DOCSLIB.ORG

The Pennsylvania State University

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Pennsylvania State University The Pennsylvania State University The Graduate School Department of Molecular Medicine IDENTIFICATION OF SOMATIC MUTATIONS IN LGL LEUKEMIA THROUGH WHOLE GENOME SEQUENCING AND CORRELATION OF STAT3 Y640F MUTATION WITH TREAMENT RESPONSE TO METHOTREXATE. A Dissertation in Molecular Medicine by Thomas Lynn Olson 2013 Thomas Lynn Olson Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy August 2013 The dissertation of Thomas L. Olson was reviewed and approved* by the following: Thomas P. Loughran Jr., Physician Director of Penn State Cancer Institute Professor of Medicine Dissertation Advisor Rosalyn B. Irby Associate Professor of Medicine Chair of Committee Richard J. Courtney Professor Emeritus Chair and Distinguished Educator of Microbiology and Immunology Thomas E. Spratt Associate Professor of Biochemistry and Microbiology Director, IBIOS-Chemical Biology Option Director, Biochemistry and Molecular Biology Program Bruce A. Stanley Director, Section of Research Resources Director, Proteomics and Mass Spectrometry Facility Co-Director, Translational Technologies, Penn State Clinical and Translational Sciences Institute Director, Basic Science Cores, Penn State Hershey Cancer Institute Special Member Charles Lang Vice Chair, Distinguished Professor of Cellular and Molecular Physiology, and Surgery. Director of Molecular Medicine Graduate Program *Signatures are on file in the Graduate School iii ABSTRACT The rare disorder, large granular lymphocyte (LGL) leukemia is characterized by a clonal expansion of cytotoxic cells. Normal LGL are critical to the removal of virally-infected or tumor cells from the body. These cells normally expand when there is an immune challenge and then rapidly undergo apoptosis when the antigen is cleared, but leukemic LGLs are resistant to apoptosis. Serological evidence indicates that LGL leukemia may be driven by chronic infection of an unknown retrovirus. Patients with LGL leukemia frequently experience autoimmune conditions such as rheumatoid arthritis and also experience cytopenias requiring medical intervention. The study of LGL leukemia is therefore important both from a patient standpoint and as a model of normal LGL. We have recently discovered and characterized multiple somatic mutations in exon 20 and 21 of STAT3 from exome sequencing of an LGL leukemia patient and confirmed this mutation to be present in both Natural Killer and T cell leukemias. A small percentage was also found to have mutations in STAT5B, with some associated with an aggressive phenotype. Concurrently, we have undertaken whole genome sequencing to determine what other mutations may contribute to LGL leukemia and to expand our knowledge of STAT3 activation. Given the antigen driven nature of LGL leukemia, it is unclear what mutations are necessary for their lack of apoptosis and whether these will resemble those seen in other leukemias. The current treatment of choice for LGL leukemia is immunosuppressive methotrexate. Firstly, we present the first large prospective trial of immunosuppressive therapy in LGL leukemia. In this trial we identify a correlation between a particular Y640F mutation in STAT3 and a favorable response to first line treatment with methotrexate. We measure and report 27 serum cytokines collected for this trial, none of which were a priori predictive of treatment response. Through microarray analysis we identified a gene expression signature indicative of response. Interestingly, this gene signature does not appear to be driven by STAT3 mutation in iv all samples. We propose that there are other methods of STAT activation in these samples and advance a model of how the Y640F mutation enforces response. Secondly, we collaboratively sequenced 6 paired, leukemic/saliva genomes from three LGL patients at a high rate of coverage. Two patient genomes contain a direct STAT3 mutation. In the third patient we observe a mutation in the 3' UTR of the IL6R that could activate JAK- STAT signaling if it is shown to regulate translation. Mutations in the histone modifiers, MLL2, MLL3 and NCOR1 are supported in the sequence reads of all three genomes. Alterations in this pathway would be a new observation for LGL leukemia. We identify and catalog numerous other mutations which may give insight into LGL biology that could allow us to better understand how symptoms arise and differ between patients and why current treatments may fail. Lastly, we present data identifying important cis-regulatory modules in LGL leukemia gathered through ChIP-Seq of genomic regions occupied by the master regulator TBET. We report evidence of binding in the promoter region of numerous constituents of lytic granules which may indicate these are important downstream effectors of TBET. Overlap between the patients used for genome work above and ChIP-Seq is allowing us to determine whether mutations can have an effect on TBET binding and subsequent transcription of nearby genes. As we collect the footprints of additional factors, this holds the promise of elucidating the potential impact of the tens of thousands of non-coding mutations observed in these samples. In summary, we present the results of large collaborative efforts to sequence mutations in LGL leukemia and determine their effect on LGL biology. We advance the knowledge of STAT3 mutations in LGL biology while identifying and stratifying other mutations for the future study of their potential impacts. Bioinformatic methods of analysis were focused on publicly available resources such as Galaxy for which new tools, file formats, and workflows were developed. This allows easy reproduction of methods by other groups and facilitates visualization and future analysis by our own. v TABLE OF CONTENTS List of Figures .......................................................................................................................... vii List of Tables ........................................................................................................................... ix Abbreviations ........................................................................................................................... x Acknowledgements .................................................................................................................. xiii Chapter 1 Introduction to LGL Leukemia ............................................................................... 1 Clinical Entity .................................................................................................................. 1 Diagnosis and Symptoms ................................................................................................. 1 Treatment ......................................................................................................................... 3 Natural Born Killers ......................................................................................................... 3 LGL Are Resistant to Apoptosis ...................................................................................... 4 LGL Are Terminal Effector Memory Cells. .................................................................... 5 Viral Antigen.................................................................................................................... 6 Survival Network of LGL Leukemia ............................................................................... 8 STAT3 Dysregulation and Somatic Mutation .................................................................. 10 Chapter 2 STAT3 Y640F Mutation Predicts Response to Immunosuppressive Therapy of LGL Leukemia: Results of a Prospective Multicenter Phase II Study of Initial Treatment with Methotrexate by the Eastern Cooperative Oncology Group (E5998) ..... 14 Introduction ...................................................................................................................... 14 Methods ............................................................................................................................ 15 Eligibility Criteria .................................................................................................... 15 Objectives ................................................................................................................. 16 Study Design ............................................................................................................ 16 Response Criteria ..................................................................................................... 17 Statistical Methods ................................................................................................... 18 Array Data ................................................................................................................ 18 Cytokine Studies ...................................................................................................... 20 STAT3 Mutational Analysis. ................................................................................... 21 STAT3 Mutagenesis ................................................................................................. 22 STAT3 Reporter Assay ............................................................................................ 22 Western Blot ............................................................................................................. 22 Results .............................................................................................................................. 23 Demographics .........................................................................................................
Load more

Recommended publications
  • 1 Evidence for Gliadin Antibodies As Causative Agents in Schizophrenia
    1 Evidence for gliadin antibodies as causative agents in schizophrenia. C.J.Carter PolygenicPathways, 20 Upper Maze Hill, Saint-Leonard’s on Sea, East Sussex, TN37 0LG [email protected] Tel: 0044 (0)1424 422201 I have no fax Abstract Antibodies to gliadin, a component of gluten, have frequently been reported in schizophrenia patients, and in some cases remission has been noted following the instigation of a gluten free diet. Gliadin is a highly immunogenic protein, and B cell epitopes along its entire immunogenic length are homologous to the products of numerous proteins relevant to schizophrenia (p = 0.012 to 3e-25). These include members of the DISC1 interactome, of glutamate, dopamine and neuregulin signalling networks, and of pathways involved in plasticity, dendritic growth or myelination. Antibodies to gliadin are likely to cross react with these key proteins, as has already been observed with synapsin 1 and calreticulin. Gliadin may thus be a causative agent in schizophrenia, under certain genetic and immunological conditions, producing its effects via antibody mediated knockdown of multiple proteins relevant to the disease process. Because of such homology, an autoimmune response may be sustained by the human antigens that resemble gliadin itself, a scenario supported by many reports of immune activation both in the brain and in lymphocytes in schizophrenia. Gluten free diets and removal of such antibodies may be of therapeutic benefit in certain cases of schizophrenia. 2 Introduction A number of studies from China, Norway, and the USA have reported the presence of gliadin antibodies in schizophrenia 1-5. Gliadin is a component of gluten, intolerance to which is implicated in coeliac disease 6.
    [Show full text]
  • Analysis of Trans Esnps Infers Regulatory Network Architecture
    Analysis of trans eSNPs infers regulatory network architecture Anat Kreimer Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2014 © 2014 Anat Kreimer All rights reserved ABSTRACT Analysis of trans eSNPs infers regulatory network architecture Anat Kreimer eSNPs are genetic variants associated with transcript expression levels. The characteristics of such variants highlight their importance and present a unique opportunity for studying gene regulation. eSNPs affect most genes and their cell type specificity can shed light on different processes that are activated in each cell. They can identify functional variants by connecting SNPs that are implicated in disease to a molecular mechanism. Examining eSNPs that are associated with distal genes can provide insights regarding the inference of regulatory networks but also presents challenges due to the high statistical burden of multiple testing. Such association studies allow: simultaneous investigation of many gene expression phenotypes without assuming any prior knowledge and identification of unknown regulators of gene expression while uncovering directionality. This thesis will focus on such distal eSNPs to map regulatory interactions between different loci and expose the architecture of the regulatory network defined by such interactions. We develop novel computational approaches and apply them to genetics-genomics data in human. We go beyond pairwise interactions to define network motifs, including regulatory modules and bi-fan structures, showing them to be prevalent in real data and exposing distinct attributes of such arrangements. We project eSNP associations onto a protein-protein interaction network to expose topological properties of eSNPs and their targets and highlight different modes of distal regulation.
    [Show full text]
  • Genetic Variation Across the Human Olfactory Receptor Repertoire Alters Odor Perception
    bioRxiv preprint doi: https://doi.org/10.1101/212431; this version posted November 1, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Genetic variation across the human olfactory receptor repertoire alters odor perception Casey Trimmer1,*, Andreas Keller2, Nicolle R. Murphy1, Lindsey L. Snyder1, Jason R. Willer3, Maira Nagai4,5, Nicholas Katsanis3, Leslie B. Vosshall2,6,7, Hiroaki Matsunami4,8, and Joel D. Mainland1,9 1Monell Chemical Senses Center, Philadelphia, Pennsylvania, USA 2Laboratory of Neurogenetics and Behavior, The Rockefeller University, New York, New York, USA 3Center for Human Disease Modeling, Duke University Medical Center, Durham, North Carolina, USA 4Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA 5Department of Biochemistry, University of Sao Paulo, Sao Paulo, Brazil 6Howard Hughes Medical Institute, New York, New York, USA 7Kavli Neural Systems Institute, New York, New York, USA 8Department of Neurobiology and Duke Institute for Brain Sciences, Duke University Medical Center, Durham, North Carolina, USA 9Department of Neuroscience, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA *[email protected] ABSTRACT The human olfactory receptor repertoire is characterized by an abundance of genetic variation that affects receptor response, but the perceptual effects of this variation are unclear. To address this issue, we sequenced the OR repertoire in 332 individuals and examined the relationship between genetic variation and 276 olfactory phenotypes, including the perceived intensity and pleasantness of 68 odorants at two concentrations, detection thresholds of three odorants, and general olfactory acuity.
    [Show full text]
  • Core Transcriptional Regulatory Circuitries in Cancer
    Oncogene (2020) 39:6633–6646 https://doi.org/10.1038/s41388-020-01459-w REVIEW ARTICLE Core transcriptional regulatory circuitries in cancer 1 1,2,3 1 2 1,4,5 Ye Chen ● Liang Xu ● Ruby Yu-Tong Lin ● Markus Müschen ● H. Phillip Koeffler Received: 14 June 2020 / Revised: 30 August 2020 / Accepted: 4 September 2020 / Published online: 17 September 2020 © The Author(s) 2020. This article is published with open access Abstract Transcription factors (TFs) coordinate the on-and-off states of gene expression typically in a combinatorial fashion. Studies from embryonic stem cells and other cell types have revealed that a clique of self-regulated core TFs control cell identity and cell state. These core TFs form interconnected feed-forward transcriptional loops to establish and reinforce the cell-type- specific gene-expression program; the ensemble of core TFs and their regulatory loops constitutes core transcriptional regulatory circuitry (CRC). Here, we summarize recent progress in computational reconstitution and biologic exploration of CRCs across various human malignancies, and consolidate the strategy and methodology for CRC discovery. We also discuss the genetic basis and therapeutic vulnerability of CRC, and highlight new frontiers and future efforts for the study of CRC in cancer. Knowledge of CRC in cancer is fundamental to understanding cancer-specific transcriptional addiction, and should provide important insight to both pathobiology and therapeutics. 1234567890();,: 1234567890();,: Introduction genes. Till now, one critical goal in biology remains to understand the composition and hierarchy of transcriptional Transcriptional regulation is one of the fundamental mole- regulatory network in each specified cell type/lineage.
    [Show full text]
  • Molecular Markers of Serine Protease Evolution
    The EMBO Journal Vol. 20 No. 12 pp. 3036±3045, 2001 Molecular markers of serine protease evolution Maxwell M.Krem and Enrico Di Cera1 ment and specialization of the catalytic architecture should correspond to signi®cant evolutionary transitions in the Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, Box 8231, St Louis, history of protease clans. Evolutionary markers encoun- MO 63110-1093, USA tered in the sequences contributing to the catalytic apparatus would thus give an account of the history of 1Corresponding author e-mail: [email protected] an enzyme family or clan and provide for comparative analysis with other families and clans. Therefore, the use The evolutionary history of serine proteases can be of sequence markers associated with active site structure accounted for by highly conserved amino acids that generates a model for protease evolution with broad form crucial structural and chemical elements of applicability and potential for extension to other classes of the catalytic apparatus. These residues display non- enzymes. random dichotomies in either amino acid choice or The ®rst report of a sequence marker associated with serine codon usage and serve as discrete markers for active site chemistry was the observation that both AGY tracking changes in the active site environment and and TCN codons were used to encode active site serines in supporting structures. These markers categorize a variety of enzyme families (Brenner, 1988). Since serine proteases of the chymotrypsin-like, subtilisin- AGY®TCN interconversion is an uncommon event, it like and a/b-hydrolase fold clans according to phylo- was reasoned that enzymes within the same family genetic lineages, and indicate the relative ages and utilizing different active site codons belonged to different order of appearance of those lineages.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Single-Cell RNA Sequencing Demonstrates the Molecular and Cellular Reprogramming of Metastatic Lung Adenocarcinoma
    ARTICLE https://doi.org/10.1038/s41467-020-16164-1 OPEN Single-cell RNA sequencing demonstrates the molecular and cellular reprogramming of metastatic lung adenocarcinoma Nayoung Kim 1,2,3,13, Hong Kwan Kim4,13, Kyungjong Lee 5,13, Yourae Hong 1,6, Jong Ho Cho4, Jung Won Choi7, Jung-Il Lee7, Yeon-Lim Suh8,BoMiKu9, Hye Hyeon Eum 1,2,3, Soyean Choi 1, Yoon-La Choi6,10,11, Je-Gun Joung1, Woong-Yang Park 1,2,6, Hyun Ae Jung12, Jong-Mu Sun12, Se-Hoon Lee12, ✉ ✉ Jin Seok Ahn12, Keunchil Park12, Myung-Ju Ahn 12 & Hae-Ock Lee 1,2,3,6 1234567890():,; Advanced metastatic cancer poses utmost clinical challenges and may present molecular and cellular features distinct from an early-stage cancer. Herein, we present single-cell tran- scriptome profiling of metastatic lung adenocarcinoma, the most prevalent histological lung cancer type diagnosed at stage IV in over 40% of all cases. From 208,506 cells populating the normal tissues or early to metastatic stage cancer in 44 patients, we identify a cancer cell subtype deviating from the normal differentiation trajectory and dominating the metastatic stage. In all stages, the stromal and immune cell dynamics reveal ontological and functional changes that create a pro-tumoral and immunosuppressive microenvironment. Normal resident myeloid cell populations are gradually replaced with monocyte-derived macrophages and dendritic cells, along with T-cell exhaustion. This extensive single-cell analysis enhances our understanding of molecular and cellular dynamics in metastatic lung cancer and reveals potential diagnostic and therapeutic targets in cancer-microenvironment interactions. 1 Samsung Genome Institute, Samsung Medical Center, Seoul 06351, Korea.
    [Show full text]
  • OR10G4 (NM 001004462) Human Tagged ORF Clone Lentiviral Particle Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for RC222878L3V OR10G4 (NM_001004462) Human Tagged ORF Clone Lentiviral Particle Product data: Product Type: Lentiviral Particles Product Name: OR10G4 (NM_001004462) Human Tagged ORF Clone Lentiviral Particle Symbol: OR10G4 Synonyms: OR11-278 Vector: pLenti-C-Myc-DDK-P2A-Puro (PS100092) ACCN: NM_001004462 ORF Size: 933 bp ORF Nucleotide The ORF insert of this clone is exactly the same as(RC222878). Sequence: OTI Disclaimer: The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene. RefSeq: NM_001004462.1, NP_001004462.1 RefSeq Size: 936 bp RefSeq ORF: 936 bp Locus ID: 390264 UniProt ID: Q8NGN3, A0A126GWS5 Protein Families: Transmembrane Protein Pathways: Olfactory transduction MW: 34.4 kDa This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 OR10G4 (NM_001004462) Human Tagged ORF Clone Lentiviral Particle – RC222878L3V Gene Summary: Olfactory receptors interact with odorant molecules in the nose, to initiate a neuronal response that triggers the perception of a smell.
    [Show full text]
  • Supplementary Table 1: Adhesion Genes Data Set
    Supplementary Table 1: Adhesion genes data set PROBE Entrez Gene ID Celera Gene ID Gene_Symbol Gene_Name 160832 1 hCG201364.3 A1BG alpha-1-B glycoprotein 223658 1 hCG201364.3 A1BG alpha-1-B glycoprotein 212988 102 hCG40040.3 ADAM10 ADAM metallopeptidase domain 10 133411 4185 hCG28232.2 ADAM11 ADAM metallopeptidase domain 11 110695 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 195222 8038 hCG40937.4 ADAM12 ADAM metallopeptidase domain 12 (meltrin alpha) 165344 8751 hCG20021.3 ADAM15 ADAM metallopeptidase domain 15 (metargidin) 189065 6868 null ADAM17 ADAM metallopeptidase domain 17 (tumor necrosis factor, alpha, converting enzyme) 108119 8728 hCG15398.4 ADAM19 ADAM metallopeptidase domain 19 (meltrin beta) 117763 8748 hCG20675.3 ADAM20 ADAM metallopeptidase domain 20 126448 8747 hCG1785634.2 ADAM21 ADAM metallopeptidase domain 21 208981 8747 hCG1785634.2|hCG2042897 ADAM21 ADAM metallopeptidase domain 21 180903 53616 hCG17212.4 ADAM22 ADAM metallopeptidase domain 22 177272 8745 hCG1811623.1 ADAM23 ADAM metallopeptidase domain 23 102384 10863 hCG1818505.1 ADAM28 ADAM metallopeptidase domain 28 119968 11086 hCG1786734.2 ADAM29 ADAM metallopeptidase domain 29 205542 11085 hCG1997196.1 ADAM30 ADAM metallopeptidase domain 30 148417 80332 hCG39255.4 ADAM33 ADAM metallopeptidase domain 33 140492 8756 hCG1789002.2 ADAM7 ADAM metallopeptidase domain 7 122603 101 hCG1816947.1 ADAM8 ADAM metallopeptidase domain 8 183965 8754 hCG1996391 ADAM9 ADAM metallopeptidase domain 9 (meltrin gamma) 129974 27299 hCG15447.3 ADAMDEC1 ADAM-like,
    [Show full text]
  • Cellular and Molecular Signatures in the Disease Tissue of Early
    Cellular and Molecular Signatures in the Disease Tissue of Early Rheumatoid Arthritis Stratify Clinical Response to csDMARD-Therapy and Predict Radiographic Progression Frances Humby1,* Myles Lewis1,* Nandhini Ramamoorthi2, Jason Hackney3, Michael Barnes1, Michele Bombardieri1, Francesca Setiadi2, Stephen Kelly1, Fabiola Bene1, Maria di Cicco1, Sudeh Riahi1, Vidalba Rocher-Ros1, Nora Ng1, Ilias Lazorou1, Rebecca E. Hands1, Desiree van der Heijde4, Robert Landewé5, Annette van der Helm-van Mil4, Alberto Cauli6, Iain B. McInnes7, Christopher D. Buckley8, Ernest Choy9, Peter Taylor10, Michael J. Townsend2 & Costantino Pitzalis1 1Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK. Departments of 2Biomarker Discovery OMNI, 3Bioinformatics and Computational Biology, Genentech Research and Early Development, South San Francisco, California 94080 USA 4Department of Rheumatology, Leiden University Medical Center, The Netherlands 5Department of Clinical Immunology & Rheumatology, Amsterdam Rheumatology & Immunology Center, Amsterdam, The Netherlands 6Rheumatology Unit, Department of Medical Sciences, Policlinico of the University of Cagliari, Cagliari, Italy 7Institute of Infection, Immunity and Inflammation, University of Glasgow, Glasgow G12 8TA, UK 8Rheumatology Research Group, Institute of Inflammation and Ageing (IIA), University of Birmingham, Birmingham B15 2WB, UK 9Institute of
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • Preclinical Characterization of Dostarlimab, a Therapeutic Anti-PD
    MABS 2021, VOL. 13, NO. 1, e1954136 (12 pages) https://doi.org/10.1080/19420862.2021.1954136 REPORTS Preclinical characterization of dostarlimab, a therapeutic anti-PD-1 antibody with potent activity to enhance immune function in in vitro cellular assays and in vivo animal models Sujatha Kumara*, Srimoyee Ghoshb, Geeta Sharmac, Zebin Wangd, Marilyn R. Kehrye, Margaret H. Marinof, Tamlyn Y. Nebenf, Sharon Lug, Shouqi Luoh, Simon Robertsi, Sridhar Ramaswamyj, Hadi Danaeek, and David Jenkinsl aTranslational Research, Immuno-Oncology, Checkmate Pharmaceuticals, Cambridge, MA, USA; bOncology Experimental Medicine Unit, GlaxoSmithKline, Waltham, MA, USA; cSynthetic Lethal Research Unit, Oncolog, GlaxoSmithKline, Waltham, MA, USA; dTranslational Strategy & Research, GlaxoSmithKline,Waltham, MA, USA; eCell Biology, AnaptysBio, Inc,San Diego, CA, USA; fProgram Management, AnaptysBio, Inc, San Diego, CA, USA; gClinical Pharmacology, Scholar Rock, Cambridge, MA, USA; hToxicology, Atea Pharmaceuticals, Boston, MA, USA; iNonclinical Development, Research In Vivo/In Vitro Translation, GlaxoSmithKline, Waltham, MA, USA; jGoogle Ventures, Cambridge, MA, USA; kTranslational Medicine, Blue Print Medicines, Cambridge, MA, USA; lExternal Innovations, IPSEN, Cambridge, MA, USA ABSTRACT ARTICLE HISTORY Inhibitors of programmed cell death protein 1 (PD-1) and its ligand (PD-L1) have dramatically changed the Received 17 December 2020 treatment landscape for patients with cancer. Clinical activity of anti-PD-(L)1 antibodies has resulted in Revised 2 July 2021 increased median overall survival and durable responses in patients across selected tumor types. To date, Accepted 7 July 2021 6 PD-1 and PD-L1, here collectively referred to as PD-(L)1, pathway inhibitors are approved by the US Food KEYWORDS and Drug Administration for clinical use.
    [Show full text]