344, multiple myeloma 55, 261, 288, 314 348, 361, 363, 383, 386 multiple sulfatase deficiency 14, 29, 257, 291, 321, 322, 333 pregnancy 31, 40, 280, 295 (MSD) proteinuria 58, 98, 184 muscle 91, 144, 274, 361 proteomics 253 mutation 12, 33, 52, 55, 58, 61, 88, 148, pseudodeficiency 107, 136 206, 264, 292, 312, 327, 333, psychosine 136 366, 390 pulmonary function 45, 165, 320, 388 natural history 3, 4, 14, 34, 41, 56, 8, 94, 97, quality of life (QOL) 7, 17, 45, 109, 131, 143, 221, 126, 141, 165, 203, 268, 276, 228, 276, 294, 302, 323, 340, 284, 319, 333, 336, 342, 349, 346, 349, 368, 388 364, 376, 377, 379, 380 quantitative 77, 87, 206, 241, 251, 337 nephropathic 58 questionnaire 71, 72, 73, 80, 101, 135, 194, nephrotic syndrome 121 199, 221, 262, 276, 280, 302, neuraminidase 12 323 neurodegeneration 4, 23, 41, 126, 223, 225, 267, recombinant enzymes 44, 222, 275, 278, 354 380 registry 34, 60, 125, 146, 156, 172, 229, neuroimaging 49, 102, 198, 251 236, 255, 295, 344, 346 neuroinflammation 9, 258, 259, 260, 279 renal 98, 161, 184, 372 neurological status 108, 328 retina 157 neuromuscular diseases 22, 159 Sandhoff disease 88, 174, 274, 379 neuronal ceroid lipofuscinosis 26, 133, 173, 182, 223, 287, screening 19, 39, 40, 5, 95, 105, 112, 128, (NCL; CLN1, CLN2) 326, 374 159, 163, 168, 212, 217, 228, neuronopathic 2, 94, 203, 328 238, 288, 317, 327, 356, 362, neuropathology 103, 183 365 neuropsychology 122, 194, 224, 323, 337, 342 sebelipase alfa 121 newborn screening 27, 28, 41, 48, 49, 51, 100, 107, severity 169 119, 173, 179, 206, 227, 230, sialidosis 274 252, 263, 309, 353, 358 sleep apnea/sleep disordered 127, 388 Niemann-Pick disease 84, 172, 222, 250, 351, 384 breathing non-neuronopathic 108 software 197, 294, 369 olipudase 351 spinal cord 131 oral delivery 181 spleen 16, 233 osteoporosis 18, 191, 300, 306 splicing 327 outcomes 6, 7, 47, 48, 72, 73, 74, 75, 124, stature 310 135, 138, 140, 153, 191, 192, stem cell transplantation 43, 129, 232, 254, 366 194, 214, 228, 236, 245, 261, structures 197, 322 265, 269, 299, 321, 336, 344, substrate reduction therapy 13, 18, 75, 122, 132, 236, 281, 375, 376, 378, 380, 383, 384 282, 320 oxidative stress 185 sulfamidase 50, 177 pain 153, 162, 199, 226, 294 sulfatase modifying factor 1 29, 321 palmitoyl protein thioesterase 1 173, 275 (SUMF 1) (PPT1) survey 5, 153, 209, 262 Parkinson disease 37, 108, 115, 258, 279, 282, survival 52, 366 381 T cell 248 pathology 96, 98, 182, 314, 351 Tay-Sachs disease 44, 174, 274, 379 pharmacogenetics 32, 38, 220 transplant 71, 129, 183 pharmacokinetics 1, 177, 181, 185, 282 tripeptidyl peptidase 1 26, 173 phenotype 10, 11, 19, 26, 70, 94, 126, 130, urinary glycosaminoglycans 97, 178, 310, 365 169, 264, 268, 277, 321, 333, velaglucerase alfa 209 334, 336, 362 visceral involvement 375 platelets 138, 233 whole exome sequencing 385 podocyte 249, 253 Wolman disease 57, 121 X-inactivation 249, 292 zinc finger nuclease (ZFN) 147, 247

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