Annual Report © 2018 Ciberned

Home , Cannabigerol, Josep Samitier

Coordination and management of content: Miguel Medina Padilla José de Arriba-Enríquez Aina Frontera Sánchez Almudena Flores Junquera Soledad Valero Rodríguez Design and editorial coordination: OnAccent.com CIBERNED 2018 ANNUAL REPORT

INDEX

3 4 CIBERNED 2018 ANNUAL REPORT

Jesús Ávila de Grado SCIENTIFIC DIRECTOR

LETTER FROM THE SCIENTIFIC DIRECTOR

As in previous years, I would like to begin by highlighting the excellent work of the research groups that make up CIBERNED and that has allowed us, once again, to be at the forefront of the Centers in biomedical research regarding the quality of publications in high- impact index scientific journals. In order to carry out this work, we have counted on the help and collaboration of the Management Office and the Steering Committee of the Center, as well as its support staff, and the help of the Carlos III Institute of Health and the Associated Institutions. A very important novelty has been the incorporation of six new research groups to CIBERNED, led by Drs. Boada, Mir, Acevedo, Paradas, Ostas and Martínez, to whom I want to welcome to our Center, hoping and wishing, as the groups that preceded them, to carry out a rigorous, efficient and quality work. During this year, 24 scientific papers have been published with an impact index greater than 20 in journals such as the New England Journal of Medicine, Nature, Science, Nature Medicine or Lancet Neurology, and 59 papers with an index greater than 10. In addition, 10 patents remain active, two clinical guidelines have been published and 85 clinical trials are in progress. In addition, courses and seminars have been organized, and the Training Plan has continued as planned. Due to the quality of the scientific activity carried out, some of our researchers have received awards and mentions at international and national level, so on behalf of the Center, I would like to congratulate them. Finally, I would like to highlight the success of our last Congress in Santiago de Composte- la, carried out with the invaluable help of Dr. Labandeira and the collaboration of the CIEN and Queen Sofia Foundations. We continue to progress properly. Thank you all.

5 INTRODUCTION

Biomedical Research Network Centers are created to promote research excellence in Bio- medicine and Health Sciences, which is carried out in the National Health System and the Science and Technology System. Behind this initiative is the purpose of generating large Translational Research Centers, of multidisciplinary and multi-institutional nature where basic, clinical and epidemiological research can be integrated, in order to develop a single common research program, focused on certain pathologies that are relevant for the National Health System due to its prevalence or that are considered strategic because of its social impact. The genesis of CIBERNED was conducted through a coordination research strategy of exploiting synergies between the different groups involved in biomedical research in these areas. It seeks to promote quality research in neurodegenerative diseases being carried out within the National Health System and the Science and Technology System through the development and promotion of a Network Research structures. The goal is to promote and finance, through the Carlos III Institute of Health, Centers for Biomedical Research in the Network (CIBER) through the association of research groups, linked to the National Health System, that contribute to scientifically substantiate the programs and policies of the National Health System in the priority areas of the National R+D+I Plan. CIBERNED was In particular, the Network Center for Biomedical Re- born with the search in Neurodegenerative Diseases (CIBERNED) was born in 2006 as the heir of the Center for Re- aim of combating search in Neurological Diseases (CIEN), which to- neurodegenerative gether with the CNIO, CNIC or CNMVIS constituted the four Centers created with the mission of fight- diseases ing the most prevalent health problems of Span- ish society: cancer, cardiovascular diseases, infectious diseases and neurodegenerative disorders. As for the other Centers, a supporting foundation named CIEN Foundation was created in 2003 as the CIEN’s management body, although unlike the other Centers, it was created as the first Network Center in Spain, eventually being renamed as CIBERNED. Since 2007 the CIEN Foundation has its headquarters in the Alzheimer Center of the Reina Sofia Foundation, created with the collaboration of CIBERNED and the CIEN Foundation, and located in Vallecas (Madrid). CIBERNED was founded under the auspices of the Carlos III Institute of Health, under a cooperation agreement signed by the Government and other participating institutions with

6 CIBERNED 2018 ANNUAL REPORT the idea of creating a Research Center in which basic, clinical and population-based studies are integrated in order to develop a single joint research program, focusing on certain pathologies of great importance for the National Health System either for its prevalence or because of its social impact. It has a multidisciplinary and multi-institutional character and it is aimed at boosting the impact of cutting research in neurodegenerative disorders through a Network Research Structure, thus contributing to scientifically substantiate the programs and policies of the National Health System in the priority areas of the Na- tional R+D+I Plan. CIBERNED is a research organism, endowed with legal status under the Article 6.5 of Law 30/1992 of November 26 on the Legal Regime of Public Administrations and Common Administrative Procedure. It is formed by the association of research groups, with no physical contiguity, belonging to different administrations, institutions and regional governments, from public and private sectors with research lines and goals focused on the specific common area of neurodegenerative diseases and being coordinated to achieve scientific objectives that could hardly be considered in a more restricted execution context. CIBERNED is governed in the internal operating rules, by way of a Regulation. Currently, the CIBERNED-CIEN Foundation CIBERNED is partnership is the only research center in made up of research Spain (and one of the few in the world) in- groups with lines and tegrated into the international network of Centres of Excellence in Neurodegeneration objectives focused on (COEN), an initiative arising from the Euro- neurodegeneration pean Union Joint Programme for Research in Neurodegenerative Diseases (JPND). This joint program constitutes an innovative collaborative research initiative created to address the growing challenges posed by this group of diseases. Its goal is to boost the impact of research by aligning existing national research programs and identifying the common objectives whose scope would benefit through joint action. Its own statutes govern CIBERNED and since the end of 2018 its activity is structured around two Scientific Programs:

• Program 1: Alzheimer’s disease and other degenerative dementias. • Program 2: Parkinson’s and Huntington’s disease and other degenerative movement disorders • Program 3: Amyotrophic Lateral Sclerosis and other neuromuscular disorders.

During 2017 CIBERNED has consisted of 50 research groups supported by different universities, hospitals and the Research Council (CSIC), each led by a principal investigator or responsible. CIBERNED is a network research center, composed of research groups belonging to different Administrations and affiliated Institutions: researcher members physically work in their parent institutions they belong to while simultaneously and actively participating on CIBERNED’s own cooperative research agenda. It should be noted that during 2018 the Carlos III Institute of Health convened, within its annual call for aid of the Strategic Action of Health (AES), the incorporation of new groups to the CIBERNED consortium. This call has allowed the incorporation of 6 new groups (a new group for Program 1, another for Program 2 and four new groups for Program 3).

7 Thus, it is the result of a collaborative partnership between different institutions and the sum of 50 research groups. Depending on the results of the scientific evaluation of the various research groups, there is the possibility to agree on the discontinuation of some groups. Depending on budget availability, it may also consider adding some new research group currently outside CIBERNED that satisfy the requirements of scientific quality and translational activity and aligns with CIBERNED priorities. The Scientific Director Office has been set since October 2011 at the Center for Molecular Biology “Severo Ochoa” (CSIC) in Madrid. The headquarters, including the General Manag- er Office is located in the Alzheimer Center of the Queen Sofia Foundation, 5 Valderrebollo Street in Madrid.

8 CIBERNED 2018 ANNUAL REPORT

AIMS

CIBERNED has as its ultimate goal the promotion of scientific and technical research of excellence in the field of human health, with the generic purpose of producing results quickly transferable to clinical practice in order to improve the health and wellbeing of patients suffering of neurodegenerative diseases, as well as their families and caregivers, providing in turn economic and social benefits. Among its specific aims, the following should be highlighted:

• Promote and develop cooperative translational research excellence in neurodegenerative diseases. • Fostering the impact of science on the health system and on the welfare of patients. • Enhance participation in coordinated actions and calls promoted by funding agencies and the international and national level. • Encourage the development of new therapeutic or/and preventive interventions. • Implement an infrastructure of strategic value for the development of research on prevention, diagnosis and treatment. • Develop plans for specialized training. • Promote support cross-cutting platforms (biobanks, brain imaging, DEGESCO...) • To involve society in the enormous medical and socioeconomic impact of neurodegenerative diseases and facilitate their involvement in the fight against neurodegeneration.

9 DIRECTORY OF RESEARCH GROUPS AND ASSOCIATED INSTITUTIONS

GROUP PRINCIPAL INVESTIGATOR INSTITUTION

301 Alberch Vie, Jordi University of Barcelona Center for Molecular Biology "Severo Ochoa" CSIC- 401 Ávila de Grado, Jesús UAM, Madrid 510 Bullido Gómez-Heras, Mª Jesús Autonomous University, Madrid 509 Calero Lara, Miguel Institute of Health Carlos III, Madrid 402 Camins Espuny, Antonio University of Barcelona 201 Canela Campos, Enric Isidre University of Barcelona 511 Cantero Lorente, José Luis Pablo de Olavide University, Sevilla 502 Carro Diaz, Eva Doce de Octubre University Hospital, Madrid 106 Ceña Callejo, Valentín University of Castilla-La Mancha 413 Comella Carnice, Joan Xavier Vall d'Hebron University Hospital, Barcelona 101 Cuadrado Pastor, Antonio Autonomous Universidad of Madrid Cajal Institute CSIC; Universidad Politécnica de 403 De Felipe Oroquieta, Javier Madrid 114 Del Río Fernández, José Antonio Catalonian Institute de Bioengineering, Barcelona 102 Fariñas Gómez, Isabel University of Valencia 606 Fernández Chacón, Rafael University of Seville 303 Fernández Ruiz, Javier Complutense University of Madrid 503 Ferrer Abizanda, Isidro Bellvitge Institute of Biomedical Research, Barcelona 103 Fuentes Rodríguez, José Manuel Universidad of Extremadura, Cáceres 113 García Verdugo, José Manuel Cavanilles Institute, University of Valencia 415 Gutiérrez Pérez, Antonia University of Málaga 305 Guzmán Pastor, Manuel Complutense University of Madrid 111 Iglesias Vacas, Teresa Institute of Biomedical Research CSIC-UAM, Madrid

10 CIBERNED 2018 ANNUAL REPORT

GROUP PRINCIPAL INVESTIGATOR INSTITUTION 601 Infante Ceberio, Jon Marqués de Valdecilla Research Institute, Santander 202 Kulisevsky Bojarski, Jaime Santa Creu i Sant Pau Hospital, Barcelona 208 Labandeira García, José Luis Universidad of Santiago de Compostela Center of Applied Medical Research, Univ. Navarra, 203 Lanciego Pérez, José Luis Pamplona 504 Lleó Bisa, Alberto Santa Creu i Sant Pau Hospital, Barcelona Virgen del Rocío University Hospital, University of 105 López Barneo, José Seville 609 López de Munain Arregui, Adolfo Biodonostia Research Institute, San Sebastian Center for Molecular Biology "Severo Ochoa" CSIC- 306 Lucas Lozano, José Javier UAM, Madrid 404 Matute Almau, Carlos University of the Basque Country, Bilbao Institute of Biomedical Research IDIBAPS-CSIC , 508 Mengod Los Arcos, Guadalupe Barcelona 204 Moratalla Villalba, Rosario Cajal Institute CSIC, Madrid 604 Muñoz Cánoves, Pura Pompeu Fabra University, ICREA, Barcelona 307 Naranjo Orovio, José Ramón National center of Biotechnology CSIC, Madrid 607 Navarro Acebes, Xavier Autonomous Universidad of Barcelona 205 Obeso Inchausti, José Ángel HM Hospitales Research Foundation 110 Pérez Castillo, Ana María Institute of Biomedical Research CSIC-UAM, Madrid 209 Pérez Tur, Jordi Institute of Biomedicine of Valencia, CSIC 406 Rodríguez Álvarez, José Autonomous University ofBarcelona 206 Rodríguez Díaz, Manuel University of La Laguna, Tenerife 407 Sáez Valero, Javier Miguel Hernández University, Elche 408 Soriano García, Eduardo University of Barcelona 207 Tolosa Sarró, Eduardo Clinic Hospital of Barcelona 409 Torres Alemán, Ignacio Cajal Institute, CSIC, Madrid Institute of Biomedical Research IDIBAPS-CSIC , 410 Trullás Oliva, Ramón Barcelona 108 Vicario Abejón, Carlos Cajal Institute, CSIC, Madrid 109 Vila Bover, Miquel Vall d'Hebron University Hospital, Barcelona 411 Vitorica Ferrández, Francisco Javier University of Seville Center for Molecular Biology "Severo Ochoa" CSIC- 412 Wandosell Jurado, Francisco UAM, Madrid

11 GEOGRAPHIC DISTRIBUTION OF CIBERNED RESEARCH GROUPS

1 1 2 1

Asturias Cantabria País Vasco Galicia Navarra La Rioja

Cataluña Castilla y León Aragón 16 17 Madrid Islas Baleares Comunidad Castilla - Valenciana 1 Extremadura La Mancha Murcia Andalucía 4 1 1 5 Comunidad Canaria

12 CIBERNED 2018 ANNUAL REPORT

ORGANIZATIONAL STRUCTURE

CIBERNED governing bodies are the Governing Council, the Permanent Commission and the Scientific Director. The Governing Council, CIBERNED highest body, consists of three representatives of the Carlos III Health Institute, that include the legal representative of the collaborating organization, the Foundation Research Centre of Neurological Diseases (CIEN) and one representative from each of the institutions participating in the consortium. The President of the Executive Council is the Director of the Carlos III Health Institute. The Secretary of the Governing Council will be the Manager of the consortium The Scientific Director and Manager of CIBERNED are also part of the Governing Council, without a right to vote. The Standing Committee is composed of the Vice President of the Executive Council or his delegate, and four members representing the consortium institutions in the Govern- ing Council. The Scientific Director of the consortium and the Manager, who acts as secretary, are also part of the Permanent Commission, without a right to vote. The Scientific Director is appointed by the President of the Governing Council for a period of four years, renewable by agreement of the parties. The current Scientific Director of CIBERNED is Dr. Jesús Avila de Grado. CIBERNED Management is entrusted to the Managing Director of the Foundation CIEN, Ms. María Ángeles Pérez Muñoz. Support and advisory bodies to governing bodies: a. The Steering Committee b. The External Scientific Advisory Committee

The Steering Committee consists of: • Dr. Jesús Ávila de Grado · Scientific Director • Dña. Mª Ángeles Pérez Muñoz · Manager • Dr. Miguel Medina Padilla · Deputy Scientific Director and the Program coordinators: • Dr. Eduardo Tolosa Sarró • Dra. Teresa Iglesias Vacas • Dr. Alberto Lleó Bisa • Dr. Rafael Fernández Chacón • Dr. José Javier Lucas Lozano • Dra. Isabel Fariñas Gómez • Dr. Adolfo López de Munaín

13 ORGANIZATION CHART

INSTITUTE OF HEALTH CARLOS III ASSOCIATED INSTITUTIONS

GOVERNING COUNCIL

STANDING COMMITTEE

Steering SCIENTIFIC DIRECTOR MANAGER Committee

DEPUTY SCIENTIFIC DIRECTOR

RESEARCH PROGRAM COORDINATORS

RESEARCH GROUPS

14 CIBERNED 2018 ANNUAL REPORT

EXTERNAL SCIENTIFIC ADVISORY COMMITTEE

• Dr. George Perry University of Texas, San Antonio, USA (Chair) • Dr. Werner Poewe Innsbruck Medical University, Austria • Dr. Ángel Cedazo-Mínguez Karolinska Institute, Stockholm, Sweden • Dra. Mary Reilly Institute of Neurology, London, United Kingdom

15 CONSORTIUM MEMBERS

The Central Administration, represented by:

• The Ministry of Economy, Industry and Competitiveness (until June 2018) and the Ministry of Science, Innovation and Universities (since June 2018) through the Carlos III Institute of Health. • The Spanish National Research Council (CSIC). The following Autonomous Regions:

• Autonomous Region of Andalusia, through the Andalusian Public Foundation for Health Research Management at Seville, University of Seville, Pablo de Olavide University, and University of Málaga. • Autonomous Region of Canary Islands, through the University of La Laguna. • Autonomous Region of Cantabria, through the Marqués de Valdecilla Founda- tion (IDIVAL). • Autonomous Region of Castilla La Mancha, through the University of Castil- la-La Mancha. • Autonomous Region of Catalonia, through the Pompeu Fabra University, Vall d’Hebron Research Institute (VHIR), Bellvitge Biomedical Research Institute (IDI- BELL), Catalonian Institute of Bioengineering (IBEC), University of Barcelona and Autonomous University of Barcelona. • Autonomous Region of Valencia, through the University of Valencia and the Miguel Hernández University at Elche. • Autonomous Region of Extremadura, through the Foundation for Research and Training of Health Professionals (FUNDESALUD). • Autonomous Region of Madrid, through the General Directorate of Planning, Research and Training (Council of Health), Autonomous University of Madrid, and Complutense University of Madrid. • Autonomous Region of the Basque Country, through the University of the Basque Country and Biodonostia Research Institute. • Autonomous Region of Galicia, through the University of Santiago de Compostela. Other centers and institutions not affiliated to the previously cited public administrations:

• Santa Creu i Sant Pau Hospital Biomedical Research Institute Foundation (Catalonia). • Hospital Clinic of Barcelona (Catalonia). • Center for Applied Medical Research (Navarra). • HM Hospitales Research Foundation (Madrid).

16 RESEARCH PROGRAMMES 18 CIBERNED 2018 ANNUAL REPORT

PROGRAM 1 ALZHEIMER’S DISEASE AND OTHER DEGENERATIVE DEMENTIAS

19 20 CIBERNED 2018 ANNUAL REPORT

The aging population and longer life expectancy in our society have led in recent decades to a significant increase in cases of people with Alzheimer’s disease (AD) in the last decades being the most common cause of dementia in the elderly and against which there is no effective treatment to date. Currently, it is estimated that between 500,000 and 1,000,000 people suffer from this disease in Spain. This figure could quadruple in the next 50 years, with devastating consequences not only for affected individuals and their families but also for the very stability of our health system. The hallmarks of AD are the presence in the patient’s brain of two aberrant structures, senile plaques and neurofibrillary tangles, as well as synapse loss (it is considered a synaptopathy), mainly between hippocampal and cortical neurons, and significant neurodegeneration. Some aspects of these pathologies can be re- produced in cellular or animal models. Currently, laboratories around the world work The number of people with great interest in identifying new causal with Alzheimer’s and risk genes involved in this pathology that disease could could help clarify its pathophysiological basis and lead to the identification of new therapeu- quadruple in the tic targets. One of the main problems in the EA next 50 years. is that at the time of clinical diagnosis, the brain has already suffered too extensive and irreparable damage. This requires urgent finding of biomarkers as a way to detect the disease much earlier, even asymptomatic, phases when any therapeutic strategy would have a greater chance of success. The 20 groups of clinical and basic researchers from this program, committed both to the diagnosis and care of patients suffering from AD and to research in the laboratory, continue to combine experiences and effort to work in a coordinated manner in the search for new genetic factors, disease biomarkers and new therapeutic strategies in AD and other degenerative dementias. The main research lines are the following: • Genetic Epidemiology • Research on disease-related biomarkers • Cellular and animal models of Alzheimer’s disease and other degenerative dementias • Molecular pathology of Alzheimer’s disease • Mechanisms of neurodegeneration, neuroprotection and design of new therapies.

21 PRINCIPAL INVESTIGATOR INSTITUTION

Ávila de Grado, Jesús Center of Molecular Biology "Severo Ochoa" CSIC-UAM, Madrid

Bullido Gómez-Heras, Mª Jesús Autonomous University of Madrid

Calero Lara, Miguel Carlos III Institute of Health, Madrid

Camins Espuny, Antonio University de Barcelona

Cantero Lorente, José Luis Pablo de Olavide University, Sevilla

Carro Díaz, Eva Doce de Octubre University Hospital, Madrid

Comella Carnice, Joan Xavier Vall d'Hebron University Hospital, Barcelona

De Felipe Oroquieta, Javier Cajal Institute CSIC, Madrid

Ferrer Abizanda, Isidro Bellvitge Biomedical Research Institute, Barcelona

Gutiérrez Pérez, Antonia University of Málaga

Lleó Bisa, Alberto Santa Creu i Sant Pau Hospital, Barcelona

Matute Almau, Carlos University of the Basque Country, Bilbao

Mengod Los Arcos, Guadalupe IDIBAPS-CSIC Biomedical Research institute, Barcelona

Rodríguez Álvarez, José Autonomous University of Barcelona

Sáez Valero, Javier Miguel Hernández University, Elche

Soriano García, Eduardo University of Barcelona

Torres Alemán, Ignacio Cajal Institute CSIC, Madrid

Trullás Oliva, Ramón IDIBAPS-CSIC Biomedical Research institute, Barcelona

Vitorica Ferrández, Francisco Javier University of Seville

Wandosell Jurado, Francisco Center of Molecular Biology "Severo Ochoa" CSIC-UAM, Madrid

Program 1 is coordinated by Drs. Alberto Lleó (Santa Creu i San Pau Hospital) and Jesús Avila de Grado (Center of Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid).

22 CIBERNED 2018 ANNUAL REPORT

401 | Jesús Ávila de Grado

CENTRO DE BIOLOGÍA MOLECULAR “SEVERO OCHOA” C/ Nicolas Cabrera, 1 (Laboratorio 208) Campus de Cantoblanco 28049 Madrid (Spain) Tel: +34 91 196 4564 Fax: +34 91 196 4420 E-mail: [email protected]

PRINCIPAL de la Flor García, Llorens Martin, Maria INVESTIGATOR Miguel. Victoria. Bachelor degree. PhD. Jesús Ávila de Grado García García, Esther. Martin-Maestro Rojas, Technician. Patricia. Bachelor degree. LIST OF García Rodríguez, Alberto. Merchán Rubira, Jesús. PERSONNEL Bachelor degree. Bachelor degree. Bolós Jurado, García-Escudero Perez Ubeda-Protugues, Marta. Barreras, Vega. Juan Ramón. PhD. PhD. Bachelor degree. Cuadros Catalán, Hernández Pérez, de la Torre Alonso, Raquel. Félix. Nuria. Technician. PhD. Graduate.

ABSTRACT After looking at the toxic effects of extracellular tau on neurons and microglia, in a process that could explain the propagation of tau pathology taking place in tauopathies, like Alzheimer disease, we have found that the effect of the presence of extracellular tau on adult hippocampal neurogenesis is similar to that found due to the absence of fractalkine receptors in microglia. On the other hand, the interaction of extracellular tau with fractalkine receptor activates the p38 kinase signaling pathway, resulting in a toxic effect for microglia cells. In a complementary study, we have found a decrease in the level, at their cerebrospinal fluid, of the levels of fractalkine in Alzheimer disease patients, compared to those of non demented subjects. Finally, in collaboration with the group of Prof Soriano (CIBERNED), we have described a novel method to look at single nucleotide polymorphism in brain cells, mainly in those present in Alzheimer disease’s patients.

23 KEYWORDS Tau, GSK3, Enfermedad de Alzheimer, Neurogénesis. Tau, GSK3, Alzheimer disease, Neurogenesis.

PUBLICATIONS 2018 Moreno-Garcia A, Kun A, Calero O, Medina M, Calero M. An overview of the role of lipofuscin in age-related neurodegeneration. Frontiers in Neuroscience. 2018;12(JUL). PMID: 30026686. Rueda N, Vidal V, Garcia-Cerro S, Narcis J.O, Llorens-Martin M, Corrales A. et al. Anti-IL17 treatment ameliorates Down syndrome phenotypes in mice. Brain, Behavior, and Immunity. 2018;73:235-251. PMID: 29758264. Engel T, Gomez-Sintes R, Alves M, Jimenez-Mateos E.M, Fernandez-Nogales M, Sanz-Rodri- guez A. et al. Bi-directional genetic modulation of GSK-3β exacerbates hippocampal neuropathology in experimental status epilepticus. Cell Death and Disease. 2018;9(10). PMID: 30237424. Kenny A, Jimenez-Mateos E.M, Calero M, Medina M, Engel T. Detecting circulating microRNAs as biomarkers in alzheimer’s disease. Methods in Molecular Biology. 2018;1779:471-484. PMID: 29886551. Kenny A, Hernández F, Avila J, Lucas JJ, Henshall DC, Prehn JH et al. Profiling of Argonaute-2-loaded microRNAs in a mouse model of frontotemporal dementia with parkinsonism-17.Interna- tional journal of physiology, pathophysiology and pharmacology. 10(6). PMID: 30697364. Medina M. An overview on the clinical development of tau-based therapeutics. International Journal of Molecular Sciences. 2018;19(4). PMID: 29641484. Cordero J.G, Garcia-Escudero R, Avila J, Gargini R, Garcia-Escudero V. Benefit of Oleuropein Agly- cone for Alzheimer’s Disease by Promoting Autophagy. Oxidative medicine and cellular longevity. 2018;2018:5010741-. PMID: 29675133. Perea J.R, Avila J, Bolos M. Dephosphorylated rather than hyperphosphorylated Tau triggers a pro-inflammatory profile in microglia through the p38 MAPK pathway. Experimental Neurolo- gy. 2018;310:14-21. PMID: 30138606. Llorens-Martín M. Exercising New Neurons to Vanquish Alzheimer Disease. Brain plasticity (Amsterdam, Netherlands). 2018;4(1). PMID: 30564550. Ritter M.L, Avila J, Garcia-Escudero V, Hernandez F, Perez M. Frontotemporal dementia-associated N279K tau mutation localizes at the nuclear compartment. Frontiers in Cellular Neurosci- ence. 2018;12. PMID: 30050413. Beamer E.H, Jurado-Arjona J, Jimenez-Mateos E.M, Morgan J, Reschke C.R, Kenny A. et al. Mi- croRNA-22 controls aberrant neurogenesis and changes in neuronal morphology after status epilepticus. Frontiers in Molecular Neuroscience. 2018;11. PMID: 30618601. Hernandez F, Llorens-Martin M, Bolos M, Perez M, Cuadros R, Pallas-Bazarra N. et al. New Begin- nings in Alzheimer’s Disease: The Most Prevalent Tauopathy. Journal of Alzheimer’s Disease. 2018;64(s1):S529-S534. PMID: 29562521. Perez M, Medina M, Hernandez F, Avila J. Secretion of full-length Tau or Tau fragments in cell culture models. Propagation of Tau in vivo and in vitro. Biomolecular Concepts. 2018;9(1):1- 11. PMID: 29509544.

24 CIBERNED 2018 ANNUAL REPORT

Perez M, Cuadros R, Medina M. Tau assembly into filaments. Methods in Molecular Biology. 2018;1779:447-461. PMID: 29886549. Fuster-Matanzo A, Hernandez F, Avila J. Tau spreading mechanisms; Implications for dysfunctional tauopathies. International Journal of Molecular Sciences. 2018;19(3). PMID: 29495325. Perea J.R, Llorens-Martin M, Avila J, Bolos M. The role of microglia in the spread of Tau: Rele- vance for tauopathies. Frontiers in Cellular Neuroscience. 2018;12. PMID: 30042659. Teixeira C.M, Pallas-Bazarra N, Bolos M, Terreros-Roncal J, Avila J, Llorens-Martin M. Untold New Beginnings: Adult Hippocampal Neurogenesis and Alzheimer’s Disease. Journal of Alzheimer’s Disease. 2018;64(s1):S497-S505. PMID: 29562522. Pose-Utrilla J, Garcia-Guerra L, Del Puerto A, Martin A, Jurado-Arjona J, De Leon-Reyes N.S. et al. Erratum: Author Correction: Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1 (Nature communications (2017) 8 1 (2275)). Nature communications. 2018;9(1):473-. PMID: 29382840. Dávila E, Targa G, Ávila J, Soriano E, Pascual M. Differential accumulation of Tau phosphorylated at residues Thr231, Ser262 and Thr205 in hippocampal interneurons and its modulation by Tau mutations (VLW) and amyloid-β peptide.Neurobiology of disease. 2018. PMID: 30553886. Picher ÁJ, Hernández F, Budeus B, Soriano E, Avila J. Human Brain Single Nucleotide Polymor- phism: Validation of DNA Sequencing.Journal of Alzheimer’s disease reports. 2018;2(1). PMID: 30480253.

RESEARCH PROJECTS 2018 Code: 2016-COHORTE BBN-NED. Title: COHORTE-BBN-NED. Busqueda de biomarcadores para la deteccion temprana de la enfermedad de Alzheimer en la cohorte del proyecto Vallecas. Principal investigator: Miguel Medina. CIBERNED’s collaboration: Yes. CIBERNED groups: G209 ; G401. Other CIBER’s collaboration: CIBER-BBN. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 150000. Duration: 2016-2018.

Code: PI2016/02. Title: Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches. Principal investigator: Jose Antonio Del Rio. CIBERNED’s collaboration: Yes. CIBERNED groups: G114 ; G401; G201. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2016-2018.

25 510 | María Jesús Bullido Gómez-Heras

CENTRO DE BIOLOGÍA MOLECULAR “SEVERO OCHOA” (CSIC-UAM) Tel: +34 91 196 4567 Fax. +34 91 196 4420 e-mail: [email protected]

CBMSO HULP

PRINCIPAL LIST OF Martín Montes, Ángel. INVESTIGATOR PERSONNEL Bachelor degree. Recuero Vicente, María. Bullido Gómez-Heras, Aldudo Soto, Jesús. PhD. María Jesús. PhD. Sastre Merlin, Isabel. Frank García, Ana. Bachelor degree. PhD. Kristen, Henrike Simone. Bachelor degree.

ABSTRACT To identify genes and mechanisms involved in the neurodegeneration process characteristic of Alzheimer’s disease (AD), we carry out functional and genomic analysis of cellular models, validating the most relevant candidates in biological samples of patients by means of genetic association and study of biomarkers. We have models of infection by the virus herpes simplex 1 (HSV 1) and oxidative stress (EO) which present characteristic markers of AD, including alterations in the metabolism of the amyloid precursor protein (APP) and in tau phosphorylation. Analysis of global gene expression of these models indicated the lysosomal pathway as the main process altered by HSV 1 and EO, and functional studies corroborated the existence of a marked effect in several stages of the route, including an increase of lysosomal burden, an inhibition of lysosomal hydrolases (cathepsins, glucosidases) activity, and alteration in the EGFR receptor trafficking. Our data suggest that lysosome dysfunction include alterations in the levels of cholesterol, very interesting fact since that failures in cholesterol homeostasis continue being consistently identified as important in the pathogenesis of AD. Altogether, the data support the hypothesis that lysosomal dysfunction is part of the mechanism of neurodegeneration in our models, so we are studying the role of several candidates of this functional route. During last year, we have found that the decrease of LAMP2 by gene si-

26 CIBERNED 2018 ANNUAL REPORT lencing or disruption partially reversed the markers of neurodegeneration induced by HSV 1 on different cell models that include mouse primary cells, and that the lysosomal related proteases CTSB and MMP14 are involved in non-canonical APP proteolysis and could be part of the mechanism of amyloidogenesis induced by EO. To replicate the neuroblastoma findings in more physiological cell models, we have generated pluripotent stem cells from AD patients and initiated its differentiation into neural phenotypes; to date, results indicate that OS modulates APP metabolism and lysosomal markers similarly to its effect in the neuroblastoma model. To assess the involvement of these candidates in the disease, we have initiated the study of biomarkers related to lysosomal function and HSV infection in AD cases at different clinical and prodromal phases. For this, we count with retrospective collections of hundreds of samples, as well as a new collection that is being recruited by the clinical team at the moment. This collection includes a longitudinal follow-up of participants, as well as the collection of CSF samples from all of them. The biomarkers that we are studying include the determination of anti-HSV antibodies in serum and CSF, and the quantification of cholesterol and several lysosomal markers on leukocytes, in addition to genetic association studies. The data obtained to date suggest a genetic association of LAMP2 with AD in a gender and APOE genotype-dependent manner, as well as an increase of anti-HSV antibody titer in serum and of lysosomal contents in leukocytes of the patients. Altogether, the results of functional studies, gene expression and biomarkers performed to date support the hypothesis that the failure of the lysosomal function could be a relevant mechanism in neurodegeneration, not only in our cell models but also in the patients. On the other hand, we have begun to study the role of microRNAs associated with frontotemporal dementia in the modulation of global gene expression in our cellular models, in the context of an intraCIBER collaborative project, and we continue participating in the research of the genetic architecture of AD, as components of the Dementia Genetics Spanish Consorti- um (DEGESCO), and of international projects and consortia such as the European Alzheimer’s Disease Bank (EADB) or the International Genomics of Alzheimer’s Project (IGAP).

KEYWORDS Alzheimer’s disease, biomarkers, Herpesvirus, neurodegeneration, Lysosomal pathway, genetics, oxidative stress.

PUBLICATIONS 2018 Llorente P, Kristen H, Sastre I, Toledano-Zaragoza A, Aldudo J, Recuero M. et al. A Free Radi- cal-Generating System Regulates Amyloid Oligomers: Involvement of Cathepsin B. Journal of Alzheimer’s Disease. 2018;66(4):1397-1408. PMID: 30400084. Kristen H, Sastre I, Munoz-Galdeano T, Recuero M, Aldudo J, Bullido M.J. The lysosome system is severely impaired in a cellular model of neurodegeneration induced by HSV-1 and oxidative stress. Neurobiology of Aging. 2018;68:5-17. PMID: 29689425. Peloso G.M, van der Lee S.J, Sims R, van der Lee S.J, Naj A.C, Bellenguez C. et al. Genetically elevated high-density lipoprotein cholesterol through the cholesteryl ester transfer protein gene does not associate with risk of Alzheimer’s disease. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2018;10:595-598.

27 RESEARCH PROJECTS 2018 Code: PI2017/01. Title: Estudio del microRNA en el compartimento exosomal del líquido cefalorraquídeo como biomarcador de la demencia frontotemporal y herramienta para el conocimiento de las bases biológicas de la enfermedad. Principal investigator: Jordi Clarimon. CIBERNED’s collaboration: Yes. CIBERNED groups: G504; G406; G510. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2017-2019.

Code: SAF2017-85747-R. Title: funcion lisosomal y homeostasis de colesterol en la neurodegeneracion inducida por HSV-1 y en la enfermedad de alzheimer: mecanismos patogenicos y biomarcadores. Principal investigator: Maria Jesus Bullido Gomez-Heras. CIBERNED’s collaboration: No. CIBERNED groups: G510. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 140000. Duration: 2018-2021.

Code: No figura codigo. Title: Role of the lysosomal protein LAMP2 in the AD-like phenotype induced by HSV-1: Study of the LAMP2 interactome in virus infected neurons. Principal investigator: Maria J Bullido Gomez-Heras. CIBERNED’s collaboration: No. CIBERNED groups: G510. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Ramon Areces. Funding: 108107,5. Duration: 2017-2019.

PHD DISSERTATIONS 2018 Author: Patricia Llorente Ginés. Title: Role of proteases implicated in the lysosomal pathway on APP processing in a cellular model of neurodegeneration. Date: 20/12/2018. Supervisor: María Jesús Bullido Gómez-Heras.

28 CIBERNED 2018 ANNUAL REPORT

509 | Miguel Calero Lara

UNIDAD DE ENCEFALOPATÍAS ESPONGIFORMES UNIDAD FUNCIONAL DE INVESTIGACIÓN DE ENFERMEDADES CRÓNICAS-CROSADIS INSTITUTO DE SALUD CARLOS III Tel.: +34 91 822 3709 E-mail: [email protected]

PRINCIPAL Calero Rueda, Olga. Kun González, Alejandra INVESTIGATOR PhD. Elizabeth. Castellote Olivito, Juan PhD. Calero Lara, Miguel. Manuel. Martínez Martín, Pablo. Bachelor degree. PhD. Damián Moreno, Moreno García, LIST OF Francisco Javier. Alexandra. PERSONNEL PhD. Bachelor degree. Alcalde Cabero, Enrique. De Pedro Cuesta, Jesús. Rodríguez Blázquez, PhD. PhD. María del Carmen. Almazán Isla, Javier. Frades Payo, Mª Belén. Bachelor degree. Technician. Bachelor degree. Ruiz Tovar, María. Avellanal Calzadilla, García López, Fernando PhD. Fuencisla. José. Technician. PhD.

ABSTRACT Public Health/aging Subgroup The subgroup activity was focused on tasks related to the preparation of research and development tasks already pointed out in the monograph Mental Health and Public Health in Spain: Epidemiological Surveillance published in 2018 (http://gesdoc.isciii.es/gesdoccon- troller?action=download&id=09/01/2018-44802ce4e8). Similarly, in the 2019’s version of the Strategy for Research and Innovation in the EU Joint Programme – Neurodegenerative Disease Research (JPND) (JPND). Regarding development initiatives, the group has contributed to the creation of the Spanish Multiple Sclerosis Registry, which promotes Pharmacovigilance tasks in collaboration with the Spanish Medicines Agency (AEMPS) and the Spanish Neurology Society. The group has contributed with a member to the Scientific Advisory group for the drafting of the new version

29 of the EU Strategy for Research and Innovation in Neurodegeneration (https://www.neu- rodegenerationresearch.eu/wp-content/uploads/2019/ 04 / Full-JPND-Research-and-Inno- vation-Strategy-3.04.pdf) that proposes a considerable extension of the work fields in the aforementioned program with respect to 2012, contributing in an important way to the elaboration of the chapter on “Origin and development of neurodegenerative disorders “. The subgroup participates in the European Project CHRODIS+ (2017-20) in relation to the coordination and execution of an activity for active aging / healthy aging that entails at the neuromotor level the trans-institutional implementation in Spain of the “Icelandic 65+ multimodal intervention Program”, focused on physical activity. Assessment and Outcomes Subgroup After the critical review of the Non-Motor Symptoms Scale for Parkinson’s Disease (2014) a new version was promoted, the Movement Disorder Society sponsored Non-Motor Symp- toms Rating Scale, whose pilot study was published in 2018 and the validation study (sponsored by the International Parkinson and Movement Disorder Society, IPMDS) has also been completed this year. Within international collaborations, studies and publications on outcomes of second-line therapies for patients with advanced Parkinson’s Disease (APD) have been carried out; development and validation of a scale (Parkinson’s Disease Composite Scale) promoted by the European Parkinson’s Disease Association and an instrument for the detection of APD; assessment scales of sleep disorders in movement disorders; gradation of severity, health-related quality of life and evaluation of urinary and pain symptoms in Parkinson’s disease, and motor and functional evaluations for Huntington’s disease. Molecular Biology Subgroup The current objectives of this subgroup are (I) molecular diagnosis, (II) basic research on genetic susceptibility factors in Alzheimer’s disease and prion diseases, and (iii) molecular basis of conformational diseases, including AD, PD and CJD. The subgroup maintains a close collaboration with different CIBERNED groups in two collaborative projects: the DEGESCO consortium, and the Spanish Prion Network (AGL2017-90665-REDT Network of Excellence). In collaboration with Dr. M. Medina (PI1 of the project and CIBERNED deputy director), we are carrying out the project entitled “miRNA and lipid metabolism markers as potential links between vascular dysfunction and Alzheimer’s pathophysiology” (MINECO). The group also collaborates with the biothech company Biocross, SL for the development of a new commer- cial test (e4Risk®) intended for the detection of the isoform apoE4 in clinical rutine by an immunoturbidimetric assay. On the other hand, the group collaborates with other institutions that provide a fundamental support, highlighting the collaboration with the CIEN Foun- dation for the early diagnosis of Alzheimer’s disease (Vallecas project, and the University of the República de Uruguay in the study of peripheral expression of CNS neurodegenerative diseases.

30 CIBERNED 2018 ANNUAL REPORT

KEYWORDS Alzheimer’s disease, Parkinson’s disease, Creutzfeldt-Jakob disease, epidemiology, neurodegeneration, disability, assessment scales, conformational diseases

PUBLICATIONS 2018 Martinez-Martin P, Ray Chaudhuri K. omprehensive grading of Parkinson’s disease using motor and non-motor assessments: addressing a key unmet need. Expert Review of Neurotherapeu- tics. 2018;18(1):41-50. PMID: 29090594. Martinez-Martin P, Kulisevsky J, Mir P, Tolosa E, García-Delgado P, Luquin MR. Validation of a simple screening tool for early diagnosis of advanced Parkinson’s disease in daily practice: the CDEPA questionnaire.NPJ Parkinson’s disease. 4. PMID: 29978014. Catalan M.J, Molina-Arjona J.A, Mir P, Cubo E, Arbelo J.M, Martinez-Martin P. Improvement of impulse control disorders associated with levodopa–carbidopa intestinal gel treatment in advanced Parkinson’s disease. Journal of Neurology. 2018;265(6):1279-1287. PMID: 29557989. Zerr I, Schmitz M, Karch A, Villar-Piqué A, Kanata E, Golanska E et al. Cerebrospinal fluid neurofilament light levels in neurodegenerative dementia: Evaluation of diagnostic accuracy in the differential diagnosis of prion diseases.Alzheimer’s & dementia : the journal of the Alzheimer’s Association. 2018. PMID: 29391125. Silverdale M.A, Kobylecki C, Kass-Iliyya L, Martinez-Martin P, Lawton M, Cotterill S. et al. A detailed clinical study of pain in 1957 participants with early/moderate Parkinson’s disease. Parkinsonism and Related Disorders. 2018;56:27-32. PMID: 29903584. Calero O, Garcia-Albert L, Rodriguez-Martin A, Veiga S, Calero M. A fast and cost-effective method for apolipoprotein e isotyping as an alternative to APOE genotyping for patient screening and stratification. Scientific Reports. 2018;8(1). PMID: 29654261. Kurtis M.M, Balestrino R, Rodriguez-Blazquez C, Forjaz M.J, Martinez-Martin P. A review of scales to evaluate sleep disturbances in movement disorders. Frontiers in Neurology. 2018;9(MAY). PMID: 29896152.

Bernabeu-Zornoza A, Coronel R, Palmer C, Calero M, Martinez-Serrano A, Cano E. et al. Aβ42 Peptide Promotes Proliferation and Gliogenesis in Human Neural Stem Cells. Molecular Neuro- biology. 2018. PMID: 30259399. Kun A, Gonzalez-Camacho F, Hernandez S, Moreno-Garcia A, Calero O, Calero M. Characteriza- tion of amyloid-β plaques and autofluorescent lipofuscin aggregates in alzheimer’s disease brain: A confocal microscopy approach. Methods in Molecular Biology. 2018;1779:497-512. PMID: 29886553. Pavy-Le Traon A, Cotterill N, Amarenco G, Duerr S, Kaufmann H, Lahrmann H. et al. Clinical Rating Scales for Urinary Symptoms in Parkinson Disease: Critique and Recommendations. Movement Disorders Clinical Practice. 2018;5(5):479-491. PMID: 30515437. Sauerbier A, Schrag A, Martinez-Martin P, Hall L.J, Parry M, Mischley L.K. et al. Dietary Variations in a Multiethnic Parkinson’s Disease Cohort and Possible Influences on Nonmotor Aspects: A Cross-Sectional Multicentre Study. Parkinson’s Disease. 2018;2018. PMID: 30662706. Martinez-Martin P, Rizos A.M, Wetmore J, Antonini A, Odin P, Pal S. et al. First comprehensive

31 tool for screening pain in Parkinson’s disease: the King’s Parkinson’s Disease Pain Question- naire. European Journal of Neurology. 2018;25(10):1255-1261. PMID: 29806962. Martinez-Martin P, Rodriguez-Blazquez C, Catalan M.J. Independent and Complementary Vali- dation of the QUIP-RS in Advanced Parkinson’s Disease. Movement Disorders Clinical Practice. 2018;5(3):341-342. PMID: 30800709. Ambrosio L, Navarta-Sanchez M.V, Meneses A, Rodriguez-Blazquez C. Living with Chronic Illness Scale: Pilot study in patients with several chronic diseases Escala de convivencia con un proceso crónico: estudio piloto en pacientes con enfermedades crónicas. Atencion Primaria. 2018. Corredor-Andres B, Munoz-Calvo M.T, Calero O, Aparicio C, Argente J, Calero M. Nephrotic syndrome associated with severe hypertriglyceridemia in a pediatric patient: Answers. Pediatric Nephrology. 2018;33(11):2075-2078. PMID: 29532233. Corredor-Andres B, Munoz-Calvo M.T, Calero O, Aparicio C, Argente J, Calero M. Nephrotic syndrome associated with severe hypertriglyceridemia in a pediatric patient: Questions. Pediatric Nephrology. 2018;33(11):2073-2074. PMID: 29532231. Dafsari H.S, Petry-Schmelzer J.N, Ray-Chaudhuri K, Ashkan K, Weis L, Dembek T.A. et al. Non-motor outcomes of subthalamic stimulation in Parkinson’s disease depend on location of active contacts. Brain Stimulation. 2018;11(4):904-912. PMID: 29655586. Dafsari H.S, Silverdale M, Strack M, Rizos A, Ashkan K, Mahlstedt P. et al. Nonmotor symptoms evolution during 24 months of bilateral subthalamic stimulation in Parkinson’s disease. Move- ment Disorders. 2018;33(3):421-430. PMID: 29465787. Antonini A, Tinazzi M, Abbruzzese G, Berardelli A, Chaudhuri K.R, Defazio G. et al. Pain in Parkinson’s disease: facts and uncertainties. European Journal of Neurology. 2018;25(7):917- 924. PMID: 29520899. Taghizadeh G, Martinez-Martin P, Fereshtehnejad S.-M, Habibi S.A, Nikbakht N, Alizadeh N.H. et al. Psychometric properties of the Berg balance scale in idiopathic Parkinson’ disease in the drug off-phase. Neurological Sciences. 2018;39(12):2175-2181. PMID: 30244299. Mestre T.A, Carlozzi N.E, Ho A.K, Burgunder J.-M, Walker F, Davis A.M. et al. Quality of Life in Hun- tington’s Disease: Critique and Recommendations for Measures Assessing Patient Health-Re- lated Quality of Life and Caregiver Quality of Life. Movement Disorders. 2018;33(5):742-749. PMID: 29570848. Wijers I.G.M, Ayala A, Rodriguez-Blazquez C, Rodriguez-Laso A, Rodriguez-Rodriguez V, Forjaz M.J. Rasch Analysis and Construct Validity of the Disease Burden Morbidity Assessment in Older Adults. Gerontologist. 2018;58(5):e302-e310. PMID: 30219906. Mestre T.A, Busse M, Davis A.M, Quinn L, Rodrigues F.B, Burgunder J.-M. et al. Rating Scales and Performance-based Measures for Assessment of Functional Ability in Huntington’s Disease: Critique and Recommendations. Movement Disorders Clinical Practice. 2018;5(4):361-372. PMID: 30363510. Mestre TA, Forjaz MJ, Mahlknecht P, Cardoso F, Ferreira JJ, Reilmann R et al. Rating Scales for Motor Symptoms and Signs in Huntington’s Disease: Critique and Recommendations.Move- ment disorders clinical practice. 5(2). PMID: 30363393. Rodriguez-Blazquez C, Forjaz M.J, Kurtis M.M, Balestrino R, Martinez-Martin P. Rating scales for movement disorders with sleep disturbances: A narrative review. Frontiers in Neurology. 2018;9(JUN). PMID: 29951032. Skorvanek M, Martinez-Martin P, Stebbins G.T, Goetz C.G. Reply: Hoehn and Yahr stage 3 and Postural stability item in the Movement Disorder Society-Unified Parkinson’s Disease Rating

32 CIBERNED 2018 ANNUAL REPORT

Scale. Movement Disorders. 2018;33(7):1189-1190. PMID: 30153386. Skorvanek M, Goldman J.G, Jahanshahi M, Marras C, Rektorova I, Schmand B. et al. Reply: MoCA for cognitive screening in Parkinson’s disease: Beware of floor effect. Movement Disorders. 2018;33(3):499-500. PMID: 29460975. Dafsari H.S, Weiss L, Silverdale M, Rizos A, Reddy P, Ashkan K. et al. Short-term quality of life after subthalamic stimulation depends on non-motor symptoms in Parkinson’s disease. Brain Stimulation. 2018;11(4):867-874. PMID: 29655587. Martinez-Martin P, Macaulay D, Jalundhwala Y.J, Mu F, Ohashi E, Marshall T. et al. The long- term direct and indirect economic burden among Parkinson’s disease caregivers in the United States. Movement Disorders. 2018. PMID: 30589953. Schmitt E, Krack P, Castrioto A, Klinger H, Bichon A, Lhommee E. et al. The Neuropsychiatric Fluctuations Scale for Parkinson’s Disease: A Pilot Study. Movement Disorders Clinical Practice. 2018;5(3):265-272. PMID: 30363450. Martinez-Martin P, Skorvanek M, Rojo-Abuin J.M, Gregova Z, Stebbins G.T, Goetz C.G. Valida- tion study of the hoehn and yahr scale included in the MDS-UPDRS. Movement Disorders. 2018;33(4):651-652. PMID: 29322548. Mata-Balaguer T, Cuchillo-Ibanez I, Calero M, Ferrer I, Saez-Valero J. Decreased generation of c-terminal fragments of apoer2 and increased reelin expression in Alzheimer’s disease. FASEB Journal. 2018;32(7):3536-3546. PMID: 29442527. Sanchez-Ferro A, Matarazzo M, Martinez-Martin P, Martinez-Avila J.C, Gomez de la Camara A, Giancardo L. et al. Minimal Clinically Important Difference for UPDRS-III in Daily Practice. Move- ment Disorders Clinical Practice. 2018;5(4):448-450. PMID: 30838303. Moreno-Garcia A, Kun A, Calero O, Medina M, Calero M. An overview of the role of lipofuscin in age-related neurodegeneration. Frontiers in Neuroscience. 2018;12(JUL). PMID: 30026686. Villar-Pique A, Schmitz M, Lachmann I, Karch A, Calero O, Stehmann C. et al. Cerebrospinal Fluid Total Prion Protein in the Spectrum of Prion Diseases. Molecular Neurobiology. 2018. PMID: 30062673. Kenny A, Jimenez-Mateos E.M, Calero M, Medina M, Engel T. Detecting circulating microRNAs as biomarkers in alzheimer’s disease. Methods in Molecular Biology. 2018;1779:471-484. PMID: 29886551. Skorvanek M, Martinez-Martin P, Kovacs N, Zezula I, Rodriguez-Violante M, Corvol J.-C. et al. Relationship between the MDS-UPDRS and Quality of Life: A large multicenter study of 3206 patients. Parkinsonism and Related Disorders. 2018;52:83-89. PMID: 29625875.

RESEARCH PROJECTS 2018 Code: PI15CIII/00037. Title: Mortalidad en poblacion que vive en residencias de ancianos de Madrid. Principal investigator: Javier Damian. CIBERNED’s collaboration: No. CIBERNED groups: G509. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: ND. Duration: 2015-2019.

33 402 | Antonio Camins Espuny

UNIVERSIDAD DE BARCELONA. Facultad de Farmacia Tel.: +34 93 402 4531 E-mail: [email protected]

PRINCIPAL Cabezón Rodríguez, Pallas Lliberia, Mercé. INVESTIGATOR Itsaso. PhD. PhD. Pelegri Gavalda, Camins Espuny, Cano Fernández, Carmen. Antonio. Amanda. PhD. Bachelor degree. Sánchez López, Elena. Ettcheto Arriola, Miren. Bachelor degree. LIST OF PhD. Vilaplana Hortensi, PERSONNEL Folch, Jaume. Jordi. Auladell Costa, Bachelor degree. PhD. Carme. García López, Mª Luisa. PhD. Bachelor degree.

ABSTRACT Pre-clinical model of Alzheimer’s disease. The results obtained during the last years with APPSwe / PS1dE9 (APP / PS1) mice produce higher levels of β-amyloid than wild type (WT) mice, treated with a high-fat diet, develop memory loss compared with mice fed with a normal diet. Although the reason is unknown, there are many studies that point out that the inactivation of the insulin receptor, at the level of the hippocampus, area of the brain related to the process of memory and learning, is the main responsible of this cognitive loos In this line, different works suggest that cognitive loss associated with metabolic disorders (type II diabetes), is a key factor in the onset of Alzheimer’s disease. Moreover, some hypotheses suggests that Alzheimer’s disease not only affects the brain, it should be considered as a whole organism disease. A key molecule involved in the regulation of the activity of the insulin receptor is JNK1, which in turn responds to different types of stress signals and, in particular, glial activation and the release of pro-inflammatory mediators.

34 CIBERNED 2018 ANNUAL REPORT

For this reason, we are working with Licochalcone A, this drug is an inhibitor of JNK1 kinase. Through this inhibition, the inactivation of the brain insulin receptor can be prevented being a potential treatment for cognitive loss in Alzheimer’s disease. General objective: To act specifically on the inhibition of JNK1, as a key target, preventing phosphorylation and inhibition of the insulin receptor in the hippocampus and preventing the cognitive loss and also the neuropathological process development of AD. Concrete objectives: 1.- Assessment of cognitive loss in the process of obesity in relation to the activity of JNK, in C57Bl6 mice of the wild strain exposed to a chronic intake during 8 months of a high-fat diet (HFD). 2.- To evaluate and characterize the impact of the chronic administration of a selective inhibitor of JNK1, licochalcone A, in APPsw / PS1E9 mice, subjected to a chronic intake of a high-fat diet (HFD), for 8 months.

Neurodegeneration, brain aging and innate immunity In recent studies we have determined that during aging, degenerative granular structures containing neo-epitopes that are recognized by natural antibodies that exist from birth in the same mice appear in the brain of mice. This same situation has been determined later in the corpora amylacea of the human brain. These facts indicate a certain relationship and possible interaction between the degenerative and aging processes on the one hand and the natural immune system on the other. Objective: Study the relationship between degenerative and aging processes on the one hand and the natural immune system on the other Concrete objectives: 1.- Characterize, in different neurodegenerative diseases and in aging, brain structures that present neo-epitopes 2.- Determine the nature of these neo-epitopes 3.- Determine the natural immune response that occurs in relation to the formation of these epitopes during aging or neurodegeneration.

Development of drugs for the treatment of Alzheimer’s disease and neurodegenerative diseases in general. Studying and clarifying the process of neuronal death after the stimulation of the ionotropic glutamate receptors is key to understanding neuronal death. We already have drugs synthesized to the Pharmaceutical Chemistry Unit of the Faculty of Pharmacy that can potentially be effective in treating Alzheimer’s disease. On the one hand it is a series of more than 400 compounds from 2008 until now (Campos et al., Bioorg Med Chem, 2008, 16, 9925-9936, Duque et al., Bioorg Med Chem, 2009, 17, 3198- 3206 and 2010, 18, 46-57, Torres et al., Bioorg Med Chem, 2012, 20, 942-948, Valverde et al., Bioorg Med Chem, 2014, 22, 2678-2683, Leiva et al., Tetrahedron. Lett, 2015, 56, 1272-1275). On the other hand, it is derived from huprins (Canudas et al., Exp. Neurol, 2003, 180, 123-130), which may have multidian activity, both on NMDA receptors and on acetylcholinesterase. General purpose To determine the activity of drugs developed as NMDA receptor antagonists and neuro-

35 toxins of marine origin, as well as their action on other targets that provide them with neuroprotective capacity. Concrete objectives 1.- To evaluate the activity of the new compounds against the increase of calcium induced by NMDA in cultures of cerebellar granule neurons 2.- Determine the activity on the monoamine oxidase (MAO) of both type A and B 3.- To evaluate the neuroprotective activity of the compounds against the induced neurodegeneration in cultures of granular calcium neurons and in animal models (APP / PS1 mice)

KEYWORDS Alzheimer’s disease, cerebral insulin resistance, ipo 3, obesity, cognition, drug development, aging.

PUBLICATIONS 2018 Kefalakes E, Boselt S, Sarikidi A, Ettcheto M, Bursch F, Naujock M. et al. Characterizing the multiple roles of FGF-2 in SOD1G93A ALS mice in vivo and in vitro. Journal of Cellular Physiology. 2019;234(5):7395-7410. epub2018. PMID: 30370540. Kuhne B.A, Puig T, Ruiz-Martinez S, Crous-Maso J, Planas M, Feliu L. et al. Comparison of migration disturbance potency of epigallocatechin gallate (EGCG) synthetic analogs and EGCG PEGylated PLGA nanoparticles in rat neurospheres. Food and Chemical Toxicology. 2019;123:195-204. epub2018. PMID: 30367911. Palomera-Avalos V, Grinan-Ferre C, Izquierdo V, Camins A, Sanfeliu C, Canudas A.M. et al. Resveratrol modulates response against acute inflammatory stimuli in aged mouse brain. Ex- perimental Gerontology. 2018;102:3-11. PMID: 29174969. Peron R, Vatanabe I.P, Manzine P.R, Camins A, Cominetti M.R. Alpha-secretase ADAM10 regulation: Insights into Alzheimer’s disease treatment. Pharmaceuticals. 2018;11(1). PMID: 29382156. Auge E, Pelegri C, Manich G, Cabezon I, Guinovart J.J, Duran J. et al. Astrocytes and neurons produce distinct types of polyglucosan bodies in Lafora disease. GLIA. 2018;66(10):2094- 2107. PMID: 30152044. Cano A, Ettcheto M, Espina M, Auladell C, Calpena A.C, Folch J. et al. Epigallocatechin-3-gallate loaded PEGylated-PLGA nanoparticles: A new anti-seizure strategy for temporal lobe epilepsy. Na- nomedicine: Nanotechnology, Biology, and Medicine. 2018;14(4):1073-1085. PMID: 29454994. Olloquequi J, Cornejo-Cordova E, Verdaguer E, Soriano F.X, Binvignat O, Auladell C. et al. Exci- totoxicity in the pathogenesis of neurological and psychiatric disorders: Therapeutic implications. Journal of Psychopharmacology. 2018;32(3):265-275. PMID: 29444621. Auge E, Duran J, Guinovart J.J, Pelegri C, Vilaplana J. Exploring the elusive composition of corpora amylacea of human brain. Scientific Reports. 2018;8(1). PMID: 30202002.

36 CIBERNED 2018 ANNUAL REPORT

Rosa P, Zerbinati C, Crestini A, Canudas AM, Ragona G, Confaloni A et al. Heme Oxygenase-1 and Brain Oxysterols Metabolism Are Linked to Egr-1 Expression in Aged Mice Cortex, but Not in Hippocampus.Frontiers in aging neuroscience. 10. PMID: 30459596. Sanguesa G, Cascales M, Grinan C, Sanchez R.M, Roglans N, Pallas M. et al. Impairment of Novel Object Recognition Memory and Brain Insulin Signaling in Fructose- but Not Glucose-Drinking Female Rats. Molecular Neurobiology. 2018;55(8):6984-6999. PMID: 29372547. Corpas R, Grinan-Ferre C, Palomera-Avalos V, Porquet D, Garcia de Frutos P, Franciscato Cozzoli- no S.M. et al. Melatonin induces mechanisms of brain resilience against neurodegeneration. Journal of Pineal Research. 2018;65(4). PMID: 29907977. Sanchez-Lopez E, Ettcheto M, Egea M.A, Espina M, Cano A, Calpena A.C. et al. Memantine loaded PLGA PEGylated nanoparticles for Alzheimer’s disease: In vitro and in vivo characterization. Journal of Nanobiotechnology. 2018;16(1). PMID: 29587747. Corpas R, Grinan-Ferre C, Rodriguez-Farre E, Pallas M, Sanfeliu C. Resveratrol Induces Brain Re- silience Against Alzheimer Neurodegeneration Through Proteostasis Enhancement. Molecular Neurobiology. 2018;:1-15. PMID: 29948950. Jara-Moreno D, Castro-Torres R.D, Ettcheto M, Auladell C, Kogan M.J, Folch J. et al. The Ethyl Acetate Extract of Leaves of Ugni molinae Turcz. Improves Neuropathological Hallmarks of Alzheimer’s Disease in Female APPswe/PS1dE9 Mice Fed with a High Fat Diet. Journal of Alzheimer’s Disease. 2018;66(3):1175-1191. PMID: 30400089. Folch J, Ettcheto M, Busquets O, Sanchez-Lopez E, Castro-Torres R.D, Verdaguer E. et al. The implication of the brain insulin receptor in late onset Alzheimer’s disease dementia. Pharma- ceuticals. 2018;11(1). PMID: 29382127. Grinan-Ferre C, Corpas R, Puigoriol-Illamola D, Palomera-Avalos V, Sanfeliu C, Pallas M. Under- standing Epigenetics in the Neurodegeneration of Alzheimer’s Disease: SAMP8 Mouse Model. Journal of Alzheimer’s Disease. 2018;62(3):943-963. PMID: 29562529. Merelli A, Rodriguez JCG, Folch J, Regueiro MR, Camins A, Alberto L. Understanding the Role of Hypoxia inducible factor during neurodegeneration for new therapeutics opportunities.Cur- rent neuropharmacology. 2018. PMID: 29318974. Ettcheto M, Sanchez-Lopez E, Gomez-Minguez Y, Cabrera H, Busquets O, Beas-Zarate C. et al. Peripheral and Central Effects of Memantine in a Mixed Preclinical Mice Model of Obesity and Fa- milial Alzheimer’s Disease. Molecular Neurobiology. 2018;55(9):7327-7339. PMID: 29404958.

RESEARCH PROJECTS 2018 Code: PI2016/01. Title: Alteraciones del metabolismo gluco-lipidico y desarrollo de la demencia de Alzheimer. Principal investigator: Ignacio Torres Aleman. CIBERNED’s collaboration: Yes. CIBERNED groups: G409 ; G402; G511; G502; G412. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

37 Code: ART. Title: Design of an age-dependent corneal membrane model for in vitro interaction studies of biodegradable polymeric nanoparticles. In vitro/ex vivo/in vivo correlation. Principal investigator: Elena Sanchez Lopez. CIBERNED’s collaboration: No. CIBERNED groups: G402. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Universitat de Barcelona. Funding: 8000. Duration: 2018-2019.

Code: SAF2016-77703-C2-1-R. Title: Estudio de la Epoxido hidrolasa soluble como una nueva diana farmacologica para la enfermedad de Alzheimer: modulacion del estres oxidativo y la funcion mitocondrial. Principal investigator: Merce Pallas. CIBERNED’s collaboration: No. CIBERNED groups: G402. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 100000. Duration: 2015-2019.

Code: BFU2016-78398-P. Title: Estudio de la presencia de neo-epitopos en estructuras degenerativas cerebrales y de la existencia en el plasma de anticuerpos naturales dirigidos contra dichos neo-epitopos. Principal investigator: Carme Pelegri Gabalda. CIBERNED’s collaboration: No. CIBERNED groups: G402. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 120000. Duration: 2016-2020.

Code: SAF2017-84283-R. Title: Modulacion de la via del receptor de insulina hipocampal como estrategia terapeutica para el tratamiento de la perdida cognitiva. Principal investigator: Antonio Camins Espuny. CIBERNED’s collaboration: No. CIBERNED groups: G402. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 133000. Duration: 2018-2020.

Code: La Marato de TV3. Title: Noves polimixines per al tractament d’infeccions causades per bacteris multiresist- ents. Principal investigator: Francesc Rabanal Anglada. CIBERNED’s collaboration: No. CIBERNED groups: G402 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 60000. Duration: 2018-2019.

38 CIBERNED 2018 ANNUAL REPORT

PHD DISSERTATIONS 2018 Author: Miren Ettcheto Arriola. Title: Efecto del síndrome metabólico provocado por una dieta rica en grasa en ratones APPswe/PS1dE9, modelo experimental de la enfermedad de Alzheimer, y posibles terapias farmacológicas. Date: 16/3/2018. Supervisor: Antonio Camins Espuny.

Author: Amanda Cano Fernandez. Title: Diseño y caracterización de nanopartículas poliméricas de Epigalocatequina-3-gala- to para el tratamiento de enfermedades del sistema nervioso central. Date: 9/11/2018. Supervisor: Antonio Camins Espuny.

39 511 | José Luis Cantero Lorente

LABORATORIO DE NEUROCIENCIA FUNCIONAL Servicios Centrales de Investigación (Edificio 21) Universidad Pablo de Olavide Ctra. de Utrera, Km. 1 41013 Sevilla (España) Tel.: +34 954 977 433 E-mail: [email protected]

PRINCIPAL Crespo García, María López Vilaret, Karel INVESTIGATOR Teresa. Mauricio. Bachelor degree. Bachelor degree. Cantero Lorente, Rodríguez Peñuela, José Luis Del Río Arronis, Paula. Sofía. Diplomado. Technician. LIST OF García Solís, David. Velasco Montero, María PERSONNEL Bachelor degree. del Mar. Bachelor degree. Atienza Ruiz, Mercedes. Gil Néciga, Eulogio. PhD. Bachelor degree.

ABSTRACT Using the cohort of the SIGNAL project, we have established the pattern of cerebral changes in asymptomatic older adults at risk for Alzheimer’s disease (AD). The Abeta+ group exhibited thickening of middle temporal regions, while p-tau+ individuals showed thinning in the superior parietal and orbitofrontal cortices. Subjects with abnormal CSF biomarkers further showed white matter atrophy and more segregated cortical networks. These results revealed a very precise pattern of cerebral changes preceding the onset of AD symptoms (Cantero et al., 2018). Further experiments performed in patients with mild cognitive impairment (MCI) showed that ApoE4 carriers are unsuccessful in re- cruiting accessory cortical regions to improve memory performance; this aspect depend- ed largely on the loss of integrity and functionality of the temporal lobe. These findings may be helpful to increase our knowledge about cerebral dysfunctions hampering neural compensation in MCI patients carrying the ApoE4 genotype (Prieto del Val et al., 2018). Given that circadian rhythmicity is progressively disrupted in senescence, we have assessed the effects of PER3 genotype, one of the most studied clock genes in humans, on aging-related changes in cognitive function and cortical integrity. Our results revealed that PER35/5 carriers had poorer cognitive performance and lower cortical integrity (structural and functional) than PER34/4. These findings may reflect cerebral vulnerabil-

40 CIBERNED 2018 ANNUAL REPORT ity associated with PER3 genotypes in late life (Dewandre et al., 2018). We have further reviewed evidence supporting the potential mediating role of chronic low-grade systemic inflammation in the complex relationship between impaired sleep, dysfunctional adiposity, and cognitive decline through the common pathway of neuroinflammation. This work led to a multi-factor model of aging-related cognitive decline that identifies sleep and adiposity as modifiable lifestyle factors that can be targeted to maximize cognitive function and quality of life in the elderly (Atienza et al., 2018).

KEYWORDS Alzheimer’s disease, aging, biomarkers, brain imaging markers.

PUBLICATIONS 2018 Dewandre D, Atienza M, Sanchez-Espinosa M.P, Cantero J.L. Effects of PER3 clock gene polymorphisms on aging-related changes of the cerebral cortex. Brain Structure and Function. 2018;223(2):597-607. PMID: 28900721. Garcia Solis D. A new stage in the diagnosis of Alzheimer’s Disease Una nueva etapa en el diagnostico de la enfermedad de Alzheimer. Revista Espanola de Medicina Nuclear e Imagen Molecular. 2018;37(6):339-340. PMID: 30471746. Maldonado-Lasuncion I, Atienza M, Sanchez-Espinosa MP, Cantero JL. Aging-Related Chang- es in Cognition and Cortical Integrity are Associated With Serum Expression of Candidate MicroRNAs for Alzheimer Disease.Cerebral cortex (New York, N.Y. : 1991). 2018. PMID: 30590432. Camacho V, Gomez-Grande A, Sopena P, Garcia-Solis D, Gomez Rio M, Lorenzo C. et al. Amy- loid PET in neurodegenerative diseases with dementia PET amiloide en enfermedades neurodegenerativas que cursan con demencia. Revista Espanola de Medicina Nuclear e Imagen Molecular. 2018;37(6):397-406. PMID: 29776894. Prieto Del Val L, Cantero JL, Baena D, Atienza M. Damage of the temporal lobe and APOE status determine neural compensation in mild cognitive impairment.Cortex; a journal devoted to the study of the nervous system and behavior. 2018;101. PMID: 29475078. Atienza M, Ziontz J, Cantero J.L. Low-grade inflammation in the relationship between sleep disruption, dysfunctional adiposity, and cognitive decline in aging. Sleep Medicine Reviews. 2018;42:171-183. PMID: 30241997. Cantero JL, Atienza M, Sanchez-Juan P, Rodriguez-Rodriguez E, Vazquez-Higuera JL, Pozueta A et al. Cerebral changes and disrupted gray matter cortical networks in asymptomatic older adults at risk for Alzheimer’s disease.Neurobiology of aging. 2018;64. PMID: 29331877.

41 RESEARCH PROJECTS 2018 Code: PI2016/01. Title: Alteraciones del metabolismo gluco-lipidico y desarrollo de la demencia de Alzheimer. Principal investigator: Ignacio Torres Aleman. CIBERNED’s collaboration: Yes. CIBERNED groups: G409 ; G402; G511; G502; G412. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

Code: PSI2016-81881-REDT. Title: Aplicaciones clinicas de la neuroimagen funcional. Principal investigator: Fernando Maestu Unturbe. CIBERNED’s collaboration: No. CIBERNED groups: G511. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 20000. Duration: 2017-2019.

Code: SAF2017-85310-R. Title: Efectos de la concentracion de beta amiloide cerebral sobre el sueno fisiologico en personas mayores sin deterioro cognitivo. Principal investigator: Jose Luis Cantero Lorente. CIBERNED’s collaboration: No. CIBERNED groups: G511. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 99999. Duration: 2018-2020.

Code: SES_2017. Title: Efectos de la concentracion de beta amiloide cerebral sobre el sueno fisiologico en personas mayores. Principal investigator: Jose Luis Cantero Lorente. CIBERNED’s collaboration: No. CIBERNED groups: G511. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Sociedad Espanola de Sueno. Funding: 10000. Duration: 2017-2018.

Code: PSI2017-85311-P. Title: La corteza prefrontal como posible mediador de la actividad fisica aguda y de la aptitud cardiorrespiratoria sobre la memoria asociativa. Principal investigator: Mercedes Atienza Ruiz. CIBERNED’s collaboration: No. CIBERNED groups: G511. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 100700. Duration: 2018-2020.

42 CIBERNED 2018 ANNUAL REPORT

502 | Eva María Carro Díaz

INSTITUTO DE INVESTIGACIÓN HOSPITAL 12 DE OCTUBRE. Avda. de Córdoba s/n 28041 Madrid Teléfono: +34 91 390 8765 Fax: +34 91 390 85 44 E-mail: [email protected]

PRINCIPAL Benito León, Julián. Llamas Velasco, Sara. INVESTIGATOR PhD. Bachelor degree. de la Cueva de Vicente, Molina Arjona, Jose Carro Díaz, Eva María. Macarena. Antonio. Bachelor degree. PhD. González Sánchez, Pérez martínez, David LIST OF Marta. Andrés. PERSONNEL Bachelor degree. Bachelor degree. Antequera Tienda, Hernández Gallego, Villarejo Galende, Desireé. Jesús. Alberto. Technician. PhD. PhD. Bartolomé Robledo, Herrero San Martín, Fernándo. Alejandro. PhD. Bachelor degree.

ABSTRACT Our scientific activity has been focused at studying pathophysiological mechanisms of neurodegenerative diseases, mainly Alzheimer’s disease and associated dementias, identifying new therapeutic targets, risk factors and biomarkers. Specific activities are: 1. Preclinical studies on new risk factors, biomarkers of disease, mechanisms and new therapeutic strategies in Alzheimer’s disease and other neurodegenerative disorders. a) Research on risk factors that highlight the relationship between alteration in blood cells (platelets) and Alzheimer’s disease (AD), through mechanisms and inflammatory responses at central and peripheral level. These findings complement and expand previous reports concerning the morphological and functional alterations in AD platelets, and provide more insights into possible mechanisms

43 that participate in the multifactorial and systemic damage in AD. b) Preclinical research searching for new therapeutic targets and design of potential drugs using cellular and animal experimental models of AD and Parkinson’s disease (PD). c) Studies on molecular pathology in neurodegenerative disorders analysing new pathophysiological mechanisms, in particular, mitochondrial physiology, and regulation on neurogenic and memory processes 2. Epidemiological research on neurodegenerative diseases, particularly risk factors. Epidemiological analysis NEDICES cohort. 3. Clinical research in dementia. Studies searching for genetic mutations associated with dementia. Determination of the clinical utility of dementia scales for the early diagnosis of AD and mild cognitive impairment (MCI).

KEYWORDS Alzheimer’s disease, Parkinson’s disease, Frontotemporal Dementia, biomarkers, systemic alterations, bioenergetics, therapeutic drugs.

PUBLICATIONS 2018 Contador I, Bermejo-Pareja F, Fernandez-Calvo B, LLamas S, Villarejo A, Vega S. et al. Disa- bility subtypes and mortality rates in older adults: A longitudinal population-based study (NEDICES). Archives of Gerontology and Geriatrics. 2019;80:88-94. epub2018. PMID: 30391685. Vida C, de Toda I.M, Garrido A, Carro E, Molina J.A, De la Fuente M. Impairment of several immune functions and redox state in blood cells of Alzheimer’s disease patients. Relevant role of neutrophils in oxidative stress. Frontiers in Immunology. 2018;8(JAN). PMID: 29375582. Quintela T, Albuquerque T, Lundkvist G, Carmine Belin A, Talhada D, Goncalves I. et al. The choroid plexus harbors a circadian oscillator modulated by estrogens. Chronobiology Inter- national. 2018;35(2):270-279. PMID: 29172740. Sanchez-Ferro A, Matarazzo M, Martinez-Martin P, Martinez-Avila J.C, Gomez de la Camara A, Giancardo L. et al. Minimal Clinically Important Difference for UPDRS-III in Daily Practice. Movement Disorders Clinical Practice. 2018;5(4):448-450. PMID: 30838303. Ettcheto M, Sanchez-Lopez E, Gomez-Minguez Y, Cabrera H, Busquets O, Beas-Zarate C. et al. Peripheral and Central Effects of Memantine in a Mixed Preclinical Mice Model of Obesity and Familial Alzheimer’s Disease. Molecular Neurobiology. 2018;55(9):7327-7339. PMID: 29404958. Graus F, Escudero D, Oleaga L, Bruna J, Villarejo-Galende A, Ballabriga J. et al. Syndrome and outcome of antibody-negative limbic encephalitis. European Journal of Neurology. 2018;25(8):1011-1016. PMID: 29667271. Figueiro-Silva J, Antequera D, Pascual C, de la Fuente Revenga M, Volt H, Acuna-Castroviejo D. et al. The Melatonin Analog IQM316 May Induce Adult Hippocampal Neurogenesis and

44 CIBERNED 2018 ANNUAL REPORT

Preserve Recognition Memories in Mice. Cell Transplantation. 2018;27(3):423-437. PMID: 29873251. del Cerro P, Alquezar C, Bartolome F, Gonzalez-Naranjo P, Perez C, Carro E. et al. Activation of the Cannabinoid Type 2 Receptor by a Novel Indazole Derivative Normalizes the Survival Pattern of Lymphoblasts from Patients with Late-Onset Alzheimer’s Disease. CNS Drugs. 2018;32(6):579-591. PMID: 29736745.

Bartolome F, De La Cueva M, Pascual C, Antequera D, Fernandez T, Gil C. et al. Amyloid β-induced impairments on mitochondrial dynamics, hippocampal neurogenesis, and memory are restored by phosphodiesterase 7 inhibition. Alzheimer’s Research and Therapy. 2018;10(1). PMID: 29458418. Bartolome F, Esteras N, Martin-Requero A, Boutoleau-Bretonniere C, Vercelletto M, Gabelle A et al. Author Correction: Pathogenic p62/SQSTM1 mutations impair energy metabolism through limitation of mitochondrial substrates.Scientific reports. 2018;8(1). PMID: 29497142. Contador I, Fernández-Calvo B, Rueda-Revé L, Olazarán J, Bermejo-Pareja F. Characterizing functional alterations in instrumental activities of daily living using latent class analysis: a population-based study (NEDICES).Aging & mental health. 2018. PMID: 30450947. Leon Ruiz M, Sospedra M, Arce Arce S, Tejeiro-Martinez J, Benito-Leon J. Current evidence on the potential therapeutic applications of transcranial magnetic stimulation in multiple sclerosis: A systematic review of the literature Evidencias actuales sobre las potenciales aplicaciones terapéuticas de la estimulación magnética transcraneal en la esclerosis múltiple: revisión sistemática de la literatura. Neurologia. 2018. Leon Ruiz M, Rodriguez Sarasa M.L, Sanjuan Rodriguez L, Benito-Leon J, Garcia-Albea Ristol E, Arce Arce S. Current evidence on transcranial magnetic stimulation and its potential usefulness in post-stroke neurorehabilitation: Opening new doors to the treatment of cerebrovascular disease Evidencias actuales sobre la estimulación magnética transcraneal y su utilidad potencial en la neurorrehabilitación postictus: Ampliando horizontes en el tratamiento de la enfermedad cerebrovascular. Neurologia. 2018;33(7):459-472. PMID: 27161423. León Ruiz M, Mitchell AJ, Benito-León J. Delusional parasitosis in multiple sclerosis: An enig- matic manifestation of a multifaceted disease.Neurologia (Barcelona, Spain). 2018. PMID: 29891338. Benito-Leon J. Epidemiology of parkinson’s disease in Spain and its contextualisation in the world Epidemiología de la enfermedad de Parkinson en España y su contextualización mun- dial. Revista de Neurologia. 2018;66(4):125-134. PMID: 29435969. Lopez-Blanco R, Velasco M.A, Mendez-Guerrero A, Romero J.P, del Castillo M.D, Serrano J.I. et al. Essential tremor quantification based on the combined use of a smartphone and a smartwatch: The NetMD study. Journal of Neuroscience Methods. 2018;303:95-102. PMID: 29481820. Lopez-Blanco R, Benito-Leon J, Llamas-Velasco S, Del Castillo M.D, Serrano J.I, Rocon E. et al. Interaction with touchscreen smartphones in patients with essential tremor and healthy individuals Interacción con pantalla táctil de smartphone en pacientes con temblor esencial y sujetos sanos. Neurologia. 2018. Leon-Ruiz M, Perez-Martinez D.A, Benito-Leon J. Major neurological complications of geni- tourinary cancers Principales complicaciones neurologicas de los canceres nefrourologicos. Revista de neurologia. 2018;67(11):441-452. PMID: 30484277. Benito-Leon J, Laurence M. Malassezia in the central nervous system and multiple sclerosis.

45 Infection. 2018. PMID: 30120719. Gonzalez-Sanchez M, Diaz T, Pascual C, Antequera D, Herrero-San Martin A, Llamas-Velas- co S. et al. Platelet Proteomic Analysis Revealed Differential Pattern of Cytoskeletal- and Immune-Related Proteins at Early Stages of Alzheimer’s Disease. Molecular Neurobiology. 2018;55(12):8815-8825. PMID: 29603091. Posa D, Martinez-Gonzalez L, Bartolome F, Nagaraj S, Porras G, Martinez A. et al. Recapit- ulation of Pathological TDP-43 Features in Immortalized Lymphocytes from Sporadic ALS Patients. Molecular Neurobiology. 2018. PMID: 30030753. Llamas-Velasco S, Garcia-Redondo A, Herrero-San Martin A, Puertas Martin V, Gonza- lez- Sanchez M, Perez-Martinez D.A. et al. Slowly progressive behavioral frontotemporal dementia with C9orf72 mutation. Case report and review of the literature. Neurocase. 2018;24(1):68-71. PMID: 29355451. Benito-Leon J, Serrano J.I, Louis E.D, Holobar A, Romero J.P, Povalej-Brzan P. et al. Tremor severity in Parkinson’s disease and cortical changes of areas controlling movement sequencing: A preliminary study. Journal of Neuroscience Research. 2018;96(8):1341-1352. PMID: 29660812. Catalan M.J, Molina-Arjona J.A, Mir P, Cubo E, Arbelo J.M, Martinez-Martin P. Improvement of impulse control disorders associated with levodopa–carbidopa intestinal gel treatment in advanced Parkinson’s disease. Journal of Neurology. 2018;265(6):1279-1287. PMID: 29557989.

RESEARCH PROJECTS 2018 Code: CAM-B2017/BMD-3700. Title: Bases metabólicas de la neurodegeneración (NEUROMETAB-CM) Programas de Ac- tividades de I+D entre Grupos de Investigacion de la Comunidad de Madrid en Tecnologias y en Biomedicina. Principal investigator: Jose Gonzalez Castano. CIBERNED’s collaboration: Yes. CIBERNED groups: G401; G502; G111; G409; G412; G205; G110. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 87499,89. Duration: 2014-2020.

Code: PI2016/01. Title: Alteraciones del metabolismo gluco-lipidico y desarrollo de la demencia de Alzheim- er. Principal investigator: Ignacio Torres Aleman. CIBERNED’s collaboration: Yes. CIBERNED groups: G409 ; G402; G511; G502; G412. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

46 CIBERNED 2018 ANNUAL REPORT

Code: DTS15/00141. Title: Evaluacion del impacto de la imagen PET de amiloide en el diagnostico de los pacientes con deterioro cognitivo evaluados por sospecha de Alzheimer. Principal investigator: Dr. Javier Arbizu. CIBERNED’s collaboration: Yes. CIBERNED groups: G504 ; G502; G609. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 59139. Duration: 2016-2018.

Code: PI15-00780. Title: Inflamacion periferica como factor de riesgo en la patologia de la enfermedad de Alzheimer: nuevas herramientas diagnosticas y dianas terapeuticas. Principal investigator: Eva Carro. CIBERNED’s collaboration: No. CIBERNED groups: G502. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 100000. Duration: 2016-2018.

47 413 | Joan Xavier Comella Carnicé

VHIR – VALL HEBRON INSTITUT DE RECERCA Passeig Vall d’Hebron 119-129 08035 Barcelona Tel.: +34 93 489 3807 Web: www.vhir.org E-mail: [email protected]

PRINCIPAL LIST OF INVESTIGATOR PERSONNEL Comella Carnice, Joan López Soriano, Joaquín. Xavier. PhD. Martínez Sirés, Anna. Bachelor degree. Pérez García, María José. PhD.

ABSTRACT Our main interest is to characterize the mechanisms controlling neuronal death induced by a group of receptors collectively known as death receptors, and also the relevance of some of their intracellular antagonists. Among those, we are most interested in the antagonists expressed in nervous system, such as FAIM-L, as well as their molecular partners. Our main objective is to characterize these proteins at the molecular level, their involvement in neuronal differentiation and physiology, and their possible role in different pathologies, mainly neurodegenerative diseases. The knowledge of the molecular basis of these receptor antagonists and the activation of survival signaling pathways could be of high relevance to understand the etiology or to open new therapeutic strategies for the treatment of neurodegenerative diseases, such as Alzheimer’s disease. We have recently characterized, in collaboration with different CIBERNED groups, the role of FAIM-L in the development of Alzheimer’s disease, both in animal models (APP/PS1) and patients (Carriba et al., 2015). At present we are further characterizing this role of FAIM-L, and with this aim we have started the characterization of the Faim knockout model, through immunohistochemistry, electrophysiological and behavior studies (Calleja-Yagüe et al, submitted). Surprisingly, this model shows an epileptic phenotype, as well as some behavior alterations, which enlarge the rele-

48 CIBERNED 2018 ANNUAL REPORT vance of FAIM-L functions in nervous system. We have also developed adenoassociated viral vectors, which will allow us to overexpress or downregulate FAIM-L levels in brain, at present in use in the murine models of Alzheimer’s disease (in the frame of a CIBERNED collaborative project), in order to study the role of FAIM-L in the development or progression of the disease, or its potential protector effect in these models. Our lab has recently characterize new unexpected physiological roles for FAIM-L, since we have observed that it is implicated in the control of some non-apoptotic effects of caspases, such as axonal degeneration and long term depression (LTD), through modulation of XIAP levels (Martínez-Mármol et al., 2016). These events of synaptic plasticity are also modulated in neurodegenerative diseases, thus positioning FAIM-L as a good candidate for the treatment of the disease. At present we are also characterizing the interaction among FAIM-L and XIAP with other proapototic partners such as Siva-1. We have verified that Siva-1, XIAP and FAIM-L interact among them. Also, Siva-1 regulates GluA2 receptors internalization, and plays and opposite role to FAIM-L and XIAP on the apoptotic and non-apoptotic functions of caspase-3 (Coccia et al., submitted), which may help better explain the role of FAIM-L in neuron physiology. We have also characterized two new isoforms of FAIM, one of them specific from neurons (Coccia et al., 2017), although their physiological role is still not fully characterized. Our main research lines at present are: 1) To study FAIM-L function in in vitro and in vivo models of AD, and its relation with TNF (its duality as a pro-apoptotic molecule and at the same time as a survival promoter) and its signaling pathways, particularly NFkB, as well in the cross-talk neuron/glia. 2) To characterize FAIM-L functional partners, particularly XIAP and Siva-, and their implications in neuron physiology (neuronal plasticity) and different neurodegenerative diseases (neuronal apoptosis, synaptic degeneration). 3) To study microRNAs that could modulate FAIM expression levels, both in physiological and pathologic states. 4) To study the role of FAIM-L in the retina, and its likely association with neurodegeneration associated to type II diabetes, as a proxy of AD neurodegeneration.

KEYWORDS Alzheimer, neuroinflammation, death receptors, FAIM, TNF, Fas.

PUBLICATIONS 2018 Gomez-Arboledas A, Davila J.C, Sanchez-Mejias E, Navarro V, Nunez-Diaz C, Sanchez-Varo R. et al. Phagocytic clearance of presynaptic dystrophies by reactive astrocytes in Alzheimer’s disease. GLIA. 2018;66(3):637-653. PMID: 29178139.

49 Simó M, Navarro X, Yuste VJ, Bruna J. Autonomic nervous system and cancer.Clinical autonomic research: official journal of the Clinical Autonomic Research Society. 2018;28(3). PMID: 29594605. Fuschini G, Cotrufo T, Ros O, Muhaisen A, Andres R, Comella J.X. et al. Syntaxin-1/TI-VAMP SNAREs interact with Trk receptors and are required for neurotrophin-dependent outgrowth. Oncotarget. 2018;9(89):35922-35940. PMID: 30542508.

RESEARCH PROJECTS 2018 Code: PI2017/04. Title: Disfunción glial en la enfermedad de Alzheimer: implicaciones patogénicas y potencial clínico. Principal investigator: Javier Vitorica. CIBERNED’s collaboration: Yes. CIBERNED groups: G411; G415; G101; G407; G413. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 240000. Duration: 2017-2019.

Code: PI2015-2/02. Title: Potencial patologico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal investigator: Joan X. Comella. CIBERNED’s collaboration: Yes. CIBERNED groups: G413 ; G415; G204; G108; G411. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350000. Duration: 2016-2018.

Code: SAF2016-80236-R. Title: Relevancia del antagonista de receptores de muerte, FAIM-L, en la enfermedad de alzheimer. Principal investigator: Joan X. Comella. CIBERNED’s collaboration: No. CIBERNED groups: G413. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 273459. Duration: 2016-2019.

50 CIBERNED 2018 ANNUAL REPORT

403 | Javier de Felipe Oroquieta

LABORATORIO CAJAL DE CIRCUITOS CORTICALES, CTB, UPM Campus de Montegancedo Pozuelo de Alarcón - 28223 Madrid Tel.: +34 91 336 4660

INSTITUTO CAJAL (CSIC). Avenida Doctor Arce 37. 28002 Madrid

PRINCIPAL Fernández Bouzo, Muñoz Céspedes, INVESTIGATOR Montserrat. Alberto. Bachelor degree. PhD. De Felipe Oroquieta, Fernández García, Ostos Campillo, Sandra. Javier. Laura. Bachelor degree. Bachelor degree. Regalado Reyes, Mari LIST OF Fernaud Espinosa, Carmen. PERSONNEL Isabel. Bachelor degree. Alonso Nanclares, Lidia. PhD. Rodriguez Cela, Yago. PhD. Flores Romero, Pilar. Technician. Alvarez Pérez, Maria del PhD. Rodriguez Sanchez, José Carmen. García Rámirez, Ana Rodrigo. Technician. Isabel. Bachelor degree. Antón Fernández, Technician. rojo Salvador, Alejandro. González Soriano, Concepción. Bachelor degree. Juncal M. PhD. Aparicio Torres, Bachelor degree. Sánchez Ponce, Diana. Guillermo. Kastanauskaite, Asta. PhD. Bachelor degree. PhD. Santuy Muñoz, Andrea. Barbado Amigo, Marta. León Espinosa, Gonzalo. Bachelor degree. Bachelor degree. PhD. Tapia Gonzalez, Silvia. Benavides Piccione, Marín Horcajadas, Bachelor degree. Ruth. Mirian. Turégano López, Marta. PhD. Technician. Bachelor degree. Blázquez Llorca, Lidia. Merchán Pérez, Ángel. Valdés Lora, Lorena. PhD. PhD. Technician. Cano Laiseca, Débora. Miguéns Vázquez, Technician. Miguel. Dominguez Alvaro, Bachelor degree. Marta. Montero Crespo, Marta. Bachelor degree. Bachelor degree.

51 ABSTRACT The two main lines of research that we have developed are: 1. Neurochemical and microanatomical analysis of the cerebral cortex in different mouse models for Alzheimer’s disease, with the idea of obtaining data about the substrate and temporal course of the microanatomical alterations that occur in Alzheimer’s disease. 2. Neurochemical and microanatomical analysis of the cerebral cortex of patients with Alzheimer’s disease. In particular, we are performing a quantitative and qualitative analysis of the synaptic circuits and neurochemical characteristics of the cerebral cortex of these patients. The aim of this objective is to try to know in detail the alterations of the neuronal circuits in relation with the senile plaques and the presence of neurofibrillary tangles.

KEYWORDS Cerebral cortex, microorganization, neuronal circuits, electron microscopy, Alzheimer.

PUBLICATIONS 2018 Merino-Serrais P, Loera-Valencia R, Rodriguez-Rodriguez P, Parrado-Fernandez C, Ismail M.A, Maioli S. et al. 27-Hydroxycholesterol Induces Aberrant Morphology and Synaptic Dysfunc- tion in Hippocampal Neurons. Cerebral cortex (New York, N.Y. : 1991). 2019;29(1):429-446. epub2018. PMID: 30395175. Luengo-Sanchez S, Fernaud-Espinosa I, Bielza C, Benavides-Piccione R, Larranaga P, DeFelipe J. 3D morphology-based clustering and simulation of human pyramidal cell dendritic spines. PLoS Computational Biology. 2018;14(6). PMID: 29897896. Aliaga Maraver J.J, Mata S, Benavides-Piccione R, DeFelipe J, Pastor L. A method for the sym- bolic representation of neurons. Frontiers in Neuroanatomy. 2018;12. PMID: 30618651. Santuy A, Turegano-Lopez M, Rodriguez J.R, Alonso-Nanclares L, Defelipe J, Merchan-Perez A. A quantitative study on the distribution of mitochondria in the neuropil of the juvenile rat somatosensory cortex. Cerebral Cortex. 2018;28(10):3673-3684.

Furcila D, Defelipe J, Alonso-Nanclares L. A Study of Amyloid-β and Phosphotau in Plaques and Neurons in the Hippocampus of Alzheimer’s Disease Patients. Journal of Alzheimer’s Disease. 2018;64(2):417-435. PMID: 29914033. Zhu F, Cizeron M, Qiu Z, Benavides-Piccione R, Kopanitsa M.V, Skene N.G. et al. Architecture of the Mouse Brain Synaptome. Neuron. 2018;99(4):781-799.e10. PMID: 30078578. Rockland KS, DeFelipe J. Editorial: Why Have Cortical Layers? What Is the Function of Layer- ing? Do Neurons in Cortex Integrate Information Across Different Layers?. Frontiers in neuroanatomy. 12. PMID: 30356892. Eyal G, Verhoog M.B, Testa-Silva G, Deitcher Y, Piccione R.B, DeFelipe J. et al. Human cortical pyramidal neurons: From spines to spikes via models. Frontiers in Cellular Neuroscience. 2018;12. PMID: 30008663.

52 CIBERNED 2018 ANNUAL REPORT

Leon-Espinosa G, Anton-Fernandez A, Tapia-Gonzalez S, DeFelipe J, Munoz A. Modifications of the axon initial segment during the hibernation of the Syrian hamster. Brain Structure and Function. 2018;223(9):4307-4321. PMID: 30219944. Varando G, Benavides-Piccione R, Munoz A, Kastanauskaite A, Bielza C, Larranaga P. et al. Mul- tiMap: A tool to automatically extract and analyse spatial microscopic data from large stacks of confocal microscopy images. Frontiers in Neuroanatomy. 2018;12. PMID: 29875639. Rodriguez J.-R, Turegano-Lopez M, DeFelipe J, Merchan-Perez A. Neuroanatomy from meso- scopic to nanoscopic scales: An improved method for the observation of semithin sections by high-resolution scanning electron microscopy. Frontiers in Neuroanatomy. 2018;12. PMID: 29568263. Leguey I, Benavides-Piccione R, Rojo C, Larranaga P, Bielza C, DeFelipe J. Patterns of Dendrit- ic Basal Field Orientation of Pyramidal Neurons in the Rat Somatosensory Cortex. eNeuro. 2018;5(6). PMID: 30656209. Roy M, Sorokina O, McLean C, Tapia-Gonzalez S, DeFelipe J, Armstrong J.D. et al. Regional diversity in the postsynaptic proteome of the mouse brain. Proteomes. 2018;6(3). PMID: 30071621. Fernandez-Cabrera M.R, Higuera-Matas A, Fernaud-Espinosa I, DeFelipe J, Ambrosio E, Miguens M. Selective effects of Δ9-tetrahydrocannabinol on medium spiny neurons in the striatum. PLoS ONE. 2018;13(7). PMID: 30048477. Santuy A, Rodriguez J.-R, DeFelipe J, Merchan-Perez A. Study of the size and shape of synapses in the juvenile rat somatosensory cortex with 3D electron microscopy. eNeuro. 2018;5(1). PMID: 29387782. Dominguez-alvaro M, Montero-Crespo M, Blazquez-Llorca L, Insausti R, DeFelipe J, Alon- so-Nanclares L. Three-dimensional analysis of synapses in the transentorhinal cortex of Alzheimer’s disease patients. Acta Neuropathologica Communications. 2018;6(1). PMID: 29499755. Mihaljevic B, Larranaga P, Benavides-Piccione R, Hill S, DeFelipe J, Bielza C. Towards a supervised classification of neocortical interneuron morphologies. BMC Bioinformatics. 2018;19(1). PMID: 30558530. Kwon T, Merchán-Pérez A, Rial Verde EM, Rodríguez JR, DeFelipe J, Yuste R. Ultrastructural, Molecular and Functional Mapping of GABAergic Synapses on Dendritic Spines and Shafts of Neocortical Pyramidal Neurons.Cerebral cortex (New York, N.Y. : 1991). 2018. PMID: 30113619.

RESEARCH PROJECTS 2018 Code: PI2016/05. Title: Dream inhibitors and Alzheimer´s Disease. Principal investigator: Jose Ramon Naranjo Orovio. CIBERNED’s collaboration: Yes. CIBERNED groups: G307; G106; G403. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2016-2018.

53 Code: Cajal Blue Brain Project. Title: Cajal Blue Brain Project. International Blue Brain Proyect. Principal investigator: Javier De Felipe. CIBERNED’s collaboration: Yes. CIBERNED groups: G204; G403. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Ecole Polytechnique Federale de Lausanne (Suiza). Funding: ND. Duration: 2009-2019.

Code: SAF2015-66603-P. Title: Estudio de la microorganizacion de la corteza cerebral en pacientes de Azheimer y del amster como modelo para estudiar la Fosforilacion de Tau. Principal investigator: Javier Defelipe. CIBERNED’s collaboration: No. CIBERNED groups: G403. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 196000. Duration: 2016-2018.

Code: ZEN-15-321663. Title: The Pyramidal Neuron in Cognition and Alzheimer’s Disease. Principal investigator: Javier Defelipe. CIBERNED’s collaboration: No. CIBERNED groups: G403. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Alzheimer’s Association. Funding: ND. Duration: 2015-2018.

PHD DISSERTATIONS 2018 Author: Andrea Santuy Muñoz. Title: Estudio tridimensional de la ultraestructura de la Estudio tridimensional de la ultraestructura de la corteza cerebral de la rata.. Date: 11/1/2018. Supervisor: Javier De Felipe Oroquieta.

Author: Alejandro Antón Fernández. Title: Estudio del aparato de Golgi y del segmento inicial del axón de neuronas corticales en el cerebro normail y en la enfermedad de Alzheimer. Date: 11/5/2018. Supervisor: Javier De Felipe Oroquieta.

54 CIBERNED 2018 ANNUAL REPORT

503 | Isidro Ferrer Abizanda

DEPARTMENT OF PATHOLOGY AND EXPERIMENTAL THERAPEUTICS, UNIVERSITY OF BARCELONA Bellvitge University Hospital-IDIBELL. Calle Feixa Llarga, sn. 08907 Hospitalet de Llobregat, Barcelona Tel.: +34 93 403 5808 E-mail: [email protected]

PRINCIPAL Andrés Benito, Pol. Garcia Esparcia, Paula. INVESTIGATOR Bachelor degree. PhD. Barrachina Castillo, Llorens Torres, Franc. Ferrer Abizanda, Isidro Marta. PhD. PhD. Povedadno Panadés, Carmona Murillo, Mónica. LIST OF Margarita. Bachelor degree. PERSONNEL Technician. Alvear Contreras, Raissa Díaz Lucena, Daniela del Camilla. Valle. Bachelor degree. Technician.

ABSTRACT As a result of the retirement of the PI in the Bellvitge University Hospital (HUB), the laboratories of the former Institute of Neuropathology (INP) at the HUB have been dismantled including the Biosecurity P3 facility used for the study of prion diseases. The Brain Bank of the INP is now a branch of the HUB-ICO-IDIBELL biobank; the management of the brain bank, including the procedures and availability of brain samples for research, is under the direction of IDIBELL. The INP laboratories of cell cultures at the Hospital Duran i Reynals now under the control of IDIBELL have been partially occupied by another group. The laboratory at the Barcelona University (UB) is maintained due to the position of the PI as a professor at the UB. An agreement has been signed between CIBERNED and IRTA/CRESA to continue the studies of prion diseases. As a consequence of pruning, the number of members of the group is reduced. Howe- ver, internal research and collaboration with national and international scientists have suffered from little modifications. We have continued with old topics and have developed new lines including: 1) the research of biomarkers in blood and CSF for the diagnosis and prognosis of several neurodegenerative diseases manifested with

55 cognitive impairment and dementia, and ALS, with the work of Marta Barrachina, PhD and Franc Llorens, PhD; 2) combined clinical and pathological studies in motor neuron diseases with the doctoral student Pol Andrés-Benito in collaboration with the reference Unit of ALS and neuromuscular diseases of the Service of Neurology at the HUB led by Dr. Mónica Povedano; 3) study of molecular neuropathology of cerebral cortex, based on the combined use of “omics” and bioinfomartics in aging, Alzhei- mer’s disease, dementia with Lewy bodies and Parkinson’s disease without and with dementia, in addition to frontotemporal lobar degeneration-TDP, ALS, and Creutz- feldt-Jakob disease, with the help of Paula García-Esparcia, PhD and the technician Margarita Carmona, and the valuable external collaborations with various national and international centres and groups; 4) study of protein-protein interactions and of the progression of neurodegenerative diseases with abnormal protein aggregates using natural and recombinant proteins in animal and cellular models in collaboration with the CIBERNED’s group led José Antonio del Rio, PhD; 5) studies dealing with the role of astrogliopathy and oligodendrogliopathy in the pathogenesis of human neurodegenerative diseases with abnormal protein aggregates; and 6) experimental procedures testing new putative treatments of Alzheimer’s disease in mouse models, mainly performed by Ester Aso, PhD. The list of the names of the valuable collabo- rators inside and outside CIBERNED, fundamental in the work of the neuropathology group, is too long to be named here. Journals of published papers in alphabetical order include: Aging (1), Alzh & Dement (1), Autophagy (2), Brain (1), Biochem Pharmacol (2), Brain Pathol (3), Cerebral cortex (1), EMBO Mol Med (1), Exp Gerontol (1), FASEB J (1), Frontiers (4), Hand Clin Neurol (1), Int J Nanomedicine (1), J Gerontol Biol (1), J Neuropathol Exp Neurol (1), Mol Brain (1), Mol Neurobiol (5), Nature Med (1), Neuroglia (1), Neuropathology (1), PLos Pathog (1), Prion (1) and Progr Neurobiol (2). An additional paper, “Sysyphus in Neverland”, was written following the kind invitation of the Journal of Alzheimer’s disease to describe my scientific trajectory over the years, a convenient remainder that my gratifying game is almost over. Finally, a major event in the history of the group has been the creation by Dr. Marta Barrachina of the Company ADMIT THERAPEUTICS, S.L. focused on the delineation and development of predictor biomarkers of Alzheimer’s disease based on her recent and robust discoveries in the laboratory. Other aspects have not been less important: the training of different students who have come to the laboratory, for variable periods to learn neuropathology techniques or to do collaborative work, from different Spanish groups but also from Austria, Germany, Italy, Greece, UK, Colombia and Venezuela. The experience of working with young people is also enriching for the persons who act as tutors during the stay of the students. In addition, different seminars and invited communications for associations of patients and relatives, and the general public, thus favoring the interaction of research with civil society have been conducted. Another interesting point has been the demand of topic-free compulsory written own studies in the third course of medicine of the UB focused on the impact of diseases in the personal life of individuals or in the history of the humankind with the aim of inviting students to have and develop a more humanistic perspective of medicine. I am also very grateful to have received the International Prize Mano Amiga for “his professional involvement and international recognition in the research and teaching on Alzheimer’s disease”. No need to say that the prize must be shared with all the people who had worked with me over time.

56 CIBERNED 2018 ANNUAL REPORT

KEYWORDS Neurodegenerative diseases with aggregates of abnormal proteins, molecular neuropathology, Alzheimer’s, synucleinopathies, taupathies, prion diseases, biomarkers.

PUBLICATIONS 2018 Tazelaar G.H.P, Dekker A.M, van Vugt J.J.F.A, van der Spek R.A, Westeneng H.-J, Kool L.J.B.G. et al. Association of NIPA1 repeat expansions with amyotrophic lateral sclerosis in a large international cohort. Neurobiology of Aging. 2019;74:234.e9-234.e15. epub2018. PMID: 30342764. Pinho R, Paiva I, Jercic K.G, Fonseca-Ornelas L, Gerhardt E, Fahlbusch C. et al. Nuclear localization and phosphorylation modulate pathological effects of alpha-synuclein. Human molecular genetics. 2019;28(1):31-50. epub2018. PMID: 30219847. Lachen-Montes M, Gonzalez-Morales A, Iloro I, Elortza F, Ferrer I, Gveric D. et al. Unveiling the olfactory proteostatic disarrangement in Parkinson’s disease by proteome-wide profiling. Neurobiology of Aging. 2019;73:123-134. epub2018. PMID: 30342273. Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Garcia-Aznar J.M. et al. Erratum to: Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons (Molec- ular Neurobiology, (2018), 55, 3, (1847-1860), 10.1007/s12035-017-0451-4). Molecular Neurobiology. 2018;55(3):1861-. Arias D.M, Sole-Bundo M, Garfi M, Ferrer I, Garcia J, Uggetti E. Integrating microalgae tertiary treatment into activated sludge systems for energy and nutrients recovery from wastewater. Bioresource Technology. 2018;247:513-519. PMID: 28972904. Tahir W, Zafar S, Llorens F, Arora AS, Thüne K, Schmitz M et al. Molecular Alterations in the Cerebellum of Sporadic Creutzfeldt-Jakob Disease Subtypes with DJ-1 as a Key Regulator of Oxidative Stress.Molecular neurobiology. 2018;55(1). PMID: 27975168. Llorens F, Villar-Piqué A, Candelise N, Ferrer I, Zerr I. Tau protein as a biological fluid biomarker in neurodegenerative dementias. Cognitive Disorders: IntechOpen; 2018. Pose-Utrilla J, Garcia-Guerra L, Del Puerto A, Martin A, Jurado-Arjona J, De Leon-Reyes N.S. et al. Erratum: Author Correction: Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1 (Nature communications (2017) 8 1 (2275)). Nature communications. 2018;9(1):473-. PMID: 29382840. Mata-Balaguer T, Cuchillo-Ibanez I, Calero M, Ferrer I, Saez-Valero J. Decreased generation of c-terminal fragments of apoer2 and increased reelin expression in Alzheimer’s disease. FASEB Journal. 2018;32(7):3536-3546. PMID: 29442527. Villar-Pique A, Schmitz M, Lachmann I, Karch A, Calero O, Stehmann C. et al. Cerebrospinal Fluid Total Prion Protein in the Spectrum of Prion Diseases. Molecular Neurobiology. 2018. PMID: 30062673. del Rio J.A, Ferrer I, Gavin R. Role of cellular prion protein in interneuronal amyloid transmission. Progress in Neurobiology. 2018;165-167:87-102. PMID: 29530723. Ranea-Robles P, Launay N, Ruiz M, Calingasan N.Y, Dumont M, Naudi A. et al. Aberrant regulation of the GSK-3β/NRF2 axis unveils a novel therapy for adrenoleukodystrophy. EMBO Molecular Medicine. 2018;10(8). PMID: 29997171. Hou X, Fiesel F.C, Truban D, Castanedes Casey M, Lin W.-L, Soto A.I. et al. Age- and disease-de-

57 pendent increase of the mitophagy marker phospho-ubiquitin in normal aging and Lewy body disease. Autophagy. 2018;14(8):1404-1418. PMID: 29947276. Andres Benito P, Dominguez Gonzalez M, Ferrer I. Altered gene transcription linked to astrocytes and oligodendrocytes in frontal cortex in Creutzfeldt-Jakob disease. Prion. 2018;12(3- 4):216-225. PMID: 30009661. Andres-Benito P, Delgado-Morales R, Ferrer I. Altered regulation of KIAA0566, and katanin signaling expression in the locus coeruleus with neurofibrillary tangle pathology. Frontiers in Cellular Neuroscience. 2018;12. PMID: 29867364. Schmitz M, Candelise N, Llorens F, Zerr I. Amplification and detection of minuscule amounts of misfolded prion protein by using the real-time quaking-induced conversion. Methods in Molecular Biology. 2018;1779:257-263. PMID: 29886538. Kanata E, Golanska E, Villar-Pique A, Karsanidou A, Dafou D, Xanthopoulos K. et al. Cerebro- spinal fluid neurofilament light in suspected sporadic Creutzfeldt-Jakob disease. Journal of Clinical Neuroscience. 2018. PMID: 30309804. Llorens F, Barrio T, Correia A, Villar-Pique A, Thune K, Lange P. et al. Cerebrospinal Fluid Prion Disease Biomarkers in Pre-clinical and Clinical Naturally Occurring Scrapie. Molecular Neurobi- ology. 2018;55(11):8586-8591. PMID: 29572672. Schmitz M, Villar-Piqué A, Llorens F, Gmitterová K, Hermann P, Varges D et al. Cerebrospinal Fluid Total and Phosphorylated α-Synuclein in Patients with Creutzfeldt-Jakob Disease and Synucleinopathy.Molecular neurobiology. 2018. PMID: 30136097. Torres P, Ramirez-Nunez O, Romero-Guevara R, Bares G, Granado-Serrano A.B, Ayala V. et al. Cryptic exon splicing function of TARDBP interacts with autophagy in nervous tissue. Auto- phagy. 2018;14(8):1398-1403. PMID: 29912613. Ermann N, Lewczuk P, Schmitz M, Lange P, Knipper T, Goebel S. et al. CSF nonphosphorylated Tau as a biomarker for the discrimination of AD from CJD. Annals of Clinical and Translational Neurology. 2018;5(7):883-887. PMID: 30009207. Ferrer A, Costas J, Labad J, Salvat-Pujol N, Segalas C, Urretavizcaya M. et al. FKBP5 polymorphisms and hypothalamic-pituitary-adrenal axis negative feedback in major depression and obsessive-compulsive disorder. Journal of Psychiatric Research. 2018;104:227-234. PMID: 30107269. Andres-Benito P, Gelpi E, Povedano M, Santpere G, Ferrer I. Gene expression profile in frontal cortex in sporadic frontotemporal lobar degeneration-TDP. Journal of Neuropathology and Experimental Neurology. 2018;77(7):608-627. PMID: 29788265. Aso E, Andres-Benito P, Ferrer I. Genetic deletion of CB1 cannabinoid receptors exacerbates the Alzheimer-like symptoms in a transgenic animal model. Biochemical Pharmacology. 2018;157:210-216. PMID: 30096288. Garcia-Esparcia P, Diaz-Lucena D, Ainciburu M, Torrejon-Escribano B, Carmona M, Llorens F. et al. Glutamate transporter GLT1 expression in Alzheimer disease and dementia with Lewy bodies. Frontiers in Aging Neuroscience. 2018;10(APR). PMID: 29755340. Vanni S, Zattoni M, Moda F, Giaccone G, Tagliavini F, Haik S. et al. Hemoglobin mRNA changes in the frontal cortex of patients with neurodegenerative diseases. Frontiers in Neuroscience. 2018;12(JAN). PMID: 29403351. López-Aguilar J, Bassi GL, Quilez ME, Martí JD, Tavares OR, Xiol EA et al. Hippocampal Damage During Mechanical Ventilation in Trendelenburg Position: A Secondary Analysis of an Exper- imental Study on the Prevention of Ventilator-Associated Pneumonia.Shock (Augusta, Ga.).

58 CIBERNED 2018 ANNUAL REPORT

2018. PMID: 30052585. Cipriani S, Ferrer I, Aronica E, Kovacs GG, Verney C, Nardelli J et al. Hippocampal Radial Glial Subtypes and Their Neurogenic Potential in Human Fetuses and Healthy and Alzheimer’s Dis- ease Adults.Cerebral cortex (New York, N.Y. : 1991). 2018;28(7). PMID: 29722804. Kruse N, Heslegrave A, Gupta V, Foiani M, Villar-Pique A, Schmitz M. et al. Interlaboratory validation of cerebrospinal fluid α-synuclein quantification in the diagnosis of sporadic Creutzfeldt-Jakob disease. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2018;10:461-470. PMID: 30294658. Diaz M, Fabelo N, Ferrer I, Marin R. Lipid raft aging in the human frontal cortex during non- pathological aging: gender influences and potential implications in Alzheimer’s disease. Neu- robiology of aging. 2018;67:42-52. PMID: 29627763. Kanata E, Thüne K, Xanthopoulos K, Ferrer I, Dafou D, Zerr I et al. MicroRNA Alterations in the Brain and Body Fluids of Humans and Animal Prion Disease Models: Current Status and Per- spectives.Frontiers in aging neuroscience. 10. PMID: 30083102. Villar-Pique A, Schmitz M, Candelise N, Ventura S, Llorens F, Zerr I. Molecular and Clini- cal Aspects of Protein Assays in Neurodegenerative Diseases. Molecular Neurobiology. 2018;55(9):7588-7605. PMID: 29429052. Ferrer I. Oligodendrogliopathy in neurodegenerative diseases with abnormal protein aggregates: The forgotten partner. Progress in Neurobiology. 2018;169:24-54. PMID: 30077775.

Silva-Abreu M, Calpena A.C, Andres-Benito P, Aso E, Romero I.A, Roig-Carles D. et al. PPARγ agonist-loaded PLGA-PEG nanocarriers as a potential treatment for Alzheimer’s disease: in vitro and in vivo studies. International journal of nanomedicine. 2018;13:5577-5590. PMID: 30271148. Fleitas C, Pinol-Ripoll G, Marfull P, Rocandio D, Ferrer I, Rampon C. et al. ProBDNF is modified by advanced glycation end products in Alzheimer’s disease and causes neuronal apoptosis by inducing p75 neurotrophin receptor processing 11 Medical and Health Sciences 1109 Neuro- sciences. Molecular Brain. 2018;11(1). PMID: 30428894. Ferrer I. Proteomics and lipidomics in the human brain. Handbook of Clinical Neurology. 2018;150:285-302. PMID: 29496147. Alonso-Andres P, Albasanz J.L, Ferrer I, Martin M. Purine-related metabolites and their converting enzymes are altered in frontal, parietal and temporal cortex at early stages of Alzheimer’s disease pathology. Brain Pathology. 2018;28(6):933-946. PMID: 29363833. Dominguez-Gonzalez M, Puigpinos M, Jove M, Naudi A, Portero-Otin M, Pamplona R. et al. Re- gional vulnerability to lipoxidative damage and inflammation in normal human brain aging. Experimental Gerontology. 2018;111:218-228. PMID: 30077575. Fernandez-Vega I, Diaz-Lucena D, Azkune Calle I, Geijo M, Juste R.A, Llorens F. et al. Sporadic Creutzfeldt–Jakob disease with glial PrPRes nuclear and perinuclear immunoreactivity. Neu- ropathology. 2018;38(5):561-567. PMID: 30123962. Petyuk V.A, Chang R, Ramirez-Restrepo M, Beckmann N.D, Henrion M.Y.R, Piehowski P.D. et al. The human brainome: network analysis identifies HSPA2 as a novel Alzheimer’s disease target. Brain : a journal of neurology. 2018;141(9):2721-2739. PMID: 30137212. Vicente-Pascual M, Rossi M, Gamez J, Llado A, Valls J, Grau-Rivera O. et al. Variably pro- tease-sensitive prionopathy presenting within ALS/FTD spectrum. Annals of Clinical and Translational Neurology. 2018;5(10):1297-1302. PMID: 30349865.

59 Andres-Benito P, Dominguez R, Colomina M.J, Llorens F, Povedano M, Ferrer I. YKL40 in sporadic amyotrophic lateral sclerosis: Cerebrospinal fluid levels as a prognosis marker of disease progression. Aging. 2018;10(9):2367-2382. PMID: 30215603. Riancho J, Bosque-Varela P, Perez-Pereda S, Povedano M, de Munain A.L, Santurtun A. The increasing importance of environmental conditions in amyotrophic lateral sclerosis. Interna- tional Journal of Biometeorology. 2018;62(8):1361-1374. PMID: 29713861. Ferrer I, Aguiló García M, López González I, Diaz Lucena D, Roig Villalonga A, Carmona Tech M et al. Aging-related tau astrogliopathy (ARTAG): Not only tau phosphorylation in astrocytes. Brain pathology (Zurich, Switzerland). 2018. PMID: 29396893. Aymerich M.S, Aso E, Abellanas M.A, Tolon R.M, Ramos J.A, Ferrer I. et al. Cannabinoid pharmacology/therapeutics in chronic degenerative disorders affecting the central nervous system. Biochemical Pharmacology. 2018;157:67-84. PMID: 30121249. Zerr I, Schmitz M, Karch A, Villar-Piqué A, Kanata E, Golanska E et al. Cerebrospinal fluid neurofilament light levels in neurodegenerative dementia: Evaluation of diagnostic accuracy in the differential diagnosis of prion diseases.Alzheimer’s & dementia : the journal of the Alzheim- er’s Association. 2018. PMID: 29391125. Sanchez-Mut J.V, Heyn H, Silva B.A, Dixsaut L, Garcia-Esparcia P, Vidal E. et al. PM20D1 is a quantitative trait locus associated with Alzheimer’s disease. Nature Medicine. 2018;24(5):598- 603. PMID: 29736028. Llorens F, Thune K, Marti E, Kanata E, Dafou D, Diaz-Lucena D. et al. Regional and subtype-dependent miRNA signatures in sporadic Creutzfeldt-Jakob disease are accompanied by alterations in miRNA silencing machinery and biogenesis. PLoS Pathogens. 2018;14(1). PMID: 29357384.

RESEARCH PROJECTS 2018 Code: PI2016/04. Title: The ALS CIBERNED Challenge: Accelerating New Drug Discovery. Principal investigator: Adolfo Lopez De Munain. CIBERNED’s collaboration: Yes. CIBERNED groups: G609 ; G303; G503; G408. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

Code: 000. Title: The part of the cloud challenge on neuroinflammation. SATIVEX. Alzheimer’s Asso- ciation. Principal investigator: Ferrer Abizanda, Isidro. CIBERNED’s collaboration: No. CIBERNED groups: G503. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Alzheimer’s Association. Funding: ND. Duration: 2017-2018.

60 CIBERNED 2018 ANNUAL REPORT

415 | Antonia Gutiérrez Pérez

DPTO. BIOLOGÍA CELULAR, GENÉTICA Y FISIOLOGÍA (ÁREA DE BIOLOGÍA CELULAR) Facultad de Ciencias, Universidad de Málaga Campus de Teatinos, 29071, Málaga, Spain Tel.: +34 952 133 344 E-mail: [email protected]

PRINCIPAL Dávila Cansino, José Mejías Ortega, Marina. INVESTIGATOR Carlos. Bachelor degree. PhD. Moreno González, Inés. Gutiérrez Pérez, Antonia Fernández Valenzuela, PhD. Juan José. Núñez Díaz, Cristina. Bachelor degree. Bachelor degree. LIST OF García León, Juan Sánchez Mejías, PERSONNEL Antonio. Elisabeth. Aneiros Ferrer, PhD. PhD. Mercedes. Gómez Arboledas, Sánchez Varo, Raquel Technician. Angela. María. Baglietto Vargas, PhD. PhD. David. Khan, Zafaruddin. Trujillo Estrada, Laura. PhD. PhD. PhD.

ABSTRACT In 2018 we have continued studying glial pathology in Alzheimer’s patients and animal models. We analyzed the microglial response in the frontal cortex of patients and, unlike the hippocampus which shows microglial degeneration, a strong activation of these immune cells was detected. This regional difference maybe due to a greater accumulation and/or Abeta aggregation in the cortex while in the hippocampus phospho-tau is prefe- rentially accumulated, therefore, we have characterized the microglial response in two models of taupathy (P301S and ThyTau22). The P301S model showed higher microglial activation and phospho-tau accumulation. Soluble phospho-tau from P301S was not toxic to the microglia whereas that from ThyTau22 produced a strong toxicity. Conse- quently, it is necessary to generate new models that integrate the complexity of human microglial pathology to guarantee success at the clinical level. Another of our objectives, part of a CIBERNED collaborative project, has been to analyze the astroglial pathology. We have described, both in patients and in amyloidogenic models, a subtype of reacti-

61 ve astrocytes that phagocytose aberrant synapses although with a limited intracellular degradation capacity. In addition, reactive astrocytes seem to have decreased their metabolic capacities. Taking into account the primordial functions of these cells in neuronal maintenance, a decrease in their functionality could lead to neuronal vulnerability. These results were presented this year as a Doctoral Thesis by a member of our team who has received the 2018 Malaga Research Award. Overall, our work supports microglial and astroglial dysfunction, with loss of their beneficial and neuroprotective capacities, as a pathogenic mechanism in Alzheimer’s disease. This novel approach opens the door to the development of therapies aimed at restoring glial functionality and modifying the course of this disease. Finally, our scientific activity includes collaboration with various national and international groups on common objectives on Alzheimer’s.

KEYWORDS Alzheimer, microglia, astroglia, pathology, neuroinflammation, transgenic models, patients, therapeutic target.

PUBLICATIONS 2018 Romero-Molina C, Navarro V, Sanchez-Varo R, Jimenez S, Fernandez-Valenzuela J.J, Sanchez-Mi- co M.V. et al. Distinct microglial responses in two transgenic murine models of TAU pathology. Frontiers in Cellular Neuroscience. 2018;12. PMID: 30487735. Navarro V, Sanchez-Mejias E, Jimenez S, Munoz-Castro C, Sanchez-Varo R, Davila J.C. et al. Microglia in Alzheimer’s disease: Activated, dysfunctional or degenerative. Frontiers in Aging Neuroscience. 2018;10(MAY). PMID: 29867449. Garcia-Bonilla M, Garcia-Martin M.L, Munoz-Hernandez M.C, Dominguez-Pinos D, Martinez-Le- on M.I, Penalver A. et al. A Distinct Metabolite Profile Correlates with Neurodegenerative Con- ditions and the Severity of Congenital Hydrocephalus. Journal of neuropathology and experimental neurology. 2018;77(12):1122-1136. PMID: 30364991. Garcia-Leon J.A, Cabrera-Socorro A, Eggermont K, Swijsen A, Terryn J, Fazal R. et al. Genera- tion of a human induced pluripotent stem cell–based model for tauopathies combining three microtubule-associated protein TAU mutations which displays several phenotypes linked to neurodegeneration. Alzheimer’s and Dementia. 2018;14(10):1261-1280. PMID: 30036493. Baglietto-Vargas D, Prieto G.A, Limon A, Forner S, Rodriguez-Ortiz C.J, Ikemura K. et al. Im- paired AMPA signaling and cytoskeletal alterations induce early synaptic dysfunction in a mouse model of Alzheimer’s disease. Aging Cell. 2018;17(4). PMID: 29877034. Dhouafli Z, Cuanalo-Contreras K, Hayouni E.A, Mays C.E, Soto C, Moreno-Gonzalez I. Inhibition of protein misfolding and by natural phenolic compounds. Cellular and Molecular Life Scienc- es. 2018;75(19):3521-3538. PMID: 30030591. García-León JA, Kumar M, Boon R, Chau D, One J, Wolfs E et al. SOX10 Single Transcription Fac- tor-Based Fast and Efficient Generation of Oligodendrocytes from Human Pluripotent Stem Cells.Stem cell reports. 2018;10(2). PMID: 29337119. Gomez-Arboledas A, Davila J.C, Sanchez-Mejias E, Navarro V, Nunez-Diaz C, Sanchez-Varo R. et al. Phagocytic clearance of presynaptic dystrophies by reactive astrocytes in Alzheimer’s disease. GLIA. 2018;66(3):637-653. PMID: 29178139.

62 CIBERNED 2018 ANNUAL REPORT

Fernandez Valenzuela JJ, Sanchez Varo R, Gutierrez A. Descifrando la contribución patogénica de Abeta y TAU en modelos transgénicos de la enfermedad de Alzheimer. In: Metabolic Re- programming as a Target for Cancer and Other Diseases. 221-234. 2018. Garcia Leon JA, Ruiz Contreras F, Gutierrez A. Identificando los mecanismos implicados en la degeneración microglial presente en la enfermedad de Alzheimer. In: Metabolic Reprogram- ming as a Target for Cancer and Other Diseases. 181-194. 2018. Mejias Ortega M, Sanchez Mejias E, Gutierrez A. Respuesta microglial en la enfermedad de Alzheimer: nuevas perspectivas y potencial terapéutico. In: Metabolic Reprogramming as a Target for Cancer and Other Diseases. 209-220. 2018. Nuñez Diaz C, Vitorica J, Gutierrez A. Toxicidad de las placas amiloides y su impacto en la pro- gresión de la enfermedad de Alzheimer. In: Metabolic Reprogramming as a Target for Cancer and Other Diseases. 195-207. 2018. Gutierrez A, Vitorica J. Toward a New Concept of Alzheimer’s Disease Models: A Perspective from Neuroinflammation. Journal of Alzheimer’s Disease. 2018;64(s1):S329-S338. PMID: 29562520.

Code: FIS PI15/00796. Title: Evaluando la disfuncion microglial y astroglial como base del proceso neurodegen- erativo y la demencia en la enfermedad de Alzheimer: nuevas aproximaciones terapeuticas. Principal investigator: Antonia Gutierrez Perez. CIBERNED’s collaboration: Yes. CIBERNED groups: G415 ; G411. Other CIBER’s collaboration: No. Type: Nacional.

63 Funding agency: Instituto de Salud Carlos III. Funding: 99220. Duration: 2016-2018.

Code: PPIT.UMA.B1.2017/26. Title: Generacion y caracterizacion de nuevos modelos murinos de Alzheimer con deficiencias en la respuesta inmune microglial. Principal investigator: Raquel Sanchez Varo. CIBERNED’s collaboration: No. CIBERNED groups: G415. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Universidad de Malaga. Funding: 8000. Duration: 2017-2019.

PHD DISSERTATIONS 2018 Author: Angela Gómez Arboledas. Title: Patología glial en el hipocampo de modelos transgénicos de la Enfermedad de Alzheimer: un enfoque ultraestructural. Date: 15/1/2018. Supervisor: Antonia Gutiérrez Pérez.

64 CIBERNED 2018 ANNUAL REPORT

504 | Alberto Lleó Bisa

SERVICIO DE NEUROLOGÍA. HOSPITAL SANTA CRUZ Y SAN PABLO C/ Sant Antoni María Claret, 167 08025 Barcelona. España Tel. +34 932 919 050 ext. 8232/8230 Fax: +34 935 565 602 E-mail: [email protected]

PRINCIPAL Dols Icardo, Oriol. Querol Vilaseca, Marta. INVESTIGATOR Bachelor degree. Bachelor degree. Estellés Pals, María Ribosa i Nogué, Roser. Lleó Bisa, Alberto. Teresa. Bachelor degree. Bachelor degree. Sala Matavera, Isabel. Fortea Ormaechea, Juan. PhD. LIST OF Bachelor degree. PERSONNEL Sánchez Saudinós, García Losada, Lidia. María Belen. Alcolea Rodriguez, Bachelor degree. Bachelor degree. Daniel Andrés. Bachelor Illán Gala, Ignacio. Sirisi Dolcet, Sonia. degree. Bachelor degree. Bachelor degree. Barroeta Espar, Isabel. Montal Blancafort, Subirana Castillo, Bachelor degree. Victor. Andrea. Belbin, Olivia. Bachelor degree. Bachelor degree. PhD. Morenas Rodriguez, Torres Alcalá, Soraya. Blesa González, Rafael. Estrella. Technician. PhD. Bachelor degree. Valldeneu Castells, Carmona Iragui, María. Muñoz Llahuna, Laia. Silvia. Bachelor degree. Bachelor degree. Bachelor degree. Cervera Carles, Laura. Nuñez Llaves, Raúl. Videla Toro, Laura. Bachelor degree. Technician. Bachelor degree. Clarimón Echavarría, Pegueroles Monllau, Vilaplana Martinez, Jordi. Jordi. Eduard. Bachelor degree. Bachelor degree. PhD.

ABSTRACT Our group is composed of a multidisciplinary team of neurologists, neuropsychologists, biologists, engineers and lab technicians. The activities of the group are focu-

65 sed on translational and clinical aspects of neurodegenerative dementias. In 2018 the group has been working on novel genetics factors and novel biomarkers of the most common neurodegenerative dementias. We have investigated new genetic risk variants in Alzheimer’s disease (AD) and frontotemporal dementia (FTD). In parallel, the group has worked on novel biomarkers in cerebrospinal fluid and plasma along the continuum of AD, dementia with Lewy bodies, FTD and Down syndrome. The group is also involved in different imaging studies, using magnetic resonance imaging and amyloid positron emission tomography (PET) to determine the structural correlates of different biomarkers along the AD continnum, Down syndrome and other neurodegenerative dementias. Finally, the group also has uncovered important aspects of the molecular basis of neurodegenerative in human brain. We have used novel microscopy techniques (Array Tomography) and Super-resolution Microscopy techniques to show the structure of human amyloid plaques in Alzheimer’s disease. The group offers a unique environment for clinicians and investigators to tackle basic and translational aspects of neurodegeneration.

KEYWORDS Alzheimer, dementia, cerebrospinal fluid, amyloid, imaging, biomarkers

PUBLICATIONS 2018 Alcolea D, Irwin D.J, Illan-Gala I, Munoz L, Clarimon J, McMillan C.T. et al. Elevated YKL-40 and low sAPPβ:YKL-40 ratio in antemortem cerebrospinal fluid of patients with pathologically confirmed FTLD. Journal of Neurology, Neurosurgery and Psychiatry. 2019;90(2):180-186. epub2018. PMID: 30297518. Delaby C, Munoz L, Torres S, Nadal A, Le Bastard N, Lehmann S. et al. Impact of CSF storage volume on the analysis of Alzheimer’s disease biomarkers on an automated platform. Clinica Chimica Acta. 2019;490:98-101. epub2018. PMID: 30579960. Peloso G.M, van der Lee S.J, Sims R, van der Lee S.J, Naj A.C, Bellenguez C. et al. Genetically elevated high-density lipoprotein cholesterol through the cholesteryl ester transfer protein gene does not associate with risk of Alzheimer’s disease. Alzheimer’s and Dementia: Diagno- sis, Assessment and Disease Monitoring. 2018;10:595-598. Blauwendraat C, Reed X, Kia D.A, Gan-Or Z, Lesage S, Pihlstrom L. et al. Frequency of Loss of Function Variants in LRRK2 in Parkinson Disease. JAMA Neurology. 2018;75(11):1416-1422. PMID: 30039155. Zhang M, Ferrari R, Tartaglia M.C, Keith J, Surace E.I, Wolf U.et al. A C6orf10/LOC101929163 locus is associated with age of onset in C9orf72 carriers. Brain : a journal of neurology. 2018;141(10):2895-2907. PMID: 30252044. Mandelli M.L, Welch A.E, Vilaplana E, Watson C, Battistella G, Brown J.A. et al. Altered topology of the functional speech production network in non-fluent/agrammatic variant of PPA. Cor- tex. 2018;108:252-264. PMID: 30292076. Strydom A, Coppus A, Blesa R, Danek A, Fortea J, Hardy J. et al. Alzheimer’s disease in Down

66 CIBERNED 2018 ANNUAL REPORT syndrome: An overlooked population for prevention trials. Alzheimer’s and Dementia: Trans- lational Research and Clinical Interventions. 2018;4:703-713. PMID: 30581976. Lewczuk P, Riederer P, O’Bryant S.E, Verbeek M.M, Dubois B, Visser P.J. et al. Cerebrospinal fluid and blood biomarkers for neurodegenerative dementias: An update of the Consensus of the Task Force on Biological Markers in Psychiatry of the World Federation of Societies of Biological Psychiatry. World Journal of Biological Psychiatry. 2018;19(4):244-328. PMID: 29076399. Illan-Gala I, Pegueroles J, Montal V, Vilaplana E, Carmona-Iragui M, Alcolea D. et al. Challenges associated with biomarker-based classification systems for Alzheimer’s disease. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2018;10:346-357. PMID: 30175226. Morenas-Rodriguez E, Sala I, Subirana A, Pascual-Goni E, Sanchez-Saudinos M.B, Alcolea D. et al. Clinical Subtypes of Dementia with Lewy Bodies Based on the Initial Clinical Presentation. Journal of Alzheimer’s Disease. 2018;64(2):505-513. PMID: 29889064. Alonso-Jimenez A, Ramon C, Dols-Icardo O, Roig C, Gallardo E, Clarimon J. et al. Corpus cal- losum agenesis, myopathy and pinpoint pupils: consider Stormorken syndrome. European Journal of Neurology. 2018;25(2):e25-e26. PMID: 29356264.

Illan-Gala I, Alcolea D, Montal V, Dols-Icardo O, Munoz L, de Luna N. et al. CSF sAPPβ, YKL- 40, and NfL along the ALS-FTD spectrum. Neurology. 2018;91(17):e1619-e1628. PMID: 30291183. Oliveira F, Leuzy A, Castelhano J, Chiotis K, Hasselbalch S.G, Rinne J. et al. Data driven diagnostic classification in Alzheimer’s disease based on different reference regions for normali- zation of PiB-PET images and correlation with CSF concentrations of Aβ species. NeuroImage: Clinical. 2018;20:603-610. PMID: 30186764. Cortes-Vicente E, Turon-Sans J, Gelpi E, Clarimon J, Borrego-Ecija S, Dols-Icardo O. et al. Distinct clinical features and outcomes in motor neuron disease associated with behavioural variant frontotemporal dementia. Dementia and Geriatric Cognitive Disorders. 2018;45(3-4):220- 231. PMID: 29886477. Borrego-Écija S, Cortés-Vicente E, Cervera-Carles L, Clarimón J, Gámez J, Batlle J et al. Does ALS-FUS without FUS mutation represent ALS-FET? Report of three cases.Neuropathology and applied neurobiology. 2018. PMID: 30375034. Menendez-Gonzalez M, Clarimon J, Rosas-Allende I, Blazquez M, San Martin E.S, Garcia-Fer- nandez C. et al. HTT gene intermediate alleles in neurodegeneration: evidence for association with Alzheimer’s disease. Neurobiology of Aging. 2018. PMID: 30583877. Broce I, Karch C.M, Wen N, Fan C.C, Wang Y, Hong Tan C. et al. Immune-related genetic en- richment in frontotemporal dementia: An analysis of genome-wide association studies. PLoS Medicine. 2018;15(1). PMID: 29315334. Baradaran-Heravi Y, Dillen L, Nguyen H.P, Van Mossevelde S, Baets J, De Jonghe P. et al. No supportive evidence for TIA1 gene mutations in a European cohort of ALS-FTD spectrum patients. Neurobiology of Aging. 2018;69:293.e9-293.e11. PMID: 29886022. Pegueroles J, Jimenez A, Vilaplana E, Montal V, Carmona-Iragui M, Pane A. et al. Obesity and Alzheimer’s disease, does the obesity paradox really exist? A magnetic resonance imaging study. Oncotarget. 2018;9(78):34691-34698. PMID: 30410669. Fortea J, Carmona-Iragui M, Benejam B, Fernández S, Videla L, Barroeta I et al. Plasma and CSF biomarkers for the diagnosis of Alzheimer’s disease in adults with Down syndrome: a cross-sectional study.The Lancet. Neurology. 2018. PMID: 30172624.

67 Gimenez S, Videla L, Romero S, Benejam B, Clos S, Fernandez S. et al. Prevalence of sleep disorders in adults with down syndrome: A comparative study of self-reported, actigraphic, and polysomnographic findings. Journal of Clinical Sleep Medicine. 2018;14(10):1725-1733. PMID: 30353801. Kalinderi K, Fragakis N, Sotiriadou M, Oriol DI, Katritsis D, Letsas K et al. PRRX1 Rs3903239 polymorphism and atrial fibrillation in a Greek population.Hellenic journal of cardiology : HJC = Hellenike kardiologike epitheorese. 2018. PMID: 29355724. Ramos De Matos M, Ferreira C, Herukka S.-K, Soininen H, Janeiro A, Santana I. et al. Quantita- tive genetics validates previous genetic variants and identifies novel genetic players influencing Alzheimer’s disease cerebrospinal fluid biomarkers. Journal of Alzheimer’s Disease. 2018;66(2):639-652. PMID: 30320580. Philtjens S, Van Mossevelde S, van der Zee J, Wauters E, Dillen L, Vandenbulcke M. et al. Rare nonsynonymous variants in SORT1 are associated with increased risk for frontotemporal dementia. Neurobiology of Aging. 2018;66:181.e3-181.e10. PMID: 29555433. Blesa R, Toriyama K, Ueda K, Knox S, Grossberg G. Strategies for continued successful treatment in patients with Alzheimer’s disease: An overview of switching between pharmacological agents. Current Alzheimer Research. 2018;15(10):964-974. PMID: 29895249. Ramos-Campoy O, Avila-Polo R, Grau-Rivera O, Antonell A, Clarimon J, Rojas-Garcia R. et al. Systematic screening of ubiquitin/p62 aggregates in cerebellar cortex expands the neuropathological phenotype of the C9orf72 Expansion Mutation. Journal of Neuropathology and Experimental Neurology. 2018;77(8):703-709. PMID: 29889265. Teunissen C.E, Otto M, Engelborghs S, Herukka S.-K, Lehmann S, Lewczuk P. et al. White paper by the Society for CSF Analysis and Clinical Neurochemistry: Overcoming barriers in biomarker development and clinical translation. Alzheimer’s Research and Therapy. 2018;10(1). PMID: 29544527. Kia DA, Sabir MS, Ahmed S, Trinh J, Bandres-Ciga S, International Parkinson’s Disease Genom- ics Consortium. et al. LRP10 in α-synucleinopathies.The Lancet. Neurology. 2018;17(12). PMID: 30507383. Lopez-Font I, Sogorb-Esteve A, Javier-Torrent M, Brinkmalm G, Herrando-Grabulosa M, Gar- cia-Lareu B. et al. Decreased circulating ErbB4 ectodomain fragments as a read-out of impaired signaling function in amyotrophic lateral sclerosis. Neurobiology of Disease. 2019;124:428- 438. epub2018. PMID: 30594809. Kun-Rodrigues C, Orme T, Carmona S, Hernandez D.G, Ross O.A, Eicher J.D. et al. A comprehensive screening of copy number variability in dementia with Lewy bodies. Neurobiology of Aging. 2019;75:223.e1-223.e10. epub2018. PMID: 30448004. Swarup V, Hinz F.I, Rexach J.E, Noguchi K.-I, Toyoshiba H, Oda A. et al. Identification of evolu- tionarily conserved gene networks mediating neurodegenerative dementia. Nature Medicine. 2019;25(1):152-164. epub2018. PMID: 30510257. Cantero JL, Atienza M, Sanchez-Juan P, Rodriguez-Rodriguez E, Vazquez-Higuera JL, Pozueta A et al. Cerebral changes and disrupted gray matter cortical networks in asymptomatic older adults at risk for Alzheimer’s disease.Neurobiology of aging. 2018;64. PMID: 29331877. Guerreiro R, Ross O.A, Kun-Rodrigues C, Hernandez D.G, Orme T, Eicher J.D. et al. Investigating the genetic architecture of dementia with Lewy bodies: a two-stage genome-wide association study. The Lancet Neurology. 2018;17(1):64-74. PMID: 29263008. Kishore A, Ashok Kumar Sreelatha A, Sturm M, von-Zweydorf F, Pihlstrom L, Raimondi F. et

68 CIBERNED 2018 ANNUAL REPORT al. Understanding the role of genetic variability in LRRK2 in Indian population. Movement Disorders. 2018. PMID: 30485545. Bergeron D, Gorno-Tempini M.L, Rabinovici G.D, Santos-Santos M.A, Seeley W, Miller B.L. et al. Prevalence of amyloid-β pathology in distinct variants of primary progressive aphasia. Annals of Neurology. 2018;84(5):729-740. PMID: 30255971. Mattsson N, Groot C, Jansen W.J, Landau S.M, Villemagne V.L, Engelborghs S. et al. Preva- lence of the apolipoprotein E ε4 allele in amyloid β positive subjects across the spectrum of Alzheimer’s disease. Alzheimer’s and Dementia. 2018;14(7):913-924. PMID: 29601787.

RESEARCH PROJECTS 2018 Code: AMEND. Title: Early Diagnosis of Alzheimer in a Multiplexed approach based on New blood biomarkers. Principal investigator: Monica Mir (CiberBBN). CIBERNED’s collaboration: Yes. CIBERNED groups: G504; G114. Other CIBER’s collaboration: CIBER-BBN. Type: Intramurales. Funding agency: CIBERBBN. Funding: ND. Duration: 2018-2018.

Code: PI2017/01. Title: Estudio del microRNA en el compartimento exosomal del líquido cefalorraquídeo como biomarcador de la demencia frontotemporal y herramienta para el conocimiento de las bases biológicas de la enfermedad. Principal investigator: Jordi Clarimon. CIBERNED’s collaboration: Yes. CIBERNED groups: G504; G406; G510. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2017-2019.

Code: PI17/01896. Title: Aplicacion de Tomografia de Array (AT) para el estudio sistematico a gran escala del dano sinaptico en las demencias neurodegenerativas. Principal investigator: Alberto Lleo. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional.

69 Funding agency: Instituto de Salud Carlos III. Funding: 93170. Duration: 2018-2020.

Code: 1 R21 AG056974-01. Title: Biological Correlates of Alzheimer in Down Syndrome. Principal investigator: Ann-Charlotte Granholm-Bentley, D.A. Paredes, A. Ledreux, M. Margittai. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Natiaonal Institute of Aging. Funding: 414425. Duration: 2017-2019.

Code: 201437 10. Title: Biomarkers profile in different phenotypes of Motor Neuron Disease. Principal investigator: Ricard Rojas. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 150625. Duration: 2015-2018.

Code: SLT006/17/00125. Title: Contractes per a la intensificacio de l’activitat investigadora de professionals facul- tatius especialistes 2017. Principal investigator: Daniel Alcolea. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 86342,79. Duration: 2018-2019.

Code: SLT006/17/00119. Title: Contractes per a la intensificacio de l’activitat investigadora de professionals facul- tatius especialistes 2017. Estudios longitudinales de RM magnetica estructural, de difu- sion y funcional en la EA preclinica esporadica y asociada al SD. Principal investigator: Juan Fortea. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 95318,14. Duration: 2018-2019.

Code: MJ-Fox-Synoligo-20180516-1. Title: Development of prototype immunoassays on the SimoA Technology for quantification in biological samples of oligomeric forms of α-Synuclein using new antibody com- binations (Acronym: SynOligo). Principal investigator: Hugo Vanderstichele. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Michael J Fox Foundation. Funding: ND. Duration: 2018-2019.

70 CIBERNED 2018 ANNUAL REPORT

Code: GBHI_ALZ-18-543740. Title: Domiciliary Alzheimer Visiting in Down syndrome. Principal investigator: Maria Carmona Iragui. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Alzheimer’s Association. Funding: 20974,19. Duration: 2017-2019.

Code: 4560/6393 La caixa-Down Alzheimer. Title: Down Alzheimer Barcelona Neuroimaging Initiative (DABNI). Principal investigator: Rafael Blesa. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Caixa. Funding: 870000. Duration: 2017-2020.

Code: EMIF-AD. Title: EMIF-AD. A biomarker platform for AD biomarkers. Principal investigator: Simon Lovestone, Pieter J. Visser. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 15000. Duration: 2013-2018.

Code: PI13/01532. Title: Enfermedad de Alzheimer y sindrome de Down. Estudios multimodales de liquido cefalorraquideo, resonancia magnetica y PET de amiloide. Principal investigator: Rafael Blesa. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 130075. Duration: 2014-2018.

Code: SLT002/16/00408. Title: Estudio de biomarcadores y desarrollo de nuevas estrategias terapeuticas en una cohorte multicentrica de Demencia Frontotemporal. Principal investigator: Alberto Lleo. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 399363. Duration: 2017-2019.

Code: PI15/00026. Title: Estudio del perfil de expresion de microRNA exosomal en biofluidos para la identificacion de biomarcadores de uso diagnostico en la demencia frontotemporal. Principal investigator: Jordi Clarimon.

71 CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 122815. Duration: 2016-2018.

Code: FUNDELA 2017. Title: Estudio del splicing alternativo en ARN de muestras con diagnostico neuropatolog- ico de ELA asociada a TDP-43. Principal investigator: Jordi Clarimon. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: FUNDELA. Funding: 30000. Duration: 2017-2018.

Code: PI16/01825. Title: Estudio del trazador de tomografia por emision de positrones (PET) para Tau [18F] THK-5351 en pacientes con diferentes Taupatias. Principal investigator: Rafael Blesa. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 122815. Duration: 2017-2019.

Code: SLT002/16/00099. Title: Evaluation and characterization of synaptic proteins as biomarkers for the synaptic damage related to AD. Principal investigator: Olivia Belbin. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 65109. Duration: 2017-2020.

Code: SGR 2017-9. Title: Grup de recerca en demències: Sant Pau (Consolided Group of Research 2017 SGR 547 “Generalitat de Catalunya”). Principal investigator: Jordi Clarimon. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 35200. Duration: 2018-2020.

Code: EU-JG1. Title: Horizon 21 Genetics Consortium (H21GC): Large-scale pooling/genotyping of individuals with Down syndrome for insight into predictive pathways for Alzheimer’s disease. Principal investigator: Cornelia Van Duijn And Tonnie Coppus. CIBERNED’s collaboration: No.

72 CIBERNED 2018 ANNUAL REPORT

CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Jerome Lejeune. Funding: 40000. Duration: 2017-2018.

Code: JEROME-LEJEUNE 2018-1. Title: Horizon 21-Clinical and trial outcome measures for dementia in individuals with Down Syndrome – the DS-1000 cognitive test study group. Principal investigator: Juan Fortea. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Jerome Lejeune. Funding: 30250. Duration: 2018-2019.

Code: PI17/00279. Title: Inflamación hipotalámica en obesidad. Papel de la dieta y efectos de la cirugia bariátrica. Principal investigator: Amanda Gimenez. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: ND. Duration: 2018-2020.

Code: PI14/01126. Title: La estructura cerebral y el metabolismo en la enfermedad de Alzheimer preclinica. Interacciones entre “nuevos y viejos” biomarcadores. Principal investigator: Juan Fortea. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 122815. Duration: 2015-2019.

Code: 201412 10. Title: La malaltia d’Alzheimer a la sindrome de Down. Estudis multimodals amb LCR, RM, EEG i PET. Principal investigator: Juan Fortea. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundació La Marato de TV3. Funding: 199000. Duration: 2015-2018.

Code: PI14/01561. Title: Marcadores sinapticos en la enfermedad de Alzheimer. Principal investigator: Alberto Lleo. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional.

73 Funding agency: Instituto de Salud Carlos III. Funding: 134915. Duration: 2015-2019.

Code: 201614 31. Title: Pre-clinical Alzheimer’s disease and Type 2 diabetes and obesity. Effects of bariatric surgery: a multimodal study. Principal investigator: Rafael Blesa, Amanda Jimenez. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: CIBEROBN. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: ND. Duration: 2017-2020.

Code: TAUDEM. Title: Study with the tau Positron Emission Tomography radiotracer [18F]THK-5351 in patients with different Tauopathies. Principal investigator: Alberto Lleo. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundación BBVA. Funding: ND. Duration: 2016-2019.

Code: 20142610. Title: Synaptic markers in preclinical Alzheimer’s disease. Principal investigator: Alberto Lleo. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 199878,75. Duration: 2015-2018.

Code: 2018-3570, NIH-PA-16-161. Title: The role of Inflammation and NGF Dysfunction in the Evolution of Alzheimer Disease Pathology in Down syndrome. Principal investigator: Juan Fortea. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: NIH. Funding: 462800,34. Duration: 2018-2022.

Code: PI15/00058. Title: Valoración y caracterización de biomarcadores en la fase preclínica de la enfermedad de Alzheimer. Principal investigator: Olivia Belbin. CIBERNED’s collaboration: No. CIBERNED groups: G504. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 68365. Duration: 2016-2018.

74 CIBERNED 2018 ANNUAL REPORT

404 | Carlos Matute Almau

DEPARTMENT OF NEUROSCIENCES FACULTY OF MEDICINE AND DENTISTRY

University of País Vasco 48940 Leioa, Spain Tel: +34 94 601 32 44 Fax: +34 94 601 50 55 E-mail: [email protected]

PRINCIPAL Gejo Crispin, Jon. Quintela López, Tania. INVESTIGATOR Bachelor degree. Bachelor degree. Manterola Juaristi, Ramos González, Paula. Matute Almau, Carlos. Andrea. Bachelor degree. Bachelor degree. Rodríguez Antigüedad Marcos Ezquerro, Saioa. Zarrantz, Alfredo. LIST OF Technician. PhD. PERSONNEL Mato Santos, Susana. Ruiz Nuñez, Asier. Alberdi Alfonso, Elena. PhD. PhD. PhD. Montilla López, Sánchez Gómez, María Ardaya Franco, María. Alejandro. Victoria. Bachelor degree. Bachelor degree. PhD. Bernal Chico, Ana. Moreno García, Álvaro. Serrano Regal, María PhD. Bachelor degree. Paz. Capetillo González de Ortiz Sanz, Carolina. Bachelor degree. Zárate, Estibaliz. Bachelor degree. Verkhrstsky, Alexei. PhD. Palma Leiva, Ana Belén. PhD. Cavaliere, Fabio. Bachelor degree. Zabala Olaizola, Alazne. PhD. Pérez Cerdá, Fernando. Bachelor degree. Chara Ventura, Juan PhD. Carlos. Pérez Samartín, Alberto. Technician. PhD. Domercq García, María. PhD.

ABSTRACT Our laboratory is focused on the study of the molecular and cellular mechanisms of neurons and glia contributing to the pathophysiology of Alzheimer disease (AD) and

75 Parkinson (PD). In 2018 we have found that morin and mangiferin strongly protect against Aβ-induced mitochondrial dysfunction and neuronal cell death. Specifically, these two natural antioxidants preserve cell respiration, promote detoxification of reactive oxygen species, protect from some forms of apoptosis, and regulate mitochondrial matrix calcium in neurons exposed to Aβ. In another study we observed that Aβ1-42 oligomers induce early, rapid, changes in the dendritic tree and spine density which is mediated by integrin β1 and downstream activation of PKC. In addition, we have analyzed the effects of mitochondrial division inhibitor-1 (mdivi-1) on mitochondrial dynamics, calcium signaling, mitochondrial bioenergetics and cell viability during neuronal excitotoxicity in vitro. We found that mdivi-1 induces a dynamin related protein 1 (Drp1)-independent protective phenotype that prevents predo- minantly NMDA receptor-mediated excitotoxicity through the modulation of mitochondrial function and intracellular calcium signaling. Moreover, we deepened on the search for signaling pathways initiating excitotoxic oligodendrocyte death and found that inhibition of casein kinase 2 protects these cells from dying via JNK/P53 activation. Finally, in an international coordinated effort we contributed to the characterization of the therapeutic potential of a molecular tweezer (CLR01) that is capable of decreasing α −synuclein aggregation in induced pluripotent stem cell-derived dopaminergic cultures. Thus, CLR01 reduces neurotoxicity induced by post-mortem brain extracts of PD patients; and in microfluidic assays decreases α-synuclein aggregation in cell somas when axonal terminals are exposed to α-synuclein oligomers.

KEYWORDS β-amiloid, α−synuclein, neurons, oligodendrocytes, oxidative stress

PUBLICATIONS 2018 Domercq M, Matute C. Excitotoxicity therapy for stroke patients still alive. EBioMedicine. 2019;39:3-4. epub2018. PMID: 30579867. Fern R, Matute C. Glutamate receptors and white matter stroke. Neuroscience Letters. 2019;694:86-92. epub2018. PMID: 30476568.

Walker C.A, Randolph L.K, Matute C, Alberdi E, Baleriola J, Hengst U. Aβ1–42 triggers the generation of a retrograde signaling complex from sentinel mRNAs in axons. EMBO Reports. 2018;19(7). PMID: 29759981. Bribian A, Perez-Cerda F, Matute C, Lopez-Mascaraque L. Clonal glial response in a multiple sclerosis mouse model. Frontiers in Cellular Neuroscience. 2018;12. PMID: 30405357. Luchena C, Zuazo-Ibarra J, Alberdi E, Matute C, Capetillo-Zarate E. Contribution of Neurons and Glial Cells to Complement-Mediated Synapse Removal during Development, Aging and in Alzheimer’s Disease. Mediators of inflammation. 2018;2018:2530414-. PMID: 30533998. Manterola A, Bernal-Chico A, Cipriani R, Canedo-Antelo M, Moreno-Garcia A, Martin-Fon- techa M. et al. Deregulation of the endocannabinoid system and therapeutic potential of ABHD6 blockade in the cuprizone model of demyelination. Biochemical Pharmacology. 2018;157:189-201. PMID: 30075103.

76 CIBERNED 2018 ANNUAL REPORT

Colas L, Domercq M, Ramos-Cabrer P, Palma A, Gomez-Vallejo V, Padro D. et al. In vivo imaging of a7 nicotinic receptors as a novel method to monitor neuroinflammation after cerebral ischemia. GLIA. 2018;66(8):1611-1624. PMID: 29528142. Martin A, Domercq M, Matute C. Inflammation in stroke: the role of cholinergic, purinergic and glutamatergic signaling. Therapeutic Advances in Neurological Disorders. 2018;11. PMID: 29774059. Canedo-Antelo M, Serrano M.P, Manterola A, Ruiz A, Llavero F, Mato S. et al. Inhibition of Casein Kinase 2 Protects Oligodendrocytes From Excitotoxicity by Attenuating JNK/p53 Sign- aling Cascade. Frontiers in Molecular Neuroscience. 2018;11. PMID: 30271323. Sanchez-Gomez M.V, Serrano M.P, Alberdi E, Perez-Cerda F, Matute C. Isolation, expansion, and maturation of oligodendrocyte lineage cells obtained from rat neonatal brain and optic nerve. Methods in Molecular Biology. 2018;1791:95-113. PMID: 30006704. Alberdi E, Sanchez-Gomez M.V, Ruiz A, Cavaliere F, Ortiz-Sanz C, Quintela-Lopez T. et al. Mangiferin and Morin Attenuate Oxidative Stress, Mitochondrial Dysfunction, and Neurocy- totoxicity, Induced by Amyloid Beta Oligomers. Oxidative medicine and cellular longevity. 2018;2018:2856063-. PMID: 30013719. Pellon A, Ramirez-Garcia A, Guruceaga X, Zabala A, Buldain I, Antoran A. et al. Microglial immune response is impaired against the neurotropic fungus Lomentospora prolificans. Cellular Microbiology. 2018;20(8). PMID: 29582549. Ruiz A, Alberdi E, Matute C. Mitochondrial division inhibitor 1 (Mdivi-1) protects neurons against excitotoxicity through the modulation of mitochondrial function and intracellular Ca2+ signaling. Frontiers in Molecular Neuroscience. 2018;11. PMID: 29386996. Zabala A, Vazquez-Villoldo N, Rissiek B, Gejo J, Martin A, Palomino A. et al. P2X4 receptor controls microglia activation and favors remyelination in autoimmune encephalitis. EMBO Molecular Medicine. 2018;10(8). PMID: 29973381. Verkhratsky A, Nedergaard M. Physiology of astroglia. Physiological Reviews. 2018;98(1):239- 389. PMID: 29351512. Zorec R, Parpura V, Verkhratsky A. Preventing neurodegeneration by adrenergic astroglial excitation. FEBS Journal. 2018;285(19):3645-3656. PMID: 29630772. Domercq M, Zabala A, Matute C. Purinergic receptors in multiple sclerosis pathogenesis. Brain Research Bulletin. 2018. PMID: 30500565. Manterola A, Bernal-Chico A, Cipriani R, Ruiz A, Perez-Samartin A, Moreno-Rodriguez M. et al. Re-examining the potential of targeting ABHD6 in multiple sclerosis: Efficacy of systemic and peripherally restricted inhibitors in experimental autoimmune encephalomyelitis. Neurophar- macology. 2018;141:181-191. PMID: 30171986. Mato S, Pilar-Cuellar F, Valdizan E.M, Gonzalez-Maeso J, Rodriguez-Puertas R, Meana J. et al. Selective up-regulation of cannabinoid CB1 receptor coupling to Go-proteins in suicide victims with mood disorders. Biochemical Pharmacology. 2018;157:258-265. PMID: 30099006. Menendez-Gonzalez M, Padilla-Zambrano H.S, Alvarez G, Capetillo-Zarate E, Tomas-Zapico C, Costa A. Targeting beta-amyloid at the CSF: A new therapeutic strategy in Alzheimer’s disease. Frontiers in Aging Neuroscience. 2018;10(APR). PMID: 29713273. Perez-Villalba A, Sirerol-Piquer M.S, Belenguer G, Soriano-Canton R, Munoz-Manchado A.B, Villadiego J. et al. Synaptic regulator α-synuclein in dopaminergic fibers is essentially required for the maintenance of subependymal neural stem cells. Journal of Neuroscience. 2018;38(4):814-825. PMID: 29217686.

77 RESEARCH PROJECTS 2018 Code: IT702-13. Title: Bases moleculares y celulares de la neurodegeneracion. Principal investigator: Carlos Matute. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 444000. Duration: 2013-2018.

Code: BIO17/ND/008. Title: Caracterizacion molecular de la mutacion E46K en la enfermedad de Parkinson: una mutacion vasca. Principal investigator: Fabio Cavaliere. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Eitb Maratoia. Funding: 50233. Duration: 2018-2020.

Code: PI_2016_1_0009. Title: Modulacion de la interaccion glia-sinapsis como diana terapeutica en la enfermedad de Alzheimer. Principal investigator: Estibaliz Capetillo. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 63224. Duration: 2016-2018.

Code: PI_2016_1_0016. Title: Plasticidad de la mielina como estrategia reparadora. Principal investigator: Maria Domercq. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 51351,9967. Duration: 2016-2018.

Code: KK-2017/00067. Title: Polifenoles bioactivos con probado efecto neuroprotector: desarrollo de ingredi- entes innovadores para la industria alimentaria vasca. Principal investigator: Estibaliz Capetillo. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 400000. Duration: 2017-2018.

78 CIBERNED 2018 ANNUAL REPORT

Code: SAF2016-75292-R. Title: Regulation of myelination and remyelination by neurotransmitters. Principal investigator: Carlos Matute. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 340000. Duration: 2017-2019.

Code: PI1800513. Title: Relevancia terapeutica de los receptores CB1 astrogliales en la esclerosis multiple. Principal investigator: Susana Mato Santos. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 99220. Duration: 2018-2021.

Code: SAF2015-74332-JIN. Title: Role of lactate and monocarboxylate transporters in oligodendrocyte progenitor cell energy metabolism and remyelination. Principal investigator: Vanja Tepavcevic. CIBERNED’s collaboration: No. CIBERNED groups: G404. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 160000. Duration: 2017-2019.

PHD DISSERTATIONS 2018 Author: Tania Quintela López. Title: Role of beta-amyloid in the oligodendrocyte lineage. Date: 18/1/2018. Supervisor: Elena Alberdi Alfonso.

79 508 | Guadalupe Mengod Los Arcos

IIBB-CSIC, IDIBAPS, CIBERNED Teléfono: +34 93 363 8323 Fax: +34 93 363 8301 E-mail: [email protected]

PRINCIPAL LIST OF INVESTIGATOR PERSONNEL Mengod Los Arcos, Cortés Colomé, Roser. Guadalupe. PhD. Martín Álvarez, Rocío. Technician. Schmidt, Natascha. Bachelor degree. Vilaró Comas , María Teresa. PhD.

ABSTRACT Our research focuses on the study of the cellular and molecular aspects of neurodegenerative diseases with an inflammatory component. The main line of research is “cAMP and neuroinflammation”. During 2018, we continued to investigate the relationship between cAMP and the cellular and molecular aspects of neurodegenerative diseases with an inflammatory component. We study the effect of specific inhibitors of phosphodiesterases PDE4, PDE7 and other drugs in clinical use or under development, acting on different targets, on the course of experimental autoimmune encephalitis (EAE), a mouse model of multiple sclerosis. We analyze the expression of inflammatory cytokines and PDEs and neuropathological changes in cellular populations of the brain and spinal cord. In primary cell cultures of bone marrow macrophages of mice and in the macrophage cell line Raw264.7 we study the effect of molecular regulation of intracellular signaling by cAMP, on the macrophage polarization towards pro-inflammatory or anti-inflammatory phenotypes. We have ended the collaboration with the group of Dr Tata of the Sapienza University of Rome with

80 CIBERNED 2018 ANNUAL REPORT the publication of a comparative study of the expression of components of the cholinergic system in the central nervous system and the spinal cord of EAE mice in two different phases of the disease: acute phase and remitting phase.

KEYWORDS Neurodegeneration, neuroinflammation, molecular neuropharmacology.

PUBLICATIONS 2018 Di Pinto G, Di Bari M, Martin-Alvarez R, Sperduti S, Serrano-Acedo S, Gatta V. et al. Com- parative study of the expression of cholinergic system components in the CNS of experimental autoimmune encephalomyelitis mice: Acute vs remitting phase. European Journal of Neuroscience. 2018;48(5):2165-2181. PMID: 30144326.

RESEARCH PROJECTS 2018 Code: PI15/00148. Title: ¿Están implicados los cambios fenotípicos de las microglia/macrofagos en los efectos beneficiosos de los inhibidores de PDEs del AMPc en la EAE crónica?. Principal investigator: Guadalupe Mengod. CIBERNED’s collaboration: No. CIBERNED groups: G508. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 101500. Duration: 2015-2018.

81 406 | José Rodríguez Álvarez

INSTITUTO DE NEUROCIENCIAS Universidad Autónoma de Barcelona. Edificio M. Campus de Bellaterra 08193 Cerdanyola del Valles. Barcelona Teléfono: +34 935 813 861 E-mail: [email protected]

PRINCIPAL LIST OF Miñano Molina, Alfredo INVESTIGATOR PERSONNEL Jesús. PhD. Rodríguez Álvarez, José. Aguilera Avila, Jose. PhD. Ortega Hernández, Laura. Catala Solsona, Judit. PhD. Bachelor degree. Parra Damas, Arnaldo. Enríquez Barreto, Lilian. PhD. PhD. Saura Antolín, Carlos A. Javier Torrent, Miriam. PhD. Bachelor degree.

ABSTRACT Synaptic dysfunction and loss of synapses are major correlates of cognitive impairment in AD but the cellular mechanisms underlying these changes are still unclear. In the past years our research efforts were focused on deciphering the molecular events underlying synaptic dysfunction at early stages of AD. Synapse-to-nucleus signaling causes synapse dysfunction, neurodegeneration and memory loss in dementia disorders, particularly in Alzheimer’s disease (AD). Using genome-wide transcriptome analysis, we found that a gene expression program regulated by the synapse-to-nucleus factor CREB-regulated transcription coactivator-1 (CRTC1) is essential for hippocampal-dependent memory and is associated with early pathological and memory changes in AD. During this year we have found that CRTC1 binds to promoter regions of neuronal excitability and plasticity genes, including glutamate receptors. Remark- ably, CRTC1 plays essential roles on long-term memory and dendritic spine morphology favorable to enhance synaptic plasticity, including long-term potentiation and depression. CRTC1-dependent synaptic potentiation involves enhancement of NMDA receptor transmission through PKC-mediated phosphorylation and recruitment of GluN1 receptors at synapses. Moreover, Nr4a2, one of the CRTC1 target genes reg-

82 CIBERNED 2018 ANNUAL REPORT ulates activity-dependent CREB/CRTC1-dependent expression of the neurotrophin BDNF in the hippocampus and BDNF-mediated expression of AMPA receptors (AM- PAR) subunits. Altogether suggests that the CRTC1-Nr4a2 axis controls the expression of several NMDA and AMPAR subunits in the hippocampus. On the other hand, several studies have shown that some miRNAs control the formation, maturation and function of synapses and alteration in their levels could underlie synaptic dysfunction in pathological states. During this year we have identified the alteration of several miRNas related to synaptic targets in plasma and brain samples from MCI and AD dementia patients. Moreover, we have obtained preliminary data to support the identification of a miRNAs signature in plasma that could be used as an early AD biomarker, predicting the progression from MCI to AD.

KEYWORDS Alzheimer disease, memory, gene regulation, glutamate receptors, CRTC1, early synaptic dysfunction, miRNAs, biomarker.

PUBLICATIONS 2018 Lopez-Font I, Sogorb-Esteve A, Javier-Torrent M, Brinkmalm G, Herrando-Grabulosa M, Garcia-Lareu B. et al. Decreased circulating ErbB4 ectodomain fragments as a read-out of impaired signaling function in amyotrophic lateral sclerosis. Neurobiology of Disease. 2019;124:428-438. epub2018. PMID: 30594809. Navarro G, Borroto-Escuela D, Angelats E, Etayo Í, Reyes-Resina I, Pulido-Salgado M et al. Re- ceptor-heteromer mediated regulation of endocannabinoid signaling in activated microglia. Role of CB1 and CB2 receptors and relevance for Alzheimer’s disease and levodopa-induced dyskinesia.Brain, behavior, and immunity. 2018;67. PMID: 28843453. Servian-Morilla E, Robles-Lanuza E, Sanchez-Hidalgo A.C, Camacho-Garcia R.J, Paez-Gomez J.A, Mavillard F. et al. Proteolytic processing of neurexins by presenilins sustains synaptic vesicle release. Journal of Neuroscience. 2018;38(4):901-917. PMID: 29229705. Franco R, Aguinaga D, Reyes I, Canela EI, Lillo J, Tarutani A et al. N-Methyl-D-Aspartate Re- ceptor Link to the MAP Kinase Pathway in Cortical and Hippocampal Neurons and Micro- glia Is Dependent on Calcium Sensors and Is Blocked by α-Synuclein, Tau, and Phospho-Tau in Non-transgenic and Transgenic APPSw,Ind Mice.Frontiers in molecular neuroscience. 11. PMID: 30233307. Reyes-Resina I, Navarro G, Aguinaga D, Canela E.I, Schoeder C.T, Zaluski M. et al. Molecular and functional interaction between GPR18 and cannabinoid CB2 G-protein-coupled receptors. Relevance in neurodegenerative diseases. Biochemical Pharmacology. 2018;157:169- 179. PMID: 29870711. Moreno-Galarza N, Mendieta L, Palafox-Sanchez V, Herrando-Grabulosa M, Gil C, Limon D.I. et al. Peripheral Administration of Tetanus Toxin Hc Fragment Prevents MPP+ Toxicity In Vivo. Neurotoxicity Research. 2018;34(1):47-61. PMID: 29460114. Van Der Jeugd A, Parra-Damas A, Baeta-Corral R, Soto-Faguas C.M, Ahmed T, Laferla F.M. et al. Reversal of memory and neuropsychiatric symptoms and reduced tau pathology by selenium in 3xTg-AD mice. Scientific Reports. 2018;8(1). PMID: 29691439.

83 RESEARCH PROJECTS 2018 Code: PI2017/01. Title: Estudio del microRNA en el compartimento exosomal del líquido cefalorraquídeo como biomarcador de la demencia frontotemporal y herramienta para el conocimiento de las bases biológicas de la enfermedad. Principal investigator: Jordi Clarimon. CIBERNED’s collaboration: Yes. CIBERNED groups: G504; G406; G510. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2017-2019.

Code: SAF2017-89271-R. Title: Implicacion de Nr4a2/Nurr1 en la disfuncion sinaptica y deficits cognitives en fases tempranas de la Enfermedad de Alzheimer. Principal investigator: Jose Rodriguez Alvarez. CIBERNED’s collaboration: No. CIBERNED groups: G406. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 145200. Duration: 2018-2020.

Code: Marato TV3 2013-343. Title: Searching new biomarkers and therapeutic targets related to cognitive deficits in early stages of Alzheimer’s Disease: Role of AKAP79/150, CPT1C and SSAO/VAP-1 in Ab-mediated AMPAR dysfunction. Principal investigator: Jose Rodriguez Alvarez. CIBERNED’s collaboration: No. CIBERNED groups: G406. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 199875. Duration: 2015-2018.

Code: SAF2016-80027-R. Title: Transcriptional mechanisms of synaptic plasticity in memory circuits in a mouse mouse of neurodegeneration. Principal investigator: Carlos A. Saura. CIBERNED’s collaboration: No. CIBERNED groups: G406. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 199650. Duration: 2017-2019.

PHD DISSERTATIONS 2018 Author: Dolores Siedlecki-Wüllich. Title: Analysis if miRNA expression in Alzheimer’s disease. Potential use as early biomarkers. Date: 16/11/2018. Supervisor: José Rodríguez Álvarez.

84 CIBERNED 2018 ANNUAL REPORT

407 | Javier Sáez Valero

INSTITUTO DE NEUROCIENCIAS DE ALICANTE

Universidad Miguel Hernández-CSIC Av. Ramón y Cajal, s/n 03550 Sant Joan d’Alacant, Spain Tel +34 96 591 9580 Fax: +34 96 591 9561 E-mail: [email protected]

PRINCIPAL Boix Rodriguez, Claudia Iborra Lázaro, INVESTIGATOR Paula. Guillermo. Bachelor degree. Bachelor degree. Sáez Valero, Javier Cuchillo Ibáñez, López Font, Inmaculada Inmaculada. Belen. LIST OF PhD. PhD. PERSONNEL García Ayllón, María Sogorb Esteve, Aitana. Alom Poveda, Jordi. PhD. Salud. Bachelor degree. PhD.

ABSTRACT Our research team has been working for years on the characterization of alterations in key proteins related with the progression Alzheimer’s disease (AD), attempting to prove mal- functions, as well interconnect β-amyloid (Aβ) and tau hyperphosphorylation (P-tau) pathologies. We have developed tools to study alteration in formation of active protein complexes, proteolytic processing by secretases, glycosylation pattern and others, all of them that could be relevant for understand the impairment of several pathways related with AD. In the past we have described an altered expression of reelin in AD. Reelin is a signaling protein that modulates synaptic function and plasticity in the brain. Our effort was focus to demonstrate a mechanism by which β-amyloid regulates reelin expression, thereby influencing its signaling cascade that ultimately controls tau phosphorylation. Intriguingly, we found that reelin expression levels are increased in the AD brain, but the β-amyloid (Aβ) protein impairs reelin activity. Reelin binding to their receptor, ApoER2, activates downstream signaling and induces the proteolytic cleavage of ApoER2, resulting in the generation of intracellular and extracellular soluble fragments. Previously, we also demonstrated that the intracellular fragment ApoER2-CTF exerts a modulatory role on reelin expression. Our recent data indicated that Aβ is able to trigger increased reelin expression through a mechanism that involved the decrease generation of ApoER2-CTF (Mata-Balaguer et al., FASEB J 2018). Moreover, to evaluate the impaired efficiency of reelin signaling in AD in vivo, we have quantified the levels the soluble

85 ectodomain fragment of ApoER2 (ectoApoER2) in the human cerebrospinal fluid (CSF). Our data suggest that the determination of decreased levels of ecto-ApoER2 in CSF could be a suitable read-out of an impaired reelin signaling in AD, but also indicate differences between sporadic AD and familiar AD (Lopez-Font et al., Clin Chem Acta 2019). We not only aim to clarify the pathological mechanisms behind these diseases, but also to define potential diagnostic tools and/or processes with therapeutic relevance. We also demonstrated that AD therapy with γ-secretase inhibitors (GSIs) affects the levels of the catalytic subunit, presenilin-1 (PS1). This rebound increase in PS1 levels after sustained γ-secretase inhibition can result in an undesirable gain of γ-secretase activity which maybe contributes to the worsening condition reported in volunteers enrolled in discontinued clinical trials (So- gorb-Esteve et al., Mol Neurobiol 2018). Furthermore, we evaluate the diagnostic potential of the disintegrin metalloproteinase 10 (ADAM10). ADAM10 is the main α-secretase acting in the non-amyloidogenic processing of the amyloid precursor protein (APP). We have demonstrated the presence of several forms of ADAM10 in CSF, including mature and immature full-length forms, as well truncated species. Our data indicated that the determination of decreased levels of mature forms of CSF- ADAM10 may be useful as a biomarker for AD (Sogorb-Esteve et al., J Neuroinflammation 2018). Finally, in a collaborative CIBERNED project we extended our experience characterizing CSF protein in the CSF to the study of ErbB4 in amyotrophic lateral sclerosis (ALS). ErbB4 is a transmembrane receptor tyrosine kinase that binds to neuregulins to activate signaling, and disruption of the neuregulin-ErbB4 pathway has been suggested to be involved in the pathogenesis of ALS. In this study we have demonstrated that soluble proteolytic fragments of the ErbB4 ectodomain (ecto-ErbB4) can be detected in CSF and plasma, and if that the levels are decreased in ALS subjects. Likewise, the ecto-ErbB4 fragments were decreased in the plasma of the two transgenic mouse models of ALS (SOD1G93A and TDP-43A315T). We conclude that the determination of circulating ecto-ErbB4 fragments could be a tool to evaluate the impairment of the ErbB4 pathway and may be a useful biomarker in ALS (Lopez-Font et al., Neurobiol Dis 2019).

KEYWORDS Alzheimer, Amyotrophic Lateral Sclerosis, ADAM10, biomarker, ApoER2, reelin, PS1, GSI.

PUBLICATIONS 2018 Lopez-Font I, Iborra-Lazaro G, Sanchez-Valle R, Molinuevo J.-L, Cuchillo-Ibanez I, Saez-Valero J. CSF-ApoER2 fragments as a read-out of reelin signaling: Distinct patterns in sporadic and autosomal-dominant Alzheimer disease. Clinica Chimica Acta. 2019;490:6-11. epub2018. PMID: 30552869. Pappolla A, Midaglia L, Boix Rodriguez C.P, Puig A.A, Lung M, Camps I.R. et al. Simultaneous CMV and Listeria infection following alemtuzumab treatment for multiple sclerosis. Neurolo- gy. 2019;92(6):296-298. epub2018. PMID: 30587519. Saez Valero J. La redacción del apartado de metodología en los estudios de biomedicina. En: Cómo elaborar un proyecto en ciencias de la salud. 40-45. 2018.

86 CIBERNED 2018 ANNUAL REPORT

Sogorb-Esteve A, Garcia-Ayllon M.-S, Gobom J, Alom J, Zetterberg H, Blennow K. et al. Levels of ADAM10 are reduced in Alzheimer’s disease CSF. Journal of Neuroinflammation. 2018;15(1). PMID: 30045733. Lopez-Font I, Sogorb-Esteve A, Javier-Torrent M, Brinkmalm G, Herrando-Grabulosa M, Garcia-Lareu B. et al. Decreased circulating ErbB4 ectodomain fragments as a read-out of impaired signaling function in amyotrophic lateral sclerosis. Neurobiology of Disease. 2019;124:428-438. epub2018. PMID: 30594809. Mata-Balaguer T, Cuchillo-Ibanez I, Calero M, Ferrer I, Saez-Valero J. Decreased generation of c-terminal fragments of apoer2 and increased reelin expression in Alzheimer’s disease. FASEB Journal. 2018;32(7):3536-3546. PMID: 29442527.

Code: PI17/00261. Title: Descifrando las alteraciones en la expresion de acetilcolinesterasa en al enfermedad de Azlheimer. Principal investigator: Maria Salud Garcia Ayllon. CIBERNED’s collaboration: No. CIBERNED groups: G407. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 99220. Duration: 2018-2020.

Code: PI15/00665. Title: Interacciones de las vias Reelina/ApoE y ligandos alternativos con el β-amiloide, su disfuncion en la enfermedad de Alzheimer. Principal investigator: Javier Saez Valero. CIBERNED’s collaboration: No. CIBERNED groups: G407 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 80465. Duration: 2016-2018.

Code: AICO/2018/090. Title: Redefiniendo nuevos marcadores para el diagnostico y terapia de la enfermedad de Alzheimer. Principal investigator: Javier Saez Valero. CIBERNED’s collaboration: No.

87 CIBERNED groups: G407. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Valencia. Funding: 40000. Duration: 2018-2018.

Code: TAUPATH. Title: Tau pathology – common and differing pathways in neurodegenerative tauopathies. Principal investigator: Javier Saez Valero y Maria Salud Garcia Ayllon. CIBERNED’s collaboration: No. CIBERNED groups: G407. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 150000. Duration: 2018-2020.

Code: APE/2017/057. Title: Tau protein – a key but highly complex protein in the pathogenesis of Alzheimer’s disease and other tauopathies. Principal investigator: Javier Saez Valero. CIBERNED’s collaboration: No. CIBERNED groups: G407. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Valencia. Funding: 4500. Duration: 2017-2020.

PHD DISSERTATIONS 2018 Author: Aitana Sogorb-Esteve. Title: Secretases as potential biomarkers and therapeutic target for Alzheimer’s Dis- ease. Date: 25/5/2018. Supervisor: Javier Sáez Valero.

88 CIBERNED 2018 ANNUAL REPORT

408 |Eduardo Soriano García

UNIVERSIDAD DE BARCELONA. FACULTAD DE BIOLOGÍA.

Departamento de Biología Celular, Fisiología e Inmunología. Av. Diagonal 643, Edificio Prevosti 1r Piso. Barcelona. 08028 Tel.: +34 93 403 71 17 Otro Tel.: +34 93 403 47 55 Tel. Fax: +34 93 403 71 16 E-mail: [email protected]

PRINCIPAL Lobón García, Irene. Roselló Busquets, INVESTIGATOR Bachelor degree. Cristina. Bachelor degree. Soriano García, Eduardo Manso Sanz, Yasmina. PhD. Rossi, Daniela. PhD. LIST OF Muhaisen, Ashraf J.M. PhD. Solis Benites, Marco PERSONNEL Antonio. Parcerisas Mosqueda, Bachelor degree. Cotrufo, Tiziana. Antoni. PhD. PhD. Ulloa Darquea, Fausto Dávila Bouziguet, Eva. Alexander. Pascual Sánchez, Bachelor degree. PhD. Marta. Hernaiz Llorens, Marc. PhD. Vilchez Acosta, Alba del Bachelor degree. Valle. Pujadas Puigdoménech, Herrero Lorenzo, Bachelor degree. Lluis. Marina. PhD. Bachelor degree.

ABSTRACT Somatic brain wiring in AD: We have deeply characterized somatic genetic wiring in Alzheimer Disease brains by determining novel somatic variations and copy number variations (CNVs) (Gómez-Ramos et al., 2017; Lobón et al., in preparation). We have also analyzed somatic genetic events in Parkinson Diseased brains, leading to the identification of brain somatic variants (Lobón et al., in preparation). Finally, a review discusses current methods to validate somatic mutations present in low percentage of neural cells (Picher et al., 2018). Reelin and Neurodegeneration: We have submitted a manuscript showing that overexpression of Reelin protects against tau pathology in AD by: a) reducing abnormal dendritic Tau translo- cation, b) reducing Tau phosphorylation, and c) rescuing Tau-associated behavioral deficits. We have completed studies determining the role of Reelin in proteinopaties transmissibility

89 in AD and in Parkinson´s Disease: Overexpression of Reelin in living mice reduced the spreading of amyloid plaques generated by the injection of J20 protein extracts. Similarly, Reelin also reduced the transmissibility of alfa-synuclein in a model of Parkinson´s Disease. These findings are important because, to our knowledge, this is the first therapeutic tool that reduces misfolded protein aggregation and prion-like propagation in living mice. Similarly, we have characterized the role in vivo of APP and Tau mutations on tau-associated pathology, in particular the accumulation of phosphorylated Tau in interneurons (Dávila et al., 2018). SNA- REs, axonal growth and regeneration: We have determined a fundamental role of the centrosomal protein Augmin in the polarization of neuronal microtubules and in the outgrowth of developing axons and dendrites (Freixo et al., 2018). Similarly, we have demonstrated a fundamental rol of SNARE proteins in neurotrophin/Trk mediated axonal growth (Fuschini et al., 2018) and that disruption of lipid rafts enhances axonal growth and regeneration in vitro and in vivo (Roselló et al., 2018). The Armcx gene cluster: We have discovered a role of Armcx protein in the control of cell cycle and in neuronal migration (Mirra et al., 2016) and have identified TDP43 (an ALS gene) and Parkin (Parkinson) as Armcx interaction proteins. The generation of CNS specific KOs (nestin-cre) for the armcx3 gene has allowed us the discovery of a phenotype associated to ALS, i.e., with spinal cord motorneuron death and severe motor deficits. In collaboration with Dr. F Villarroya we have discovered that Armcx3 inactivation (conditional KO) dramatically reduces liver tumor formation by controlling cell proliferation, cell death and lipid accumulation.

KEYWORDS Reelin, Synapses, Alzheimer Disease, Somatic Mutations.

PUBLICATIONS 2018 Dávila E, Targa G, Ávila J, Soriano E, Pascual M. Differential accumulation of Tau phosphorylated at residues Thr231, Ser262 and Thr205 in hippocampal interneurons and its modulation by Tau mutations (VLW) and amyloid-β peptide.Neurobiology of disease. 2018. PMID: 30553886. Picher ÁJ, Hernández F, Budeus B, Soriano E, Avila J. Human Brain Single Nucleotide Poly- morphism: Validation of DNA Sequencing.Journal of Alzheimer’s disease reports. 2018;2(1). PMID: 30480253. Ros O, Barrecheguren P.J, Cotrufo T, Schaettin M, Rosello-Busquets C, Vilchez-Acosta A. et al. A conserved role for Syntaxin-1 in pre- and post-commissural midline axonal guidance in fly, chick, and mouse. PLoS Genetics. 2018;14(6). PMID: 29912942. Freixo F, Martinez Delgado P, Manso Y, Sanchez-Huertas C, Lacasa C, Soriano E. et al. NEK7 regulates dendrite morphogenesis in neurons via Eg5-dependent microtubule stabilization. Nature Communications. 2018;9(1). PMID: 29899413. Garzon F, Coimbra D, Parcerisas A, Rodriguez Y, Garcia J.C, Soriano E. et al. NeuroEPO Pre- serves Neurons from Glutamate-Induced Excitotoxicity. Journal of Alzheimer’s Disease. 2018;65(4):1469-1483. PMID: 30175978.

90 CIBERNED 2018 ANNUAL REPORT

Ulloa F, Cotrufo T, Ricolo D, Soriano E, Araujo S.J. SNARE complex in axonal guidance and neuroregeneration. Neural Regeneration Research. 2018;13(3):386-392. PMID: 29623913. Mata A, Gil V, Pérez-Clausell J, Dasilva M, González-Calixto MC, Soriano E et al. New functions of Semaphorin 3E and its receptor PlexinD1 during developing and adult hippocampal formation.Scientific reports. 2018;8(1). PMID: 29358640. Fuschini G, Cotrufo T, Ros O, Muhaisen A, Andres R, Comella J.X. et al. Syntaxin-1/TI-VAMP SNAREs interact with Trk receptors and are required for neurotrophin-dependent outgrowth. Oncotarget. 2018;9(89):35922-35940. PMID: 30542508.

RESEARCH PROJECTS 2018 Code: PI2016/04. Title: The ALS CIBERNED Challenge: Accelerating New Drug Discovery. Principal investigator: Adolfo Lopez De Munain. CIBERNED’s collaboration: Yes. CIBERNED groups: G609; G303; G503; G408. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

Code: 2017SGR1280 (Solicitado). Title: Ayudas para apoyar las actividades de los grupos de investigacion de Cataluna SGR 2017-2019. Principal investigator: Eduardo Soriano Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G408. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 70500. Duration: 2017-2019.

Code: SAF2016-76340-R. Title: Novel approaches to understand Alzheimers Disease pathogenesis and therapeutics. Principal investigator: Eduardo Soriano Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G408. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 380010. Duration: 2017-2019.

Code: (PI17/02285) (Solicitado). Title: Potencial de los Inhibidores de proteinas SNARE como agentes terapeuticos antitu- morales. Principal investigator: Fausto Ulloa. CIBERNED’s collaboration: No. CIBERNED groups: G408. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 151250. Duration: 2018-2020.

91 Code: La Marató TV3 2016 (Denegado). Title: Synergistic effect of cholesterol depletion and semaphorin blockade in peripheral and central nervous system regeneration. Principal investigator: Ignacio Alfonso Rodriguez / Eduardo Soriano. CIBERNED’s collaboration: No. CIBERNED groups: G408. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundació La Marató de TV3. Funding: 200000. Duration: 2018-2020.

Code: 15/C/2014. Title: The role of reelin at the crossroads of alzheimer’s disease mechanisms: tauopathy, amyloid toxicity and transmissibility. Principal investigator: Lluis Pujadas Puigdomènech. CIBERNED’s collaboration: No. CIBERNED groups: G408. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundació La Marató de TV3. Funding: 187635,5. Duration: 2016-2018.

92 CIBERNED 2018 ANNUAL REPORT

409 | Ignacio Torres Alemán

INSTITUTO CAJAL

Avda. Dr. Arce 37, Madrid Tel. de contacto: +34 91 585 47 23 Fax: +34 91 585 47 54 E-mail de contacto: [email protected]

PRINCIPAL LIST OF Martínez Rachadell, INVESTIGATOR PERSONNEL Laura. Bachelor degree. Torres Alemán, Ignacio. Fernández de Sevilla Pignatelli Garrigós, García-Agenjo, María Jaime. Estrella. PhD. Bachelor degree. Santi Miño, Andrea. Fernández García, Ana. PhD. PhD. Zegarra Valdivia, García García, Miguel. Jonathan Adrián. Technician. Bachelor degree. Herrero Labrador, Raquel. Bachelor degree.

ABSTRACT Ever since its inception, the laboratory works in the neurobiology of mammalian insulin factors (ILPs). During 2018 we have analyzed the role of insulin in brain metabolism, linking it to neurovascular coupling and exploring underlying mechanisms. The latter may be of great significance in vascular brain aging. We also bred transgenic mice with impaired insulin or IGF-I signaling and brain Alzheimer´s disease pathology to explore the impact of these signaling pathways in disease progression. The line of research that we started last year related to IGF-I crosstalk with the hypothalamic orexinergic system, is yielding observations that will help us better understand the neurobiolology of ILPs. We focused on this multifunctional circuit because shares many of the multiple actions exerted by IGF-I on the brain. We are finding that this relatively small hypothalamic nucleus is involved in the effects of IGF-I on sleep, amyloidosis, mood and response to exercise, and even unveiled a striking impact on peripheral glucose metabolism. Further, we are analyzing mechanisms underlying

93 the neuron-astrocyte dialog in glucose metabolism and its potential impact on Abeta handling. Also, we are completing the characterization of an interaction between IGF-I and ApoE in Abeta metabolism.

KEYWORDS Insulin factors, neuronal plasticity, and degenerative diseases/ Therapeutic targets in neurodegeneration/ the aging brain and cognition/ Blood brain barrier/ Mood homeostasis/Orexin circuitry.

PUBLICATIONS 2018 Santi A, Bot M, Aleman A, Penninx B.W.J.H, Aleman I.T. Circulating insulin-like growth factor I modulates mood and is a biomarker of vulnerability to stress: from mouse to man. Transla- tional Psychiatry. 2018;8(1). PMID: 30068974. Fernandez AM, Santi A, Torres Aleman I. Insulin Peptides as Mediators of the Impact of Life Style in Alzheimer’s disease.Brain plasticity (Amsterdam, Netherlands). 2018;4(1). PMID: 30564544.

Code: PI2016/01. Title: Alteraciones del metabolismo gluco-lipidico y desarrollo de la demencia de Alzheimer. Principal investigator: Ignacio Torres Aleman. CIBERNED’s collaboration: Yes. CIBERNED groups: G409; G402; G511; G502; G412. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

94 CIBERNED 2018 ANNUAL REPORT

Code: SAF2016-76462-C2-1-P. Title: El sistema IGF-I/orexina/acetilcolina como nexo de union entre co-morbilidad. Principal investigator: Ignacio Torres Aleman. CIBERNED’s collaboration: No. CIBERNED groups: G409. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 260000. Duration: 2017-2020.

PHD DISSERTATIONS 2018 Author: Andrea Santi Miño. Title: Vulnerabilidad a estrés del cerebro lesionado: el IGF-I como modulador de la sensibilidad del eje HPA. Date: 21/2/2018. Supervisor: Ignacio Torres Alemán.

95 410 | Ramón Trullás Oliva

UNIDAD DE NEUROBIOLOGÍA. IIBB-CSIC. IDIBAPS c/ Rosselló 161, 7ª Planta, Lab. 711 08036 Barcelona Tel.de contacto: +34 659 696 187 Fax: +34 93 363 83 01 E-mail de contacto: [email protected]

PRINCIPAL LIST OF Rodríguez Allue, Manuel INVESTIGATOR PERSONNEL José. PhD. Trullás Oliva, Ramón. Colell Riera, Anna. PhD. Serra Barea, Nuria. Technician. Podlesniy, Petar. PhD. Puigrós Serra, Margalida. Bachelor degree.

ABSTRACT During the year 2018, we have characterized a new digital PCR method to measure directly the concentration of circulating extracellular mitochondrial DNA in cerebrospinal fluid. This new method allows the detection and absolute quantification of the amount of mitochondrial DNA with high analytical sensitivity, specificity and precision. Using this new technique, we have initiated studies to determine whether the content of mitochondrial DNA in the cerebrospinal fluid of patients with Alzheimer’s disease is related with disease progress. Other studies from our group have shown that high cholesterol levels prevent the degradation by autophagy of beta amyloid and tau while promoting their secretion. Specifically, we have shown that high cholesterol levels increase the formation of autophagosomes, but at the same time inhibit the fusion of autophagosomes with lysosomal vesicles. In addition, treatment with 2-hydroxypropyl-β-cyclodextrin reduces cholesterol levels and completely reverses the alterations evoked by cholesterol, which may represent a new therapeutic strategy in the treatment of Alzheimer’s disease. Another line of research of our group has studied the role of the ATP-sensitive potassium channel (KATP) in the pathology of amyotrophic lateral sclerosis (ALS). The results indicate that the expression of the KATP channel is increased in post-mortem human tissue samples from the motor cor-

96 CIBERNED 2018 ANNUAL REPORT tex and the cervical and thoracic spinal cord of people with sporadic ALS. Likewise, the results indicate that polymorphisms of genes that encode subunits of KATP are related to the progression of the disease, which suggests that this channel plays a key role in the pathophysiology of ALS.

KEYWORDS Mitochondrial DNA. Synaptic Neurodegeneration. Biomarkers of neurodegenerative diseases. Mitochondrial dynamics. Mitochondrial transport. Neuronal Death. Neuronal Pentraxins. Molecular mechanisms of neurodegeneration.

PUBLICATIONS 2018 Menal M.J, Jorba I, Torres M, Montserrat J.M, Gozal D, Colell A. et al. Alzheimer’s disease mutant mice exhibit reduced brain tissue stiffness compared to wild-type mice in both nor- moxia and following intermittent hypoxia mimicking sleep apnea. Frontiers in Neurology. 2018;9(JAN). PMID: 29403429. Podlesniy P, Trullas R. Biomarkers in cerebrospinal fluid: Analysis of cell-free circulating mitochondrial dna by digital PCR. Methods in Molecular Biology. 2018;1768:111-126. PMID: 29717440. Barbero-Camps E, Roca-Agujetas V, Bartolessis I, de Dios C, Fernandez-Checa J.C, Mari M. et al. Cholesterol impairs autophagy-mediated clearance of amyloid beta while promoting its secretion. Autophagy. 2018;14(7):1129-1154. PMID: 29862881. Vidal-Taboada J.M, Pugliese M, Salvado M, Gamez J, Mahy N, Rodriguez M.J. KATP Chan- nel Expression and Genetic Polymorphisms Associated with Progression and Survival in Amyotrophic Lateral Sclerosis. Molecular Neurobiology. 2018;55(10):7962-7972. PMID: 29492846. Figueiro-Silva J, Antequera D, Pascual C, de la Fuente Revenga M, Volt H, Acuna-Castroviejo D. et al. The Melatonin Analog IQM316 May Induce Adult Hippocampal Neurogenesis and Preserve Recognition Memories in Mice. Cell Transplantation. 2018;27(3):423-437. PMID: 29873251.

RESEARCH PROJECTS 2018 Code: PI2016/06. Title: Identificacion de vias fisiopatologicas y biomarcadores candidatos en la fase pre-di- agnostica de la enfermedad de Parkinson. Principal investigator: Miquel Vila Bover. CIBERNED’s collaboration: Yes. CIBERNED groups: G109 ; G601; G207; G410. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 196000. Duration: 2016-2018.

97 Code: SAF2017-89791-R. Title: Replication and transcription of mitochondrial DNA as a central mechanism of the pathological sequence leading to neurodegeneration. Principal investigator: Ramon Trullas. CIBERNED’s collaboration: No. CIBERNED groups: G410. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 242000. Duration: 2018-2020.

98 CIBERNED 2018 ANNUAL REPORT

411 | Javier Vitorica Ferrández

DPTO. BIOQUÍMICA Y BIOLOGÍA MOLECULAR · FACULTAD DE FARMACIA · UNIVERSIDAD DE SEVILLA. INSTITUTO DE BIOMEDICINA DE SEVILLA · HOSPITAL UNIVERSITARIO VIRGEN DEL ROCÍO. SEVILLA.

Tél.: +34 954 556 770 Fax: +34 955 923 053 E-mail: [email protected]

PRINCIPAL LIST OF Romero Molina, Carmen. INVESTIGATOR PERSONNEL Bachelor degree. Sánchez Micó, María Vitorica Ferrández, Jiménez Muñoz, Virtudes. Francisco Javier. Sebastián. Bachelor degree. Bachelor degree. Vizuete Chacón, María Muñoz Castro, Clara. Luisa. Becario. Bachelor degree. Navarro Garrido, Victoria. Bachelor degree.

ABSTRACT In 2018, we continued with the characterization of the microglial and astroglial response in murine models of Alzheimer’s disease, Abeta and Tau, and in AD patient samples. Within this characterization, we are developing new models to mimic the microglial dysfunction observed in Alzheimer’s patients. Specifically, we are developing Abeta and Tau models, conditional knockouts of the Csf1R gene specifically in microglial cells with the idea of diminishing their ability to survive and function. These models will allow us to establish the function of microglial cells in the development of Abeta and /or tau pathology. On the other hand, we have also transcriptionally characterized the microglia and isolated astrocytes of Abeta (APPsl) and Tau (P301S) models. In this sense, the microglial cells have been isolated by cell sorting using both classical markers, CD11b and CD45, and the activation marker DAM Clec7a (see attached figure). Astrocytes from the same animals were isolated using ACSA2 (see figure). This approach was carried out at two ages, prepatology (3 months) or established pathology (12 or 9 months for APP and Tau, respectively). Among the most relevant results, and contrary to what might be expected based on their morphology, the astrocytes of both models present a clear pattern of metabolic deficiencies. We are currently completing this analysis for publication. On the other hand, the microglia in both models also presents a clear profile of activation to the

99 phenotype called DAM (disease associated microglia). Interestingly, although APP and Tau proteinopities are completely different, we did not observe large differences in their transcriptional profile. We are currently characterizing its function in pathology. This objective is encompassed within an intramural CIBERNED project (IP Javier Vitorica) and has been carried out in collaboration with Dr. E. Galea (Marató de TV3). Finally, we have established collaborations to determine, on the one hand, the role of hypoxia in the microglial response in the pathology of Alzheimer’s and, on the other hand, we are studying the role of possible ligands of Trem2 in the microglial activation in this pathology.

KEYWORDS Alzheimer, degeneration, inflammation, Abeta, tau, oligomers, soluble, transgenic models, neuropathology.

PUBLICATIONS 2018 Gutierrez A, Vitorica J. Toward a New Concept of Alzheimer’s Disease Models: A Perspective from Neuroinflammation. Journal of Alzheimer’s Disease. 2018;64(s1):S329-S338. PMID: 29562520. Gomez-Arboledas A, Davila J.C, Sanchez-Mejias E, Navarro V, Nunez-Diaz C, Sanchez-Varo R. et al. Phagocytic clearance of presynaptic dystrophies by reactive astrocytes in Alzheimer’s disease. GLIA. 2018;66(3):637-653. PMID: 29178139. Romero-Molina C, Navarro V, Sanchez-Varo R, Jimenez S, Fernandez-Valenzuela J.J, Sanchez-Mi- co M.V. et al. Distinct microglial responses in two transgenic murine models of TAU pathology. Frontiers in Cellular Neuroscience. 2018;12. PMID: 30487735. Navarro V, Sanchez-Mejias E, Jimenez S, Munoz-Castro C, Sanchez-Varo R, Davila J.C. et al. Microglia in Alzheimer’s disease: Activated, dysfunctional or degenerative. Frontiers in Aging Neuroscience. 2018;10(MAY). PMID: 29867449.

100 CIBERNED 2018 ANNUAL REPORT

101 412 | Francisco Wandosell Jurado

CENTRO DE BIOLOGÍA MOLECULAR “SEVERO OCHOA”, CSIC-UAM, UNV. AUTÓNOMA DE MADRID C/ Nicolás Cabrera nº 1, 28049 Cantoblanco, Madrid Tel: +34 91 196 45 61 / 45 91 Fax: +34 91 196 44 20 E-mail: [email protected], [email protected] Web: www.ciberned.es/ grupofranciscowandosell.aspx

PRINCIPAL LIST OF Ordóñez Gutiérrez, Lara. INVESTIGATOR PERSONNEL PhD. Wandosell Jurado, Antón Gutiérrez, Inés. Pérez Álvarez , María Francisco. PhD. José. PhD. Benítez Moreno, María José. Rivas Muñoz, Sergio. PhD. Bachelor degree. Garrido Jurado, Juan Villa, Mario. Bachelor degree. Jose. PhD.

ABSTRACT Our group, “Molecular mechanisms of Neurodegeneration“, is generated from an established line in the CBM over several years. At present, this CIBERNED line is formally composed by three sub-groups (IP´s-F Wandosell (CBM), I. Anton (CNB) and J. J. Gar- rido, Cajal Inst.). Our group is mostly devoted to the analysis of neurodegenerative disorders. Now is focusing in a group of brain disorders associated with aging, such as Alzheimer, disease, Stroke and some type of brain tumours. We are interested in the analysis of which cellular signals are altered throughout the aging process and how the modification of some of them triggers a pathological process. Thus one open question is ... are there common molecular mechanisms underlying the age-de- pendency of several disorders?. Data from different animal models strongly support that idea that -PI3K-Akt pathway is deregulated in several brain pathologies. First, we are interested in the analysis of the molecular mechanisms of both brain disorders, mainly focusing in the role of PI3K-Akt and some of the Akt substrates. Our previous work has led us to analyze in more detail some Akt downstream element such as, mTORC1. This protein complex controls general aspects of protein synthesis

102 CIBERNED 2018 ANNUAL REPORT and autophagy; whereas its dysfunction has been considering a key factor of beta amyloid generation and/or degradation in Alzheimer disease (AD). We just analyze the role of mTORC1 activity in the generation of amyloid in AD transgenic models, and we are interested in the analysis of early stages of this pathology. (CoIPs: Dr. J.J.Garrido & F Wandosell). Our second goal is the analysis of a new oncogenic pathway that we recently described: Akt/WIP/YAP/TAZ. Our data indicated that Akt and WIP are responsible of the cell division of cancer-stem cells and the maintenance of its stem-like phenotype. Our work is about defining the elements of this path that are between Akt and WIP and WIP and YAP/TAZ that regulate the conversion from astrocyte-astrocytoma-glioma (CoIPs: Dr. I. Anton & F. Wandosell). In summary, we are analysing some signalling and elements that control physiological process, from cell division to differentiation, and how some of these proteins are modified in pathology.diferenciación, y cómo de PI3K-Akt algunas de estas proteínas se modifican en patología.

KEYWORDS Neurodegeneration, Cell signaling, Neuroprotection, Estrogens, Alzheimer’s disease, Neuritogenesis, Axonal polarity, glia growth, Astrocyte / glioma conversion, Cytoskeleton, Actin.

PUBLICATIONS 2018 Antón IM, Gómez-Oro C, Rivas S, Wandosell F. Crosstalk between WIP and Rho family GTPases. Small GTPases. 2018. PMID: 29172947. Ordonez-Gutierrez L, Benito-Cuesta I, Abad J.L, Casas J, Fabrias G, Wandosell F. Dihydrocer- amide Desaturase 1 Inhibitors Reduce Amyloid-β Levels in Primary Neurons from an Alzheim- er’s Disease Transgenic Model. Pharmaceutical Research. 2018;35(3). PMID: 29411122. Herrera J.L, Ordonez-Gutierrez L, Fabrias G, Casas J, Morales A, Hernandez G. et al. Ovarian function modulates the effects of long-chain polyunsaturated fatty acids on the mouse cerebral cortex. Frontiers in Cellular Neuroscience. 2018;12. PMID: 29740285. Rivas S, Gómez-Oro C, Antón IM, Wandosell F. Role of Akt Isoforms Controlling Cancer Stem Cell Survival, Phenotype and Self-Renewal.Biomedicines. 2018;6(1). PMID: 29518912. Perez-Alvarez M.J, Gonzalez M.V, Benito-Cuesta I, Wandosell F.G. Role of mTORC1 controlling proteostasis after brain ischemia. Frontiers in Neuroscience. 2018;12(FEB). PMID: 29497356. Sanz-Rodriguez M, Gruart A, Escudero-Ramirez J, de Castro F, Delgado-Garcia J.M, Wandosell F. et al. R-ras1 and r-ras2 are essential for oligodendrocyte differentiation and survival for cor- rect myelination in the central nervous system. Journal of Neuroscience. 2018;38(22):5096- 5110. PMID: 29720552. Céspedes Rubio ÁE, Pérez-Alvarez MJ, Lapuente Chala C, Wandosell F. Sex steroid hormones as neuroprotective elements in ischemia models.The Journal of endocrinology. 2018;237(2). PMID: 29654072.

103 Rivas S, Anton I.M, Wandosell F. WIP-YAP/TAZ as a new pro-oncogenic pathway in glioma. Cancers. 2018;10(6). PMID: 29890731. Pose-Utrilla J, Garcia-Guerra L, Del Puerto A, Martin A, Jurado-Arjona J, De Leon-Reyes N.S. et al. Erratum: Author Correction: Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1 (Nature communications (2017) 8 1 (2275)). Nature communications. 2018;9(1):473-. PMID: 29382840.

Code: PI2016/01. Title: Alteraciones del metabolismo gluco-lipidico y desarrollo de la demencia de Alzheimer. Principal investigator: Ignacio Torres Aleman. CIBERNED’s collaboration: Yes. CIBERNED groups: G409 ; G402; G511; G502; G412. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

Code: SAF2015-70368-R. Title: Analisis de la senalizacion mediada por akt en neurodegeneracion y en prolifera- cion, migracion/invasion celulares. Principal investigator: Francisco Wandosell, Ines M. Anton Gutierrez. CIBERNED’s collaboration: No. CIBERNED groups: G412. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 220000. Duration: 2016-2018.

Code: OC-2015-1-19840. Title: European Network of multidisciplinary research and translation of autophagy knowledge. Principal investigator: Caty Casas. CIBERNED’s collaboration: No. CIBERNED groups: G412 .Other CIBER’s collaboration: No. Type: Europeo.

104 CIBERNED 2018 ANNUAL REPORT

Funding agency: Comision Europea. Funding: ND. Duration: 2016-2020.

Code: Fundacion RAMON ARECES. Title: Interactoma diferencial de WIP: Actividad oncogenica versus supresora de tumores”. Principal investigator: Francisco Wandosell, Ines Anton. CIBERNED’s collaboration: No. CIBERNED groups: G412. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Ramon Areces. Funding: 40000. Duration: 2017-2019.

Code: PCIN-2016-108. Title: Regulacion del reflejo de miccion despues de lesionmedular: abolicion por silenciam- iento de los aferentes de las fibras C de la vejiga hiper-excitados mediante terapia genica para restaurar la continencia y la miccion”. Principal investigator: Francisco Wandosell. CIBERNED’s collaboration: No. CIBERNED groups: G412. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 150000. Duration: 2017-2019.

105 106 CIBERNED 2018 ANNUAL REPORT

PROGRAM 2 PARKINSON’S AND HUNTINGTON’S DISEASE AND OTHER DEGENERATIVE MOVEMENT DISORDERS

107 108 CIBERNED 2018 ANNUAL REPORT

This Program brings together 25 basic and clinical research groups with a mainly translational character, joining forces to study neurodegenerative diseases of different etiology that cause important problems in patient’s mobility. Among this group of diseases we can find, by decreasing prevalence: Parkinson’s disease, Huntington’s chorea, and different kinds of ataxias among other movement disorders.

RESEARCH LINE 1: PARKINSON’S DISEASE Parkinson’s disease (PD) is mainly characterized by neuronal loss, the formation of Lewy bodies and neurites in the substantia nigra and the consequent loss of striatal dopamine (DA). However, it is currently well known that PD is a multisystem neurodegenerative process, in which, as the neurodegenerative process evolves, many areas of the nervous system are affected and there exists a deficit in several neurotransmission and neuromodu- It is estimated lation systems. It is estimated to affect around 160,000 people in Spain, a figure that is ex- to affect around pected to be increasing due to the progressive 160,000 people aging of the population. in Spain, a figure Although preventing and correcting DA deficit are still important goals, they cannot be that is expected to considered the ultimate challenge in PD now- be increasing due adays. Within this area, it is considered of vital to the progressive importance to make progress on defining the following issues: aging of the a) Key aspects related to the ethiopatho- population. genesis in PD. b) Physiopatological mechanisms related to disease onset and progression. c) Development of symptomatic treatments, especially neuroprotective and curative. Thus, a truly translational research is sought, having the disease and the patients as the main targets. Thus, this line of research intends to achieve a true translational research nature, whose main objectives are the disease and the patient. The main research topics in this line are the following:

• Cognitive impairment and non-motor problems in PD.

109 • Biomarkers in Parkinson’s disease. • Problems related to symptomatic treatment: diskynesias. • New targets and novel therapeutic strategies in Parkinson’s disease. • Circuits and physiopathology of basal ganglia. • Neuronal stress, cell protection and death in Parkinson’s disease. • Neurogenesis and cell therapy in Parkinson’s disease. • Early biomarkers in Parkinson’s disease.

RESEARCH LINE 2: HUNTINGTON’S DISEASE AND ATAXIAS This program also focuses on the research into other movement disorders such as Hun- tington’s disease (HD) and ataxias. HD is characterized by the initial loss of spiny interneurons of the striatum. It is an autosomal dominant neurodegenerative pathology with complete penetrance produced by polyglutamine expansion in the huntingtin N-ter- minus. HD has no treatment and leads to death in around 10-20 years depending on the number of polyglutamines, the age of onset, some unknown environmental factors and the modulation of some genes, some of which have been located but remain unidentified. HD has a much lower prevalence than AD or PD, 10 cases/100,000 inhabitants, and it is estimated that there are about 4,000 patients in Spain and about 50,000 in the Europe- an Union. Its social health cost is considerable because of the importance of cognitive and motor deficits, as well as the severe behavioral problems that patients present. ... it is estimated There is a large number of studies with neu- there are about 4,000 roprotective drugs which modify the supposed pathogenic mechanisms or that are used in patients in Spain and other neurodegenerative diseases. These about 50,000 in the drugs include inhibitors of neuronal excitation, European Union. coenzymes of the respiratory chain, vitamins, antioxidants, co-adjuvants in energy production, etc. Some of these products offer encouraging results in experimental models of the disease but, unfortunately, not confirmed in the clinic. The study of HD is important because is the best studied and most prevalent model of neurodegenerative diseases caused by triplet expansions, which also includes some ataxias. Discovering the pathogenic mechanisms of HD and finding an effective, either neuroprotective or curative, would have immediate implications on any of the other neurodegenerative pathologies caused by triplet expansions. The main research topics in this line are the following:

• Identification of molecular and cellular basis of Huntington’s disease. • Experimental studies in animal models of Huntington’s disease. • Clinic, genetics and neuropathology of Huntington’s disease.

110 CIBERNED 2018 ANNUAL REPORT

PRINCIPAL INVESTIGATOR INSTITUTION

Alberch Vie, Jordi University of Barcelona

Canela Campos, Enric Isidre University of Barcelona

Ceña Callejo, Valentín University of Castilla La Mancha, Albacete

Cuadrado Pastor, Antonio Autonomous Universidad of Madrid

Del Río Fernández, José Antonio Catalonian Institute de Bioengineering, Barcelona

Fariñas Gómez, Isabel University of Valencia

Fernández Ruiz, Javier Complutense University of Madrid

Fuentes Rodríguez, José Manuel Universidad of Extremadura, Cáceres

García Verdugo, José Manuel Cavanilles Institute, University of Valencia

Guzmán Pastor, Manuel Complutense University of Madrid

Iglesias Vacas, Teresa Institute of Biomedical Research CSIC-UAM, Madrid

Kulisevsky Bojarski, Jaime Santa Creu i Sant Pau Hospital, Barcelona

Labandeira García, José Luis Universidad of Santiago de Compostela

Lanciego Pérez, José Luis Center of Applied Medical Research, Univ. Navarra, Pamplona

López Barneo, José Virgen del Rocío University Hospital, University of Seville

Lucas Lozano, José Javier Center for Molecular Biology "Severo Ochoa" CSIC-UAM, Madrid

Moratalla Villalba, Rosario Cajal Institute CSIC, Madrid

Naranjo Orovio, José Ramón National center of Biotechnology CSIC, Madrid

Obeso Inchausti, José Ángel Hospital Madrid Foundation

Pérez Castillo, Ana María Institute of Biomedical Research CSIC-UAM, Madrid

Pérez Tur, Jordi Institute of Biomedicine of Valencia, CSIC

Rodríguez Díaz, Manuel University of La Laguna, Tenerife

Tolosa Sarró, Eduardo Clinic Hospital of Barcelona

Vicario Abejón, Carlos Cajal Institute, CSIC, Madrid

Vila Bover, Miquel Vall d'Hebron University Hospital, Barcelona

Program 2 is coordinated by Drs. José J. Lucas (Center for Molecular Biology, CSIC, and Eduardo Tolosa (Clinic Hospital, Barcelona). Teresa Iglesias Vacas (Biomedical Research Institute CSIC-UAM, Madrid), and Isabel Fariñas (University of Valencia).

111 301 | Jordi Alberch Vié

DEPARTAMENTO DE BIOMEDICINA. FACULTAT DE MEDICINA, UNIVERSITAT DE BARCELONA INSTITUT D’INVESTIGACIONS BIOMÈDIQUES AUGUST PI I SUNYER (IDIBAPS)

Tel. +34 93 403 52 58 / 37252 Fax +34 93 402 19 07 [email protected]

PRINCIPAL LIST OF López Alonso, Ana INVESTIGATOR PERSONNEL María. Technician. Alberch Vié, Jordi. Conde Berriozábal, Sara. Pérez Navarro, Esther. Bachelor degree. PhD. Ginés Padrós, Silvia. Perpiña Martin, Unai. PhD. PhD.

ABSTRACT The lines of research during 2018 have focused on identifying new therapeutic targets for Huntington’s disease (HD) and in the study of the intracellular mechanisms activated by mutant huntingtin that produce neuronal dysfunction. We have also continued our studies on the role of mutant huntingtin during development and in the differentiation of iPSCs to striatal neuronal phenotypes. Following our interest in the contribution of neurotrophic factors and their receptors in the pathology of HD, we have validated the imbalance between TrkB / p75NTR receptors as a mechanism involved in the appearance of alterations in the motor coordination of this disease (Suelves et al. al, 2018). In addition, we have demonstrated the participation of the ARMS protein in the deregulation of BDNF secretion in the hippocampus of HD models, which could be contributing to cognitive deficits (Lopez-Benito et al, 2018). We have also described a new molecule with therapeutic potential for HD as the pituitary adenylate cyclase activating polypeptide (PACAP). This factor enhances the expression of BDNF and reduces the formation of aggregates. Treatment with PACAP produced an improvement in cognitive alterations in models of HD. During this year we have extended the studies on the implication of reduced levels of STEP phosphatase in the pathophysiology of HD. For this we obtained a model of the HD and deficient in STEP, and we also inhibited the activity of STEP through pharmacological treatment. The lack of expression of STEP delays the onset of motor symptoms and prevents cognitive dysfunction. In addition, the acute inhibition of its activity improves cognitive function in a

112 CIBERNED 2018 ANNUAL REPORT model of HD. Therefore, the inhibition of STEP would be a good therapeutic strategy for HD (Garcia-Forn, Martínez-Torres et al., 2018). We also analyzed the expression of STEP during postnatal development in control mice, and its modulation by BDNF. The results obtained show that STEP levels decrease with age in the striatum and their levels are regulated by BDNF in the striatum and in the cerebral cortex during postnatal development (Cases et al., 2018). We have also observed that the activity of the mTORC2 kinase is increased in the striatum of HD mice due to an increase in Rictor levels. Our results indicate that it is a compensatory mechanism to avoid the toxicity of mutant huntingtin, suggesting the modulation of Rictor levels in the striatum as a new therapeutic target for HD (Creus-Muncunill et al., 2018). For the study of the contribution of Cdk5 in the depressive aspects in HD, we have created a new murine model that expresses mutant huntingtin and is heterozygous for the phosphorylation site of DARPP32 controlled by Cdk5. In these mice we have shown that the depressive phenotype manifested at two months of age was not observed, suggesting a critical role of the Cdk5 / DARPP32 pathway in the development of psychiatric disorders such as depression in HD. In our studies on how the development of the striatum affects the progression of HD, we have demonstrated that the regulation of the expression of Kv7 channels are crucial for the functional maturation of striatal neurons in the mouse and in human iPSC striatal neurons (Telezhkin et al., 2018).

KEYWORDS Neuronal plasticity - Neurotrophic factors - Depression - Neurodevelopment - Huntington’s disease.

PUBLICATIONS 2018 Siegert A.-M, Garcia Diaz-Barriga G, Esteve-Codina A, Navas-Madronal M, Gorbenko del Blanco D, Alberch J. et al. A FBN1 3′UTR mutation variant is associated with endoplasmic reticulum stress in aortic aneurysm in Marfan syndrome. Biochimica et Biophysica Acta - Molecular Ba- sis of Disease. 2019;1865(1):107-114. epub2018. PMID: 30385411. Blazquez G, Castane A, Saavedra A, Masana M, Alberch J, Perez-Navarro E. Social memory and social patterns alterations in the absence of striatal-enriched protein tyrosine phosphatase. Frontiers in Behavioral Neuroscience. 2019;12epub2018. PMID: 30760987. Fernandez-Flores F, Garcia-Verdugo J.M, Martin-Ibanez R, Herranz C, Fondevila D, Canals J.M. et al. Characterization of the canine rostral ventricular-subventricular zone: Morphological, immunohistochemical, ultrastructural, and neurosphere assay studies. Journal of Compara- tive Neurology. 2018;526(4):721-741. PMID: 29205371. Cases S, Saavedra A, Tyebji S, Giralt A, Alberch J, Perez-Navarro E. Age-related changes in STri- atal-Enriched protein tyrosine Phosphatase levels: Regulation by BDNF. Molecular and Cellular Neuroscience. 2018;86:41-49. PMID: 29122705.

113 Suelves N, Miguez A, López-Benito S, Barriga GG, Giralt A, Alvarez-Periel E et al. Early Down- regulation of p75NTR by Genetic and Pharmacological Approaches Delays the Onset of Motor Deficits and Striatal Dysfunction in Huntington’s Disease Mice.Molecular neurobiology. 2018. PMID: 29804232. Masana M, Westerholz S, Kretzschmar A, Treccani G, Liebl C, Santarelli S. et al. Expression and glucocorticoid-dependent regulation of the stress-inducible protein DRR1 in the mouse adult brain. Brain Structure and Function. 2018;223(9):4039-4052. PMID: 30121783. Cabezas-Llobet N, Camprubi S, Garcia B, Alberch J, Xifro X. Human alpha 1-antitrypsin protects neurons and glial cells against oxygen and glucose deprivation through inhibition of inter- leukins expression. Biochimica et Biophysica Acta - General Subjects. 2018;1862(9):1852- 1861. PMID: 29857082. Saavedra A, Garcia-Diaz Barriga G, Perez-Navarro E, Alberch J. Huntington’s disease: novel therapeutic perspectives hanging in the balance. Expert Opinion on Therapeutic Targets. 2018;22(5):385-399. PMID: 29671352. Creus-Muncunill J, Rue L, Alcala-Vida R, Badillos-Rodriguez R, Romani-Aumedes J, Marco S. et al. Increased Levels of Rictor Prevent Mutant Huntingtin-Induced Neuronal Degeneration. Molecular Neurobiology. 2018;55(10):7728-7742. PMID: 29460266. Telezhkin V, Straccia M, Yarova P, Pardo M, Yung S, Vinh N.-N. et al. Kv7 channels are upregulated during striatal neuron development and promote maturation of human iPSC-derived neurons. Pflugers Archiv European Journal of Physiology. 2018;470(9):1359-1376. PMID: 29797067. Garcia-Forn M, Martinez-Torres S, Garcia-Diaz Barriga G, Alberch J, Mila M, Azkona G. et al. Pharmacogenetic modulation of STEP improves motor and cognitive function in a mouse model of Huntington’s disease. Neurobiology of Disease. 2018;120:88-97. PMID: 30176350. Cabezas-Llobet N, Vidal-Sancho L, Masana M, Fournier A, Alberch J, Vaudry D. et al. Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) Enhances Hippocampal Synaptic Plas- ticity and Improves Memory Performance in Huntington’s Disease. Molecular Neurobiology. 2018;55(11):8263-8277. PMID: 29526016. Saavedra A, Ballesteros JJ, Tyebji S, Martínez-Torres S, Blázquez G, López-Hidalgo R et al. Proteolytic Degradation of Hippocampal STEP61 in LTP and Learning.Molecular neurobiology. 2018. PMID: 29948948. Giralt A, de Pins B, Cifuentes-Diaz C, Lopez-Molina L, Farah A.T, Tible M. et al. PTK2B/Pyk2 overexpression improves a mouse model of Alzheimer’s disease. Experimental Neurology. 2018;307:62-73. PMID: 29803828. Lopez-Benito S, Sanchez-Sanchez J, Brito V, Calvo L, Lisa S, Torres-Valle M. et al. Regulation of BDNF release by ARMS/Kidins220 through modulation of synaptotagmin-IV levels. Journal of Neuroscience. 2018;38(23):5415-5428. PMID: 29769266. Kretzschmar A, Schulke J.-P, Masana M, Durre K, Muller M.B, Bausch A.R. et al. The stress-inducible protein DRR1 exerts distinct effects on actin dynamics. International Journal of Mo- lecular Sciences. 2018;19(12). PMID: 30545002.

114 CIBERNED 2018 ANNUAL REPORT

RESEARCH PROJECTS 2018 Code: COMRDI15-1-0013. Title: ADVANCECAT: acceleradora pel desenvolupament de teràpies avançades a Catalun- ya (ADVANCECAT: accelerating development for advanced therapies in Catalonia). Principal investigator: Josep M. Canals. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: CIBER-BBN. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 969352. Duration: 2016-2019.

Code: RD16/0011. Title: Red de Terapia Celular. Principal investigator: Isabel Farinas. CIBERNED’s collaboration: Yes. CIBERNED groups: G607 ; G301; G102; G113; G208; G105; G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 284999. Duration: 2016-2020.

Code: SAF2016-08573-R. Title: Alteraciones de la lamina nuclear y de la traduccion proteica como nuevos mecanismos patogenicos en la enfermedad de Huntington. Principal investigator: Esther Perez Navarro. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 181500. Duration: 2016-2019.

Code: 813851. Title: ASCTN-Training: Training on Advanced Stem Cell Technologies in Neurology. Principal investigator: Josep M. Canals (Coordinador general: Josep M. Canals). CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 3749403. Duration: 2018-2022.

Code: SAF2015-67474-R. Title: Cdk5 como nueva diana terapeutica y biomarcador del trastorno depresivo en la enfermedad de Huntington. Principal investigator: Silvia Gines. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 217800. Duration: 2016-2018.

115 Code: 713140. Title: Custom architecturally defined 3D stem cell derived functional human neural networks for transformative progress in neuroscience and medicine (MESO_BRAIN). Principal investigator: Jordi Soriano. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 3225890. Duration: 2016-2019.

Code: 307272. Title: Desarrollo experimental del tratamiento con AAT-1 como terapia antiinflamatoria en la isquemia neonatal (el estudio A) y ensayo de la AAT-1 en la lesion aguda de la medula espinal (Estudio B). Principal investigator: Jordi Alberch Vie. CIBERNED’s collaboration: No. CIBERNED groups: G30 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Instituto Grifols. Funding: 50254. Duration: 2013-2018.

Code: SAF2015-66505-R. Title: Desarrollo, diferenciacion y maduracion neuronal en la enfermedad de Huntington. Principal investigator: Josep M. Canals. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 181500. Duration: 2016-2018.

Code: H2020-MSCA-ITN-2016. Title: European Training Network for Cell-based Regenerative Medicine. Principal investigator: Jordi Alberch. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 247873. Duration: 2016-2020.

Code: 309965. Title: Identification of chorein function in neurodegeneration of the basal ganglia for developing new therapeutical approaches in Chorea-Acanthocytosis. Principal investigator: Jordi Alberch Vie. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion ChAc. Funding: 37200. Duration: 2018-2021.

116 CIBERNED 2018 ANNUAL REPORT

Code: A14079. Title: In vitro Study of Neurodevelopment in Huntington’s disease. Principal investigator: Josep M. Canals. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: CHDI. Funding: 1731009. Duration: 2018-2021.

Code: RL000776. Title: Mitochondrial outcomes measures in fibroblasts of HD patients. Principal investigator: Silvia Gines. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: HDSA. Funding: 75000. Duration: 2017-2018.

Code: 20140130/1. Title: Modulacion del deficit de la plasticidad sinaptica como estrategia terapeutica en la enfermedad de Hunttington. Principal investigator: Jordi Alberch. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 299375. Duration: 2015-2018.

Code: FBG 308408. Title: Obtaining, processing an/or preserving non-human tissues, cells and related products. Principal investigator: Josep M. Canals. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: Cytes Biotechnologies. Funding: 450000. Duration: 2015-2020.

Code: FBG 308841. Title: Servicio de Salas Blancas para produccion de vacuna de HIV en condiciones GMP. Principal investigator: Josep M. Canals, Raquel Martin Ibanez. CIBERNED’s collaboration: No. CIBERNED groups: G301. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: IDIBAPS. Funding: 360000. Duration: 2016-2018.

Code: A12076. Title: Studying Human MSN Differentiation from PSC using Single-Cell RNAseq and Ro- dent Chimeric Models.

117 Principal investigator: Josep M. Canals. CIBERNED’s collaboration: No. CIBERNED groups: G301 . Other CIBER’s collaboration: No. Type: Internacional. Funding agency: CHDI. Funding: 553417. Duration: 2016-2018.

PHD DISSERTATIONS 2018 Author: Elena Alvarez Periel. Title: Dual role of Cdk5 in cogntive deficits and striatal vulnerability in Huntington´s disease. Date: 5/6/2018. Supervisor: Silvia Ginés Padrós.

Author: Nuria Suelves Caballol. Title: Evaluation of therapeutic targets for the treatment of behavioral alterations and neuropathology in Huntington´s disease. Date: 19/6/2018. Supervisor: Silvia Ginés Padrós.

Author: Alfonso Gerardo García Díaz Barriga. Title: Therapeutic strategies based on neuroprotective mechanisms of neurotrophic factors in Huntington’s Disease models. Date: 17/7/2018. Supervisor: Jordi Alberch Vié.

Author: Andrea Comella Bolla. Title: Neuronal differentiation and maturation of human pluripotent stem cells for modeling Huntington’s disease. Date: 27/7/2018. Supervisor: Josep Mª. Canals Coll.

Author: Jordi Creus Muncunill. Title: Dual role of mTOR in Huntington’s disease: contribution to striatal neuronal survival and dysfunction. Date: 24/10/2018. Supervisor: Esther Pérez Navarro.

118 CIBERNED 2018 ANNUAL REPORT

106 | Valentín Ceña Callejo

UNIDAD ASOCIADA NEURODEATH UNIVERSIDAD DE CASTILLA-LA MANCHA FACULTAD DE MEDICINA

Avda. Almansa, 14 02006 Albacete Teléfono: 680222322 Correo electrónico: [email protected]

PRINCIPAL LIST OF Játiva Carbajal, Pablo. INVESTIGATOR PERSONNEL Bachelor degree. Manzanares Sandoval, Ceña Callejo, Valentín. De la Torre Martínez, Darío. Cristina. Bachelor degree. Bachelor degree. Posadas Mayo, Gallego Yerga, Laura. Inmaculada Concepción. Bachelor degree. Bachelor degree. García Saez, Ana Belen Inmaculada. Technician.

ABSTRACT The research of the Neurodeath group has been focused on 3 different areas: a) the use of nanoparticles, mainly dendrimers (but also polymers derived from cyclodextrin) to transfect genetic material, mainly siRNA, into different cell types to knock down specific proteins involved in tumoral progression and neurodegenerative diseases; b) to show that certain dendrimers, having themselves anti-inflammatory activity, are able to prevent, in mice, the development of experimental allergic encephalitis, a generally accepted animal model for multiple sclerosis, and c) to study the mechanisms involved in nanoparticle blood-brain-barrier crossing and to investigate the different approaches used to facilitate such a crossing. We have developed two nanoparticles that are able to transport fluorescent siRNA to the brain following injection in the mouse tail vein.The use of siRNA as therapeutic agent will produce, in a short period of time, new generations of drugs, based on siRNA, with the potential to revolutionize therapeutics in different areas. In addition, our group has focused on the development of dendrimers to transfect neuronal cultures with the aim of targeting the nanoparticles and their therapeutic cargo to the brain in a near future.

119 KEYWORDS Gene therapy, dendrimers, nanoparticles, neurodegeneration, siRNA, transfection, glioblastoma, multiple sclerosis, hematoebcephalic barrier.

PUBLICATIONS 2018 Cena V. Nanomedicine: A New Approach to Glioblastoma Therapy?. Basic & Clinical Pharmacol- ogy & Toxicology. Vol. 123. 111 River St, Hoboken 07030-5774, NJ USA: Wiley, 2018. Stenstrom P, Manzanares D, Zhang Y, Cena V, Malkoch M. Evaluation of amino-functional polyester dendrimers based on Bis-MPA as nonviral vectors for siRNA delivery. Molecules. 2018;23(8). PMID: 30110914. Cena V, Jativa P. Nanoparticle crossing of blood-brain barrier: A road to new therapeutic approaches to central nervous system diseases. Nanomedicine. 2018;13(13):1513-1516. PMID: 29998779. Gallego-Yerga L, Benito J.M, Blanco-Fernandez L, Martinez-Negro M, Velaz I, Aicart E. et al. Plasmid-Templated Control of DNA–Cyclodextrin Nanoparticle Morphology through Molecular Vector Design for Effective Gene Delivery. Chemistry - A European Journal. 2018;24(15):3825- 3835. PMID: 29341305. de la Torre C, Cena V. The delivery challenge in neurodegenerative disorders: The nanoparticles role in alzheimer’s disease therapeutics and diagnostics. Pharmaceutics. 2018;10(4). PMID: 30336640.

RESEARCH PROJECTS 2018 Code: PI2016/05. Title: Dream inhibitors and Alzheimer´s Disease. Principal investigator: Jose Ramon Naranjo Orovio. CIBERNED’s collaboration: Yes. CIBERNED groups: G307 ; G106; G403. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2016-2018.

120 CIBERNED 2018 ANNUAL REPORT

101 | Antonio Cuadrado Pastor

DEPARTAMENTO DE BIOQUÍMICA E INSTITUTO DE INVESTIGACIONES BIOMÉDICAS FACULTAD DE MEDICINA, UNIVERSIDAD AUTÓNOMA DE MADRID

Arzobispo Morcillo 4, 28029 Madrid Tel: +34 91 585 43 83 Fax: +34 91 585 44 01 E-mail: [email protected]

PRINCIPAL LIST OF Lastra Martínez, Diego. INVESTIGATOR PERSONNEL Bachelor degree. Lastres Becker, Isabel. Cuadrado Pastor, Blanco García, Ruth. PhD. Antonio. Bachelor degree. Pajares Cabetas, Marta. Escoll Guerrero, María PhD. Isabel. PhD. Fernández Ginés, Raquel. Bachelor degree.

ABSTRACT Our team studies the neuropathological processes associated with Parkinson’s disease (PD) and Alzheimer’s disease (AD) in order to identify new therapeutic targets to prevent or stop the development of these diseases. Studies in animal models and post-mortem brain tissues of patients indicate that many pathological changes in the brain derive from various types of local stresses, such as proteotoxic stress, closely related to oxidative and inflammatory stress. Therefore, in our group we studied the role of the transcription factor NRF2, master regulator of several neuroprotection pathways, as a new therapeutic target. During the last year we have carried out two main lines of research: 1) We are studying the role of the transcription factor NRF2 in the protection against stimuli that induce neurodegeneration. NRF2 is a protein that regulates the expression of about 250 genes that participate in adaptive responses to oxidative, inflammatory and proteotoxic stress and in the regulation of enzymes involved in biotransformation and metabolic reprogramming. Using genetically modified rodents, during 2018 we studied the contribution of this transcription factor to the protection against neuroinflammation and proteinopathy in transgenic mice that possess amyloidopathy (APPV717I) and tauopathy (TauP301L), which are characteristic of AD. In these mice the deficiency in NRF2 correlated with exacerbated astrogliosis and microgliosis, as evidenced by the increase

121 in the levels of the GFAP, IBA1 and CD11b markers. We have also described that chaperone-mediated autophagy, responsible for the degradation of alpha-synuclein and other neurotoxic proteins, is regulated by NRF2 at the level of the expression of the LAMP2 gene, which encodes the autophagy receptor LAMP2A. La pérdida sináptica está relacionada con la presencia de péptido soluble de Abeta en las sinapsis. Dado que en estudios previos mostramos que la proteína Dickkopf-1 (Dkk1), activadora de la vía Wnt-PCP (Wnt-planar cell polarity) conduce a tauopatía y muerte neuronal, hemos comparado el efecto de Abeta y Dkk1 en la morfología sináptica y en alteraciones cognitivas mediante la modulación de elementos clave de la vía Wnt-PCP. Hemos descrito que la sinaptotoxicidad inducida por Abeta es dependiente de Dkk1 y de la vía Wnt-PCP, a nivel de la dinámica del citoesqueleto por vía de Daam1, RhoA and ROCK. Estos resultados muestran la relevancia de la alteración de la vía Wnt en la neuropatolo- gía asociada a la EA. 2) Through a novel approach based on systems medicine related to chronic diseases, we have analyzed possible drugs that could activate NRF2 in the brain and provide a therapeutic benefit. We have described that the immunomodulatory effect of dimethyl fumarate (DMF), a drug currently used for the treatment of multiple sclerosis, improves cognition and reduces motor complications in the mouse model with amyloidopathy and tauopathy. Dimethyl fumarate also decreases the inflammatory response in this model and, therefore, provides a novel strategy to reposition this drug in the treatment of AD. As previously mentioned, the pathway (Wnt-PCP) induces synaptic retraction and is activated by soluble Abeta peptides. We have observed that the inhibitor of ROCK, fasudil, which inhibits the Abeta-Dkk1 pathway, prevents the loss of dendritic spines in vitro, and reduces the Abeta load in mice with advanced amyloidopathy. These results suggest the importance of the Wnt pathway in AD and propose the repositioning of fasudil, currently used for the treatment of cerebral vasospasm, for the treatment of AD.

KEYWORDS NRF2, Wnt-PCP, dimethyl fumarate, fasudil, amyloidopathy, tauopathy, neuroinflammation, autophagy.

PUBLICATIONS 2018 Egea J, Fabregat I, Frapart YM, Ghezzi P, Görlach A, Kietzmann T et al. Corrigendum to Eu- ropean contribution to the study of ROS: A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS) [Redox Biol. 13 (2017) 94-162].Redox biology. 2018;14. PMID: 29107648. Cuadrado A, Kugler S, Lastres-Becker I. Pharmacological targeting of GSK-3 and NRF2 provides neuroprotection in a preclinical model of tauopathy. Redox Biology. 2018;14:522- 534. PMID: 29121589.

Sellers KJ, Elliott C, Jackson J, Ghosh A, Ribe E, Rojo AI et al. Amyloid β synaptotoxicity is Wnt- PCP dependent and blocked by fasudil.Alzheimer’s & dementia : the journal of the Alzheim- er’s Association. 2017. PMID: 29055813.

122 CIBERNED 2018 ANNUAL REPORT

Elliott C, Rojo A.I, Ribe E, Broadstock M, Xia W, Morin P. et al. A role for APP in Wnt signalling links synapse loss with β-amyloid production. Translational Psychiatry. 2018;8(1). PMID: 30232325. Rojo A.I, Pajares M, Garcia-Yague A.J, Buendia I, Van Leuven F, Yamamoto M. et al. Deficien- cy in the transcription factor NRF2 worsens inflammatory parameters in a mouse model with combined tauopathy and amyloidopathy. Redox Biology. 2018;18:173-180. PMID: 30029164. Pajares M, Cuadrado A, Engedal N, Jirsova Z, Cahova M. The role of free radicals in autophagy regulation: Implications for ageing. Oxidative Medicine and Cellular Longevity. 2018;2018. PMID: 29682156. Pajares M, Rojo A.I, Arias E, Diaz-Carretero A, Cuervo A.M, Cuadrado A. Transcription factor NFE2L2/NRF2 modulates chaperone-mediated autophagy through the regulation of LAM- P2A. Autophagy. 2018;14(8):1310-1322. PMID: 29950142. Cuadrado A, Manda G, Hassan A, Alcaraz M.J, Barbas C, Daiber A. et al. Transcription factor NRF2 as a therapeutic target for chronic diseases: A systems medicine approach. Pharmaco- logical Reviews. 2018;70(2):348-383. PMID: 29507103. Rius-Pérez S, Pérez S, Martí-Andrés P, Finamor I, Taléns-Visconti R, Cuadrado A, Toledano MB, Sastre J. Time-course of thiol oxidation of protein phosphatases during cerulein-induced acute pancreatitis. Free Radical Biology and Medicine. 2018;120:S143. Castro-Sanchez S, Garcia-Yague A.J, Lopez-Royo T, Casarejos M, Lanciego J.L, Lastres-Beck- er I. Cx3cr1-deficiency exacerbates alpha-synuclein-A53T induced neuroinflammation and neurodegeneration in a mouse model of Parkinson’s disease. GLIA. 2018;66(8):1752-1762. PMID: 29624735.

Code: PCIN-2016-071. Title: Advanced theranostic approach in cancer combining photodynamic therapy and nanoparticles. Principal investigator: Luis Felipe Ferreira. CIBERNED’s collaboration: No. CIBERNED groups: G101. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 49000. Duration: 2016-2019.

123 Code: P_37_732. Title: Knowledge transfer in redox biology for developing advanced molecular tools in neurodegenerative diseases. Principal investigator: Antonio Cuadrado. CIBERNED’s collaboration: No. CIBERNED groups: G101. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 1915000. Duration: 2016-2020.

Code: SAF2016-76520-R. Title: Papel de NRF2 en la función y el destino del cerebro con alzheimer. Principal investigator: Antonio Cuadrado. CIBERNED’s collaboration: No. CIBERNED groups: G101. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 340000. Duration: 2017-2019.

Code: BMD2017/BMD3813. Title: Diseno y desarrollo de farmacos innovadores para el tratamiento de la esclerosis laterial amiotrofica. Principal investigator: Eva de Lago. CIBERNED’s collaboration: Yes. CIBERNED groups: G101; G303. Other CIBER’s collaboration: CIBERER. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 767395. Duration: 2018-2021.

PHD DISSERTATIONS 2018 Author: Marta Pajares Cabetas. Title: Transcription Factor NRF2 regulates the expression of autophagy genes. Date: 19/10/2018. Supervisor: Antonio Cuadrado Pastor.

124 CIBERNED 2018 ANNUAL REPORT

102 | Isabel Fariñas Gómez

DEPARTAMENTO DE BIOLOGÍA CELULAR. UNIVERSIDAD DE VALENCIA

Doctor Moliner 50 46100 Burjassot Tel.: +34 96 354 37 84 (despacho/ office) 32 46 (lab) Fax: +34 96 354 34 04 E-mail: [email protected]

PRINCIPAL Carrillo Barberá, Pau. Morante Redolat, José INVESTIGATOR Bachelor degree. Manuel. Domingo Muelas, Ana. PhD. Fariñas Gómez, Isabel. Bachelor degree. Palop Benlloch, María Duart Abadia, Pere. José. LIST OF Bachelor degree. Technician. PERSONNEL Durupt , Fabrice Claude. Pérez Sánchez, Francisco. Andrés Carbonell, Bachelor degree. PhD. Cristina. Gil Sanz, Cristina. Technician. PhD. Pérez Villalba, Ana. PhD. Barberá Ferragud, Elba. Kirstein Ruiz, Martina. Rodriguez Ferrón, Bachelor degree. PhD. Sacramento. PhD. Belenguer Sanzchez, Lozano Ureña, Anna. German. Bachelor degree. Sirerol Piquer, María Salomé. PhD. Marí Zaragoza, Miguel. PhD. Blasco Chamarro, Laura. Bachelor degree. Bachelor degree. Montalbán Loro, Raquel. Bachelor degree.

ABSTRACT Our laboratory develops two lines of research in neurodegeneration and cell therapy: 1) the study of cellular and molecular alterations underlying dopaminergic neurodegeneration associated with Parkinson’s disease (PD) through the analysis of alpha-synuclein effects, and 2) the study of neural stem cell (NSC) regulation. We use experimental mouse models in which to identify and evaluate cellular and molecular mechanisms that may underlie dopaminergic neurodegeneration or that can be used for the manipulation of NSC with therapeutic purposes. In the context of the CIBERNED program, we work on aspects that combine both of these lines, i.e. those related with the regulation of adult

125 neurogenesis in parkinsonism and during aging. During 2018, we continued to characterize the role of alpha-synuclein and dopamine as regulators of NSCs. In collaboration with groups of CIBERSAM and with the group of Miquel Vila (Hospital Vall d’Hebrón) in CIBERNED, we have analyzed the potential of a strategy based on the use of specific oligonucleotides administered intra-nasally for the reduction of alpha-synuclein levels in the brain. In parallel, and in collaboration with the group from the University of Barcelona led by Oriol Bachs, we have shown that negative regulator of the cell cycle of the Cip/Kip family of inhibitors of cyclin-dependent kinases, p27, is a transcriptional modulator of alpha-synuclein, opening the possibility of manipulating its levels of expression. On the other hand, we have initiated studies to analyze the interaction of microglia with toxic and aggregated forms of alpha-synuclein and how this interaction can influence the pathological transmission of this molecule during aging.

KEYWORDS Neural stem cells, adult neurogenesis, Parkinson´s disease, cell therapy.

PUBLICATIONS 2018 Gallastegui E, Domuro C, Serratosa J, Larrieux A, Sin L, Martinez J. et al. p27Kip1 regulates alpha-synuclein expression. Oncotarget. 2018;9(23):16368-16379. PMID: 29662651. Bachs O, Gallastegui E, Orlando S, Bigas A, Morante-Redolat J.M, Serratosa J. et al. Role of p27Kip1 as a transcriptional regulator. Oncotarget. 2018;9(40):26259-26278. PMID: 29899857. Soares M.L, Edwards C.A, Dearden F.L, Ferron S.R, Curran S, Corish J.A. et al. Targeted deletion of a 170-kb cluster of LINE-1 repeats and implications for regional control. Genome Research. 2018;28(3):345-356. PMID: 29367313. Perez-Villalba A, Sirerol-Piquer M.S, Belenguer G, Soriano-Canton R, Munoz-Manchado A.B, Villadiego J. et al. Synaptic regulator α-synuclein in dopaminergic fibers is essentially required for the maintenance of subependymal neural stem cells. Journal of Neuroscience. 2018;38(4):814-825. PMID: 29217686.

RESEARCH PROJECTS 2018 Code: RD16/0011. Title: Red de Terapia Celular. Principal investigator: Isabel Farinas. CIBERNED’s collaboration: Yes. CIBERNED groups: G607 ; G301; G102; G113; G208; G105; G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 284999. Duration: 2016-2020.

126 CIBERNED 2018 ANNUAL REPORT

Code: RD16/0011. Title: Red de Terapia Celular. Principal investigator: Isabel Farinas. CIBERNED’s collaboration: Yes. CIBERNED groups: G607 ; G301; G102; G113; G208; G105; G207. Other CIBER's collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 284999. Duration: 2016-2020.

Code: RD16/0011/0016. Title: Combined protective/restorative cell-mediated strategies for neurodegenerative diseases. Principal investigator: Jose Luis Labandeira Garcia. CIBERNED’s collaboration: Yes. CIBERNED groups: G208 ; G102; G113; G105; G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 240245. Duration: 2017-2021.

Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: Yes. CIBERNED groups: G604 ; G102; G606; G111; G306. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350000. Duration: 2016-2018.

Code: PROMETEO/2017/030. Title: Efectos directos y remotos de la respuesta inflamatoria sobre las celulas madre. Principal investigator: Isabel Farinas. CIBERNED’s collaboration: No. CIBERNED groups: G102. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 384957. Duration: 2017-2021.

Code: Convenio Botin-UV. Title: Estudio de celulas madre en el ambito de las investigaciones basicas en terapia celular. Principal investigator: Isabel Farinas. CIBERNED’s collaboration: No. CIBERNED groups: G102. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Botin-Banco Santander. Funding: 625000. Duration: 2014-2018.

Code: SAF2016-78845-R. Title: Papel de la impronta genomica y su regulacion epigenetica en celulas madre neu-

127 rales: relacion con la formacion de tumores. Principal investigator: Sacramento Rodriguez Ferron. CIBERNED’s collaboration: No. CIBERNED groups: G102. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 181500. Duration: 2017-2019.

Code: SAF2017-86690-R. Title: Regulacion del comportamiento de celulas madre neurales por el medio sistemico: el nicho extendido. Principal investigator: Isabel Farinas. CIBERNED’s collaboration: No. CIBERNED groups: G102. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 423016. Duration: 2018-2020.

128 CIBERNED 2018 ANNUAL REPORT

303 | Javier Fernández Ruiz

DEPARTAMENTO DE BIOQUÍMICA Y BIOLOGÍA MOLECULAR, FACULTAD DE MEDICINA. UNIVERSIDAD COMPLUTENSE

Avda. Complutense s/n, Pab. IV 4ª Planta, Laboratorios 9-12, 28040-Madrid, Spain Tel. +34 91 394 14 50 Fax +34 91 394 16 91 E-mails: [email protected]; [email protected]

PRINCIPAL García Movellán, Mena Gómez, María INVESTIGATOR Yolanda. Angeles. Auxiliar Administrativo. PhD. Fernández Ruiz, Javier García Toscano, Laura. Ramos Atance, José Bachelor degree. Antonio. LIST OF Gómez Cañas, Maria. PhD. PERSONNEL PhD. Rodriguez Cueto, Alonso Gómez, Cristina. Gómez Ruiz, María Carmen Aurora. Bachelor degree. Sagrario. PhD. Burgaz García de PhD. Sagredo Ezkioga, Oteyza, Sonia. Hernández Gálvez, Onintza. Bachelor degree. María Luz. PhD. Ceprian Costoso, María. PhD. Santos-Garcia Sanz, Bachelor degree. Lago Femia, Eva de. PhD. Irene. García García, María Luna Piñel, Eva. Bachelor degree. Concepción. Bachelor degree. PhD.

ABSTRACT The activity of the group has been again concentrated in the development of cannabinoid-based medicines with neuroprotective potential useful for several chronic neurodegenerative disorders, as well as in the identification of cellular and molecular mechanisms that underlie this potential. Again, during this year, the work has been concentrated in Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD) and spinocerebellar ataxia type-3 (SCA-3). We have enhanced our collaboration with groups of Medicinal Chemistry working in different directions: development of selective agonists of the CB2 receptor, allosteric modulators of this receptor, CB2/PPAR-γ hybrid compounds, GPR55 agonists, which have been investigated in in vitro studies of receptor binding and activity, as well as in cell-based models. We have

129 also increased our collaboration with companies in this activity, adding Roche, Emerald Heal- th Pharmaceuticals and Symrise to the previous collaborations already in progress with GW Pharmaceuticals and VivaCell Biotechnology-Spain. With these companies, we are working in the development and evaluation of cannabinoids with CB2 and/or PPAR-γ activity as novel neuroprotectants. One of the new compounds in whose evaluation we have participated, EHP- 101, just entried in a clinical trial due to its anti-inflammatory potential in skin disorders, so we expect it can be soon in clinical evaluation for some of the neurodegenerative disorders that we investigate. In the case of PD, the work has consisted in investigating the advantages of the combination of cannabidiol and Δ9-THCV (GW Pharmaceuticals) for the symptomatic and neuroprotective treatment of this disease, including their antidyskinetic effects which are studied in collaboration with the CIBERNED group leaded by Rosario Moratalla, results that we are presently preparing for publication. Other line of research has consisted in determining the potential of cannabigerol-quinones (they are PPAR-γ agonists) and cannabidiol-quinones (CB2 and PPAR-γ agonists) in experimental models of this disease, for which we have identified two derivatives particularly active (VivaCell Biotechnology-Spain and Emerald Health Pharmaceuticals). We have extended our research to a novel endocannabinoid-related receptor, so-called GPR55, for which we are developing selective ligands in collaboration with a CSIC group of Medicinal Che- mistry. In the case of ALS, the work has continued the evaluation of CBDA and other phytocannabinoids for their neuroprotective effects (GW Pharmaceuticals), and of different phytocannabinoid derivatives (Vivacell Biotechnology and Emerald Health Pharmaceuticals), using SOD-1 mutant and TDP-43 transgenic mice. The targets we are investigating include mainly CB2 receptors, but also PPAR-γ and GPR55. receptors as potential targets. All this work has been carried out as the final part of our project funded by the MINEICO-Biomedicine National Program, concentrated in ALS and FTD, which is a joint project with a group of the CSIC Medical Chemistry Institute, and addressed to the development of new therapies based on targeting GPR55, GPR18, CB2 receptors using allosteric modulators, and CB2/PPAR-γ using hybrid compounds, as well as within a CIBERNED project with the groups of Adolfo López de Munaín (coordinator), Isidre Ferrer and Eduardo Soriano concentrated in the development of new therapies for ALS. Lastly, in the case of SCA-3 (Machado-Joseph disease), we have finished the biochemical and histopathological studies addressed to demonstrate the occurrence of an endocannabinoid dysregulation in different CNS areas during the course of the disease, and now we are using SCA-3 transgenic mice to investigate potential pharmacological treatments addressed to co- rrect such dysregulation and to elicit neuroprotective effects, including CB2 and GPR55 as new therapeutic targets.

KEYWORDS Cannabinoids, CB1 receptors, CB2 receptors, GPR55, PPAR-γ, neuroprotection, Parkinson’s disease, amyotrophic lateral sclerosis/frontotemporal dementia, spinocerebellar ataxias

PUBLICATIONS 2018 Barata L, Arruza L, Rodriguez M.-J, Aleo E, Vierge E, Criado E. et al. Neuroprotection by cannabidiol and hypothermia in a piglet model of newborn hypoxic-ischemic brain damage. Neuropharmacol- ogy. 2019;146:1-11. epub2018. PMID: 30468796.

130 CIBERNED 2018 ANNUAL REPORT

Espejo-Porras F, Fernández-Ruiz J, de Lago E. Analysis of endocannabinoid receptors and enzymes in the post-mortem motor cortex and spinal cord of amyotrophic lateral sclerosis patients.Amyo- trophic lateral sclerosis & frontotemporal degeneration. 2018. PMID: 29334787. Murineddu G, Deligia F, Ragusa G, Garcia-Toscano L, Gomez-Canas M, Asproni B. et al. Novel sulfenamides and sulfonamides based on pyridazinone and pyridazine scaffolds as CB1 receptor ligand antagonists. Bioorganic and Medicinal Chemistry. 2018;26(1):295-307. PMID: 29229226. Fernández Ruiz J, de Lago Femia E, Sagredo Ezkioga O, Ramos Atance JA. Introducción a la farma- cología del sistema nervioso central: neurotransmisores, receptores y otros elementos sinápticos. En: Farmacología Básica y Clínica 19ª edición. 179-208. 2018. Aymerich M.S, Aso E, Abellanas M.A, Tolon R.M, Ramos J.A, Ferrer I. et al. Cannabinoid pharmacology/therapeutics in chronic degenerative disorders affecting the central nervous system. Biochem- ical Pharmacology. 2018;157:67-84. PMID: 30121249. García C, Gómez-Cañas M, Burgaz S, Palomares B, Gómez-Gálvez Y, Palomo-Garo C et al. Benefits of VCE-003.2, a cannabigerol quinone derivative, against inflammation-driven neuronal deterioration in experimental Parkinson’s disease: possible involvement of different binding sites at the PPARγ receptor.Journal of neuroinflammation. 2018;15(1). PMID: 29338785. Rodriguez-Cueto C, Santos-Garcia I, Garcia-Toscano L, Espejo-Porras F, Bellido M, Fernandez-Ruiz J. et al. Neuroprotective effects of the cannabigerol quinone derivative VCE-003.2 in SOD1G93A transgenic mice, an experimental model of amyotrophic lateral sclerosis. Biochemical Pharmacolo- gy. 2018;157:217-226. PMID: 30076846. Espejo-Porras F, García-Toscano L, Rodríguez-Cueto C, Santos-García I, de Lago E, Fernandez-Ruiz J. Targeting glial cannabinoid CB2 receptors to delay the progression of the pathological phenotype in TDP-43 (A315T) transgenic mice, a model of amyotrophic lateral sclerosis.British journal of pharmacology. 2018. PMID: 29574689. Fernandez-Ruiz J. The biomedical challenge of neurodegenerative disorders: An opportunity for cannabinoid-based therapies to improve on the poor current therapeutic outcomes. British Journal of Pharmacology. 2018. PMID: 29856067. del Rio C, Cantarero I, Palomares B, Gomez-Canas M, Fernandez-Ruiz J, Pavicic C. et al. VCE-004.3, a cannabidiol aminoquinone derivative, prevents bleomycin-induced skin fibrosis and inflammation through PPARγ- and CB2 receptor-dependent pathways. British Journal of Pharmacology. 2018;175(19):3813-3831. PMID: 30033591. Castro-Sanchez S, Garcia-Yague A.J, Lopez-Royo T, Casarejos M, Lanciego J.L, Lastres-Becker I. Cx- 3cr1-deficiency exacerbates alpha-synuclein-A53T induced neuroinflammation and neurodegeneration in a mouse model of Parkinson’s disease. GLIA. 2018;66(8):1752-1762. PMID: 29624735. Sagredo O, Palazuelos J, Gutierrez-Rodriguez A, Satta V, Galve-Roperh I, Martinez-Orgado J. Can- nabinoid signalling in the immature brain: Encephalopathies and neurodevelopmental disorders. Biochemical Pharmacology. 2018;157:85-96. PMID: 30118663.

131 Type: Intramurales. Funding agency: CIBERNED. Funding: 200000. Duration: 2016-2018.

Code: NEUROLATAM. Title: Unraveling the neurobiological sustrate of protective cannabinoid actions in the brain. Principal investigator: Ismael Galve Roperh, Jose A. Ramos Atance. CIBERNED’s collaboration: Yes. CIBERNED groups: G303 ; G305. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Unión Iberoamericana de Universidades. Funding: 24225. Duration: 2017-2018.

Code: SAF2015-68580-C2-1-R. Title: Dianas del sistema endocannabinoide para el desarrollo de terapias frente a la neurodegeneracion: enfasis en la ELA y otras enfermedades neurodegenerativas. Principal investigator: Javier Fernandez Ruiz/Eva de Lago Femia. CIBERNED’s collaboration: No. CIBERNED groups: G303. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 302500. Duration: 2016-2019.

Code: PR26/16-18B. Title: Estudio de nuevas dianas en demencias neurodegenerativas basadas en tratamien- tos neuroprotectoras y neurogenicos. Principal investigator: Eva De Lago, Maria Gomez Ruiz, Felipe Ortega De La O. CIBERNED’s collaboration: No. CIBERNED groups: G303 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Banco Santander - UCM. Funding: 30000. Duration: 2017-2018.

Code: 2017/VIVACELL/JFR. Title: Investigation in the anti-inflammatory and neuroprotective properties of the phytocannabinoid derivative VCE003.2 in Parkinson´s disease using LPS-lesioned alpha-synuclein transgenic mice. Principal investigator: Javier Fernandez Ruiz, Maria Concepcion Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G303 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: VivaCell Biotechnology Espana. Funding: 80000. Duration: 2017-2018.

Code: EMERALD. Title: Investigation in the anti-inflammatory and neuroprotective properties of the phytocannabinoid derivatives VCE-004.8 and VCE-003.2 (and its analogs CBG-Q-Salt and CB- GA-Q) in Parkinson’s disease using 6-hydroxydopamine-lesioned mice. Principal investigator: Javier Fernandez Ruiz y Mª Concepcion Garcia Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G303. Other CIBER’s collaboration: No.

132 CIBERNED 2018 ANNUAL REPORT

Type: Privado. Funding agency: EMERALD. Funding: 30000. Duration: 2018-2019.

Code: EME5.1ALS. Title: Investigation in the anti-inflammatory and neuroprotective properties of VCE- 005.1, in experimental models of amyotrophic lateral sclerosis. Principal investigator: Javier Fernandez Ruiz y Eva de Lago Femia. CIBERNED’s collaboration: No. CIBERNED groups: G303. Other CIBER’s collaboration: No. Type: Privado. Funding agency: EMERALD. Funding: 30000. Duration: 2018-2019.

Code: GW2015. Title: Preclinical development of phytocannabinoid-based therapies for the treatment of disease progression in amyotrophic lateral sclerosis/frontotemporal dementia using TDP-43 transgenic mice. Principal investigator: Javier Fernandez Ruiz, Eva De Lago Femia. CIBERNED’s collaboration: No. CIBERNED groups: G303 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: GW Research Ltd. (UK). Funding: 126544,5. Duration: 2015-2018.

Code: VVC BINDING. Title: Validation of the CB2 receptor binding of VCE-004.8 using membranes obtained from mouse cells and tissues. Principal investigator: Javier Fernandez Ruiz. CIBERNED’s collaboration: No. CIBERNED groups: G303. Other CIBER’s collaboration: No. Type: Privado. Funding agency: VivaCell Biotechnology Espana. Funding: 10000. Duration: 2018-2018.

PHD DISSERTATIONS 2018 Author: Francisco Espejo Porras. Title: Relevancia del receptor cannabinoide CB2 en la esclerosis lateral amiotrófica (ELA). Date: 17/4/2018. Supervisor: Javier Fernández Ruiz.

133 201 | Rafael Franco Fernández

DEPARTAMENTO DE BIOQUÍMICA Y BIOLOGÍA MOLECULAR FACULTAD DE BIOLOGÍA. UNIVERSIDAD DE BARCELONA

Diagonal 643, planta -2 08028 Barcelona. Catalunya. Spain. Tel.: +34 93 402 12 11 Fax: +34 93 402 15 59 E-mail: [email protected]

PRINCIPAL Casadó Anguera, Mallol Montero, Josefa. INVESTIGATOR Veronica. PhD. Bachelor degree. Medrano Moya, Mireia. Franco Fernández, Casadó Burillo, Vicente. Bachelor degree. Rafael. PhD. Moreno Guillén, Cortés Tejedor, Antonio. Estefanía. LIST OF PhD. PhD. PERSONNEL Etayo Labiano, Iñigo Navarro Brugal, Gemma. Aguinaga Andrés, Javier. PhD. David. Bachelor degree. Reyes Resina, Irene. PhD. Homar Ruano, Patricia. Bachelor degree. Angelats Canals, Edgar. Bachelor degree. Rodriguez Ruiz, Mar. Bachelor degree. Jiménez Cano, Jasmina. Bachelor degree. Canela Campos, Enric Technician. Sánchez Soto, Marta. Isidre. Lluis Biset, Carmen. PhD. Bachelor degree. Principal investigator.

ABSTRACT The research in the Molecular Neurobiology (NBM) group at the University of Barcelona is eminently translational and focused on receptors coupled to G proteins (GPCRs) involved in the pathophysiology of neurodegenerative diseases. The laboratory has identified new therapeutic targets and new molecules with potential to combat these diseases have been designed and developed in collaboration with medicinal chemistry groups. During 2018 there have been several research projects developed in parallel. Within the framework of projects funded by the Fundació la Marató de TV3, one related to Huntington’s disease and another related to Parkinson’s disease, therapies have been proposed based on molecular studies focused on receptors that are expressed in both neurons and glia.

134 CIBERNED 2018 ANNUAL REPORT

At the molecular level, the functional peculiarities of complexes formed by two or more GPCRs have been characterized and structural models have been proposed that would explain all the results obtained in the laboratory as well as those described in the literature. Complexes are often formed by two homodimers of two GPCRs (tetramers) that are coupled to two different G proteins. Two highly relevant results, which have been published in journals with a high impact factor, reflect a pre-coupling of G proteins to inactive GPCRs (i.e. not yet activated by their corresponding neurotransmitters) and discover additional / unexpected functionalities of GPCR heteromers coming from long C-terminal domains (>80 amino acids). The length of terminal C end of GPCRs is very diverse and our data shows, for a specific case, the reason. It is the “tail” of the A2A adenosine receptor that is responsible for adenosine increasing or decreasing the release of glutamate from cortical neurons according to the concentration. The structure of the adenosine “concentration sensor” that is regulating glutamatergic neurotransmission is unique. The GPCRs upregulated in reactive microglia and their functional role have been the focus of a glia-centered research theme. The expression of cannabinoid CB2 receptors is increased in activated microglia and the endocannabinoids go from being molecules that prevent nonspecific activation to be regulators of activation. Complexes formed by CB2 receptors and other GPCRs, such as A2A adenosine receptors, constitute potential targets to ensure that the microglia skewing towards the M2 or neuroprotective phenotype. In animal models of neurodegenerative diseases, primary cultures, obtained from neonates, show that the microglia displays an activated phenotype. Given that the newborn animals do not present neuronal death or cognitive deficits, the hypothesis that we contemplate is that activated microglia is exerting a neuroprotective effect (M2 phenotype). A detailed study of the pharmacology of the CB2 receptor has shown that cannabidiol, which is currently approved for the therapy of epilepsy, is a negative allosteric modulator (a NAM according to the current nomenclature). Based on the mutants developed in the laboratory and the recent elucidation of the three-dimensional structure of the receptor, we have identified the allosteric center with high reliability. Looking to the future there are two strategies in relation to this discovery: i) design more potent allosteric compounds and ii) test the neuroprotective potential of cannabidiol and the new compounds, some of which are already synthesized. The efficacy of the allosteric modulators of the CB2 receptor will be tested in animal models of Parkinson’s, Alzheimer’s and stroke. The potential of the compounds to regulate the M1/M2 conversion in microglia will also be determined.

KEYWORDS Targeting G-protein-coupled receptors (GPCRs) for neuroprotection, GPCR heteromers as targets to combat neurodegenerative diseases, GPCR heteromers in the pathophysiology of Parkinson’s, Huntington’s and Alzheimer’s disease, Natural and synthetic cannabinoids for neuroprotection, Strategies to convert reactive M1 microglia into the neuroprotective (M2) phenotype.

135 PUBLICATIONS 2018 Casado-Anguera V, Moreno E, Mallol J, Ferre S, Canela E.I, Cortes A. et al. Reinterpreting anomalous competitive binding experiments within G protein-coupled receptor homodimers using a dimer receptor model. Pharmacological Research. 2019;139:337-347. epub2018. PMID: 30472462. Borroto-Escuela DO, de la Mora MP, Zoli M, Benfenati F, Narvaez M, Rivera A, Díaz-Cabiale Z, Beg- giato S, Ferraro L, Tanganelli S, Ambrogini P, Filip M, Liu F, Franco R, Agnati LF, Fuxe K Analysis and Quantification of GPCR Allosteric Receptor–Receptor Interactions Using Radioligand Binding Assays: The A2AR-D2R Heteroreceptor Complex Example. Receptor-Receptor Interactions in the Central Nervous System: Springer; 2018. p. 1-14. Borroto-Escuela D O, Narvaez M, Valladolid-Acebes I, Shumilov K, Di Palma M, Wydra K, et al. De- tection, Analysis, and Quantification of GPCR Homo-and Heteroreceptor Complexes in Specific Neuronal Cell Populations Using the In Situ Proximity Ligation Assay. In Receptor-Receptor Inter- actions in the Central Nervous System (pp. 299-315). Humana Press, New York, NY. 2018. Reyes-Resina I, Martínez-Pinilla E, Borroto-Escuela DO, Fuxe K, Navarro G, Franco R. Methods to Identify the Signature of Trimers Formed by Three G Protein-Coupled Receptors or by Two G Pro- tein-Coupled and One Ionotropic Receptor with Special Emphasis in the Functional Role in the Central Nervous System. In Receptor-Receptor Interactions in the Central Nervous System (pp. 187-203). Humana Press, New York, NY. 2018. Moreno E, Canet J, Gracia E, Lluís C, Mallol J, Canela EI, Cortés A, Casadó V. Molecular evidence of adenosine deaminase linking adenosine A2A receptor and CD26 proteins. Frontiers in Pharma- cology. 2018;9:106. Navarro G, Martínez-Pinilla E, Ortiz R, Noé V, Ciudad CJ, Franco R. Resveratrol and Related Stilbe- noids, Nutraceutical/Dietary Complements with Health-Promoting Actions: Industrial Production, Safety, and the Search for Mode of Action. Comprehensive reviews in food science and food safety. 2018;17(4):808-26. Navarro G, Borroto-Escuela DO, Fuxe K, Franco R. Adenosine Receptors as a Paradigm to Identi- fy Dimer/Oligomers of G-Protein-Coupled Receptors and as Targets in Parkinson’s Disease and Schizophrenia. In The Adenosine Receptors (pp. 239-258). Humana Press, Cham. 2018. Zamarbide M, Gil-Bea F.J, Bannenberg P, Martinez-Pinilla E, Sandoval J, Franco R. et al. Maternal imprinting on cognition markers of wild type and transgenic Alzheimer’s disease model mice. Scientific Reports. 2018;8(1). PMID: 29691440. Rivera-Oliver M, Moreno E, Álvarez-Bagnarol Y, Ayala-Santiago C, Cruz-Reyes N, Molina-Castro GC et al. Adenosine A1-Dopamine D1 Receptor Heteromers Control the Excitability of the Spinal Motoneuron. Molecular neurobiology. 2018. PMID: 29797183. Franco R, Navarro G. Adenosine A2A Receptor Antagonists in Neurodegenerative Diseases: Huge Potential and Huge Challenges.Frontiers in psychiatry. 9. PMID: 29593579. Hinz S, Navarro G, Borroto-Escuela D, Seibt BF, Ammon YC, de Filippo E et al. Adenosine A2A receptor ligand recognition and signaling is blocked by A2B receptors.Oncotarget. 2018;9(17). PMID: 29568380. Franco R, Aguinaga D, Jimenez J, Lillo J, Martinez-Pinilla E, Navarro G. Biased receptor functionality versus biased agonism in G-protein-coupled receptors. Biomolecular Concepts. 2018;9(1):143- 154. PMID: 30864350. Borroto-Escuela DO, Perez De La Mora M, Manger P, Narváez M, Beggiato S, Crespo-Ramírez M et al. Brain Dopamine Transmission in Health and Parkinson’s Disease: Modulation of Synaptic Trans- mission and Plasticity Through Volume Transmission and Dopamine Heteroreceptors.Frontiers in synaptic neuroscience. 10. PMID: 30042672.

136 CIBERNED 2018 ANNUAL REPORT

Navarro G, Reyes-Resina I, Rivas-Santisteban R, Sanchez de Medina V, Morales P, Casano S. et al. Cannabidiol skews biased agonism at cannabinoid CB1 and CB2 receptors with smaller effect in CB1-CB2 heteroreceptor complexes. Biochemical Pharmacology. 2018;157:148-158. PMID: 30194918. Navarro G, Varani K, Reyes-Resina I, Sánchez de Medina V, Rivas-Santisteban R, Sánchez-Carnerero Callado C et al. Cannabigerol Action at Cannabinoid CB1 and CB2 Receptors and at CB1-CB2 Het- eroreceptor Complexes.Frontiers in pharmacology. 9. PMID: 29977202. Galindo L, Moreno E, López-Armenta F, Guinart D, Cuenca-Royo A, Izquierdo-Serra M et al. Canna- bis Users Show Enhanced Expression of CB1-5HT2A Receptor Heteromers in Olfactory Neuroepi- thelium Cells.Molecular neurobiology. 2018. PMID: 29294249. Aguinaga D, Medrano M, Cordomi A, Jimenez-Roses M, Angelats E, Casanovas M. et al. Cocaine Blocks Effects of Hunger Hormone, Ghrelin, Via Interaction with Neuronal Sigma-1 Receptors. Mo- lecular Neurobiology. 2018;:1-15. PMID: 29876881. Aguinaga D, Medrano M, Vega-Quiroga I, Gysling K, Canela E.I, Navarro G. et al. Cocaine effects on dopaminergic transmission depend on a balance between sigma-1 and sigma-2 receptor expression. Frontiers in Molecular Neuroscience. 2018;11. PMID: 29483862. Navarro G, Cordomí A, Brugarolas M, Moreno E, Aguinaga D, Pérez-Benito L et al. Cross-communication between Gi and Gs in a G-protein-coupled receptor heterotetramer guided by a receptor C-terminal domain.BMC biology. 2018;16(1). PMID: 29486745. Pulido D, Casado-Anguera V, Perez-Benito L, Moreno E, Cordomi A, Lopez L. et al. Design of a True Bivalent Ligand with Picomolar Binding Affinity for a G Protein-Coupled Receptor Homodimer. Journal of Medicinal Chemistry. 2018;61(20):9335-9346. PMID: 30257092. Navarro G, Medrano M, Aguinaga D, Vega-Quiroga I, Lillo A, Jiménez J et al. Differential effect of amphetamine over the corticotropin-releasing factor CRF2 receptor, the orexin OX1 receptor and the CRF2-OX1 heteroreceptor complex.Neuropharmacology. 2018. PMID: 30465812. Ferré S, Bonaventura J, Zhu W, Hatcher-Solis C, Taura J, Quiroz C et al. Essential Control of the Func- tion of the Striatopallidal Neuron by Pre-coupled Complexes of Adenosine A2A-Dopamine D2 Receptor Heterotetramers and Adenylyl Cyclase.Frontiers in pharmacology. 9. PMID: 29686613. Navarro G, Cordomi A, Casado-Anguera V, Moreno E, Cai N.-S, Cortes A. et al. Evidence for functional pre-coupled complexes of receptor heteromers and adenylyl cyclase. Nature Communications. 2018;9(1). PMID: 29593213. Moreno E, Canet J, Gracia E, Lluís C, Mallol J, Canela EI et al. Molecular Evidence of Adenosine Deam- inase Linking Adenosine A2A Receptor and CD26 Proteins.Frontiers in pharmacology. 9. PMID: 29497379. Angelats E, Requesens M, Aguinaga D, Kreutz MR, Franco R, Navarro G. Neuronal Calcium and cAMP Cross-Talk Mediated by Cannabinoid CB1 Receptor and EF-Hand Calcium Sensor Interac- tions.Frontiers in cell and developmental biology. 6. PMID: 30073165. Ferre S, Quiroz C, Guitart X, Rea W, Seyedian A, Moreno E. et al. Pivotal role of adenosine neurotransmission in Restless Legs Syndrome. Frontiers in Neuroscience. 2018;11(JAN). PMID: 29358902. Sánchez-Soto M, Yano H, Cai NS, Casadó-Anguera V, Moreno E, Casadó V et al. Revisiting the Func- tional Role of Dopamine D4 Receptor Gene Polymorphisms: Heteromerization-Dependent Gain of Function of the D4.7 Receptor Variant.Molecular neurobiology. 2018. PMID: 30387076. Borroto-Escuela D.O, Hinz S, Navarro G, Franco R, Muller C.E, Fuxe K. Understanding the role of adenosine A2AR heteroreceptor complexes in neurodegeneration and neuroinflammation. Fron- tiers in Neuroscience. 2018;12(FEB). PMID: 29467608.

137 Sanchez-Soto M, Casado-Anguera V, Yano H, Bender B.J, Cai N.-S, Moreno E. et al. α2A- and α2C-Adrenoceptors as Potential Targets for Dopamine and Dopamine Receptor Ligands. Molecu- lar Neurobiology. 2018;55(11):8438-8454. PMID: 29552726. Navarro G, Borroto-Escuela D, Angelats E, Etayo Í, Reyes-Resina I, Pulido-Salgado M et al. Recep- tor-heteromer mediated regulation of endocannabinoid signaling in activated microglia. Role of CB1 and CB2 receptors and relevance for Alzheimer’s disease and levodopa-induced dyskinesia. Brain, behavior, and immunity. 2018;67. PMID: 28843453. Franco R, Aguinaga D, Reyes I, Canela EI, Lillo J, Tarutani A et al. N-Methyl-D-Aspartate Receptor Link to the MAP Kinase Pathway in Cortical and Hippocampal Neurons and Microglia Is Dependent on Calcium Sensors and Is Blocked by α-Synuclein, Tau, and Phospho-Tau in Non-transgenic and Transgenic APPSw,Ind Mice.Frontiers in molecular neuroscience. 11. PMID: 30233307. Reyes-Resina I, Aguinaga D, Labandeira-García JL, Lanciego JL, Navarro G, Franco R. Usefulness of identifying G-protein-coupled receptor dimers for diagnosis and therapy of neurodegenerative diseases and of gliomas.Histology and histopathology. 2018. PMID: 29336473. García-Gutiérrez MS, Navarrete F, Navarro G, Reyes-Resina I, Franco R, Lanciego JL et al. Alterations in Gene and Protein Expression of Cannabinoid CB2 and GPR55 Receptors in the Dorsolateral Prefrontal Cortex of Suicide Victims.Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics. 2018. PMID: 29435814. Franco R, Sanchez-Arias J.A, Navarro G, Lanciego J.L. Glucocerebrosidase Mutations and Synucle- inopathies. Potential Role of Sterylglucosides and Relevance of Studying Both GBA1 and GBA2 Genes. Frontiers in Neuroanatomy. 2018;12. PMID: 30002620. Reyes-Resina I, Navarro G, Aguinaga D, Canela E.I, Schoeder C.T, Zaluski M. et al. Molecular and functional interaction between GPR18 and cannabinoid CB2 G-protein-coupled receptors. Rel- evance in neurodegenerative diseases. Biochemical Pharmacology. 2018;157:169-179. PMID: 29870711.

RESEARCH PROJECTS 2018 Code: PI2016/02. Title: Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches. Principal investigator: Jose Antonio Del Rio. CIBERNED’s collaboration: Yes. CIBERNED groups: G114 ; G401; G201. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2016-2018.

Code: CI18-00045. Title: Fighting pain with cannabis avoiding side-effects. Principal investigator: Maldonado. CIBERNED’s collaboration: No. CIBERNED groups: G201 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Caixa. Funding: 70000. Duration: 2018-2019.

138 CIBERNED 2018 ANNUAL REPORT

Code: SAF2017-87629-R. Title: Validacion de los heteromeros entre receptores de dopamina D1 y de adenosina A1 como una nueva diana terapeutica para el tratamiento de disfunciones motoras medu- lares y cerebrales. Principal investigator: ENRIC I. CANELA CAMPOS. CIBERNED’s collaboration: No. CIBERNED groups: G201 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 229900. Duration: 2018-2020.

PHD DISSERTATIONS 2018 Author: Iñigo Javier Etayo Labiano. Title: Heterómeros de receptores CB1, CB2 Y GPR55 y su implicación en enfermedades neurodegenerativas y alcoholismo. Date: 15/5/2018. Supervisor: Gemma Navarro Brugal.

Author: Irene Reyes Resina. Title: Heterómeros de receptores CB1, CB2, GPR55 y GPR18: señalización celular, farma- cología y análisis de su potencial como dianas terapéuticas de enfermedades neurodegenerativas. Date: 11/6/2018. Supervisor: Rafael Franco Fernández.

Author: Verònica Casadó Anguera. Title: Allosteric interactions between catecholamine receptors and other G protein-coupled receptors: pharmacological and functional characterization. Date: 14/6/2018. Supervisor: Enric Isidre Canela Campos.

Author: Edgar Angelats Canals. Title: Relevant molecular and functional G-protein coupled receptors interactions in neu- roinflamation and addiction.. Date: 7/9/2018. Supervisor: Rafael Franco Fernández.

Author: Mar Rodríguez Ruiz. Title: Heterómeros de receptores de dopamina D1 y de histamina H3 como potenciales dianes terapéuticas en transtornos adictivos y enfermedades neurodegenerativas. Date: 10/9/2018. Supervisor: Vicente Casadó Burillo.

139 103 | José Manuel Fuentes Rodríguez

DEPTO. BIOQUÍMICA Y BIOLOGÍA MOLECULAR Y GENÉTICA F. ENFERMERÍA Y T.O. UNIVERSIDAD DE EXTREMADURA

Avda de la Universidad s/n 10003 Cáceres Tel. +34 927 257 450 (ext 51286) Fax +34 927 257 451 E-mail: [email protected]

PRINCIPAL Delgado Luceño, Niso Santano, Mireia. INVESTIGATOR María Pura. PhD. Technician. Rodriguez Arribas, Fuentes Rodríguez, José Diop, Sokhana M. S. Mario. Manuel Yakhine. Bachelor degree. PhD. Uribe Carretero, LIST OF González Polo, Rosa Elisabet. PERSONNEL Ana. Bachelor degree. PhD. Casado Naranjo, Ignacio. Bachelor degree. Martínez Chacón, Guadalupe. Technician.

ABSTRACT Our lines of work during 2018 have continued focused on the study of basic molecular mechanisms related to the etiopathogenesis of Parkinson’s disease, regulation of autophagy and role of proteins derived from PARK genes in both processes. On the other hand we have also continued our collaboration and provision of services to the Extremadura Health Service in the field of molecular diagnosis of Parkinson’s disease, as well as we have initiated contacts to establish scientific projects with the Isabel Gemio Foundation on the role of autophagy in certain neuromuscular pathologies As a continuation of the work we have been doing since 2009 in collaboration with the group of Dr. Adolfo López de Munain (CIBERNED), we have continued with characterization of the basal deregulation of macroautophagy in fibroblasts from Par- kinson’s patients carriers of the G2019S pathogenic mutation and R1441G in LRRK2. In 2018 this work has focused on the role of the histone acetyltransferase (HATs) and histone deacetylases (HDACs) activities on mitophagy. For this, fibroblasts from patients with PE (with or without the LRRK2 G2019S mutation) and control subjects were used to evaluate the different phenotypes between idiopathic and genetic EP.

140 CIBERNED 2018 ANNUAL REPORT

The group G2019S shows an increase in mitophagy due to the activation of HDAC class III, while the idiopathic group shows a down regulation of the elimination of defective mitochondria. This reduction in mitophagy is accompanied by an increase in oxygen species (ROS). In parallel, protein acetylation levels of idiopathic individuals and carriers of the G2019S mutation are different due to a positive regulation in HDAC class I and II. Despite this upregulation, total HDAC activity decreases in idiopathic PE and total HAT activity does not vary significantly. The up-regulation of mitophagy is beneficial in reducing the damage induced by ROS in genetic EP. Defective mitophagy in idiopathic PE is inherent in the decrease of class III HDACs. Therefore, there is an imbalance between the activities of total HAT and HDAC in idiopathic PD, which increases cell death. The inhibition of HAT in PD idiopathic cells shows a cyto- protective effect. As an intraCiberned collaboration we continue working on the results of a Coopera- tive Project (PI2015 / 03) together with the Groups of Dr. Pérez Tur and Dr. Pérez Castillo (with the participation of Dr. López de Munain) in which we have addressed the study of Differential metabolic profiles in Parkinson’s disease. During 2018 we completed the process of statistical analysis of results, which we can be published in 2019 Also in the framework of collaboration between groups of CIBERNED (Dres Kulisews- ki, Gutierrez and Vicario) and led by Dr. Moratalla we have described in samples of Parkinson’s patients carriers of the N370P mutation in the gene GBA1 (β-glucocerebrosidase 1) produces a reduction of this enzyme both at the level of protein and activity and its accumulation at the level of the endoplasmic reticulum generating reticulum stress (UPR response) and Golgi apparatus fragmentation, accumulation of autophagosomes and alteration of the autophagic flow with lysosomal dysfunction and cholesterol accumulation. This phenotype modifies the capacity for mitochondrial turnover, generation of oxidative stress and cell death. This work has been a continuation of the work carried out in 2017. In relation to our collaboration with the Extremadura Health Service, we emphasize that we maintain the agreement to provide services in relation to the molecular diagnosis of Parkinson’s and Alzheimer’s disease. We have also been a promoter of the creation of the Institute of Biosanitary Research of Extremadura (INUBE), which has the support of the Ministry of Health and Social Policy of the Junta de Extremadura and the Extremeño Health Service itself. During 2018 the definitive approval of the same has taken place by the university and autonomous government bodies, waiting for its effective starts in 2019.

KEYWORDS Pesticides and Parkinson’s Disease. Roles of apoptosis and autophagy in Parkinson’s Disease. Detection of mutations in PARK genes. Functional roles of PARK gene-derived proteins (synuclein, parkin, DJ-1, PINK1 and LRRK2). Nrf2/keap1 axis. Metabolism.

141 PUBLICATIONS 2018 Yakhine-Diop S, Martinez-Chacon G, Uribe-Carretero E, Niso-Santano M, Gonzalez-Polo R, Fuentes J. The paradigm of protein acetylation in Parkinson’s disease. Neural Regeneration Research. 2019;14(6):975-976. epub2018. PMID: 30762005. Lopez-Espuela F, Gonzalez-Gil T, Amarilla-Donoso J, Cordovilla-Guardia S, Portilla-Cuenca J.C, Casa- do-Naranjo I. Critical points in the experience of spouse caregivers of patients who have suffered a stroke. A phenomenological interpretive study. PLoS ONE. 2018;13(4). PMID: 29617425. Gomez-Suaga P, Bravo-San Pedro J.M, Gonzalez-Polo R.A, Fuentes J.M, Niso-Santano M. ER-mitochondria signaling in Parkinson’s disease review-article. Cell Death and Disease. 2018;9(3). Rodriguez-Hernandez M.A, Gonzalez R, de la Rosa A.J, Gallego P, Ordonez R, Navarro-Villaran E. et al. Molecular characterization of autophagic and apoptotic signaling induced by sorafenib in liver cancer cells. Journal of Cellular Physiology. 2018. PMID: 30132846. Marti-Fabregas J, Medrano-Martorell S, Merino E, Prats-Sanchez L, Marin R, Delgado-Mederos R. et al. Statins do not increase Markers of Cerebral Angiopathies in patients with Cardioembolic Stroke. Scientific Reports. 2018;8(1). Sevilla RR, Naranjo IC, Cuenca JCP, de San Juan BD, Rodriguez JMF, Espuela FL et al. Vascular risk factors and white matter hyperintensities as predictors of progression to dementia in patients with mild cognitive impairment.Current Alzheimer research. 2018. PMID: 29357793. García-Sanz P, Orgaz L, Fuentes JM, Vicario C, Moratalla R. Cholesterol and multilamellar bodies: Lyosomal dysfunction in GBA-Parkinson disease.Autophagy. 2018. PMID: 29368986. Yakhine-Diop S.M.S, Rodriguez-Arribas M, Martinez-Chacon G, Uribe-Carretero E, Gomez-Sanchez R, Aiastui A. et al. Acetylome in human fibroblasts from Parkinson’s disease patients. Frontiers in Cellular Neuroscience. 2018;12. PMID: 29719501. Yakhine-Diop S.M.S, Niso-Santano M, Rodriguez-Arribas M, Gomez-Sanchez R, Martinez-Chacon G, Uribe-Carretero E. et al. Impaired Mitophagy and Protein Acetylation Levels in Fibroblasts from Parkinson’s Disease Patients. Molecular Neurobiology. 2018. PMID: 30032424.

RESEARCH PROJECTS 2018 Code: PI15/00034. Title: Alteraciones del perfil metabolico inducidas por mutaciones patogenicas de LRRK2 como biomarcadores de la enfermedad de Parkinson. Principal investigator: Jose Manuel Fuentes Rodriguez. CIBERNED’s collaboration: No. CIBERNED groups: G103. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 98615. Duration: 2016-2018.

Code: GR18063. Title: Ayudas para la realización de actividades de investigación y desarrollo tecnológico, de divulgación y de transferencia de conocimiento por los grupos de investigación de Extremadura. Principal investigator: Jose Manuel Fuentes Rodriguez. CIBERNED’s collaboration: No. CIBERNED groups: G103. Other CIBER’s collaboration: No.

142 CIBERNED 2018 ANNUAL REPORT

Type: CCAA. Funding agency: Junta de Extremadura. Funding: 49481,25. Duration: 2018-2020.

Code: PI16/01840. Title: Influencia del tratamiento anticoagulante sobre la funcion cognitiva de los pacientes con fibrilacion auricular no valvular. Principal investigator: Ignacio Casado Naranjo. CIBERNED’s collaboration: No. CIBERNED groups: G103. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 55055. Duration: 2017-2019.

Code: Rol patogenico de la disfuncion autofagica/lisosomal en enfermedades neuromusculares. Title: Rol patogenico de la disfuncion autofagica/lisosomal en enfermedades neuromusculares. Principal investigator: Jose Manuel Fuentes Rodriguez. CIBERNED’s collaboration: No. CIBERNED groups: G103. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Isabel Gemio. Funding: 150000. Duration: 2018-2021.

PHD DISSERTATIONS 2018 Author: Mario Rodriguez Arribas. Title: Caracterización de autofagia en células procedentes de enfermos de Párkinson. Date: 6/7/2018. Supervisor: José Manuel Fuentes Rodríguez.

143 113 | José Manuel García Verdugo

LABORATORIO DE NEUROBIOLOGÍA COMPARADA (SS7). INSTITUTO CAVANILLES DE BIODIVERSIDAD Y BIOLOGÍA EVOLUTIVA. UNIVERSIDAD DE VALENCIA

c/ Catedrático José Beltrán, nº 2 46980 Paterna (Valencia) e-mail: [email protected] Tel.: +34 96 354 35 87 Fax: +34 96 354 36 70

PRINCIPAL LIST OF González Granero, INVESTIGATOR PERSONNEL Susana María. Bachelor degree. García Verdugo, José Cebrian Silla, Arantxa. Herranz Pérez, Vicente. Manuel PhD. PhD. González Calixto, María Ulloa Navas, María José. del Carmen. Bachelor degree. Bachelor degree.

ABSTRACT Our group has traditionally been focused on the study of regions with possible adult neurogenesis, analyzing them in different groups of vertebrates, including the human species. We also analyze the behavior of neural stem cells in various pathological conditions of the central nervous system. During the last year we have advanced in the characterization of the neurogenic regions from a comparative point of view, including the study, both at a morphological and molecular level, of these areas in the nervous system of different species. For example, in the ventricular-subventricular zone (V-SVZ) of the canine brain we have described characteristics at the organizational level that are more similar to the human brain than that of the mouse brain. Following this line, we also performed a work on amphibians comparing the spinal cord channel of tadpoles and adult frogs. Using molecular markers of cell proliferation and electron microscopy we were able to describe the regionalization of said channel in a dorsal zone, a ventral zone and two lateral ones, with different proliferative activity throughout the different life stages of the animal. From the point of view of the behavior of the neural stem cells, we wanted to deepen into the mechanism of self-renewal of the murine V-SVZ neural stem cells. Neural stem cells, known as B1 cells, are mainly responsible for neurogenesis in V-SVZ. With age, it is shown that this neurogenic activity decreases, as it does the number of B1

144 CIBERNED 2018 ANNUAL REPORT cells. The stem cells of different somatic tissues are capable of self-renewal, either by asymmetric division, or by symmetric division and subsequent differentiation. In our work, we describe by clonal labeling and lineage tracing that most of these B1 cells divide symmetrically, remaining 20-30% of them in the neurogenic niche, while 70- 80% end up differentiating and, therefore, resulting in the depletion of B1 cells over time. Also using the mouse as an experimental model, we have participated in two works in which we try to improve functional recovery in brain injury models, both in the neonatal and adult stages. For this, we relied on the implantation of soluble hydrogels combined with chemoattractant molecules, which improved the migration of new neurons to the injury region. This strategy allowed us to observe significant improvements at functional level in the treated animals. Finally, another of the points of interest of our group is the study of neurogenic processes in the human brain. After recently describing three routes of migration of new neurons in infant stages from the V-SVZ to the ventral prefrontal cortex, the olfactory bulb and the dorsal prefrontal cortex, we decided to address this analysis in the dentate gyrus of the hippocampus, the other main neurogenic region described in mammals. Given that previous studies suggested that the dentate gyrus incorporated hundreds of new neurons daily in a process affected to some extent by stress and physical exercise, we wanted to delve into aspects about the nature of the stem cells as its rate of proliferation, or the presence of migrating cells. However, in the samples examined, what we observed was a dramatic decrease in the number of immature neurons of this region during the first year of life, being already very scarce in juvenile stages. These results raised important questions about the neurogenic capacity of this region in the adult human. However, these observations have generated an interesting debate due to the contributions of different groups that describe diverging results. Therefore, it is necessary to further dig into this field through the combination of different techniques and the study of quality samples to clarify this dilemma.

KEYWORDS Adult neurogenesis, V-SVZ, neural stem cells, dentate gyrus, hippocampus.

PUBLICATIONS 2018 Jinnou H, Sawada M, Kawase K, Kaneko N, Herranz-Perez V, Miyamoto T. et al. Radial Glial Fib- ers Promote Neuronal Migration and Functional Recovery after Neonatal Brain Injury. Cell Stem Cell. 2018;22(1):128-137.e9. PMID: 29276142.

Francés-Soriano L, Zakharko MA, González-Béjar M, Panchenko PA, Herranz-Pérez V, Pritmov DA, Grin MA, Mironov AF, García-Verdugo JM, Fedorova OA, Perez Prieto J. Nanohybrid for Pho- todynamic Therapy and Fluorescence Imaging Tracking without Therapy. Chemistry of Mate- rials. 2018;30(11):3677-82.

Obernier K, Cebrian-Silla A, Thomson M, Parraguez J.I, Anderson R, Guinto C. et al. Adult Neuro- genesis Is Sustained by Symmetric Self-Renewal and Differentiation. Cell Stem Cell. 2018;:221- 234.e8. PMID: 29395056.

145 Edwards-Faret G, Cebrian-Silla A, Mendez-Olivos E.E, Gonzalez-Pinto K, Garcia-Verdugo J.M, Lar- rain J. Cellular composition and organization of the spinal cord central canal during metamor- phosis of the frog Xenopus laevis. Journal of Comparative Neurology. 2018;526(10):1712- 1732. PMID: 29603210.

Fujikake K, Sawada M, Hikita T, Seto Y, Kaneko N, Herranz-Perez V. et al. Detachment of chain-forming neuroblasts by Fyn-mediated control of cell– cell adhesion in the postnatal brain. Journal of Neuroscience. 2018;38(19):4598-4609. PMID: 29661967.

Paredes M.F, Sorrells S.F, Cebrian-Silla A, Sandoval K, Qi D, Kelley K.W. et al. Does Adult Neu- rogenesis Persist in the Human Hippocampus?. Cell Stem Cell. 2018;23(6):780-781. PMID: 30526879.

Sorrells S.F, Paredes M.F, Cebrian-Silla A, Sandoval K, Qi D, Kelley K.W. et al. Human hippocampal neurogenesis drops sharply in children to undetectable levels in adults. Nature. 2018;555(7696):377-381. PMID: 29513649.

Díaz LR, Saavedra-López E, Romarate L, Mitxitorena I, Casanova PV, Cribaro GP et al. Imbalance of immunological synapse-kinapse states reflects tumor escape to immunity in glioblastoma. JCI insight. 2018;3(18). PMID: 30232280.

Morais G.B, Viana D.A, Verdugo J.M, Rosello M.G, Porcel J.O, Rocha D.D. et al. Morphological characterization of ckd in cats: Insights of fibrogenesis to be recognized. Microscopy Research and Technique. 2018;81(1):46-57. PMID: 29024123.

Kaneko N, Herranz-Perez V, Otsuka T, Sano H, Ohno N, Omata T. et al. New neurons use slit-ro- bo signaling to migrate through the glial meshwork and approach a lesion for functional regeneration. Science Advances. 2018;4(12). PMID: 30547091.

Kirwan P, Kay R.G, Brouwers B, Herranz-Perez V, Jura M, Larraufie P. et al. Quantitative mass spectrometry for human melanocortin peptides in vitro and in vivo suggests prominent roles for β-MSH and desacetyl α-MSH in energy homeostasis. Molecular Metabolism. 2018;17:82- 97. PMID: 30201275.

Atienzar-Aroca S, Serrano-Heras G, Freire Valls A, Ruiz de Almodovar C, Muriach M, Barcia J.M. et al. Role of retinal pigment epithelium-derived exosomes and autophagy in new blood ves- sel formation. Journal of Cellular and Molecular Medicine. 2018;22(11):5244-5256. PMID: 30133118.

Lucia F.S, Pacheco-Torres J, Gonzalez-Granero S, Canals S, Obregon M.-J, Garcia-Verdugo J.M. et al. Transient hypothyroidism during lactation arrests myelination in the anterior commissure of rats. A magnetic resonance image and electron microscope study. Frontiers in Neuroanato- my. 2018;12. PMID: 29755326.

Fernandez-Flores F, Garcia-Verdugo J.M, Martin-Ibanez R, Herranz C, Fondevila D, Canals J.M. et al. Characterization of the canine rostral ventricular-subventricular zone: Morphological, immunohistochemical, ultrastructural, and neurosphere assay studies. Journal of Comparative Neurology. 2018;526(4):721-741. PMID: 29205371.

Mata A, Gil V, Pérez-Clausell J, Dasilva M, González-Calixto MC, Soriano E et al. New functions of Semaphorin 3E and its receptor PlexinD1 during developing and adult hippocampal formation. Scientific reports. 2018;8(1). PMID: 29358640.

146 CIBERNED 2018 ANNUAL REPORT

Code: BFU2015-64207-P. Title: Estudio del cerebro humano: caracterizacion de nichos neurogenicos y nuevas vias de migracion. Principal investigator: Jose Manuel Garcia Verdugo. CIBERNED’s collaboration: No. CIBERNED groups: G113 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 166012. Duration: 2016-2018.

147 305 | Manuel Guzmán Pastor

DEPARTAMENTO DE BIOQUÍMICA Y BIOLOGÍA MOLECULAR FACULTAD DE CIENCIAS QUÍMICAS, UNIVERSIDAD COMPLUTENSE

28040 Madrid Tel.: +34 91 394 46 68 Fax: +34 91 394 46 72 E-mail: [email protected]

PRINCIPAL de Salas Quiroga, Adan. Palazuelos Diego, Javier. INVESTIGATOR Bachelor degree. PhD. Galve Roperh, Ismael. Paraíso Luna, Juan. Guzmán Pastor, Manuel PhD. Bachelor degree. García de Yébenes Resel Mariné, Eva. LIST OF Prous, Justo. Bachelor degree. PERSONNEL PhD. Rodríguez Crespo, José García Rincón, Daniel. Ignacio. Aguareles Gorines, José. Bachelor degree. PhD. Bachelor degree. García Taboada, Elena. Ruiz Calvo, Andrea. Blázquez Ortiz, Cristina. Bachelor degree. Bachelor degree. Bachelor degree. Maroto Martínez, Irene Velasco Díez, Guillermo. Costas Insua, Carlos. Berenice. Bachelor degree. Bachelor degree. Bachelor degree.

ABSTRACT Our research focuses globally on the study of the molecular and cellular mechanisms underlying the control of neural cell physiopathology by the endocannabinoid system. Thus, in 2018 we have kept on studying how this system tunes neural progenitor proliferation, differentiation and survival. Specifically, in the context of our CIBERNED program, we have evaluated the role of the endocannabinoid system in Huntington’s disease. As a matter of fact, Huntington’s disease constitutes, in our opinion, the best currently available model disease to assess the pathophysiological relevance and therapeutic potential of the endocannabinoid system in neurodegenerative diseases. This is due to several reasons: (1) CB1 is the most abundant G protein-coupled receptor in the brain; specifically, it is highly expressed in the basal ganglia at synapses established by neurons containing GABA [especially medium spiny neurons (MSNs), the cells that primarily degenerate in Huntington’s disease] or glutamate (especially corticos-

148 CIBERNED 2018 ANNUAL REPORT triatal projecting neurons, which critically control MSN function) as transmitters, and play a key role in the control of motor behaviour (one of the processes that is most characteristically affected in Huntington’s disease). (2) A remarkable and dorsolate- rally-selective down-regulation of the CB1 receptor has been documented in the basal ganglia of Huntington’s disease patients and animal models, and, of interest, this loss of CB1 receptors seems to reflect, at least in part, the neuron-damage pattern characteristic of the disease. (3) This loss of CB1 receptors occurs at early stages of Hunting- ton’s disease and prior to the appearance of overt clinical symptoms, neurodegeneration and bulk changes in other neurochemical parameters. In 2018, our studies on the endocannabinoid system in Huntington´s disease have aimed at defining (1) whether stimulation of the endocannabinoid system in Huntington’s disease models promotes neuroprotection and therefore delays the onset and/or attenuates the progression of the pathology, (2) whether the alterations of CB1 cannabinoid receptors observed in MSNs and/or corticostriatal terminals at early stages of the disease are involved in the pathogenesis of Huntington’s disease, and (3) whether the induction of CB2 cannabinoid receptors in reactive microglial cells modulates the excitotoxicity processes associated with Huntington’s disease . We aim at unravelling the molecular and cellular bases as well as the potential clinical relevance of those processes.

KEYWORDS Endocannabinoid system; Huntington’s disease; neuroprotection; neurogenesis; cell signalling; experimental therapeutics

PUBLICATIONS 2018 Estevez-Fraga C, Lopez-Sendon Moreno J.L, Martinez-Castrillo J.C, Perez-Perez J, Matarazzo M, Espiga P.G.-R. et al. Phenomenology and disease progression of chorea-acanthocytosis patients in Spain. Parkinsonism and Related Disorders. 2018;49:17-21. PMID: 29317187.

Sagredo O, Palazuelos J, Gutierrez-Rodriguez A, Satta V, Galve-Roperh I, Martinez-Orgado J. Cannabinoid signalling in the immature brain: Encephalopathies and neurodevelopmental disorders. Biochemical Pharmacology. 2018;157:85-96. PMID: 30118663.

Blasco-Benito S, Seijo-Vila M, Caro-Villalobos M, Tundidor I, Andradas C, Garcia-Taboada E. et al. Appraising the “entourage effect”: Antitumor action of a pure cannabinoid versus a bo- tanical drug preparation in preclinical models of breast cancer. Biochemical Pharmacology. 2018;157:285-293. PMID: 29940172.

Guzman M. Cannabis for the management of cancer symptoms: THC version 2.0?. Cannabis and Cannabinoid Research. 2018;3(1):117-119. PMID: 29789813.

Bhattacharjee S, Alarcon F, Cedeno Y, Garcia Yebenes J. Movement disorders associated with neurocysticercosis. Parkinsonism and Related Disorders. 2018;51:111-112. PMID: 29506821.

Lopez-Valero I, Torres S, Salazar-Roa M, Garcia-Taboada E, Hernandez-Tiedra S, Guzman M. et al. Optimization of a preclinical therapy of cannabinoids in combination with temozolomide against glioma. Biochemical Pharmacology. 2018;157:275-284. PMID: 30125556.

149 Lopez-Valero I, Saiz-Ladera C, Torres S, Hernandez-Tiedra S, Garcia-Taboada E, Rodriguez-Fornes F. et al. Targeting Glioma Initiating Cells with A combined therapy of cannabinoids and temozolomide. Biochemical Pharmacology. 2018;157:266-274. PMID: 30195736.

RESEARCH PROJECTS 2018 Code: NEUROLATAM. Title: Unraveling the neurobiological sustrate of protective cannabinoid actions in the brain. Principal investigator: Ismael Galve Roperh, Jose A. Ramos Atance. CIBERNED’s collaboration: Yes. CIBERNED groups: G303 ; G305. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Unión Iberoamericana de Universidades. Funding: 24225. Duration: 2017-2018.

Code: SAF2017-83516. Title: Diseccionando el papel de los receptores CB1 en el desarrollo y la regeneracion de celulas oligodendrogliales. Principal investigator: Javier Palazuelos Diego. CIBERNED’s collaboration: No. CIBERNED groups: G305 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 112500. Duration: 2018-2020.

Code: SAF2015-64945-R. Title: Identificacion y caracterizacion de subpoblaciones del receptor CB1 cannabinoide con actividad neuroprotectora. Principal investigator: Manuel Guzman Pastor. CIBERNED’s collaboration: No. CIBERNED groups: G305 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 330000. Duration: 2016-2018.

Code: PI15/00339. Title: Papel de la autofagia en cancer: mecanismos de muerte mediada por autofagia en celulas tumorales y participacion de genes reguladores de la autofagia en el control de la tumorigenesis. Principal investigator: Guillermo Velasco Diez. CIBERNED’s collaboration: No. CIBERNED groups: G305 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 202000. Duration: 2016-2018.

150 CIBERNED 2018 ANNUAL REPORT

Code: PI15/00310. Title: Papel del sistema endocannabinoide en malformaciones del desarrollo cortical aso- ciadas a epilepsia refractaria. Principal investigator: Ismael Galve Roperh. CIBERNED’s collaboration: No. CIBERNED groups: G305 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 110500. Duration: 2016-2018.

Code: 2016-T1-BMD-1060. Title: The role of the endocannabinoid system in oligodendrocyte development and regeneration during CNS myelination and myelin repair. Principal investigator: Javier Palazuelos Diego. CIBERNED’s collaboration: No. CIBERNED groups: G305 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 421000. Duration: 2018-2021.

Code: EUR.TRAIN16. Title: Tribbles Research and Innovation Network. Principal investigator: Guillermo Velasco Diez. CIBERNED’s collaboration: No. CIBERNED groups: G305 . Other CIBER’s collaboration: CIBERONC. Type: Europeo. Funding agency: Comision Europea. Funding: 452545,92. Duration: 2016-2020.

PHD DISSERTATIONS 2018 Author: Sandra Blasco Benito. Title: El receptor cannabinoide CB2 como diana terapéutica y herramienta pronóstico/ predictiva en cáncer de mama HER2+. Date: 16/11/2018. Supervisor: Manuel Guzmán Pastor.

151 111 | Teresa Iglesias Vacas

INSTITUTO DE INVESTIGACIONES BIOMÉDICAS “ALBERTO SOLS” (CSIC-UAM) Y CIBERNED (ISCIII) DEPARTAMENTO DE FISIOPATOLOGÍA ENDOCRINA Y DEL SISTEMA NERVIOSO

C/ ArturoDuperier, nº 4 28029 Madrid E-mail: [email protected] Tél.: +34 91 585 44 87 Fax: +34 91 585 44 01

PRINCIPAL LIST OF Prudencio Sánchez- INVESTIGATOR PERSONNEL Carralero, Marina. Technician. Iglesias Vacas, Teresa Clares Pedrero, Irene. Simón García, Ana. Bachelor degree. Bachelor degree. Pose Utrilla, Julia. Bachelor degree.

ABSTRACT During the last years our research group has been interested in the study of molecules, processes and mechanisms involved in neuronal death taking place in neuropathologies that produce acute neurodegeneration (such as stroke, traumatic brain injury or epilepsy) and in chronic neurodegenerative diseases (such as amyotrophic lateral sclerosis, Parkinson´s disease, Huntington disease and Alzheimer´s disease). Excitotoxicity is a type of neuronal death that takes place in numerous human neuropathologies as a consequence of an excess of the excitatory amino acid glutamate. It is evident that knowing the molecular mechanisms of excitotoxicity will facilitate to design therapeutic strategies that may result neuroprotective for a wide range of acute or chronic neurodegenerative conditions. Our studies also aim to identify molecules and pathways that participate in neuronal survival, searching how to potentiate their activity and confer neuroprotection. In this sense, we have deepen into the study of two molecules PKD1 (Protein Kinase D1) and its substrate Kidins220 (Kinase D interacting substrate of 220 kDa), and demonstrated that both play a key role enhancing neuronal viability. Last year we publised that PKD1 activity participates in the elmination of mitochondrial free radicals in healthy neurons, decreasing this way neuronal death under conditions of high oxidative stress. Excitotoxicity provokes PKD1 inactivation, increases reactive oxigen species, neuronal damage and neurodegeneration. These harming processes can be rescued using lentivirus for the expression of a constitutively active mutant of PKD1 that confers strong neuroprotection in animal models (Pose-Utrilla & García-Guerra et

152 CIBERNED 2018 ANNUAL REPORT al, Nat Commun, 2017). More recently, we have designed adenoviral vectors for the neurospecific expression of this mutant in preclinical studies of neuroprotection using different animal models of neurodegenerative diseases. In the context of cooperative project CIBERNED 2013/07 we have discovered the differential regulation of two Kidins220 isoforms in Huntington´s disease, that show significant changes from early presyntomatic stages. We will continue our research to determine the contribution of the observed modifications in Kidins220 and PKD1 to the etiopathology of different neurodegenerative diseases, and to design novel neuroprotective strategies. Finally, we have generated conditional PKD1 or Kidins220 deficient mice in different cell lineages and are analysing their phenotype. Our studies this last year show alterations in brain homeostasis, neuroinflammation, and mitochondrial, metabolic, synaptic and neurogenic dysfunctions, accompained by behavioral and memory deficiencies. At present, we continue dilucidating the molecular mechanism involved in the appearance of the observed phenotypes highly related to neurodegeneration.

KEYWORDS Alzheimer, Excitotoxicity, Huntington, Kidins220, Neuroprotection, Oxidative Stress, Protein Kinase D1 (PKD1)

PUBLICATIONS 2018 de Diego García L, Sebastián-Serrano Á, Hernández IH, Pintor J, Lucas JJ, Díaz-Hernández M. The regulation of proteostasis in glial cells by nucleotide receptors is key in acute neuroinflammation.FASEB journal : official publication of the Federation of American Societies for Experi- mental Biology. 2018. PMID: 29401585.

Jimenez-P. R, Martin-Cortazar C, Kourani O, Chiodo Y, Cordoba R, Dominguez-Franjo M.P. et al. CDCA7 is a critical mediator of lymphomagenesis that selectively regulates anchorage-independent growth. Haematologica. 2018;103(10):1669-1678. PMID: 29880607.

Sebastian-Serrano A, de Diego-Garcia L, Henshall D.C, Engel T, Diaz-Hernandez M. Haploinsuf- ficient TNAP mice display decreased extracellular ATP levels and expression of Pannexin-1 channels. Frontiers in Pharmacology. 2018;9(MAR). PMID: 29551976.

Sebastian-Serrano A, Diego-Garcia L.D, Diaz-Hernandez M. The neurotoxic role of extracellular tau protein. International Journal of Molecular Sciences. 2018;19(4). PMID: 29584657.

Pose-Utrilla J, Garcia-Guerra L, Del Puerto A, Martin A, Jurado-Arjona J, De Leon-Reyes N.S. et al. Erratum: Author Correction: Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1 (Nature communications (2017) 8 1 (2275)). Nature communications. 2018;9(1):473-. PMID: 29382840.

153 RESEARCH PROJECTS 2018 Code: CAM-B2017/BMD-3700. Title: Bases metabólicas de la neurodegeneración (NEUROMETAB-CM) Programas de Ac- tividades de I+D entre Grupos de Investigacion de la Comunidad de Madrid en Tecnologias y en Biomedicina. Principal investigator: Jose Gonzalez Castano. CIBERNED’s collaboration: Yes. CIBERNED groups: G401; G502; G111; G409; G412; G205; G110. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 87499,89. Duration: 2014-2020.

Code: SAF2017-88885-R. Title: Mecanismos moleculares implicados en dano cerebral y neurodegeneracion cau- sados por deficiencias en Kidins220 o por eliminacion selectiva de PKD1 en neuronas y astrocitos. Principal investigator: Teresa Iglesias Vacas. CIBERNED’s collaboration: No. CIBERNED groups: G111. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 242000. Duration: 2018-2020.

154 CIBERNED 2018 ANNUAL REPORT

202 | Jaime Kulisevsky Bojarsky

UNIDAD DE TRASTORNOS DE MOVIMIENTO, SERVICIO DE NEUROLOGÍA, HOSPITAL DE LA SANTA CREU I SANT PAU.

Sant Antoni M Claret 167, 08025 Barcelona Teléfono +34 93 553 76 13 Fax +34 93 553 78 72 E-mail: [email protected]

PRINCIPAL Garcia Sanchez, Carmen. Pascual Sedano, Berta. INVESTIGATOR PhD. PhD. Gironell Carrero, Pérez González, Rocío. Kulisevsky Bojarski, Alexandre. PhD. Jaime Bachelor degree. Pérez Pérez, Jesús. LIST OF Horta Barba, Andrea. Bachelor degree. Bachelor degree. PERSONNEL Sampedro Santaló, Marín Lahoz, Juán. Frederic. Aracil Bolaños, Ignacio. Bachelor degree. PhD. Bachelor degree. Martinez Horta, Saul Bejr-Kasem Marco, Indra. Helena. Bachelor degree. Bachelor degree. Pagonabarraga Mora, Campolongo Perillo, Javier. Antonia. PhD. Diplomado.

ABSTRACT Directed by Jaime Kulisevsky MD, PhD (Movement Disorders Unit, Sant Pau Hospital, Biomedical Research Institute), we focus on research of Parkinson’s Disease (PD) and other Movement Disorders. During 2018 we initiated new projects and achieved different objectives:

1. PARKINSON’S DISEASE (PD) A. General • COPPADIS: COhort of Patients with PArkinson’s DIsease in Spain (national collaborative study).

155 B. Cognition • FIS PI14 / 02058: Blood and neurophysiological markers of progression of cognitive impairment. • DuoCog: randomized, double blind and crossed study (immediate levodopa vs. intraduodenal in cognition and mood). • Ciberned intramural project: randomized, double-blind, placebo-controlled trial of efficacy and safety of candesartan in mild cognitive impairment. • Diagnostic accuracy and neuronal correlates of the Buschke memory test in mild cognitive impairment. • Study of perception of colors and visuospatial functions in PD. • Collaboration in the study of Normative data of the PD-CRS in Brazilian population, with Pedro Brandao, University of Brazilia. C. Neuropsychiatric disorders • Marató TV3 (nº 477, 2013): Prediction of impulse control disorders (TCI) and apathy. • FIS (PI15 / 00962): Anatomical-functional correlates of TCI and apathy • Minor hallucinations: analysis of functional correlates (rEEG) and neuroimaging (structural MR, fMRI). • Prospective study on the efficacy of music therapy in apathy and depression of PD. • Collaborative study of Normative Data of the PD-CRS scale (Pedro Brandao, University of Brasilia). D. Advanced PD • Analysis of nutritional status in patients with intraduodenal levodopa. • Prospective cognitive, behavioral, sleep and quality of life study in patients with deep brain stimulation and intraduodenal levodopa. • Prospective study of patient satisfaction after functional surgery. 2. ENFERMEDAD DE HUNTINGTON (EH) A. Coordination of the “Cognitive Phenotype Working Group” (European Huntington’s Dis- ease Network). B. Spanish coordination of Enroll-HD: A Prospective Registry Study in a Global Huntington’s Disease Cohort (PI in Spain: J. Kulisevsky) C. Validation of cognition scales (HD-CRS) and functionality (HD-CFRS). D. Project FIS PI17/01885. Longitudinal study of clinical and neuroradiological correlates associated with levels of huntingtin and other biomarkers in CSF and plasma in HD (BCN- HD project) (PI: Jesús Pérez) E. Collaboration in: a) FIS PI14 / 00834: Longitudinal changes of functional and structural connectivity in pre-symptomatic patients; b) FIS PI15 / 02227: Phenotype and haplo- types associated with intermediate alleles of the EH gene; c) Overcoming Depression in HD: Cdk5 as a potential biomarker and pharmacological target (HD’s disease Society of America); d) The nuclear lamina in HD: physiopathology and therapeutic applications (Ramón Areces Foundation). F. Oxytocin study, plasma levels and modifications in its receptor, and association with apathy and impairment of social cognition in HD (Financing: Regional Health Management and Ministry of Health, Junta de Castilla y León). G. Identification of molecular signatures in exosomes associated with HD.

156 CIBERNED 2018 ANNUAL REPORT

3. TREMOR A. Development of a hardware-software system for a portable neurophysiological study. Collaborative project (SEIDOR company and University of Vic). B. Development of diagnostic system through mobile App and software. Collaborative project with Mediktor company. C. Perampanel in primary orthostatic tremor (pilot study) Collaboration with Association of Patients TOP (Paris, France). 4. OTHER NEURODEGENERATIVE DISEASES A. Neuronal correlates of hypofrontality in subtypes of progressive supranuclear palsy (PSP). B. Neuropsychiatric and behavioral symptoms in subtypes of PSP. C. Multicenter study to validate the diagnosis of neurodegenerative diseases through OS- CANN oculographic registry (Alzheimer’s, PD and frontotemporal dementia vs. controls). D. Lewy Bodies Dementia: participation in an international multi-center study integrated in the “Joint Program for Neurodegenerative Diseases” (European Commission). E. MSA: Collaborative project within the Catalan MSA Registry (CMSAR). F. FXTAS: Collaborative research with Genetics Hospital Clinic (Barcelona) and Associació Catalana X Fràgil (ACXF). G. Collaboration with GEMT-CAT (Group of Movement Disorders of the Catalan Society Neu- rology): Document Consensus on genetics in movement disorders. 5. NEUROIMAGING AND DATA ENGINEERING A. Functional and structural cerebral correlates in HD: cognitive deterioration, apathy, depression and irritability. B. Neuronal correlates of the classification of cognitive impairment of PD according to the MDS. C. Dynamic connectivity models in cognitive impairment of PD (collaboration with Center for Brain and Cognition, Pompeu Fabra University). D. Effects of dopaminergic treatment of PE on neurocognitive networks. E. Analysis of public databases (“big-data”): ENROLL (HD) and PPMI (PD).

KEYWORDS Parkinson - Cognition - Mild Cognitive Impairment - Hallucinations - Huntington - FxTAS - PSP - Essential Tremor – Nauroimaging

PUBLICATIONS 2018 Garcia-Gorro C, Garau-Rolandi M, Escrichs A, Rodriguez-Dechicha N, Vaquer I, Subira S. et al. An active cognitive lifestyle as a potential neuroprotective factor in Huntington’s disease. Neuropsychologia. 2019;122:116-124. epub2018. PMID: 30563619.

157 Sampedro F, Marin-Lahoz J, Martinez-Horta S, Pagonabarraga J, Kulisevsky J. Dopaminergic degeneration induces early posterior cortical thinning in Parkinson’s disease. Neurobiology of Disease. 2019;124:29-35. epub2018. PMID: 30408592.

Lopez Mora D.A, Sampedro F, Camacho V, Fernandez A, Fuentes F, Duch J. et al. Selection of reference regions to model neurodegeneration in huntington disease by 18F-FDG PET/ CT using imaging and clinical parameters. Clinical Nuclear Medicine. 2019;44(1):e1-e5. epub2018. PMID: 30325816.

Compta Y, Giraldo D.M, Munoz E, Antonelli F, Fernandez M, Bravo P. et al. Cerebrospinal fluid levels of coenzyme Q10 are reduced in multiple system atrophy. Parkinsonism and Related Disorders. 2018;46:16-23. PMID: 29107645.

Garcia-Gorro C, de Diego-Balaguer R, Martinez-Horta S, Perez-Perez J, Kulisevsky J, Rodri- guez-Dechicha N. et al. Reduced striato-cortical and inhibitory transcallosal connectivity in the motor circuit of Huntington’s disease patients. Human Brain Mapping. 2018;39(1):54- 71. PMID: 28990240.

Kishore A, Ashok Kumar Sreelatha A, Sturm M, von-Zweydorf F, Pihlstrom L, Raimondi F. et al. Understanding the role of genetic variability in LRRK2 in Indian population. Movement Disorders. 2018. PMID: 30485545.

Rodriguez-Perez A.I, Sucunza D, Pedrosa M.A, Garrido-Gil P, Kulisevsky J, Lanciego J.L. et al. Angiotensin Type 1 Receptor Antagonists Protect Against Alpha-Synuclein-Induced Neu- roinflammation and Dopaminergic Neuron Death. Neurotherapeutics. 2018;15(4):1063- 1081. PMID: 29987762.

Diago E.B, Martinez-Horta S, Lasaosa S.S, Alebesque A.V, Perez-Perez J, Kulisevsky J. et al. Circadian rhythm, cognition, and mood disorders in huntington’s disease. Journal of Hun- tington’s Disease. 2018;7(2):193-198. PMID: 29843249.

Sampedro F, Marin-Lahoz J, Martinez-Horta S, Pagonabarraga J, Kulisevsky J. Cortical Thin- ning Associated with Age and CSF Biomarkers in Early Parkinson’s Disease Is Modified by the SNCA rs356181 Polymorphism. Neurodegenerative Diseases. 2018;:233-238. PMID: 30336481.

Bejr-kasem H, Pagonabarraga J, Martinez-Horta S, Sampedro F, Marin-Lahoz J, Horta-Barba A. et al. Disruption of the default mode network and its intrinsic functional connectivity underlies minor hallucinations in Parkinson’s disease. Movement Disorders. 2018. PMID: 30536829.

Sampedro F, Marín-Lahoz J, Martínez-Horta S, Pagonabarraga J, Kulisevsky J. Early Gray Matter Volume Loss in MAPT H1H1 de Novo PD Patients: A Possible Association With Cog- nitive Decline.Frontiers in neurology. 9. PMID: 29899731.

Prange S, Pagonabarraga J, Krack P, Kulisevsky J, Sgambato V, Tremblay L. et al. Histor- ical crossroads in the conceptual delineation of apathy in Parkinson’s disease. Brain. 2018;141(2):613-619. PMID: 29329378.

Marrero-Gonzalez P, Pascual-Sedano B, Martinez-Domeno A, Diaz-Polo L.E, Granell E, Rod- riguez R. et al. Isolated Propriospinal Myoclonus as a Presentation of Cervical Myelopa- thy. Tremor and other hyperkinetic movements (New York, N.Y.). 2018;8:598-. PMID: 30622837.

Cattaneo C, Kulisevsky J, Tubazio V, Castellani P. Long-term Efficacy of Safinamide on Parkin- son’s Disease Chronic Pain. Advances in Therapy. 2018;:1-8. PMID: 29542008.

Bellosta Diago E, Perez-Perez J, Santos Lasaosa S, Viloria Alebesque A, Martinez-Horta S,

158 CIBERNED 2018 ANNUAL REPORT

Kulisevsky J. et al. Neurocardiovascular pathology in pre-manifest and early-stage Hun- tington’s disease. European Journal of Neurology. 2018;25(7):956-962. PMID: 29537687.

Marín-Lahoz J, Pagonabarraga J, Martinez-Horta S, Fernandez de Bobadilla R, Pascual-Se- dano B, Pérez-Pérez J et al. Parkinson’s Disease: Impulsivity Does Not Cause Impulse Control Disorders but Boosts Their Severity.Frontiers in psychiatry. 9. PMID: 30323775.

Gardoni F, Morari M, Kulisevsky J, Brugnoli A, Novello S, Pisano C.A. et al. Safinamide modulates striatal glutamatergic signaling in a rat model of levodopa-induced dyskinesia. Journal of Pharmacology and Experimental Therapeutics. 2018;367(3):442-451. PMID: 30291173.

Martinez-Horta S, Perez-Perez J, Sampedro F, Pagonabarraga J, Horta-Barba A, Carceller-Sin- dreu M. et al. Structural and metabolic brain correlates of apathy in Huntington’s disease. Movement Disorders. 2018;33(7):1151-1159. PMID: 29676484.

Martinez-Horta S, Moreu A, Perez-Perez J, Sampedro F, Horta-Barba A, Pagonabarraga J. et al. The impact of bilingualism on brain structure and function in Huntington’s disease. Parkinsonism and Related Disorders. 2018. PMID: 30249427.

Gironell A, Pascual-Sedano B, Aracil I, Marin-Lahoz J, Pagonabarraga J, Kulisevsky J. Tremor Types in Parkinson Disease: A Descriptive Study Using a New Classification. Parkinson’s Dis- ease. 2018;2018. PMID: 30363956.

Kulisevsky J, Oliveira L, Fox S.H. Update in therapeutic strategies for Parkinson’s disease. Current Opinion in Neurology. 2018;31(4):439-447. PMID: 29746402.

Kia DA, Sabir MS, Ahmed S, Trinh J, Bandres-Ciga S, International Parkinson’s Disease Genom- ics Consortium. et al. LRP10 in α-synucleinopathies.The Lancet. Neurology. 2018;17(12). PMID: 30507383.

Estevez-Fraga C, Lopez-Sendon Moreno J.L, Martinez-Castrillo J.C, Perez-Perez J, Matarazzo M, Espiga P.G.-R. et al. Phenomenology and disease progression of chorea-acanthocytosis patients in Spain. Parkinsonism and Related Disorders. 2018;49:17-21. PMID: 29317187.

Martinez-Martin P, Kulisevsky J, Mir P, Tolosa E, García-Delgado P, Luquin MR. Validation of a simple screening tool for early diagnosis of advanced Parkinson’s disease in daily practice: the CDEPA questionnaire.NPJ Parkinson’s disease. 4. PMID: 29978014.

Toledo M, Carnero-Pardo C, Carreno-Martinez M, Escudero-Torrella J, Gaig C, Garcia-Ribas G et al. Apuntes en Neurologia» (Notes in Neurology): a synthesis of the evidence on common paroxysmal neurological disorders and on neurodegenerative disorders].Revista de neurologia. 2018;67(s01). PMID: 30779106.

RESEARCH PROJECTS 2018 Code: PI2017/02. Title: Glucocerebrosidasa y Proteinopatías Neurodegenerativas. Principal investigator: Jose L. Lanciego. CIBERNED’s collaboration: Yes. CIBERNED groups: G203; G208; G202. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2017-2019.

159 Code: MARATO 20142910. Title: Blood-based and neurophysiological markers of cognitive deterioration and dementia in Parkinson’s disease. Principal investigator: Javier Pagonabarraga. CIBERNED’s collaboration: No. CIBERNED groups: G202 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 199486,25. Duration: 2015-2018.

Code: PI15/00962. Title: Correlatos anatomico-funcionales de los trastornos del control de impulsos y apatia en la enfermedad de parkinson. Principal investigator: Jaime Kulisevsky. CIBERNED’s collaboration: No. CIBERNED groups: G202 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 74415. Duration: 2016-2018.

Code: PI17/01885. Title: Estudio longitudinal de los asociados a los niveles de huntingtina y otros biomarcadores en LCR y plasma en la enfermedad de Huntington. Principal investigator: Jesus Perez Perez. CIBERNED’s collaboration: No. CIBERNED groups: G202 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 117370. Duration: 2018-2020.

Code: MARATO 20142410. Title: Prediction of apathy and impulse control disorders in Parkinson’s disease based on Feedback related negativity. Principal investigator: Jaime Kulisevsky. CIBERNED’s collaboration: No. CIBERNED groups: G202 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 199607,5. Duration: 2015-2018.

160 CIBERNED 2018 ANNUAL REPORT

208 | José Luis Labandeira García

CENTER FOR RESEARCH IN MOLECULAR MEDICINE AND CHRONIC DISEASES (CIMUS)

Av. Barcelona, 22 Universidad de Santiago de Compostela. Santiago de Compostela, Spain Tel.: +34 88 181 22 23, +34 88 181 54 71 Fax: +34 88 181 23 78

PRINCIPAL Garrido Gil, Pablo. Rodríguez Pallarés, INVESTIGATOR Bachelor degree. Jannette. Guerra Seijas, María PhD. Labandeira García, José José. Rodríguez Pérez, Ana Luis. PhD. Isabel. Bachelor degree. LIST OF López López, Andrea. Bachelor degree. Soto Otero, Ramón. PERSONNEL Méndez Álvarez, PhD. Costa Besada, María Estefanía. Valenzuela Limiñana, Alicia. PhD. Rita. Bachelor degree. Muñoz Patiño, Ana Bachelor degree. Diaz Ruiz, Carmen. María. Villar Cheda, María Bachelor degree. PhD. Begoña. García Garrote, María. Pedrosa Sánchez, Mª PhD. Bachelor degree. Ángeles. Bachelor degree.

ABSTRACT In 2018, we have investigated mechanisms involved in progression of Parkinson’s disease (PD) and higher vulnerability of dopaminergic neurons with aging. We investigated the role of mechanisms involved in the protective effect of physical exercise on aging-related dopaminergic vulnerability, including effects of exercise on levels of nigral angiotensin, IGF-1, SIRT1, SIRT3 or VEGF (J. Gerontol. A Biol. Sci. Med. 10, 1594-1601, 2018). We have also investigated the role of brain renin-angiotensin system (RAS) on neuroinflammation and dopaminergic degeneration, using new animal models in which dopaminergic degeneration was induced by over-expression of alpha-synuclein in dopaminergic neurons using adeno-associated viral vectors (AAV9) (Neurotherapeutics 15, 1063-1081, 2018). Furthermore, we have identified an Ang1-7/MAS receptor axis in dopaminergic neurons of rodents and primates and

161 clarified its functional role (Mol. Neurobiol. 58-47-5867, 2018). We have studied new mechanisms that counteract oxidative stress induced by activation of angiotensin-type 1 receptors (AT1) in dopaminergic neurons that involve the compensatory over-regulation of the NFR2/KLF9 pathway (Free Radic Biol Med 129, 394-406, 2018; Data in Brief 21: 934-942, 2018). We also observed that prostaglandin EP2 receptors may be a new target for dopaminergic neuroprotection, and that these receptors mediate mesenchymal stromal cell-neuroprotective effects on dopaminergic neurons (Mol Neurobiol 55: 4763-4776, 2018). There is an interesting interaction between Parkinson´s disease and gastrointestinal (GI) dysfunction. In 2018, we have published two studies on this question, in which we observed that a nigral dopamine depletion induces changes in intestinal neurotransmiters that may lead to GI motility dysfunction, as well as an increase in GI inflammatory markers. Conversely, GI inflammation led to early changes in dopaminergic components and inflammatory markers in the nigra that may induce an increase in dopaminergic neuron vulnerability (Mol Neurobiol 7297-7316, 2018; Oncotarget 9: 10834-10846, 2018). Finally, we have also collaborated with other groups in their research lines on neurodegeneration. This included studies on relevance of CB1/CB2 receptors in Alzheimer disease (AD) or L-DOPA-induced dyskinesias (Brain Behav Immun 139-151, 2018), Glucocerebrosidase expression in primates as possible new target for neuroprotection in PD (Brain Struct Func 343-355, 2018), or Alzheimer´s disease DNA methylome of pyramidal layers in human frontal cortex (Epigenomics 1365-1382, 2018). An important part of the above-mentioned studies and experiments undertaken in 2018 have been performed in collaboration with different Ciberned research groups: Dr. Lanciego, Dr. R. Franco, Dr. Kulisevsky, Dra. Moratalla.

KEYWORDS Neurodegeneration, neuroprotection, neuroinflammation, Parkinson, aging, angiotensin, cell therapy.

PUBLICATIONS 2018 Diaz-Ruiz C, Villar-Cheda B, Dominguez-Meijide A, Garrido-Gil P, Guerra MJ, Labandei- ra-Garcia JL. Aging-Related Overactivity of the Angiotensin/AT1 Axis Decreases Sirtu- in 3 Levels in the Substantia Nigra, which Induces Vulnerability to Oxidative Stress and Neurodegeneration.The journals of gerontology. Series A, Biological sciences and medical sciences. 2018. PMID: 30412227.

Hernandez H.G, Sandoval-Hernandez A.G, Garrido-Gil P, Labandeira-Garcia J.L, Zelaya M.V, Bayon G.F. et al. Alzheimer’s disease DNA methylome of pyramidal layers in frontal cortex: laser-assisted microdissection study. Epigenomics. 2018;10(11):1365-1382. PMID: 30324800.

Parga J.A, Rodriguez-Perez A.I, Garcia-Garrote M, Rodriguez-Pallares J, Labandeira-Gar- cia J.L. Angiotensin II induces oxidative stress and upregulates neuroprotective signaling from the NRF2 and KLF9 pathway in dopaminergic cells. Free Radical Biology and Medi- cine. 2018;129:394-406. PMID: 30315936.

162 CIBERNED 2018 ANNUAL REPORT

Garrido-Gil P, Rodriguez-Perez A.I, Dominguez-Meijide A, Guerra M.J, Labandeira-Gar- cia J.L. Bidirectional Neural Interaction Between Central Dopaminergic and Gut Lesions in Parkinson’s Disease Models. Molecular Neurobiology. 2018;55(9):7297-7316. PMID: 29404956.

Cruces-Sande A, Rodriguez-Perez A.I, Herbello-Hermelo P, Bermejo-Barrera P, Mendez-Al- varez E, Labandeira-Garcia J.L. et al. Copper Increases Brain Oxidative Stress and Enhances the Ability of 6-Hydroxydopamine to Cause Dopaminergic Degeneration in a Rat Model of Parkinson’s Disease. Molecular Neurobiology. 2018. PMID: 30066305.

Parga J.A, Rodriguez-Perez A.I, Garcia-Garrote M, Rodriguez-Pallares J, Labandeira-Garcia J.L. Data on the effect of Angiotensin II and 6-hydroxydopamine on reactive oxygen species production, antioxidant gene expression and viability of different neuronal cell lines. Data in Brief. 2018;21:934-942. PMID: 30426047.

Munoz A, Correa C.L, Lopez-Lopez A, Costa-Besada M.A, Diaz-Ruiz C, Labandeira-Garcia J.L. Physical Exercise Improves Aging-Related Changes in Angiotensin, IGF-1, SIRT1, SIRT3, and VEGF in the Substantia Nigra. The journals of gerontology. Series A, Biological sciences and medical sciences. 2018;73(12):1594-1601. PMID: 29659739.

Navarro G, Borroto-Escuela D, Angelats E, Etayo Í, Reyes-Resina I, Pulido-Salgado M et al. Receptor-heteromer mediated regulation of endocannabinoid signaling in activated microglia. Role of CB1 and CB2 receptors and relevance for Alzheimer’s disease and levodopa-induced dyskinesia.Brain, behavior, and immunity. 2018;67. PMID: 28843453.

Reyes-Resina I, Aguinaga D, Labandeira-García JL, Lanciego JL, Navarro G, Franco R. Use- fulness of identifying G-protein-coupled receptor dimers for diagnosis and therapy of neurodegenerative diseases and of gliomas.Histology and histopathology. 2018. PMID: 29336473.

Garrido-Gil P, Dominguez-Meijide A, Moratalla R, Guerra M.J, Labandeira-Garcia J.L. Ag- ing-related dysregulation in enteric dopamine and angiotensin system interactions: Im- plications for gastrointestinal dysfunction in the elderly. Oncotarget. 2018;9(13):10834- 10846. PMID: 29541380.

163 Code: RD16/0011/0016. Title: Combined protective/restorative cell-mediated strategies for neurodegenerative diseases. Principal investigator: Jose Luis Labandeira Garcia. CIBERNED’s collaboration: Yes. CIBERNED groups: G208 ; G102; G113; G105; G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 240245. Duration: 2017-2021.

Code: PI2017/02. Title: Glucocerebrosidasa y Proteinopatías Neurodegenerativas. Principal investigator: Jose L. Lanciego. CIBERNED’s collaboration: Yes. CIBERNED groups: G203; G208; G202. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2017-2019.

Code: GRC2014/002. Title: Consolidacion y estructuracion de grupos de referencia competitiva. Principal investigator: Jose Luis Labandeira Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G208. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Xunta de Galicia. Funding: 320000. Duration: 2015-2018.

Code: PI17/00828. Title: Estudio experimental y clinico para identificacion de marcadores de mecanismos de progresion de la Enfermedad de Parkinson y posibles dianas terapeuticas para neuropro- teccion. Principal investigator: Ana Isabel Rodriguez Perez. CIBERNED’s collaboration: No. CIBERNED groups: G208. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 105270. Duration: 2018-2020.

Code: BFU2015-70523. Title: Sistema renina-angiotensina cerebral en neuroinflamacion y degeneracion dopaminergica. Mas alla del eje AII/AT1/NADPH-oxidase. Principal investigator: Jose Luis Labandeira Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G208. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 296500. Duration: 2016-2018.

164 CIBERNED 2018 ANNUAL REPORT

Code: R2014/050. Title: Red Gallega de Terapia celular. Principal investigator: Jose Luis Labandeira Garcia. CIBERNED’s collaboration: No. CIBERNED groups: G208. Other CIBER’s collaboration: CIBEROBN. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 120000. Duration: 2017-2018.

PHD DISSERTATIONS 2018 Author: Maria Alicia Costa Besada. Title: Sistema renina-angiotensina mitocondrial y nuc lear en neuronas. Implicaciones en la enfermedad de Parkinson. Date: 27/4/2018. Supervisor: José Luis Labandeira García.

165 203 | José Luis Lanciego Pérez

CFUNDACIÓN PARA LA INVESTIGACIÓN MÉDICA APLICADA (FIMA). CENTRO DE INVESTIGACIÓN MÉDICA APLICADA (CIMA). DEPARTAMENTO DE NEUROCIENCIAS. LABORATORIO DE ANATOMÍA FUNCIONAL DE LOS GANGLIOS BASALES.

Avda. Pio XII, 55 - Edificio CIMA Tel.: +34 948 194 700 x 2002 Fax: +34 948 194 715 E-mail: [email protected]

PRINCIPAL LIST OF Rico Martín, Alberto INVESTIGATOR PERSONNEL José. PhD. Lanciego Pérez, José Gonzalez Aseguinolaza, Roda Recalde, Elvira. Luis. Gloria. Technician. PhD. Sucunza Guibert, Diego. Gonzalez-Dopeso Reyes, Bachelor degree. Iria Maria. PhD. Pignataro Lopez, Jose Diego. Bachelor degree.

ABSTRACT 1. Studies on heteromers made of G protein-coupled receptors (GPCRs). It might be argued that GPCRs have a natural tendency to form heteromeric complexes that represent novel molecular entities with properties distinct from those of each component receptor, when considered separately. GPCR heteromers represent novel targets for pharmacological developments in the field of Parkinson’s disease. At present, our main activities focus on GPCR heteromers made of dopaminergic receptors (D1 and D2), as well as those formed by endocannabinoid receptors (CB1, CB2 and GPR55). 2. Gene therapy for Parkinson’s disease. We are currently using adeno-associated viral vectors for modeling a number of synucleinopathies in non-human primates, these including Parkinson’s disease, demetia with Lewy bodies (DLB) and multiple system atrophy (MSA). - NHP models of PD with AAV9-SynA53T. - NHP models of DLB with AAV2-retro-SynA53T - NHP models of MSA with AAV-Olig001-SynA53T

166 CIBERNED 2018 ANNUAL REPORT

The ultimate goal is to use gene therapy approaches to further increase the activity of glucocerebrosidase (GCase). GCase enhanced activity is expected to induce a safe and efficient clearance of aggregated alpha-synuclein, thus slowing-down (and ideally arrest), the progressive course that typically characterizes these neurodegenerative disorders. - Gene therapy with AAV9-GBA1 for PD - Gene therapy with AAV2-retro-GBA1 for DLB - Gene therapy with AAV-Olig001-GBA1 for MSA

KEYWORDS GPCR, dopamine, cannabis, basal ganglia, gene therapy, adeno-associated viral vectors, alpha-synuclein, glucocerebrosidase, Gaucher disease.

PUBLICATIONS 2018 Blandini F, Cilia R, Cerri S, Pezzoli G, Schapira A.H.V, Mullin S. et al. Glucocerebrosidase mutations and synucleinopathies: Toward a model of precision medicine. Movement Disorders. 2019;34(1):9-21. epub2018. PMID: 30589955.

Castro-Sanchez S, Garcia-Yague A.J, Lopez-Royo T, Casarejos M, Lanciego J.L, Lastres-Beck- er I. Cx3cr1-deficiency exacerbates alpha-synuclein-A53T induced neuroinflammation and neurodegeneration in a mouse model of Parkinson’s disease. GLIA. 2018;66(8):1752- 1762. PMID: 29624735.

García-Gutiérrez MS, Navarrete F, Navarro G, Reyes-Resina I, Franco R, Lanciego JL et al. Alterations in Gene and Protein Expression of Cannabinoid CB2 and GPR55 Receptors in the Dorsolateral Prefrontal Cortex of Suicide Victims.Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics. 2018. PMID: 29435814.

Franco R, Sanchez-Arias J.A, Navarro G, Lanciego J.L. Glucocerebrosidase Mutations and Synucleinopathies. Potential Role of Sterylglucosides and Relevance of Studying Both GBA1 and GBA2 Genes. Frontiers in Neuroanatomy. 2018;12. PMID: 30002620.

Guridi J, Rodriguez-Rojas R, Carmona-Abellan M, Parras O, Becerra V, Lanciego J.L. History and future challenges of the subthalamic nucleus as surgical target: Review article. Move- ment Disorders. 2018;33(10):1540-1550. PMID: 30288779.

Navarrete F, Garcia-Gutierrez M.S, Aracil-Fernandez A, Lanciego J.L, Manzanares J. Cannab- inoid CB1 and CB2 Receptors, and Monoacylglycerol Lipase Gene Expression Alterations in the Basal Ganglia of Patients with Parkinson’s Disease. Neurotherapeutics. 2018;:1-11. PMID: 29352424.

Navarro G, Borroto-Escuela D, Angelats E, Etayo Í, Reyes-Resina I, Pulido-Salgado M et al. Receptor-heteromer mediated regulation of endocannabinoid signaling in activated micro-

167 glia. Role of CB1 and CB2 receptors and relevance for Alzheimer’s disease and levodopa-induced dyskinesia.Brain, behavior, and immunity. 2018;67. PMID: 28843453.

Lanciego JL. El Conectoma. Colección National Geographic Las Fronteras de la Ciencia. Na- tional Geographic España. 27. 2018. ISBN: 978-84-473-9367-1.

Lanciego JL. Los Trastornos Cerebrales. Colección National Geographic Las Fronteras de la Ciencia. National Geographic España. 27. 2018. ISBN: 978-84-473-9367-1.

168 CIBERNED 2018 ANNUAL REPORT

105 | José López Barneo

INSTITUTO DE BIOMEDICINA DE SEVILLA. HOSPITAL UNIVERSITARIO VIRGEN DEL ROCÍO / CSIC/ UNIVERSIDAD DE SEVILLA

Tel.: +34 955 923 007 Fax: +34 955 923 101 E-mail: [email protected]

PRINCIPAL D´anglemont de Moreno Domínguez, INVESTIGATOR Tassiony, Xavier. Alejandro. PhD. PhD. López Barneo, José. Gao Chen, Lin. Muñoz Cabello, Ana PhD. María. LIST OF García Flores, Gloria PhD. PERSONNEL Paula. Ortega Sáenz, Patricia. Bonilla Henao, Victoria Bachelor degree. PhD. Eugenia. Gómez Garre, María Rho, Hee-Sool. PhD. Pilar. PhD. Cabello Rivera, Daniel. PhD. Sarmiento Soto, Helia. Bachelor degree. López Barneo, José. Bachelor degree. Cabeza Fernandez, José Principal investigator. Toledo Aral, Juan José. María. López López, Ivette. PhD. PhD. Technician. Villadiego Luque, Colinas Miranda, Olalla. Francisco Javier. PhD. PhD.

ABSTRACT In 2018 our group has continued with the work in the two main research lines: a) Mo- lecular nature of oxygen sensors in cells, especially in the glomus cells of the carotid body (CB); and b) Neuroproptection and pathogenesis in Parkinson’s disease (PD), especially the early modifications of the dopaminergic neurons of the nigrostriatal pathway. a) In this line of work we have developed several models of genetically modified KO animals for NDUFS2 (a component of mitochondrial complex I) and other mitochondrial proteins The results derived from this study support our proposal of the mitochondrial-cell membrane interaction, which explains the sensitivity to hypoxia of the CB chemoreceptor cells. In addition, based on the fact that the CB is a chemoreceptor

169 and knowing that hypoxia produces an increase in blood lactate, we have analyzed the role of the olfactory receptor Olfr78, expressed atypically in CB, in this process. The results obtained in the KO mice for the atypical olfactory receptor Olfr78 indicate that this receptor does not participate in the activation of the glomus cells by hypoxia or by lactate. The CB is not only a peripheral chemoreceptor, but acts as a sensor of the global metabolic state of the organism, capable of integrating multiple stimuli such as hypoxemia, hypoglycemia, hypercapnia, acidosis and lactatemia. b) Neuroprotection and pathogenesis of Parkinson’s disease b.1. To carry out this line of research we are using several animal models with aboli- tion of the gene encoding GDNF. We have studied the physiological function of GDNF on the protection of central catecholaminergic neurons. In addition, we have searched for potential pharmacological targets to increase the endogenous production of GDNF. We have confirmed the neuroprotective effect of GDNF on the dopaminergic nigrostriatal neurons. We have also been able to establish a possible model of regulation of GDNF through different intracellular signaling pathways and activation of certain receptors, which would allow the pharmacological stimulation of GDNF production. In collaboration with the group of Dr. José Obeso, we continue with the characterization of the PV+ interneurons producing GDNF in the monkey, using different histological and molecular biology techniques. In parallel, with this work, we continue our collaboration with the group of Dr. James Surmeier from Northwestern University (Chicago, USA) to study in detail the role of mitochondrial dysfunction on PD pathogenesis. b.2. Cell therapy in PD. We continue our collaboration with Dr. Juan José Toledo-Aral´s group on the trophic effects of carotid body (CB) transplants in models of PD. We have analyzed several factors (i.e. age and sex of the human donors) that may influence the action exerted by the transplanted CB tissue. In this regard, age is represents a limiting factor that reduces the therapeutic efficacy of carotid tissue. Currently, we are further studying the molecular mechanisms involved in this phenomenon.

KEYWORDS Hypoxia, mitochondria, carotid body (CB), ion channels, glial cell line derived neurotrophic factor (GDNF), Parkinson’s disease (PD), nigorestriatal pathway, neurodegeneration, parvalbumin positive interneurons (PV+).

PUBLICATIONS 2018 Regidor I, Santos-Garcia D, Catalan M.J, Puente V, Valldeoriola F, Grandas F. et al. Impact of disease duration in effectiveness of treatment with levodopa-carbidopa intestinal gel and factors leading to discontinuation. Journal of Parkinson’s Disease. 2019;9(1):173-182. epub2018. PMID: 30562907.

Santos-Garcia D, Catalan M.J, Puente V, Valldeoriola F, Regidor I, Mir P. et al. Continuous intestinal infusion of levodopa-carbidopa in patients with advanced Parkinson’s disease in Spain: Subanalysis by autonomous community Uso de la infusión intestinal continua de levodopa-carbidopa en pacientes con enfermedad de Parkinson avanzada en España. Subanálisis por comunidades autóno- mas. Neurologia. 2018.

Ray NJ, Bradburn S, Murgatroyd C, Toseeb U, Mir P, Kountouriotis GK et al. In vivo cholinergic basal forebrain atrophy predicts cognitive decline in de novo Parkinson’s disease.Brain : a journal of neurology. 2018;141(1). PMID: 29228203.

170 CIBERNED 2018 ANNUAL REPORT

Corral-Juan M, Serrano-Munuera C, Rabano A, Cota-Gonzalez D, Segarra-Roca A, Ispierto L. et al. Clinical, genetic and neuropathological characterization of spinocerebellar ataxia type 37. Brain. 2018;141(7):1981-1997. PMID: 29939198.

Arias-Mayenco I, González-Rodríguez P, Torres-Torrelo H, Gao L, Fernández-Agüera MC, Bonil- la-Henao V et al. Acute O2 Sensing: Role of Coenzyme QH2/Q Ratio and Mitochondrial ROS Com- partmentalization.Cell metabolism. 2018;28(1). PMID: 29887397.

Gao L, Ortega-Saenz P, Lopez-Barneo J. Acute oxygen sensing—Role of metabolic specifications in peripheral chemoreceptor cells. Respiratory Physiology and Neurobiology. 2018. PMID: 30172779.

Anttila V, Bulik-Sullivan B, Finucane H.K, Walters R.K, Bras J, Duncan L. et al. Analysis of shared her- itability in common disorders of the brain. Science. 2018;360(6395). PMID: 29930110.

Martin-Rodriguez J.F, Mir P. Automatic and voluntary motor inhibition: Intact processes for tic sup- pression?. Movement Disorders. 2018;33(11):1667-1669. PMID: 30306617.

Trillo-Contreras J.L, Ramirez-Lorca R, Hiraldo-Gonzalez L, Sanchez-Gomar I, Galan-Cobo A, Suarez-Luna N. et al. Combined effects of aquaporin-4 and hypoxia produce age-related hydrocephalus. Biochim- ica et Biophysica Acta - Molecular Basis of Disease. 2018;1864(10):3515-3526. PMID: 30293570.

Sobrino V, Gonzalez-Rodriguez P, Annese V, Lopez-Barneo J, Pardal R. Fast neurogenesis from carotid body quiescent neuroblasts accelerates adaptation to hypoxia. EMBO Reports. 2018. PMID: 29335248.

Macias D, Cowburn A.S, Torres-Torrelo H, Ortega-Saenz P, Lopez-Barneo J, Johnson R.S. HIF-2α is essential for carotid body development and function. eLife. 2018;7. PMID: 29671738.

Barrientos-Moreno M, Murillo-Pineda M, Munoz-Cabello A.M, Prado F. Histone depletion prevents telomere fusions in pre-senescent cells. PLoS Genetics. 2018;14(6). PMID: 29879139.

Villadiego J, Romo-Madero S, Garcia-Swinburn R, Suarez-Luna N, Bermejo-Navas A, Echevarria M. et al. Long-term immunosuppression for CNS mouse xenotransplantation: Effects on nigrostriatal neurodegeneration and neuroprotective carotid body cell therapy. Xenotransplantation. 2018;25(6). PMID: 29932254.

Munoz-Cabello A.M, Torres-Torrelo H, Arias-Mayenco I, Ortega-Saenz P, Lopez-Barneo J. Monitor- ing functional responses to hypoxia in single carotid body cells. Methods in Molecular Biology. 2018;1742:125-137. PMID: 29330796.

Lopez-Barneo J. Oxygen sensing and stem cell activation in the hypoxic carotid body. Cell and Tis- sue Research. 2018;:1-9. PMID: 29368257.

Martin-Rodriguez J.F, Mir P. Short-afferent inhibition and cognitive impairment in Parkinson’s disease: A quantitative review and challenges. Neuroscience Letters. 2018. PMID: 29960056.

Ortega-Saenz P, Caballero C, Gao L, Lopez-Barneo J. Testing acute oxygen sensing in genetically modified mice: Plethysmography and amperometry. Methods in Molecular Biology. 2018;1742:139- 153. PMID: 29330797.

Torres-Torrelo H, Ortega-Saenz P, Macias D, Omura M, Zhou T, Matsunami H. et al. The role of Olfr78 in the breathing circuit of mice. Nature. 2018;561(7724):E33-E40. PMID: 30258151.

Tejera-Parrado C, Jesus S, Lopez-Ruiz A, Buiza-Rueda D, Bonilla-Toribio M, Bernal-Bernal I. et al. TMEM230 in Parkinson’s disease in a southern Spanish population. PLoS ONE. 2018;13(5). PMID: 29771939.

Perez-Villalba A, Sirerol-Piquer M.S, Belenguer G, Soriano-Canton R, Munoz-Manchado A.B, Villadie- go J. et al. Synaptic regulator α-synuclein in dopaminergic fibers is essentially required for the

171 maintenance of subependymal neural stem cells. Journal of Neuroscience. 2018;38(4):814-825. PMID: 29217686.

Catalan M.J, Molina-Arjona J.A, Mir P, Cubo E, Arbelo J.M, Martinez-Martin P. Improvement of impulse control disorders associated with levodopa–carbidopa intestinal gel treatment in advanced Parkin- son’s disease. Journal of Neurology. 2018;265(6):1279-1287. PMID: 29557989.

Garcia-Ruiz P.J, Sanz-Cartagena P, Martinez-Castrillo J.C, Ares-Pensado B, Aviles-Olmos I, Blazquez-Estrada M. et al. Myths and evidence on the use of botulinum toxin: Neuropharmacology and dystonia Mitos y evidencias en el empleo de la toxina botulínica: Neurofarmacología y dis- tonías. Revista de Neurologia. 2018;66(5):163-172.

Code: PI-0125-2016. Title: Complicaciones cardiovasculares y Metabolicas del Sindrome de Apnea del Sueno. Patogrnia y modulacion farmacologica de la actividad del eje cuerpo carotideo/medula adrenal. Principal investigator: Gracia Patricia Ortge Saenz. CIBERNED’s collaboration: No. CIBERNED groups: G105. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Junta de Andalucia. Funding: 49909,91. Duration: 2017-2019.

Code: Retos-Colaboracion 15.

172 CIBERNED 2018 ANNUAL REPORT

Title: Desarrollo de una terapia para el tratamiento de variantes geneticas de alfa-sinu- cleina en la enfermedad de Parkinson. Principal investigator: Pablo Mir Rivera. CIBERNED’s collaboration: No. CIBERNED groups: G105. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 317528. Duration: 2016-2018.

Code: FIS 16 (PI16/01575). Title: Estudio integral de los biomarcadores implicados en el desarrollo y evolución de la enfermedad de Parkinson. Principal investigator: Pablo Mir Rivera. CIBERNED’s collaboration: No. CIBERNED groups: G105. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 160325. Duration: 2017-2019.

Code: ERC 2014. Title: Molecular mechanisms of acute oxygen sensing (OXYGENSENSING). Principal investigator: Jose Lopez Barneo. CIBERNED’s collaboration: No. CIBERNED groups: G105 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 2843750. Duration: 2016-2020.

Code: SAF2016-74990-R. Title: Sensibilidad al oxigeno y Neurodegeneracion. Principal investigator: Jose Lopez Barneo, Lin Gao Chen. CIBERNED’s collaboration: No. CIBERNED groups: G105. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 484000. Duration: 2016-2019.

Code: PROPAG-AGEING 2014. Title: The continuum between healthy ageing and idiopathic Parkinson Disease within a propagation perspective of inflammation and damage: the search for new diagnostic, prognostic and therapeutic targets. Principal investigator: Pablo Mir Rivera. CIBERNED’s collaboration: No. CIBERNED groups: G105. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 671167,5. Duration: 2015-2019.

173 306 | José Javier Lucas Lozano

CENTRO DE BIOLOGÍA MOLECULAR “SEVERO OCHOA” CSIC/UAM

C/ Nicolás Cabrera,1. 28049 Madrid Tels. +34 91 196 4552 / +34 91 196 4582 (Lab) Fax +34 91 196 4420 http://www.cbm.uam.es/lineas/ joselucas.htm http://www.ciberned.es/grupo- lucas-lozano.html

PRINCIPAL LIST OF Ollà, Ivana. INVESTIGATOR PERSONNEL Bachelor degree. Parras Rodriguez, Lucas Lozano, José Cobeña Díaz, Sara. Alberto. Javier. Technician. Bachelor degree. Hernandez Hernandez, Picó del Pino, Sara. Ivo. Bachelor degree. Bachelor degree. Santos Galindo, Maria. Lucas Santamaría, PhD. Miriam. Technician.

ABSTRACT Huntington’s disease (HD) is the most prevalent genetic neurodegenerative disease, caused by an expansion of the trinucleotide CAG in the huntingtin gene. In our lab we study the molecular basis of HD through in vitro and in vivo (generating and characterizing transgenic models that could mimic as well as revert the disease) approaches. This way, we discovered an isoform imbalance in tau, a protein related with Alzheimer’s disease and other dementias, and a new histopathological hallmark (the Tau Nuclear Rods or TNRs) in HD. Furthermore, the splicing factor SRSF6, involved in tau splicing and capable to binding CAG-repeats, is altered in HD and sequestered in the mutant huntingtin inclusion bodies (Nat Med. 20(8):881-5 2014). To characterize if tau alteration is enough for TNR appearance, we have tested a mouse model of FTDP17, a disease caused by altered tau. The TNR appearance in this model let us conclude that TNR detection could be useful as marker of tau imbalance (Brain Pa- thol. 27(3):314-322 2017). We have also studied levels and isoforms of MAP2, another microtubule-associated protein which is related to tau. Its levels drop, its isoforms are imbalanced and its subcellular localization is aberrant in HD. In cell models, this splicing can be perfor-

174 CIBERNED 2018 ANNUAL REPORT med by SRSF6 (Brain Pathol. 27(2):181-189 2017). Another research line that we have been exploring for several years involves UPS (J. Neurosci. 23:11653-61 2003, 24:9361-71 2004 and 30:3675-88 2010; Trends Neurosci. 27:66-69 2004; J. Neu- rochem. 98:1585-159 2006, PNAS 106:3986-91 2009) and ERstress (Neurobiol Dis. 41(1):23-32 2011). Related to the latter, we described for the first time the expression of ATF5 in adult neurons (Brain 136(Pt4):1161-76 2013) and its induction with neuroprotective effects in an epilepsy model. More recently, we were able to demonstrate ATF5 expression in adult human neurons and found decreased levels and sequestration into huntingtin inclusion bodies in HD, rendering neurons more vulnerable to mutant huntingtin-induced apoptosis (Acta Neuropathol 134(6):839-850 2017). Our last important finding has relied in a serendipitous finding which has related HD with idiopathic Autism Spectrum Disorder (ASD). Studying the polyadenylation-related proteins CPEBs in HD, we found that CPEB4 binds transcripts of ASD-risk genes. In ASD cases, CPEB4 splicing is imbalanced and an equivalent alteration of CPEB4 isoforms in mice mimics the polyadenylation and protein alterations found in ASD and gives rise to ASD-like phenotype (electrophysiological, neuroanatomical and behavioral), identifying CPEB4 as regulator of ASD risk genes (Nature 560(7719):441-446 2018).

KEYWORDS Huntington’s disease, autism, ASD, splicing, ATF5, CPEBs.

PUBLICATIONS 2018 Parras A, Anta H, Santos-Galindo M, Swarup V, Elorza A, Nieto-Gonzalez J.L. et al. Au- tism-like phenotype and risk gene mRNA deadenylation by CPEB4 mis-splicing. Nature. 2018;560(7719):441-446. PMID: 30111840.

Engel T, Gomez-Sintes R, Alves M, Jimenez-Mateos E.M, Fernandez-Nogales M, Sanz-Rodri- guez A. et al. Bi-directional genetic modulation of GSK-3β exacerbates hippocampal neuropathology in experimental status epilepticus. Cell Death and Disease. 2018;9(10). PMID: 30237424.

Kenny A, Hernández F, Avila J, Lucas JJ, Henshall DC, Prehn JH et al. Profiling of Argo- naute-2-loaded microRNAs in a mouse model of frontotemporal dementia with parkinsonism-17.International journal of physiology, pathophysiology and pharmacology. 10(6). PMID: 30697364. de Diego García L, Sebastián-Serrano Á, Hernández IH, Pintor J, Lucas JJ, Díaz-Hernández M. The regulation of proteostasis in glial cells by nucleotide receptors is key in acute neuroinflammation.FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 2018. PMID: 29401585.

175 RESEARCH PROJECTS 2018 Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: Yes. CIBERNED groups: G604 ; G102; G606; G111; G306. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350000. Duration: 2016-2018.

176 CIBERNED 2018 ANNUAL REPORT

204 | Rosario Moratalla Villalba

INSTITUTO CAJAL (CSIC)

Avenida Doctor Arce, 37 28002 Madrid Tel. +34 915 854 705 Fax +34 915 854 754 E-mail: [email protected]

PRINCIPAL LIST OF Orgaz Gordillo, Lorena. INVESTIGATOR PERSONNEL Bachelor degree. Ruiz de Diego, Irene. Moratalla Villalba, Alberquilla Martínez, PhD. Rosario. Samuel. Solis Castrejon, Oscar. Bachelor degree. Bachelor degree. García Sanz, Patricia Suárez González, Luz Julia. María. PhD. PhD. Granado Martínez, Noelia. PhD.

ABSTRACT During 2018, we have studied the functional and structural synaptic remodeling in Parkinson’s disease (PD) and in L-DOPA-induced dyskinesia in a genetic model of the disease, the Pitx3-/- or Aphakia mouse. Pitx3-/- mice exhibit a parkinsonian phenotype because their dopaminergic neurons of the substantia nigra do not develop normally and therefore, the dorsal striatum (related to motor performance) lacks dopamine. In addition, Pitx3-/- mice are the only genetic model of PD that develops dyskinesias after L-DOPA treatment. We have demonstrated that in Pitx3-/- mice this lack of dopamine reduces the density of the dendritic spines in the main striatal neurons, the striatal projection neurons (SPNs) of both pathways, direct and indirect, which control motor function The spine pruning concurred with an increase in the firing rate of the SPNs, probably due to compensatory mechanisms to maintain overall neuronal activity. After L-DOPA treatment, which induces dyskinesias, the spine density and the firing rate selectively recover in the indirect SPNs. By contrast, the direct-SPNs remain with lower spines and higher firing rate. All these synaptic alterations are similar to those observed in other well-establish models of PD and in human patients, and therefore, our results indicate that Pitx3−/− mice are an excellent mice

177 model to study the synaptic remodeling in PD. In addition, our results demonstra- tes that dopamine-mediated synaptic remodeling and plasticity is independent of dopamine innervation during SPN development (Suarez et al., 2018 J Neurosci). In addition, using 6-OHDA-lesion mice, we have further confirm that L-DOPA increases the activation of direct-SPNs despite the spine pruning whereas indirect-SPNs recover their activation. Therefore, L-DOPA produces an imbalance of striatal function, in favor of direct-SPNs, (Gomez et al., 2019 Mol Neurobiol). In this sense, we have also shown that reducing the activation of direct-SPNs with an mGluR5 antagonist, dis- kinesia were also reduced (García-Montes et al., 2018 Mol Neurobiol). Moreover, we have demonstrated that in dyskinetic mice, L-DOPA produces a maximal activation of the Ras-ERK signaling mediated by D1R stimulation and that upregulation of its intracellular signaling does not further increase dyskinesias while its down-regulation reduce them (Ruiz deDiego et al., 2018 Sci Rep). We have also studied the detailed morphology and distribution of dopaminergic fibers in mice treated with methamphetamine which induces regeneration after the degeneration (demonstrated with silver staining) of dopaminergic fibers in the striatum. We have demonstrated that striatal regeneration was associated with an increase in GAP-43, a sprouting marker, and the presence of growth cone-like TH-ir structures astroglia, indicative of a new terminal generation. Striatal re-generation was associated with an increase in astroglia and decrease in microglia expression, suggesting a possible role for the neuroimmune system in regenerative processes induced by methamphetamine (Granado et al., 2018 Neurotox Res). Finally we have studied the association between alteration in cholesterol pathways and PD. We have demonstrated that the presence of the N370S mutation in the GBA genes (related with autophagy and lysosomal function) produces an accumulation of cholesterol, which alters autophagy-lysosome function with the appearance of multilamellar bodies (MLBs), rendering the cell more vulnerable and sensitive to apoptosis (García-Sanz et al., 2018 Autophagy).

KEYWORDS Parkinson’s disease, dyskinesia, neurotoxicity, dopamine cell death, addiction and autophagy.

PUBLICATIONS 2018 García-Sanz P, Orgaz L, Fuentes JM, Vicario C, Moratalla R. Cholesterol and multilamellar bodies: Lyosomal dysfunction in GBA-Parkinson disease.Autophagy. 2018. PMID: 29368986.

Garrido-Gil P, Dominguez-Meijide A, Moratalla R, Guerra M.J, Labandeira-Garcia J.L. Aging-related dysregulation in enteric dopamine and angiotensin system interactions: Implications for gastrointestinal dysfunction in the elderly. Oncotarget. 2018;9(13):10834-10846. PMID: 29541380.

Suarez L.M, Alberquilla S, Garcia-Montes J.R, Moratalla R. Differential synaptic remodeling by dopamine in direct and indirect striatal projection neurons in pitx3-/- mice, a genetic model of parkinson’s disease. Journal of Neuroscience. 2018;38(15):3619-3630. PMID: 29483281.

178 CIBERNED 2018 ANNUAL REPORT

Solís O, Moratalla R, Moratalla R. Dopamine receptors: homomeric and heteromeric complexes in L-DOPA-induced dyskinesia.Journal of neural transmission (Vienna, Austria : 1996). 2018. PMID: 29417335.

Ruiz-DeDiego I, Fasano S, Solis O, Garcia-Montes J.-R, Brea J, Loza M.I. et al. Genetic enhancement of Ras-ERK pathway does not aggravate L-DOPA-induced dyskinesia in mice but prevents the decrease induced by lovastatin. Scientific reports. 2018;8(1):15381-. PMID: 30337665.

Garcia-Montes J.-R, Solis O, Enriquez-Traba J, Ruiz-DeDiego I, Drucker-Colin R, Moratalla R. Ge- netic Knockdown of mGluR5 in Striatal D1R-Containing Neurons Attenuates l-DOPA-Induced Dyskinesia in Aphakia Mice. Molecular Neurobiology. 2018. PMID: 30259400.

Granado N, Ares-Santos S, Tizabi Y, Moratalla R. Striatal Reinnervation Process after Acute Methamphetamine-Induced Dopaminergic Degeneration in Mice. Neurotoxicity Research. 2018;34(3):627-639. PMID: 29934756.

Garcia-Sanz P, Moratalla R. The importance of cholesterol in Parkinson’s disease. Movement Disorders. 2018;33(2):343-344. PMID: 29315826.

RESEARCH PROJECTS 2018 Code: PI2015-2/02. Title: Potencial patologico de los astrocitos: una nueva perspectiva en la enfermedad de Alzheimer. Principal investigator: Joan X. Comella. CIBERNED’s collaboration: Yes. CIBERNED groups: G413 ; G415; G204; G108; G411. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350000. Duration: 2016-2018.

Code: Cajal Blue Brain Project. Title: Cajal Blue Brain Project. International Blue Brain Proyect. Principal investigator: Javier De Felipe. CIBERNED’s collaboration: Yes. CIBERNED groups: G204 ; G403. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Ecole Polytechnique Federale de Lausanne (Suiza. Funding: ND. Duration: 2009-2019.

Code: Fundacion Ramon Areces 2017. Title: Efecto de las mutaciones del gen glucocerebrosidasa-1 en neuronas derivadas de celulas iPS de enfermos de Parkinson. Rescate del fenotipo y trasplante celular. Area: Ter- apia celular en enfermedades neurodegenerativas. Principal investigator: Carlos Vicario Abejon. CIBERNED’s collaboration: Yes. CIBERNED groups: G204 ; G108. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Ramon Areces. Funding: 120000. Duration: 2017-2020. Code: SAF2016-48532-R.

179 Title: Bases moleculares de la plasticidad sinaptica estriatal en las disquinesias y el trastorno de control de impulsos inducidos por L-DOPA en la enfermedad de Parkinson. Principal investigator: Rosario Moratalla. CIBERNED’s collaboration: No. CIBERNED groups: G204 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 350000. Duration: 2017-2019.

Code: PCIN-2015-098. Title: Development of a new in vivo radiotracer for alpha-synuclein. Principal investigator: Mireille Dumoilin. CIBERNED’s collaboration: No. CIBERNED groups: G204. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 110000. Duration: 2015-2018.

Code: PNSD 2016I033. Title: Estudio del consumo de metanfetamina en la adolescencia como factor de riesgo para la adicion y la vulnerabilidad dopaminergica en el adulto: papel de la glia y del glu- tamato. Principal investigator: Rosario Moratalla. CIBERNED’s collaboration: No. CIBERNED groups: G204. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MSSSI. Funding: 68943. Duration: 2017-2019.

Code: MHE-200017. Title: Mecanismos involucrados en la generacion de disquinesiaspor L-DOPA en unmodelo experimental de la enfermedad de Parkinson. Principal investigator: Rosario Moratalla. CIBERNED’s collaboration: No. CIBERNED groups: G204. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: CSIC. Funding: 25500. Duration: 2016-2018.

180 CIBERNED 2018 ANNUAL REPORT

307 | José Ramón Naranjo Orovio

CSIC CENTRO NACIONAL DE BIOTECNOLOGÍA

Mol. Cell. Biol. C/ Darwin, 3 28049 Madrid E-mail: [email protected] Tél.: +34 915 854 682

PRINCIPAL LIST OF Naranjo Sánchez, María INVESTIGATOR PERSONNEL del Rocío. Bachelor degree. Naranjo Orovio, José Dopazo Santos, José Rábano Gutierrez del Ramón. Manuel. Arroyo, Alberto. Bachelor degree. PhD. González Pérez, Paz. Bachelor degree. López Hurtado, Alejandro. Bachelor degree.

ABSTRACT We investigate changes in neuronal calcium and protein homeostasis that are associated with neurodegenerative pathologies, like Alzheimer’s (AD), Down syndrome (DS) and Huntington’s disease (HD). Dysfunction of calcium and/or protein homeostasis trigger early compensatory changes in transcriptional programs and in synaptic function that precede and, with time, are responsible of disease symptoms characteristic of these pathologies. In particular, our work is focused on the protein DREAM (Downstream Responsive Element Antagonist Modulator), a Ca2+-dependent transcriptional repressor, also known as calsenilin due to its interaction with presenilins. DREAM may have an important role in neurodegenerative diseases (NDD) through the control of Ca2+ and protein homeostasis. Importantly, an early reduction in DREAM levels is found in mouse models for these pathologies (AD, DS and HD) and, genetic experiments show that this could be part of a neuroprotective mechanism operating in HD. The mechanism involves the protein-protein interaction between DREAM and ATF6, a key protein in the early activation of the unfolded protein response (UPR), as well as in the regulation at the transcriptional level of several cytoskeletal proteins that are im-

181 portant for axonal transport and synaptic function. Our results point out the DREAM- ATF6 interaction as a new target for pharmacological intervention in HD. During 2018, we have developed an ELISA assay to check the DREAM-ATF6 interaction in the test tube in a reproducible and quantitative manner. This new ELISA assay will allow for high through-put screening of chemical libraries to identify new active compounds to displace the DREAM-ATF6 interaction and to modify the functional properties of DREAM. We aim to identify new DREAM inhibitors for the treatment of neurodegenerative pathologies. Also during this past year we have reexamined the DREAM-Presenilin interaction to evaluate new therapeutical opportunities. From these studies we have found that i) DREAM increases Presenilin-2 (PS2) endoproteolysis in vivo, in mouse brain and, ii) DREAM inhibition with repaglinide reduces basal Pre- senilin-2 endoproteolysis in N2a mouse neuroblastoma cells. Ongoing experiments are investigating the effect of new DREAM inhibitors on PS-2 endoproteolysis and g-secretase activity. Special attention will be devoted to the analysis of potential inhibition of g-secretase activity on APP cleavage without compromising Notch processing, an unwanted secondary effect that trigger neuronal death and that has blocked the use of previous g-secretase inhibitors for AD treatment.

KEYWORDS DREAM, Alzheimer, ATF6, UPR, DREAM inhibitors, γ−secretase, presenilins.

PUBLICATIONS 2018 Boyano I, Ramos A, Lopez-Alvarez J, Mendoza-Rebolledo C, Osa-Ruiz E, Rodriguez I. et al. Cere- bral Microbleeds in Advanced Dementia: Clinical and Pathological Correlates. American Journal of Alzheimer’s Disease and other Dementias. 2018;33(6):362-372. PMID: 29734821.

Lopez-Hurtado A, Burgos D.F, Gonzalez P, Dopazo X.M, Gonzalez V, Rabano A. et al. Inhibition of DREAM-ATF6 interaction delays onset of cognition deficit in a mouse model of Huntington’s disease. Molecular Brain. 2018;11(1). PMID: 29523177.

Naranjo R, González P, Lopez-Hurtado A, Dopazo XM, Mellström B, Naranjo JR. Inhibition of the Neuronal Calcium Sensor DREAM Modulates Presenilin-2 Endoproteolysis.Frontiers in molecular neuroscience. 11. PMID: 30559648.

Cantero-Recasens G, Butnaru C.M, Valverde M.A, Naranjo J.R, Brouwers N, Malhotra V. KChIP3 coupled to ca2+ oscillations exerts a tonic brake on baseline mucin release in the colon. eLife. 2018;7. PMID: 30272559.

Momtazi G, Lambrecht B, Naranjo JR, Schock BC. Regulators of A20 (TNFAIP3) - new drug-able targets in inflammation.American journal of physiology. Lung cellular and molecular physiology. 2018. PMID: 30543305.

182 CIBERNED 2018 ANNUAL REPORT

Code: SAF2017-89554-R. Title: INHIBIDORES DE DREAM, PROCESAMIENTO DE ATF6 Y PERDIDA COGNITIVA EN AD. Principal investigator: Jose R Naranjo Orovio. CIBERNED’s collaboration: No. CIBERNED groups: G307. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 217800. Duration: 2018-2020.

183 205 | Jose Ángel Obeso Inchausti

CENTRO INTEGRAL DE NEUROCIENCIAS AC (CINAC). HOSPITAL PUERTA DEL SUR Y UNIVERSIDAD SAN PABLO CEU.

Avda. Carlos V, nº 70. 28938. Móstoles (Madrid) Tel.: +34 91 267 32 10. E-mail: [email protected]

PRINCIPAL Del Alamo de Pedro, Mata Marin, David. INVESTIGATOR Marta. Bachelor degree. Bachelor degree. Obeso Martín, Ignacio. Obeso Inchausti, José Etienne , Stephanie. PhD. Ángel. Bachelor degree. Pineda Pardo, José Fernández Hernández, Angel. LIST OF María Ledia. PhD. PERSONNEL PhD. Reinares Sebastian, Foffani, Guglielmo. Alejandro. Alonso Frech, Fernando. PhD. Bachelor degree. Bachelor degree. Gasca Salas, Carmen. Riederer, Gabriela. Arenas Moreno, Laura. Bachelor degree. Technician. Bachelor degree. Hernández Fernández, Rodríguez Rojas, Rafael. Ávila González, María. Frida. Bachelor degree. Bachelor degree. Diplomado. Sánchez Ferro, Alvaro. Blesa De Los Mozos, López González del Rey, Bachelor degree. Francisco Javier. Natalia. PhD. Trigo Damas, Inés. Bachelor degree. PhD. Calvo Gonzalez, Cristina. Oficial Administrativo. Márquez López, Raquel. Vela Desojo, Lydia. Technician. Bachelor degree. Castela Muñoz, Iván. Bachelor degree. Martínez Fernández, Raul. Bachelor degree.

184 CIBERNED 2018 ANNUAL REPORT

ABSTRACT Dr. Obeso heads HM CINAC, a movement disorders research center, which is mainly focused on the study of Parkinson´s disease (PD) from a multidisciplinary approach. The principal objective is the study of the onset and progression of the disease using basic and clinical experiments aiming to define the etiopathogenesis of the disease, optimizing the diagnosis and advancing in the treatment of neurodegenerative and neuropsiquiatric diseases. ONSET OF THE DISEASE: COMPENSATORY MECHANISMS AND SELECTIVE VULNERABILITY So far, treatments for PD have not been successful and one of the reasons is because diagnosis tends to be delayed respect to the onset of the neurodegenerative process. At CINAC, patients are treated with magnetic stimulation (TMS) and neuroimaging directed to define the onset of the disease. Interestingly, PD is principally characterized by the progressive degeneration of dopaminergic neurons of the substantia nigra pars compacta (SNc), which are crucial during learning and habit acquisition. Additionally, since the dopaminergic loss is previous to the diagnosis it is assumed that there might be a series of compensatory mechanisms which delay the rise of the symptoms. At CINAC, in vivo neuronal activity re- cordings in PD animals models are used to define these mechanisms. Surmeier, D. J., Obeso, J. A., & Halliday, G. M. (2017). Parkinson’s Disease Is Not Simply a Prion Disorder. The Journal of Neuroscience : The Official Journal of the Society for Neuroscience EVOLUTION AND TREATMENT OF THE DISEASE HIFU (High Intensity Focused Ultrasound) is a novel technique which allows performing controlled and focused brain lesions without surgery. Thus, mobility, mortality and economic costs are greatly reduced compared to current surgical treatments. This technique has demonstrated efficacy and safety in the performance of thalamotomies aimed to cease essential tremor and PD tremor. HM CINAC has been the first center in perform subthala- motomies to treat cardinal manifestations of Parkinson Disease (rigidity, akinesia, tremor). This study has been published in The Lancet Neurology Journal (IF= 26,28). Martínez-Fernández R, Rodríguez-Rojas R, Del Álamo M, Hernández-Fernández F, Pineda-Par- do JA, Dileone M, Alonso-Frech F, Foffani G, Obeso I, Gasca-Salas C, de Luis-Pastor E, Vela L, Obeso JA. Focused ultrasound subthalamotomy in patients with asymmetric Parkinson’s disease: a pilot study. Lancet Neurol. 2018 Jan;17(1):54-6. PHARMACOLOGICAL TREATMENT OF PARKINSON´S DISEASE AND MOTOR COMPLICATIONS Dopamine replacement therapy has been largely used when treating PD. Both dopamine agonists and levodopa (LDOPA) are the main treatments. After long term use, this therapy is also associated not only with involuntary movements and dyskinesias, but also with with motor complications, which make reduce patients. At CINAC animal models showing progressive dopamine depletion are utilized in order to determine the molecular base and mechanisms involved in the onset of LDOPA dyskinesias (LID) using recording and optogenetic techniques which allow a more specific and selective neuronal modulation. Picconi B, Hernández LF, Obeso JA, Calabresi P. Motor complications in Parkinson’s disease: Striatal molecular and electrophysiological mechanisms of dyskinesias. Mov Disord. 2018 Jul;33(6):867-876.

KEYWORDS Parkinson’s disease, Substantia nigra pars compacta, Basal Ganglia, Dopamine, Vulnerability, Dyskinesias, Compensatory mechanisms

185 PUBLICATIONS 2018 Martinez-Fernandez R, Rodriguez-Rojas R, del Alamo M, Hernandez-Fernandez F, Pineda-Pardo J.A, Dileone M. et al. Focused ultrasound subthalamotomy in patients with asymmetric Par- kinson’s disease: a pilot study. The Lancet Neurology. 2018;17(1):54-63. PMID: 29203153.

Garcia-Ruiz P.J, Sanz-Cartagena P, Martinez-Castrillo J.C, Ares-Pensado B, Aviles-Olmos I, Blazquez-Estrada M. et al. Myths and evidence on the use of botulinum toxin: Neurophar- macology and dystonia Mitos y evidencias en el empleo de la toxina botulínica: Neurofarma- cología y distonías. Revista de Neurologia. 2018;66(5):163-172.

Caballol N, Navarro-Otano J, Avila A, Balague-Marmana M, Martinez-Fernandez R, de Fabregas O. et al. Myoclonus-Dominant Corticobasal Degeneration. Movement Disorders Clinical Prac- tice. 2018;5(6):649-652. PMID: 30637289.

Obeso I, Moisa M, Ruff C.C, Dreher J.-C. A causal role for right temporo-parietal junction in signaling moral conflict. eLife. 2018;7. PMID: 30561334.

Foffani G, Obeso J.A. A Cortical Pathogenic Theory of Parkinson’s Disease. Neuron. 2018;99(6):1116-1128. PMID: 30236282.

Antonini A, Obeso J.A. DBS for Parkinson’s disease with behavioural disturbances. The Lancet Neurology. 2018;17(3):195-197. PMID: 29452675.

Arroyo-Gallego T, Ledesma-Carbayo M.J, Butterworth I, Matarazzo M, Montero-Escribano P, Puertas-Martin V. et al. Detecting Motor Impairment in Early Parkinson’s Disease via Natural Typing Interaction With Keyboards: Validation of the neuroQWERTY Approach in an Uncon- trolled At-Home Setting. Journal of Medical Internet Research. 2018;20(3). PMID: 29581092.

Martinez-Fernandez R, Kibleur A, Chabardes S, Fraix V, Castrioto A, Lhommee E. et al. Different effects of levodopa and subthalamic stimulation on emotional conflict in Parkinson’s disease. Human Brain Mapping. 2018;39(12):5014-5027. PMID: 30259598.

Jimenez S, Mordillo-Mateos L, Dileone M, Campolo M, Carrasco-Lopez C, Moitinho-Ferreira F. et al. Effects of patterned peripheral nerve stimulation on soleus spinal motor neuron excitability. PLoS ONE. 2018;13(2). PMID: 29451889.

Arlotti M, Marceglia S, Foffani G, Volkmann J, Lozano A.M, Moro E. et al. Eight-hours adaptive deep brain stimulation in patients with Parkinson disease. Neurology. 2018;90(11):e971-e976. PMID: 29444973.

Giancardo L, Sanchez-Ferro A, Arroyo-Gallego T, Butterworth I, Mendoza C.S, Montero P. et al. Erratum to: Computer keyboard interaction as an indicator of early Parkinson’s disease (Sci- entific Reports, (2016), 6, 1, (34468), 10.1038/srep34468). Scientific Reports. 2018;8(1). PMID: 30327480.

Sánchez-León CA, Sánchez-López Á, Ammann C, Cordones I, Carretero-Guillén A, Márquez-Ruiz J et al. Exploring new transcranial electrical stimulation strategies to modulate brain function in animal models.Current opinion in biomedical engineering. 2018;8. PMID: 30272042.

Martinez J.H, Lopez M.E, Ariza P, Chavez M, Pineda-Pardo J.A, Lopez-Sanz D. et al. Functional brain networks reveal the existence of cognitive reserve and the interplay between network topology and dynamics. Scientific Reports. 2018;8(1). PMID: 30002460.

Hansen C, Sanchez-Ferro A, Maetzler W. How mobile health technology and electronic health records will change care of patients with Parkinson’s disease. Journal of Parkinson’s Disease. 2018;8(s1):S41-S45. PMID: 30584169.

Humphries M.D, Obeso J.A, Dreyer J.K. Insights into Parkinson’s disease from computational

186 CIBERNED 2018 ANNUAL REPORT models of the basal ganglia. Journal of Neurology, Neurosurgery and Psychiatry. 2018. PMID: 29666208.

Dileone M, Mordillo-Mateos L, Oliviero A, Foffani G. Long-lasting effects of transcranial static magnetic field stimulation on motor cortex excitability. Brain Stimulation. 2018;11(4):676- 688. PMID: 29500043.

Bourquard A, Pablo-Trinidad A, Butterworth I, Sanchez-Ferro A, Cerrato C, Humala K. et al. Non-invasive detection of severe neutropenia in chemotherapy patients by optical imaging of nailfold microcirculation. Scientific Reports. 2018;8(1). PMID: 29593221.

Trigo-Damas I, Del Rey N.L.-G, Blesa J. Novel models for Parkinson’s disease and their impact on future drug discovery. Expert opinion on drug discovery. 2018;13(3):229-239. PMID: 29363335.

Monje M.H.G, Sanchez-Ferro A. Tor1a gene in GABApre interneurons: The new player in the “impaired inhibition” game of dystonia?. Movement Disorders. 2018;33(9):1408-. PMID: 30216539.

Sánchez-León CA, Ammann C, Medina JF, Márquez-Ruiz J, Obeso Inchausti José Ángel. Using animal models to improve the design and application of transcranial electrical stimulation in humans.Current behavioral neuroscience reports. 2018;5(2). PMID: 30013890.

Del Rey N.L.-G, Quiroga-Varela A, Garbayo E, Carballo-Carbajal I, Fernandez-Santiago R, Monje M.H.G. et al. Advances in parkinson’s disease: 200 years later. Frontiers in Neuroanatomy. 2018;12. PMID: 30618654.

Sanchez-Ferro A, Matarazzo M, Martinez-Martin P, Martinez-Avila J.C, Gomez de la Camara A, Giancardo L. et al. Minimal Clinically Important Difference for UPDRS-III in Daily Practice. Move- ment Disorders Clinical Practice. 2018;5(4):448-450. PMID: 30838303.

Mestre T.A, Pont-Sunyer C, Kausar F, Visanji N.P, Ghate T, Connolly B.S. et al. Clustering of motor and nonmotor traits in leucine-rich repeat kinase 2 G2019S Parkinson’s disease nonparkinso- nian relatives: A multicenter family study. Movement Disorders. 2018;33(6):960-965. PMID: 29665080.

Guridi J, Rodriguez-Rojas R, Carmona-Abellan M, Parras O, Becerra V, Lanciego J.L. History and future challenges of the subthalamic nucleus as surgical target: Review article. Movement Disorders. 2018;33(10):1540-1550. PMID: 30288779.

Diez-Fairen M, Benitez B.A, Ortega-Cubero S, Lorenzo-Betancor O, Cruchaga C, Lorenzo E. et al. Pooled-DNA target sequencing of Parkinson genes reveals novel phenotypic associations in Spanish population. Neurobiology of Aging. 2018;70:325.e1-325.e5. PMID: 29887346.

187 CIBERNED’s collaboration: Yes. CIBERNED groups: G401; G502; G111; G409; G412; G205; G110. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 87499,89. Duration: 2014-2020.

Code: CAM-B2017/BMD-3700. Title: Bases metabólicas de la neurodegeneración (NEUROMETAB-CM) Programas de Ac- tividades de I+D entre Grupos de Investigacion de la Comunidad de Madrid en Tecnologias y en Biomedicina. Principal investigator: Jose Gonzalez Castano. CIBERNED’s collaboration: Yes. CIBERNED groups: G401; G502; G111; G409; G412; G205; G110. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 87499,89. Duration: 2014-2020.

Code: PEJ-2017-AL/BMD-7337. Title: Ayuda para la contratacion de ayudantes de investigacion. Principal investigator: Ines Trigo Damas. CIBERNED’s collaboration: No. CIBERNED groups: G205 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 22500. Duration: 2017-2018.

Code: BES2016-077493. Title: Ayudas para contratos predoctorales para la formacion de doctores. Principal investigator: Jose A. Obeso. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 26750. Duration: 2017-2019.

Code: CD15/00092. Title: Contrato Posdoctoral de Perfeccionamiento Sara Borrell. Grupo habitual. Principal investigator: Ignacio Obeso Martin. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 80598. Duration: 2016-2019.

Code: PEJD-2017-PRE/HUM-4034. Title: Convocatoria de ayudas de investigadores predoctorales. Principal investigator: Ignacio Obeso. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No.

188 CIBERNED 2018 ANNUAL REPORT

Type: CCAA. Funding agency: Comunidad de Madrid. Funding: 25000. Duration: 2018-2018.

Code: EULACH16/T01-0047. Title: DASYN - Early detection and cellular disintegration of α-synuclein aggregates using nanobodies. Principal investigator: Prof. Pedro Chana. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 44978,12. Duration: 2018-2021.

Code: SAF2017-86246-R. Title: Estimulacion transcraneal por campo magnetico estatico en la Enfermedad de Par- kinson. Principal investigator: Guglielmo Foffani. CIBERNED’s collaboration: No. CIBERNED groups: G20 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 18150. Duration: 2018-2020.

Code: AC16/00044. Title: Harmonisation metabolic FDG brain pattern characteristics. Principal investigator: Jose Angel Obeso Inchausti. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 47069. Duration: 2017-2019.

Code: Sin codigo_Tatiana. Title: Objetivo α-synucleina: entender la vulnerabilidad celular y detener la progresion en la Enfermedad de Parkinson. Principal investigator: Javier Blesa De Los Mozos, Jose Angel Obeso Inchausti. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Tatiana Perez de Guzman el Bueno. Funding: 84500. Duration: 2017-2019.

Code: Sin codigo. Title: Selective Vulnerability, progression and Synuclein Toxicity in Parkinson`s disease. Principal investigator: Jose Angel Obeso Inchausti. CIBERNED’s collaboration: No. CIBERNED groups: G205. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion BBVA. Funding: 125000. Duration: 2016-2019.

189 110 | Ana María Pérez Castillo

INSTITUTO DE INVESTIGACIONES BIOMÉDICAS (CSIC-UAM). DPTO. MODELOS EXPERIMENTALES DE ENFERMEDADES HUMANAS

C/ Arturo Duperier 4 28029 Madrid Tel. +34 91 585 44 36 Fax. +34 91 585 44 01 E-mail: [email protected]

PRINCIPAL LIST OF Sanz San Cristóbal, INVESTIGATOR PERSONNEL Marina. Technician. Morales García, José Pérez Castillo, Ana Sierra Magro, Ana. Ángel. María Bachelor degree. PhD. Van Bulck, Michiel. Santos Montes, Ángel. Bachelor degree. Bachelor degree.

ABSTRACT Our research is focused on identification of new therapeutic targets for the treatment of neurodegenerative diseases. To that purpose our work is based on the use of several preclinical models that mimics some of the aspects that characterize Alzheimer’s and Par- kinson’s diseases. Using these models we identify and analyze potential cellular targets in order to develop new drugs for the treatment of these diseases. During the year 2017 we have continued working on this topic, studying the processes that characterize these pathologies. In that sense, we have studied the involvement of several genes in different brain disorders such as C/EBPβ, whose inhibition has been shown to have a potent neuroprotector in a Parkinson’s model. Some other gen of interest we have analyzed is PDE7, which codes for an enzyme involved in the degradation of cAMP. Our results suggest that this gene is expressed early in degenerative processes that affect the dopaminergic neurons of the substantia nigra, as well as promotes the appearance of pro-inflammatory phenomena. Also, a main focus of the lab concerns research on neurogenesis and aging. In this regard we are expanding our previous observations that describe the neurogenic effect of certain components of the brew known as Ayahuasca. In this sense, we are currently working in the role of different new cellular targets which can expand our knowledge of the processes that lead to improved neurogenesis and that can be of use for a better understanding and new treatments of aging-related disorders.

190 CIBERNED 2018 ANNUAL REPORT

KEYWORDS Alzheimer, APP, Ayahuasca, C/EBPβ, LRRK2, neurogenesis, neuroinflammation, neuroprotection, Parkinson.

PUBLICATIONS 2018 Estrada Valencia M, Herrera-Arozamena C, de Andres L, Perez C, Morales-Garcia J.A, Perez-Cas- tillo A. et al. Neurogenic and neuroprotective donepezil-flavonoid hybrids with sigma-1 affinity and inhibition of key enzymes in Alzheimer’s disease. European Journal of Medicinal Chemistry. 2018;156:534-553. PMID: 30025348.

Code: SAF2017-85199-P. Title: CCAAT/Enhancer binding protein β (C/EBPβ) como modulador de la neuroinflamacion. Una nueva diana terapeutica en la enfermedad de Parkinson. Principal investigator: Ana Maria Perez Castillo. CIBERNED’s collaboration: No. CIBERNED groups: G110. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 217558. Duration: 2018-2020.

Code: MARIE SKŁODOWSKA-CURIE ACTIONS. H2020-MSCA-ITN-2016 Title: Blood Biomarker-based Diagnostic Tools for Early-stage Alzheimer’s Disease. Principal investigator: Ana Perez Castillo. CIBERNED’s collaboration: No. CIBERNED groups: G110. Other CIBER’s collaboration: CIBERER. Type: Europeo. Funding agency: Comision Europea. Funding: 3465749,52. Duration: 2016-2020.

191 209 | Jordi Pérez Tur

UNITAT DE GENÈTICA MOLECULAR INSTITUT DE BIOMEDICINA DE VALÈNCIA-CSIC

C/ Jaume Roig, 11 · 46010 València Tél.: +34 963 391 755 Fax: +34 963 393 774 E-mail: [email protected]

PRINCIPAL LIST OF Rubio Granero, INVESTIGATOR PERSONNEL Concepción. Bachelor degree. Cardona Serrate, Pérez Tur, Jordi Szymanski, Jacek Pawel. Fernando. Bachelor degree. PhD.

ABSTRACT Throughout 2018, our group has continued with the characterization of Mendelian forms of neurological diseases. Thus we have determined the presence of several mutations related to Alzheimer’s disease, primary lateral sclerosis, frontotemporal dementia or paroxysmal kinesigenic dystonia. In this area, we have determined the segregation of the mutations found in the families, although the small size of these does not allow to reach definitive conclusions about this, as well as the population frequency of the same. In addition, we have begun the study of two other family forms with rare neurological disorders. On the one hand, an atypical form of what appears to be frontotemporal dementia and, on the other, a form of disease that presents as optic atrophy with or without ataxia, mental retardation and epileptic seizures but that is not related to mutations in RTN4IP. In a second phase of this same project, we are advancing in the description of the mechanisms through which some of these mutations could be causing the pathological process with which they are related, developing cellular models of the respec- tive diseases on which to study the degradation of the mRNA of the mutated allele and the possible haploinsufficiency that said degradation produces. This seems to be the mechanism involved in the appearance of paroxysmal kinesigenic dystonia. However, in the case of Alzheimer’s disease, our data, even preliminary, suggest that the mutation we have found affects intracellular signalling pathways. On the other hand, we have begun the study of non-Mendelian forms of Alzhei- mer’s disease through the genomic analysis of part of the Vallecas cohort, collected by the CIEN Foundation. This is a collaborative study with the group of Dr. Máñez (CiberBBN) in which we will combine information on metabolites present in

192 CIBERNED 2018 ANNUAL REPORT these same patients with genetic information to determine characteristics of the disease that help to know its appearance and progression. In parallel, we continue analyzing the results of the study of expression in patients with Parkinson’s disease of genetic origin, combining them with metabolomic data on the same patients. Finally, we have initiated a collaboration with groups of other entities in which it is sought to determine if it is possible to advance in a very early diagnosis of the appearance of cognitive deficits by combining neuropsychological, genetic and lifestyle data. This project, has the particularity of that the population that is intended to study is collected in community pharmacies. This is a novel approach that takes as reference the trust that is established between a person and their pharmacist when there is a continuous treatment between them.

KEYWORDS Alzheimer’s, Parkinson’s, Frontotemporal dementia, Mendelian diseases, Neurodegeneration, Neurogenetics, ALS

PUBLICATIONS 2018 Vazquez-Costa J.F, Carratala-Bosca S, Tembl J.I, Fornes-Ferrer V, Perez-Tur J, Marti-Bonmati L. et al. The width of the third ventricle associates with cognition and behaviour in motor neuron disease. Acta Neurologica Scandinavica. 2019;139(2):118-127. epub2018. PMID: 30183086.

Sanchez-Mut J.V, Heyn H, Silva B.A, Dixsaut L, Garcia-Esparcia P, Vidal E. et al. PM20D1 is a quantitative trait locus associated with Alzheimer’s disease. Nature Medicine. 2018;24(5):598- 603. PMID: 29736028.

Navarro-Sánchez L, Águeda-Gómez B, Aparicio S, Pérez-Tur J. Epigenetic Study in Parkinson’s Disease: A Pilot Analysis of DNA Methylation in Candidate Genes in Brain.Cells. 2018;7(10). PMID: 30261625.

193 114 | José Antonio del Río Fernández

MOLECULAR AND CELLULAR NEUROBIOTECHNOLOGY INSTITUTE FOR BIOENGINEERING OF CATALONIA (IBEC).

Baldiri Reixac 10-15 08028 Barcelona, Spain Tfno.: +34 93 4035923 / 20296 E-mail: [email protected] E-mail: [email protected]

PRINCIPAL Gil Fernández, Vanesa. Matamoros i Angles, INVESTIGATOR PhD. Andreu. Hervera Abad, Arnau. Bachelor degree. Del Río Fernández, José PhD. Mesquida Veny, Antonio Lidón Gil, Laia. Francina. Bachelor degree. Bachelor degree. LIST OF López Mengual, Ana. Segura Feliu, Miriam. PERSONNEL Bachelor degree. Technician. Urrea Zazurca, Laura. Gavín Marín, Rosalina. Mata Rodriguez, Agata. Bachelor degree. PhD. Bachelor degree.

ABSTRACT 1) Development of new lab on chip devices for Neurobiology. One of our focus is to mimic the developing and neurodegenerating nervous system in lab on chip devices. We believe that combining several different stimuli in the chip resembles a more realistic environment that nerve cells will encounter in the living animal in normal and disease conditions. Thus, we developed last year a new device able to reproduce the formation of the neuromuscular junction (NMJ) in lab on chip devices (Sala et al., submitted). Members of our lab used biophysics and image analysis to study changes in muscle contraction in particular diseases (Figure 1). In addition, a device designed to analyze axon lesions of cortical neurons was also developed (Mesquida et al., submitted). Current experiments of our group in collaboration with several groups of Ciberned and other labs aimed at developing new lab on chip devices to mimics and modulate particular neurodegenerative processes. For example: I) on chip lab platform to monitor drug delivery and network connectivity between different neuronal populations; II) cortico-spinal chips to develop axon regenerative studies of new drug formulation (in collaboration with Imperial College; UK) or, IV) in silico 3D modeling for neurodegenerative diseases (Alzheimer and Parkinson chip) (Figure 2) 2) New strategies to monitor and overcome a-synuclein and tau transport in neurons and

194 CIBERNED 2018 ANNUAL REPORT glial cells during neurodegeneration. α-Synuclein is a key player in the pathogenesis of synucleinopathies, including Parkinson’s disease (PD), Dementia with Lewy Bodies (DLB), and multiple system atrophy (MSA). Tau is present as neurofibrillary tangles (NFT) in different neurodegenerative diseases including Alzheimer´s disease (AD) or Frontotem- poral dementia-tau (FTD-tau). Transmission of synthetic α-synuclein or Tau aggregates has been demonstrated in several cellular and animal models. Several groups have reported that insoluble α-synuclein or Tau shows “prion-like” propagation in wild-type mice. However, the basis of the spreading process remains poorly understood although cell to cell transport via classical exocytosis, exosomes, nanotubules or receptor mediated endocytosis has been proposed. We described in 2018 that the cellular prion protein PrPC is a new receptor for α-synuclein involved in their seeding and propagation (Urrea et al., 2018a; 2018b) reviewed in Progress in Neurobiology (Del Rio, Ferrer et al., 2018). Our new objectives aimed to block this interaction to reduce the neuropathological transport of α-synuclein. Similar experiments are also developed in the case of Tau since recent data of our group showed that Tau also binds to PrPC during its inter-neuronal propagation. In addition, we also described a feed-back neuroprotective answer to affected neurons since intracellular soluble Tau fractions also trigger PrPC production at its promoter level enhancing neuronal survival at early neurodegenerative stages of different tauopathies including AD (Lidón et al., submitted). 3) Understanding axon regeneration: from reactive oxygen species to epigenetics. Our group in a fruitful collaboration with Prof. Simone di Giovanni (Imperial College, UK) published in Nat. Cell Biology we demonstrated that the generation of ROS and NOX2 activity is mandatory to enhance axon regeneration in the lesioned spinal injury in vivo (Hervera et al., 2018). In fact, exosomes containing NOX2 are released from macrophages and incorporated into injured axons via endocytosis and transported to the cell body through an importin-b1-dynein-dependent mechanism. Endosomal NOX2 oxidizes PTEN stimula- ting PI3K-Akt inducing axon regeneration (Hervera et al., 2018). These data have been reviewed recently in Trends in cell biology (Hervera et al., 2019). Recently accepted publication points to epigenetic changes in the lesioned neurons that might enhance axon regeneration (Hervera et al., 2019a; Hervera et al., EMBOJ in press).

KEYWORDS α-synuclein, tauopathies, interneuronal spreading, axon regeneration, lab-on-chip.

PUBLICATIONS 2018 Mata A, Gil V, Pérez-Clausell J, Dasilva M, González-Calixto MC, Soriano E et al. New functions of Semaphorin 3E and its receptor PlexinD1 during developing and adult hippocampal formation. Scientific reports. 2018;8(1). PMID: 29358640.

Ferrer I, Aguiló García M, López González I, Diaz Lucena D, Roig Villalonga A, Carmona Tech M et al. Aging-related tau astrogliopathy (ARTAG): Not only tau phosphorylation in astrocytes. Brain pathology (Zurich, Switzerland). 2018. PMID: 29396893.

Llorens F, Thune K, Marti E, Kanata E, Dafou D, Diaz-Lucena D. et al. Regional and subtype-dependent miRNA signatures in sporadic Creutzfeldt-Jakob disease are accompanied by alterations in miRNA silencing machinery and biogenesis. PLoS Pathogens. 2018;14(1). PMID: 29357384.

195 Badiola-Mateos M, Hervera A, Del Río JA, Samitier J. Challenges and Future Prospects on 3D in-vitro Modeling of the Neuromuscular Circuit.Frontiers in bioengineering and biotechnology. 6. PMID: 30622944.

Tomas-Roig J, Piscitelli F, Gil V, Quintana E, Ramió-Torrentà LL, Del Río JA et al. Effects of repeated long-term psychosocial stress and acute cannabinoid exposure on mouse corticostriatal circuitries: Implications for neuropsychiatric disorders.CNS neuroscience & therapeutics. 2018. PMID: 29388323.

Hervera A, De Virgiliis F, Palmisano I, Zhou L, Tantardini E, Kong G et al. Publisher Correction: Reactive oxygen species regulate axonal regeneration through the release of exosomal NADPH oxidase 2 complexes into injured axons.Nature cell biology. 2018;20(9). PMID: 29520084.

Hervera A, De Virgiliis F, Palmisano I, Zhou L, Tantardini E, Kong G et al. Reactive oxygen species regulate axonal regeneration through the release of exosomal NADPH oxidase 2 complexes into injured axons.Nature cell biology. 2018. PMID: 29434374.

Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Garcia-Aznar J.M. et al. Erratum to: Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons (Molec- ular Neurobiology, (2018), 55, 3, (1847-1860), 10.1007/s12035-017-0451-4). Molecular Neurobiology. 2018;55(3):1861-.

Franco R, Aguinaga D, Reyes I, Canela EI, Lillo J, Tarutani A et al. N-Methyl-D-Aspartate Re- ceptor Link to the MAP Kinase Pathway in Cortical and Hippocampal Neurons and Microglia Is Dependent on Calcium Sensors and Is Blocked by α-Synuclein, Tau, and Phospho-Tau in Non-transgenic and Transgenic APPSw,Ind Mice.Frontiers in molecular neuroscience. 11. PMID: 30233307.

del Rio J.A, Ferrer I, Gavin R. Role of cellular prion protein in interneuronal amyloid transmission. Progress in Neurobiology. 2018;165-167:87-102. PMID: 29530723.

Code: INTRAMURAL CIBERBBN-CIBERNED. Title: Gamma-peptides as anti–alzheimer drugs with BBB crossing properties (GAM- MA-AD). Principal investigator: Miriam Royo Exposito. CIBERNED’s collaboration: No. CIBERNED groups: G114 . Other CIBER’s collaboration: CIBER-BBN. Type: Intramurales. Funding agency: CIBERBBN. Funding: ND. Duration: 2018-2018.

196 CIBERNED 2018 ANNUAL REPORT

Code: PI2016/02. Title: Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches. Principal investigator: Jose Antonio Del Rio. CIBERNED’s collaboration: Yes. CIBERNED groups: G114; G401; G201. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210000. Duration: 2016-2018.

Code: TEC2015-72718-EXP. Title: Robots biologicos basados en el control de la union neuromuscular. Principal investigator: Josep Samitier. CIBERNED’s collaboration: No. CIBERNED groups: G114. Other CIBER’s collaboration: CIBER-BBN. Type: Nacional. Funding agency: MICINN. Funding: 55000. Duration: 2016-2018.

Code: BFU2015-6777-R. Title: Funciones de genes implicados en angiogenesis y remodelacion vascular durante el desarrollo cortical y en neurodegeneracion. Principal investigator: Jose Antonio Del Rio. CIBERNED’s collaboration: No. CIBERNED groups: G114. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 279259,2593. Duration: 2016-2018.

Code: SGR2017-648. Title: Grupo de investigación de calidad de la Generalitat de Catalunya. Principal investigator: Jose A Del Rio. CIBERNED’s collaboration: No. CIBERNED groups: G114. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 20000. Duration: 2018-2019.

PHD DISSERTATIONS 2018 Author: Laura Urrea Zazurca. Title: Funciones de la proteína prionica celular, a-sinucleína y reclina en enfermedades neurodegenerativas. Date: 16/3/2018. Supervisor: José Antonio Del Río Fernández.

197 206 | Manuel Rodríguez Díaz

DEPARTAMENTO DE FISIOLOGÍA, FACULTAD DE MEDICINA UNIVERSIDAD DE LA LAGUNA

La Laguna Tél.: +34 922 319 361 Fax: +34 922 319 397 E-mail: [email protected]

PRINCIPAL LIST OF Rodríguez Sabaté, Clara. INVESTIGATOR PERSONNEL Bachelor degree. Sabaté Bel, Magdalena. González Hernández, Rodríguez Díaz, Manuel PhD. Tomás. PhD. Sánchez Fernández, Alberto. Montón Álvarez, Bachelor degree. Fernando. PhD. Morales Pérez, Ingrid. PhD.

ABSTRACT We studied the action of astrocytes in Parkinson’s disease. During the process of degeneration of the dopaminergic terminals in the striatum dopaminergic, spheroids of 1-8 microns accumulate the intracellular detritus of these neurons, detritus involving proteins such as the APP, the TH and DAT and organelles such as mitochondria. This accumulation prevents scattered detritus that would facilitate the inflammatory response and would alter the functioning of the tissue surrounding the dopaminergic degeneration. In addition, the clustering of detritus in spheroids facilitates disposal. Dopaminergic spheroids present autophagosomes (LC3) but not lysosomes (Lamp1 or Lamp2), which suggests that autophagy stars but does not end in the spheroids. We found evidence suggesting that the autophagy that begins at dopaminergic terminals ends in surrounding astrocytes, and that astrocytes are keys to the cleaning of the dopaminergic debris generated in the normal brain with the age. We also observed microglial activation, but activated microglia does not present phagocyte characteristics (e.g. it does not increase the expression of CD68). These data suggest that, under basal conditions, the detritus cleaning is

198 CIBERNED 2018 ANNUAL REPORT done by astrocytes rather than by the microglia. Last year, we have proceeded to assess the evolution of the dopaminergic organelles which are also accumulated in spheroids during the dopaminergic degeneration. Studies with different transgenic mice suggest that astrocytes are also involved in the recovery/metabolization of these organelles. These studies were conducted in animal models of Parkinson’s disease, and now we are proceeding to validate these findings in striatal samples of of parkinsonian patient which were provided by the Brain Bank of the CIEN Fun- dation. Finally, these studies are also being verified through additional techniques such as electron microscopy. During the year 2018, we have expanded our studies on the functional connectivity of the basal ganglia of normal human brain, proceeding to evaluate the changes in connectivity caused by Parkinson’s disease. We had previously studied the basal ganglia by means of neuroimaging techniques based on the correlation of BOLD signals. These studies showed a circular cortico-subcortical interaction which, as a whole, was similar to that described in experimental animals. However the correlation techniques have some important limitations, since they assume normality in the sample and a linear relationship between basal ganglia that does not occur in reality (in previous studies we showed non-linear activity in centers like the substantia nigra). In addition, correlation only provide information about the relationship between two centers at the same time, and basal ganglia present multiple simultaneous interactions which cannot be properly assessed with this technique. For this reason we developed a new analytical procedure based on the Multiple Correspondence Analysis, a procedure that can be used for the study of multiple non-linear interactions. This method identified up to seven functional groupings in the cortico-subcortical motor circuit of the basal ganglia, clusters that showed significant changes in the parkinsonian brain. However, the Multiple Corresponden- ce Analysis also has limitations. It does not offer the possibility of carrying out a statistical evaluation of the results (a circumstance that it shares with other techniques such as independent component analysis), and can only identify a limited number of functional groupings (7 groupings in our study). For this reason we have developed a new technique that allows the identification of multiple functional interactions (without the limit showed by the Multiple Correspondence Analysis) in brain centers with non-linear relationships, and that check these interactions through a proper statistical contrast. With this new technique we have identified more than 16 functional groupings in the activity of the basal ganglia. We are presently proceeding to determine the effect of Parkinson’s disease in these functional groupings.

KEYWORDS Basal ganglia, astrocytes, dopaminergic vulnerability, neuroimflammation, functional neuroimaging.

199 PUBLICATIONS 2018 Nieto A, Hernandez-Torres A, Perez-Flores J, Monton F. Depressive symptoms in Friedreich ataxia Síntomas depresivos en la ataxia de Friedreich. International Journal of Clinical and Health Psychology. 2018;18(1):18-26.

Luis-Ravelo D, Estevez-Silva H, Barroso-Chinea P, Afonso-Oramas D, Salas-Hernandez J, Rodri- guez-Nunez J. et al. Pramipexole reduces soluble mutant huntingtin and protects striatal neurons through dopamine D3 receptors in a genetic model of Huntington’s disease. Experimental Neurology. 2018;299:137-147. PMID: 29056363.

200 CIBERNED 2018 ANNUAL REPORT

207 | Eduard Tolosa Sarró

INSTITUT DE NEUROCIENCIES. UNITAT DE PARKINSON I TRASTORNS ANORMALS DEL MOVIMENT.

c/ Villarroel 171, esc 8 - 4 planta 08036 Barcelona. España. Tel: +34 932 275 785 Fax: +34 932 275 783 E-mail: [email protected]

PRINCIPAL Fernández Sánchez, José. INVESTIGATOR Manuel. PhD. Technician. Navarro Otano, Judith. Tolosa Sarró, Eduard Gaig Ventura, Carlos. Bachelor degree. PhD. Simonet Hernández, LIST OF Garrido Pla, Alicia. PhD. Cristina. PERSONNEL Iranzo de Riquer, Bachelor degree. Alejandro. Valldeoriola Serra, Compta Hirnyj, PhD. Francesc. Yaroslau. PhD. PhD. Junqué Plaja, Carmen. PhD. Vilas Rolán, Dolores. Ezquerra Trabalón, Bachelor degree. Mario. Maragall Moreno, Laura. PhD. Auxiliar Administrativo. Martí Doménech, María

ABSTRACT In 2018 we continued our research on clinical and biological aspects of LRRK2-associated Parkinson‘s disease (LRRK2-PD) which include results on microRNA alterations in iPSC-derived dopaminergic neurons from LRRK2-PD and sporadic PD (Neurobiol Aging; Tolosa et al., 2018), characterization of mitochondrial function in a a cell model of LRRK2-PD (J Transl Med, Juárez-Flores et al., 2018), a study of candidate genetic modifiers of age at onset in LRRK2-PD (Mov Disord, Fernandez-Santiago et al., 2018).We have described the lack of central and peripheral nervous system synuclein pathology in R1441G LRRK2-PD, unlike the case of G2019S LRRK2 mutations (JNNP, Vilas et al., 2018) and studies on the traits associated to healthy carriers of LRRK2 mutations (Mov Disord; Mestre et al., 2018). We are currently working towards deepening our knowledge of existing models of PD epigenetic alterations by using iPSC-DAn models and patient blood samples, work on miRNA/RNA prognostic and diagnostic biomarkers in blood, serum and cultured pri-

201 mary fibroblast of parkinsonisms (MSA, idiopathic PD, LRRK2-PD, healthy controls) (work in progress). We have described reduced levels of Q10 in multisystem atrophy (MSA) (Compta et al., Park Rel Dis 2018) and initiated recruitment of patients for the collection of CSF and brain MRI images in MSA patients. Experiments with the protein cyclic amplification method RTQuIC for synuclein have been initiated in our lab (IPs: Dr. Y. Compta y Dra. M.J. Martí) using both CSF and brain tissue. We have also contributed to the International DLB consortium (Lancet Neurol, Guerreiro et al., 2018) and, in the field of experimental therapeutics described the effects of subthalamic and substantia nigra pars reticulata deep brain stimulation upon the gait of patients with PD (Park Rel Dis., Valldeoriola et al., 2018). The research line MRI of the neuroanatomical and neurofunctional bases of cognitive impairment in PD, led by Dr C. Junque and supported by two grants (MINECO PSI2017-86930 and Marato201423.10), provided evidence about the utility of MRI techniques to identify subtle degenerative changes in PD over time. MRI data-driven analyses techniques proved useful to detect subtypes of cortical atrophy even in de novo patients. (Park Re Dis, Uribe et al 2018) and we described that the gray/white matter contrast is an excellent MRI marker of ageing effects. Moreover, searching for variables sensitive to PD progression, we detected a differential vulnerability to hippocampal subfields atrophy in PD compared to normal ageing. (Frontiers in Aging Neuroscience, Uribe et al. 2018). In addition, we showed functional connectivity deficits in PD and found that connectivity threshold-free network-based statistics (TFNBS) is an appropriate technique for the statistical assessment of brain graphs (Hum Brain Mapping 2018 Baggio et al). We reporte don the neuroanatomical correlates of several visuospatial/visuoperceptual tests, as markers of cognition (J Intern Neuropsychol; Garcia Diaz et al. 2018; Park Rel Dis; Garcia Diaz et al, 2018) and the psychometric characteristics of the Spanish version of the UPSIT test (Arch Clin Neuropsychol; Campabadal et al. 2018,). In studies published in 2018 year in journals including Lancet Neurology, Brain and Sleep, we shown that idiopathic REM sleep behavior disorder represents the prodromal phase of Parkinson disease, alpha-synuclein can be found in living subjects in the minor labial salivary glands and in the parotid gland, and we assessed the factors that determine its early conversion. We described in in conjunction with the PET center in Aahrus, extrastriatal monoaminergic involvement as well as microglial activation in the subjects with prodromal PD (Neurobiol Dis. 2018). In addition, we have continued characterizing anti-IgLON5 disease, a newly described tauopathy with prominent RBD and movement disorders.

KEYWORDS Prodromal Parkinson; RBD, LRRK2; biomarkers, CSF, neuroimaging, multisystem atrophy, progressive supranuclear palsy

202 CIBERNED 2018 ANNUAL REPORT

PUBLICATIONS 2018 Kun-Rodrigues C, Orme T, Carmona S, Hernandez D.G, Ross O.A, Eicher J.D. et al. A comprehensive screening of copy number variability in dementia with Lewy bodies. Neurobiology of Aging. 2019;75:223.e1-223.e10. epub2018. PMID: 30448004.

Iranzo A, Stefani A, Hogl B, Santamaria J. Sleep and sleep disorders in Franz Kafka’s narrative works. Sleep Medicine. 2019;55:69-73. epub2018. PMID: 30772696.

Guerreiro R, Ross O.A, Kun-Rodrigues C, Hernandez D.G, Orme T, Eicher J.D. et al. Investigating the genetic architecture of dementia with Lewy bodies: a two-stage genome-wide association study. The Lancet Neurology. 2018;17(1):64-74. PMID: 29263008.

Vilas D, Sharp M, Gelpi E, Genis D, Marder K.S, Cortes E. et al. Clinical and neuropathological features of progressive supranuclear palsy in Leucine rich repeat kinase (LRRK2) G2019S mutation carriers. Movement Disorders. 2018;33(2):335-338. PMID: 29119599.

Garcia-Diaz A.I, Segura B, Baggio H.C, Uribe C, Campabadal A, Abos A. et al. Cortical thinning correlates of changes in visuospatial and visuoperceptual performance in Parkinson’s disease: A 4-year follow-up. Parkinsonism and Related Disorders. 2018;46:62-68. PMID: 29132765.

Iranzo A. Parasomnias and Sleep-Related Movement Disorders in Older Adults. Sleep Medicine Clinics. 2018;13(1):51-61. PMID: 29412983.

Mestre T.A, Pont-Sunyer C, Kausar F, Visanji N.P, Ghate T, Connolly B.S. et al. Clustering of motor and nonmotor traits in leucine-rich repeat kinase 2 G2019S Parkinson’s disease nonpar- kinsonian relatives: A multicenter family study. Movement Disorders. 2018;33(6):960-965. PMID: 29665080.

Kim H, Calatayud C, Guha S, Fernandez-Carasa I, Berkowitz L, Carballo-Carbajal I. et al. Cor- rection to: The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neu- ron Function and Survival Against α-Synuclein-Induced Toxicity (Molecular Neurobiology, (2018), 55, 9, (7533-7552), 10.1007/s12035-018-0881-7). Molecular Neurobiology. 2018;55(9):7553-7554.

Kim H, Calatayud C, Guha S, Fernández-Carasa I, Berkowitz L, Carballo-Carbajal I et al. The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neuron Function and Survival Against α-Synuclein-Induced Toxicity.Molecular neurobiology. 2018. PMID: 29429047. Martin-Trias P, Lanteaume L, Solana E, Casse-Perrot C, Fernandez-Cabello S, Babiloni C. et al. Adaptability and reproducibility of a memory disruption rTMS protocol in the PharmaCog IMI European project. Scientific Reports. 2018;8(1). PMID: 29921865.

Ottino-Gonzalez J, Jurado M.A, Garcia-Garcia I, Segura B, Marques-Iturria I, Sender-Palacios M.J. et al. Allostatic load and disordered white matter microstructure in overweight adults. Scientific Reports. 2018;8(1). PMID: 30367110.

Fernandez-Arcos A, Vilaseca I, Aldecoa I, Serradell M, Tolosa E, Santamaria J. et al. Alpha-synuclein aggregates in the parotid gland of idiopathic REM sleep behavior disorder. Sleep Med-

203 icine. 2018;52:14-17. PMID: 30195197.

Muller T, Tolosa E, Badea L, Asgharnejad M, Grieger F, Markowitz M. et al. An observational study of rotigotine transdermal patch and other currently prescribed therapies in patients with Parkinson’s disease. Journal of Neural Transmission. 2018;125(6):953-963. PMID: 29484495.

Markopoulou K, Compta Y. Cerebrospinal fluid levels of alpha-synuclein in PARKINSON’S disease: Another long and winding road. Parkinsonism and Related Disorders. 2018;49:1-3. PMID: 29548634.

Uribe C, Segura B, Baggio H.C, Abos A, Garcia-Diaz A.I, Campabadal A. et al. Cortical atrophy patterns in early Parkinson’s disease patients using hierarchical cluster analysis. Parkinson- ism and Related Disorders. 2018;50:3-9. PMID: 29449187.

Sanz-Granda A, Marti M.J, Catalan M.J. Cost-utility analysis of two formulations of botulinum toxin type A in the treatment of blepharospasm and cervical dystonia in Spain Analisis coste-utilidad de dos formulaciones de toxina botulinica de tipo A en el tratamiento del ble- faroespasmo y la distonia cervical en España. Revista de neurologia. 2018;67(12):465-472. PMID: 30536360.

Campabadal A, Segura B, Baggio HC, Abos A, Uribe C, Garcia-Diaz AI et al. Diagnostic Accuracy, Item Analysis and Age Effects of the UPSIT Spanish Version in Parkinson’s Disease.Archives of clinical neuropsychology : the official journal of the National Academy of Neuropsychologists. 2018. PMID: 30007334.

Uribe C, Segura B, Baggio H.C, Campabadal A, Abos A, Compta Y. et al. Differential progression of regional hippocampal atrophy in aging and Parkinson’s disease. Frontiers in Aging Neuro- science. 2018;10(OCT). PMID: 30364338.

Iranzo A. Dissecting premotor Parkinson’s disease with multimodality neuroimaging. The Lancet Neurology. 2018;17(7):574-576. PMID: 29866442.

Juárez-Flores DL, González-Casacuberta I, Ezquerra M, Bañó M, Carmona-Pontaque F, Catalán- García M et al. Exhaustion of mitochondrial and autophagic reserve may contribute to the development of LRRK2 G2019S-Parkinson’s disease.Journal of translational medicine. 2018;16(1). PMID: 29884186.

Zhang Z, Mayer G, Dauvilliers Y, Plazzi G, Pizza F, Fronczek R. et al. Exploring the clinical features of narcolepsy type 1 versus narcolepsy type 2 from European Narcolepsy Network database with machine learning. Scientific Reports. 2018;8(1). PMID: 30006563.

Stokholm M.G, Iranzo A, Ostergaard K, Serradell M, Otto M, Bacher Svendsen K. et al. Extra- striatal monoaminergic dysfunction and enhanced microglial activation in idiopathic rapid eye movement sleep behaviour disorder. Neurobiology of Disease. 2018;115:9-16. PMID: 29522818.

Gamez-Valero A, Iranzo A, Serradell M, Vilas D, Santamaria J, Gaig C. et al. Glucocerebrosidase gene variants are accumulated in idiopathic REM sleep behavior disorder. Parkinsonism and Related Disorders. 2018;50:94-98. PMID: 29487000.

Uribe C, Segura B, Baggio H.C, Abos A, Garcia-Diaz A.I, Campabadal A. et al. Gray/White matter contrast in Parkinson’s disease. Frontiers in Aging Neuroscience. 2018;10(MAR). PMID: 29636679.

Blauwendraat C, Kia D.A, Pihlstrom L, Gan-Or Z, Lesage S, Gibbs J.R. et al. Insufficient evidence for pathogenicity of SNCA His50Gln (H50Q) in Parkinson’s disease. Neurobiology of Aging. 2018;64:159.e5-159.e8. PMID: 29398121.

204 CIBERNED 2018 ANNUAL REPORT

Iranzo A, Stefani A, Hogl B, Santamaria J. Kafkas’ insomnia and narrative works. Sleep Medi- cine. 2018;52:233-. PMID: 30327164.

Vilas D, Gelpi E, Aldecoa I, Grau O, Rodriguez-Diehl R, Jauma S. et al. Lack of central and peripheral nervous system synuclein pathology in R1441G LRRK2 -associated Parkinson’s disease. Journal of Neurology, Neurosurgery and Psychiatry. 2018.

Mayer G, Plazzi G, Iranzo A, Ortega-Albas J, Quinnell T, Pesch H. et al. Long-term compliance, safety, and tolerability of sodium oxybate treatment in patients with narcolepsy type 1: A postauthorization, noninterventional surveillance study. Sleep. 2018;41(9). PMID: 29986085.

Tolosa E, Botta-Orfila T, Morato X, Calatayud C, Ferrer-Lorente R, Marti M.-J. et al. MicroRNA alterations in iPSC-derived dopaminergic neurons from Parkinson disease patients. Neurobi- ology of Aging. 2018;69:283-291. PMID: 29935433.

Martin-Flores N, Fernandez-Santiago R, Antonelli F, Cerquera C, Moreno V, Marti M.J. et al. MTOR Pathway-Based Discovery of Genetic Susceptibility to L-DOPA-Induced Dyskinesia in Parkinson’s Disease Patients. Molecular Neurobiology. 2018;:1-9. PMID: 29992529.

Weibel D, Sturkenboom M, Black S, de Ridder M, Dodd C, Bonhoeffer J. et al. Narcolepsy and adjuvanted pandemic influenza A (H1N1) 2009 vaccines – Multi-country assessment. Vac- cine. 2018;36(41):6202-6211. PMID: 30122647.

Rodriguez-Vazquez A, Vicente-Pascual M, Maya-Casalprim G, Valldeoriola F. Neuroimaging in hypoglycaemic encephalopathy diagnosis and prognosis: A case report Neuroimagen en el diagnóstico y pronóstico de la encefalopatía hipoglucémica: A propósito de un caso. Neuro- logia. 2018.

Fabbri M, Ferreira J.J, Lees A, Stocchi F, Poewe W, Tolosa E. et al. Opicapone for the treatment of Parkinson’s disease: A review of a new licensed medicine. Movement Disorders. 2018;33(10):1528-1539. PMID: 30264443.

Dauvilliers Y, Schenck C.H, Postuma R.B, Iranzo A, Luppi P.-H, Plazzi G. et al. REM sleep behaviour disorder. Nature Reviews Disease Primers. 2018;4(1). PMID: 30166532.

Stefani A, Heidbreder A, Brandauer E, Guaita M, Neier L.-M, Mitterling T. et al. Screening for idiopathic REM sleep behavior disorder: Usefulness of actigraphy. Sleep. 2018;41(6). PMID: 29554362.

Brittain C, McCarthy A, Irizarry M.C, McDermott D, Biglan K, Hoglinger G.U. et al. Severity dependent distribution of impairments in PSP and CBS: Interactive visualizations. Parkinsonism and Related Disorders. 2018. PMID: 30201421.

Valldeoriola F, Munoz E, Rumia J, Roldan P, Camara A, Compta Y. et al. Simultaneous low-frequency deep brain stimulation of the substantia nigra pars reticulata and high-frequency stimulation of the subthalamic nucleus to treat levodopa unresponsive freezing of gait in Par- kinson’s disease: A pilot study. Parkinsonism and Related Disorders. 2018. PMID: 30241951.

Baggio H.C, Abos A, Segura B, Campabadal A, Garcia-Diaz A, Uribe C. et al. Statistical infer- ence in brain graphs using threshold-free network-based statistics. Human Brain Mapping. 2018;39(6):2289-2302. PMID: 29450940.

Razquin C, Ortega-Cubero S, Rojo-Bustamante E, Diez-Fairen M, Lorenzo E, Alonso E. et al. Target-enriched sequencing of chromosome 17q21.31 in sporadic tauopathies reveals no candidate variants. Neurobiology of Aging. 2018;66:177.e7-177.e10. PMID: 29398119.

Borrego-Ecija S, Grau-Rivera O, Colom-Cadena M, Molinuevo J.L, Tolosa E, Sanchez-Valle R. et

205 al. Tauopathy with Hippocampal 4-Repeat Tau Immunoreactive Spherical Inclusions in a Pa- tient with PSP. Brain Pathology. 2018;28(2):284-286. PMID: 29516660.

Pizza F, Antelmi E, Vandi S, Meletti S, Erro R, Baumann C.R. et al. The distinguishing motor features of cataplexy: A study from video-recorded attacks. Sleep. 2018;41(5). PMID: 29425380.

Valldeoriola F, Salvador A, Gomez-Arguelles J.M, Marey J, Moya M, Ayuga A. et al. The effects of transdermal rotigotine on nonmotor symptoms of Parkinson’s disease: A multicentre, observational, retrospective, postmarketing study. International Journal of Neuroscience. 2018;128(4):369-375. PMID: 29249180.

Iranzo A, Stefani A, Hogl B, Santamaria J. The insomnia of Franz Kafka. Sleep Medicine. 2018;50:24-28. PMID: 29982086.

Marek K, Chowdhury S, Siderowf A, Lasch S, Coffey CS, Caspell-Garcia C et al. The Parkinson’s progression markers initiative (PPMI) - establishing a PD biomarker cohort.Annals of clinical and translational neurology. 2018;5(12). PMID: 30564614.

Iranzo A. The REM sleep circuit and how its impairment leads to REM sleep behavior disorder. Cell and Tissue Research. 2018;373(1):245-266. PMID: 29846796.

Gaig C, Iranzo A, Santamaria J, Graus F. The Sleep Disorder in Anti-lgLON5 Disease. Current Neurology and Neuroscience Reports. 2018;18(7). PMID: 29796717.

Gonzalez-Casacuberta I, Moren C, Juarez-Flores D.-L, Esteve-Codina A, Sierra C, Catalan-Garcia M. et al. Transcriptional alterations in skin fibroblasts from Parkinson’s disease patients with parkin mutations. Neurobiology of Aging. 2018;65:206-216. PMID: 29501959.

Cano-Lopez I, Calvo A, Boget T, Carreno M, Donaire A, Setoain X. et al. Typical asymmetry in the hemispheric activation during an fMRI verbal comprehension paradigm is related to better performance in verbal and non-verbal tasks in patients with epilepsy. NeuroImage: Clinical. 2018;20:742-752. PMID: 30238918.

Compta Y, Munoz E, Marti M.J. Ubiquinone, ubiquinol, 4-hydroxybenzoic acid… What ‘coenzyme Q10’ should we care about in multiple system atrophy?. Parkinsonism and Related Disorders. 2018;50:117-118. PMID: 29433994.

Iranzo A, Borrego S, Vilaseca I, Marti C, Serradell M, Sanchez-Valle R. et al. α-Synuclein aggregates in labial salivary glands of idiopathic rapid eye movement sleep behavior disorder. Sleep. 2018;41(8). PMID: 29790977.

Kia DA, Sabir MS, Ahmed S, Trinh J, Bandres-Ciga S, International Parkinson’s Disease Genom- ics Consortium. et al. LRP10 in α-synucleinopathies.The Lancet. Neurology. 2018;17(12). PMID: 30507383.

Regidor I, Santos-Garcia D, Catalan M.J, Puente V, Valldeoriola F, Grandas F. et al. Impact of disease duration in effectiveness of treatment with levodopa-carbidopa intestinal gel and factors leading to discontinuation. Journal of Parkinson’s Disease. 2019;9(1):173-182. epub2018. PMID: 30562907.

Santos-Garcia D, Catalan M.J, Puente V, Valldeoriola F, Regidor I, Mir P. et al. Continuous intestinal infusion of levodopa-carbidopa in patients with advanced Parkinson’s disease in Spain: Subanalysis by autonomous community Uso de la infusión intestinal continua de levodo-

206 CIBERNED 2018 ANNUAL REPORT pa-carbidopa en pacientes con enfermedad de Parkinson avanzada en España. Subanálisis por comunidades autónomas. Neurologia. 2018.

Compta Y, Dias S, Pulido-Salgado M, Giraldo D, Perez-Soriano A, Fernandez M, Camara A, Bra- vo P, Munoz E, Saura J, Marti MJ. Cerebrospinal fluid levels of cytokines in multiple system atrophy: A cross-sectional study of the atalan msa registry (CMSAR). 33:S446-S447. 2018.

Marek K, Chowdhury S, Siderowf A, Lasch S, Coffey CS, Caspell-Garcia C, Simuni T, Jennings D, Tanner CM, Trojanowski JQ. The Parkinson’s progression markers initiative (PPMI)–establishing a PD biomarker cohort. Annals of clinical and translational neurology. 2018;5(12):1460- 77.

Borm C.D.J.M, Krismer F, Wenning G.K, Seppi K, Poewe W, Pellecchia M.T. et al. Axial motor clues to identify atypical parkinsonism: A multicentre European cohort study. Parkinsonism and Related Disorders. 2018;56:33-40. PMID: 29910157.

Hoogland J, van Wanrooij L.L, Boel J.A, Goldman J.G, Stebbins G.T, Dalrymple-Alford J.C. et al. Detecting Mild Cognitive Deficits in Parkinson’s Disease: Comparison of Neuropsychological Tests. Movement Disorders. 2018;33(11):1750-1759. PMID: 30216541.

Blauwendraat C, Reed X, Kia D.A, Gan-Or Z, Lesage S, Pihlstrom L. et al. Frequency of Loss of Function Variants in LRRK2 in Parkinson Disease. JAMA Neurology. 2018;75(11):1416-1422. PMID: 30039155.

Fernandez-Santiago R, Garrido A, Infante J, Gonzalez-Aramburu I, Sierra M, Fernandez M. et al. α-synuclein (SNCA) but not dynamin 3 (DNM3) influences age at onset of leucine-rich repeat kinase 2 (LRRK2) Parkinson’s disease in Spain. Movement Disorders. 2018;33(4):637- 641.

207 Code: RD16/0011/0016. Title: Combined protective/restorative cell-mediated strategies for neurodegenerative diseases. Principal investigator: Jose Luis Labandeira Garcia. CIBERNED’s collaboration: Yes. CIBERNED groups: G208 ; G102; G113; G105; G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 240245. Duration: 2017-2021.

Code: PI2016/06. Title: Identificacion de vias fisiopatologicas y biomarcadores candidatos en la fase pre-di- agnostica de la enfermedad de Parkinson. Principal investigator: Miquel Vila Bover. CIBERNED’s collaboration: Yes. CIBERNED groups: G109 ; G601; G207; G410. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 196000. Duration: 2016-2018.

Code: 2017 SGR/1502. Title: 2017 SGR/1502. Principal investigator: Maria jose Marti Domenech. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 35200. Duration: 2017-2019.

Code: PI17/00096. Title: Amplification of alpha-synuclein & 4R-tau auto-aggregation by RTQuIC in brain tissue and CSF as a differential biomarker of degenerative parkinsonisms. Principal investigator: Maria Jose Marti/ Yaroslau Compta. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 90901,25. Duration: 2018-2020.

Code: 288/C/2014. Title: Catalan Network of Multiple System Atrophy: Biomarkers and Pathophysiology’. Principal investigator: Maria Jose Marti Domenech. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 183316. Duration: 2014-2018.

Code: EudraCT 2015 003679-31. Title: FAIR-PARK-II A multicentre, parallel group, randomized, placebo-controlled trial of

208 CIBERNED 2018 ANNUAL REPORT deferiprone (DFP) 15 mg/kg BID. Principal investigator: Yaroslau Compta Hirnjy. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 282452. Duration: 2016-2021.

Code: MJFF _DYSK_2014. Title: Gene-gene interaction analysis of the mTOR pathway in L-DOPA induced dyskinesia’. Principal investigator: Maria Jose Marti Domènech, Cristina Malagelada. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Michael J. Fox Foundation. Funding: 125000. Duration: 2015-2018.

Code: ID11849. Title: LRRK2 peripheral blood mononuclear cells and urine biosample collection’. Principal investigator: Maria Jose Marti Domenech, Eduardo Tolosa. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Michael J Fox Foundation. Funding: 138506. Duration: 2016-2018.

Code: FUNDTATIANA16_PROY03. Title: MicroRNAs as biomarker of conversion of REM behaviour disorder to Parkinson’s disease. Principal investigator: Mario Ezquerra Trabalon. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacion Tatiana Perez de Guzman el Bueno. Funding: 77000. Duration: 2018-2018.

Code: 0002018. Title: MicroRNAs AS BIOMARKERS OF THE PRODROMAL STAGE OF PARKINSON DISEASE(EX- PANSSION OF microRNA FINDINGS TO ASHKENAZI JEWS PD COHORTS). Principal investigator: Mario Ezquerra Trabalon; Ruben Fernandez Santiago. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Michael J Fox Foundation. Funding: 185803,06. Duration: 2018-2019.

Code: TAPP - Telematic Applications in Medicine. Title: Prediction of cognitive properties of new drug candidates for neurodegenerative diseases in early clinical development. Acrònim: PHARMACOG. Principal investigator: Carme Junque.

209 CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: Comision Europea. Funding: 660250. Duration: 2018-2018.

Code: SAF2015-73508-JIN. Title: The beginnings of Parkinson Disease: a study of early disease mechanisms in a humanized in-vitro model. Principal investigator: Ruben Fernandez-Santiago. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 170000. Duration: 2016-2018.

Code: 00000. Title: The Parkinson’s Progression Markers Initiative (PPMI). Principal investigator: Eduardo Tolosa. CIBERNED’s collaboration: No. CIBERNED groups: G207. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Michael J Fox Foundation. Funding: 1830210. Duration: 2013-2018.

210 CIBERNED 2018 ANNUAL REPORT

108 | Carlos Vicario Abejón

CSIC

Avda. Doctor Arce, 37 28002 Madrid Tél.: +34 915 854 721 Fax: +34 915 854 754 E-mail: [email protected]

PRINCIPAL Crespo Galán, Nieto Estévez, Vanesa. INVESTIGATOR Inmaculada. PhD. PhD. Oueslati Morales, Carlos Vicario Abejón, Carlos Defterali, Cagla. Bachelor Omar. degree. Bachelor degree. LIST OF Diaz Guerra, Eva. Román Alonso, María PERSONNEL PhD. José. Bachelor degree. Arribas González, Fernández Rodríguez, Esther. Pablo. PhD. Bachelor degree.

ABSTRACT 1. My research group has deposited in the Spanish National Bank of Stem Cell Lines (BNLC) of the ISCIII, 9 lines of induced pluripotent stem cells (iPS cells or iPSCs) derived from Par- kinson’s disease patients (4 lines), from Alzheimer’s disease patients (2 lines) and from healthy subjects (3 lines). The generation (by reprogramming human fibroblasts) and characterization of all the iPSC lines has required an extraordinary effort, as each deposit document consists of 7-8 figures with results. I would like to emphasize that these results have been evaluated by the Technical Committee of the BNLC, which has considered that our iPS cells comply with all the quality parameters required nationally and internationally. The documents justifying the generation and characterization of the 9 lines of iPSCs have been published on the WEB page http://www.isciii.es/ISCIII/es/contenidos/fd-el-instituto/fd-organizacion/fd-estructura-directiva/fd-subdireccion-general-investigacion-terapia-celular-medicina-regenerativa/fd-centros-unidades/fd-banco-nacional-lineas-celulares/fd-lineas-celulares-disponi- bles/lineas-de-celulas-iPS.shtml We are currently writing the corresponding articles (manuscripts in preparation) (E. Ro- dríguez-Traver, ..., C. Vicario; E. Díaz-Guerra, ..., C. Vicario). 2. We have made significant progress in the study of the effects of N370S and L444P

211 mutations in the GBA1 gene on dopaminergic neurons derived from iPS cells of Parkin- son’s patients. The results obtained with the participation of Dr. Rosario Moratalla´s research group (Cajal Institute-CSIC and CIBERNED), have been presented (as both oral communication and poster) in the VI International Congress of Research and Innovation in Neurodegenerative Diseases CIIIEN (E. Rodríguez-Traver, ..., C. Vicario, 2018) and we are preparing the corresponding article. In addition and in collaboration with Dr. Moratalla, we have published a commentary-article in the journal Autophagy (P. García-Sanz, ..., C. Vicario, R. Moratalla, 2018). 3. We have also made great progress in studying the effects of familial Alzheimer’s and sporadic Alzheimer’s on astrocytes and neurons obtained by differentiating our iPSCs. Specifically, we are studying the phenotypes caused by the APOE gene alleles and by the G206D mutation in the PRESEN1 gene, in collaboration with Dr. Moratalla. Some of the new results have been presented (as a poster) in the VI International Congress of Re- search and Innovation in Neurodegenerative Diseases CIIIEN (E. Díaz-Guerra, ..., C. Vicario, 2018) and we are preparing the corresponding article. A Master Project (TFM) has been successfully defended. We participate in the European project ADAPTED, funded in the IMI call, in which we collaborate with Dr. Agustín Ruiz (Fundació ACE), among other researchers. The generation of isogenic iPSCs has been presented at the IMI 10th Anniversary Scientific Symposium held in Brussels under the title “An iPSC technology development collaboration - a critical component to enable ADAPTED” (P. Reinhardt and B. Schmid, ..., C. Vicario, ..., A. Ruiz, 2018). 4. We have completed the characterization of local neural stem / progenitor cells of the adult mouse olfactory bulb and demonstrated its neurogenic potential in vivo (Defterali et al., manuscript in preparation). 5. We have completed a study on the effects of the neurotrophins BDNF and NT-3 on the differentiation and maturation of olfactory bulb interneurons (Nieto-Estévez et al., manuscript in preparation). 6. We are studying the role of IGF-I factor (specifically, that synthesized in the brain) in the induction of hippocampal Long-Term Potentiation and in the behavior of a conditional IGF-I knockout mouse, in collaboration with Drs. Gertrudis Perea (Cajal Institute-CSIC) and Ignacio Torres Alemán (Cajal Institute-CSIC and CIBERNED). In order to find possible molecular targets of IGF-1, we are conducting proteomic analyzes in the olfactory bulb and hippocampus, in collaboration with Dr. Enrique Santamaría (Navarrabiomed, Pamplona)

KEYWORDS iPS cells, neural stem cells, Parkinson´s disease, Alzheimer´s disease, adult neurogenesis, olfactory bulb, hippocampus, IGF-1.

PUBLICATIONS 2018 García-Sanz P, Orgaz L, Fuentes JM, Vicario C, Moratalla R. Cholesterol and multilamellar bodies: Lyosomal dysfunction in GBA-Parkinson disease.Autophagy. 2018. PMID: 29368986.

212 CIBERNED 2018 ANNUAL REPORT

Code: 115975-2. Title: Alzheimer´s Disease Apolipoprotein Pathology for Treatment Elucidation and De- velopment. Principal investigator: Carlos Vicario Abejon. CIBERNED’s collaboration: No. CIBERNED groups: G108 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 307010,12. Duration: 2016-2019.

Code: SAF2016-80419-R. Title: Regulacion de la multipotencialidad de celulas madre neurales humanas y la neurogenesis en modelos de Alzheimer esporadico y Alzheimer familiar. Principal investigator: Carlos Vicario Abejon. CIBERNED’s collaboration: No. CIBERNED groups: G108 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 157300. Duration: 2016-2019.

PHD DISSERTATIONS 2018 Author: Manuel Antonio Oria Muriel. Title: Caracterización de una línea de iPSCs humanas con una mutación en Presenilina1 como modelo celular para el estudio de la enfermedad de Alzheimer familiar. Date: 24/9/2018. Supervisor: Carlos Vicario Abejón.

213 109 | Miquel Vila Bover

NEURODEGENERATIVE DISEASES RESEARCH GROUP VALL D’HEBRON RESEARCH INSTITUTE

Mediterranean Building, First Floor, Lab. 102 Pg. Vall d’Hebron, 119-129 08035 Barcelona Tel. +34 93 489 45 43 Fax +34 93 489 40 15 E-mail: [email protected] www.vilalab.org

PRINCIPAL De Fabregues-Boixar Parent, Annabelle INVESTIGATOR Nebot, Oriol. Suzanne Chantal. PhD. Technician. Vila Bover, Miquel Franco Iborra, Sandra. Peñuelas Peñarroya, Bachelor degree. Nuria. LIST OF Galiano Landeira, Jordi. Bachelor degree. PERSONNEL Bachelor degree. Perier, Celine. González Sepúlveda, PhD. Bove Badell, Jordi. PhD. Marta. Rodríguez Galván, Carballo Carbajal, Iria. PhD. Beatriz. PhD. Laguna Tuset, Ariadna. Technician. Cuadros Arasa, Thais. PhD. Romero Giménez, Jordi. PhD. Martinez Vicente, Bachelor degree. Marta. Torra i Talavera, Albert. PhD. Bachelor degree.

ABSTRACT In 2018 we have developed a new therapeutic strategy to selectively decrease the brain levels of α-synuclein in vivo selectively within monoaminergic neurons, the type of neurons most susceptible to neurodegeneration in Parkinson’s disease (PD). This technique is based on the intranasal administration of chemically-modified oligonucleotides. In addition, we have shown that PD-derived peripheral α-synuclein aggregates are not pathogenic when injected into the brains of experimental animals. We have also described a neuroprotective role of transcription factor EB (TFEB) in a neurotoxic model of PD and demonstrated that the beneficial effect of TFEB goes beyond the activation of autophagy and implies a neurotrophic effect on dopaminergic neurons.

214 CIBERNED 2018 ANNUAL REPORT

KEYWORDS Parkinson’s disease, α-synuclein, TFEB

PUBLICATIONS 2018 Perez-Villalba A, Sirerol-Piquer M.S, Belenguer G, Soriano-Canton R, Munoz-Manchado A.B, Vil- ladiego J. et al. Synaptic regulator α-synuclein in dopaminergic fibers is essentially required for the maintenance of subependymal neural stem cells. Journal of Neuroscience. 2018;38(4):814- 825. PMID: 29217686. Franco-Iborra S, Cuadros T, Parent A, Romero-Gimenez J, Vila M, Perier C. Defective mitochondrial protein import contributes to complex I-induced mitochondrial dysfunction and neurodegeneration in Parkinson’s disease. Cell Death and Disease. 2018;9(11). PMID: 30405116. Recasens A, Carballo-Carbajal I, Parent A, Bove J, Gelpi E, Tolosa E. et al. Lack of pathogenic potential of peripheral α-synuclein aggregates from Parkinson’s disease patients. Acta Neuropatholog- ica Communications. 2018;25(1). PMID: 29422109. Franco-Iborra S, Vila M, Perier C. Mitochondrial quality control in neurodegenerative diseases: Fo- cus on Parkinson’s disease and Huntington’s disease. Frontiers in Neuroscience. 2018;12(MAY). PMID: 29875626. Torra A, Parent A, Cuadros T, Rodriguez-Galvan B, Ruiz-Bronchal E, Ballabio A. et al. Overexpres- sion of TFEB Drives a Pleiotropic Neurotrophic Effect and Prevents Parkinson’s Disease-Related Neurodegeneration. Molecular Therapy. 2018;26(6):1552-1567. PMID: 29628303. De Fabregues O, Ferré A, Romero O, Quintana M, Álvarez-Sabin J. Sleep Quality and Levodopa Intestinal Gel Infusion in Parkinson’s Disease: A Pilot Study.Parkinson’s disease. 2018. PMID: 30515291. Lassot I, Mora S, Lesage S, Zieba B.A, Coque E, Condroyer C. et al. The E3 Ubiquitin Ligases TRIM17 and TRIM41 Modulate α-Synuclein Expression by Regulating ZSCAN21. Cell Reports. 2018;25(9):2484-2496.e9. PMID: 30485814. Compta Y, Giraldo D.M, Munoz E, Antonelli F, Fernandez M, Bravo P. et al. Cerebrospinal fluid levels of coenzyme Q10 are reduced in multiple system atrophy. Parkinsonism and Related Disorders. 2018;46:16-23. PMID: 29107645. Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Garcia-Aznar J.M. et al. Er- ratum to: Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons (Molecular Neurobiology, (2018), 55, 3, (1847-1860), 10.1007/s12035-017-0451-4). Molecular Neurobi- ology. 2018;55(3):1861-. Del Rey N.L.-G, Quiroga-Varela A, Garbayo E, Carballo-Carbajal I, Fernandez-Santiago R, Mon- je M.H.G. et al. Advances in parkinson’s disease: 200 years later. Frontiers in Neuroanatomy. 2018;12. PMID: 30618654. Kim H, Calatayud C, Guha S, Fernandez-Carasa I, Berkowitz L, Carballo-Carbajal I. et al. Correction to: The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neuron Function and Survival Against α-Synuclein-Induced Toxicity (Molecular Neurobiology, (2018), 55, 9, (7533- 7552), 10.1007/s12035-018-0881-7). Molecular Neurobiology. 2018;55(9):7553-7554. Kim H, Calatayud C, Guha S, Fernández-Carasa I, Berkowitz L, Carballo-Carbajal I et al. The Small GTPase RAC1/CED-10 Is Essential in Maintaining Dopaminergic Neuron Function and Survival Against sy -Synuclein-Induced Toxicity.Molecular neurobiology. 2018. PMID: 29429047.

215 RESEARCH PROJECTS 2018 Code: PI2016/06. Title: Identificacion de vias fisiopatologicas y biomarcadores candidatos en la fase pre-di- agnostica de la enfermedad de Parkinson. Principal investigator: Miquel Vila Bover. CIBERNED’s collaboration: Yes. CIBERNED groups: G109 ; G601; G207; G410. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 196000. Duration: 2016-2018.

Code: SAF2016-77541-R. Title: Diagnostic and therapeutic potential of neuromelanin in Parkinson’s disease using novel humanized preclinical in vivo model. Principal investigator: Miquel Vila Bover. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 363000. Duration: 2017-2020.

Code: COEN4016. Title: Focused ultrasound modulation of neuromelanin accumulation in a humanized rat model of Parkinson’s disease. Principal investigator: Miquel Vila Bover. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 456488. Duration: 2018-2019.

Code: 2017 SGR 1806. Title: Grup de Recerca Consolidat-Malalties Neurodegeneratives. Principal investigator: Miquel Vila Bover. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Generalitat de Catalunya. Funding: 35200. Duration: 2017-2019.

Code: PI17/00496. Title: La disfuncion autofagica/lisosomal en la enfermedad de Parkinson: caracterizacion y desarrollo de nuevas estrategias traslacionales con el lisosoma como diana terapeutica. Principal investigator: Marta Martinez Vicente. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 99219,99. Duration: 2018-2020.

216 CIBERNED 2018 ANNUAL REPORT

Code: Fundacio Marato de TV3-Malalties del cor (320/U/2015). Title: New molecular functions of apoptotic genes in cardiac development and stress. Principal investigator: Daniel Sanchis Morales. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 191656. Duration: 2015-2018.

Code: SAF2015-73997-JIN. Title: The Gut-Brain axis in Prodromal Parkinson’s disease: new possibilities for risk/diagnostic biomarkers and therapeutic interventions. Principal investigator: Ariadna Laguna Tuset. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 170000. Duration: 2016-2018.

Code: GCASE_MJFF-SILVERSTEIN2018. Title: Therapeutic target of GCase enzyme in Parkinson’s disease with novel pharmacological chaperones. Principal investigator: Marta Martinez Vicente. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Michael J Fox Foundation. Funding: 498698. Duration: 2018-2020.

Code: PMP15/00025. Title: A precise approach for nucleoside-based therapy of neuromuscular disorders with defects in mitochondrial dna. Principal investigator: Miguel Angel Martin Casanueva. CIBERNED’s collaboration: No. CIBERNED groups: G109 . Other CIBER’s collaboration: CIBERER. Type: Nacional. Funding agency: MICINN. Funding: 806223. Duration: 2016-2019.

PHD DISSERTATIONS 2018 Author: Sandra Franco Iborra. Title: Mitochondrial quality control in neurodegenerative diseases: focus on Parkinson’s disease and Huntington’s disease. Date: 19/4/2018. Supervisor: Miquel Vila Bover.

217 218 PROGRAM 3 AMYOTROPHIC LATERAL SCLEROSIS AND OTHER NEUROMUSCULAR DISORDERS

CIBERNED 2018 ANNUAL REPORT

Neuromuscular diseases (NMDs) form a heterogeneous group of pathologies affecting the spinal cord and its tracts, nerve roots as well as motor and sensitive peripheral nerves, the neuromuscular junction and the muscles. Diagnosis involves using sophisti- cated methods covering: neurophysiological studies, muscle or nerve biopsy with the use of immunohistochemical techniques and some others, metabolic analysis and tests, MRI studies, quantitative muscle assessment, and in many cases, genetic studies. Just a few epidemiological studies are available in Spain, from which it can be the prevalence of chronic inferred that the prevalence of chronic neurodegenerative and/ neurodegenerative and/or hereditary pathology would account for around 60,000 or hereditary pathology patients throughout the whole country. would account for The main research topics in this line focus around 60,000 patients on the following aspects: throughout the whole • Clinical characterization of neuro- country. muscular pathologies and correlation between the clinical phenotype and their genotype (or proteomic characterization). • Development of animal models based on the genotypically identified dystrophies. • Research on the physiopathology of neuromuscular diseases. • Development of new therapeutic strategies.

PRINCIPAL INVESTIGATOR INSTITUTION

Fernández Chacón, Rafael University of Seville

Infante Ceberio, Jon Marqués de Valdecilla Research Institute, Santander

López de Munain Arregui, Adolfo Biodonostia Research Institute, San Sebastian

Muñoz Cánoves, Pura Pompeu Fabra University , ICREA, Barcelona

Navarro Acebes, Xavier Autonomous University of Barcelona

Program 3 is coordinated by Drs. Rafael Fernández Chacón (University of Seville) and Adolfo López de Munain Arregui (Biodonostia Research Institute, San Sebastián).

221 606 | Rafael Fernández-Chacón

INSTITUTO DE BIOMEDICINA DE SEVILLA (IBIS)

Hosp.Univ. Virgen del Rocío/CSIC/ Universidad de Sevilla Dep. de Fisiología Médica y Biofísica Avda. Manuel Siurot s/n 41013 Sevilla Teléfono:+34-955923047 (office)/955923045 (laboratory) Fax:+34-955923101 E-mail: [email protected] web: www.ibis-sevilla.es

PRINCIPAL Cabrera Romero, Reina Castillo, María INVESTIGATOR Casandra. Isabel. Technician. Bachelor degree. Fernández Chacón, García-Junco Clemente, Rivero Mena, María del Rafael Pablo Luis. Carmen. PhD. Technician. LIST OF Muñoz Bravo, José Luis. Rubio Pastor, Fátima. PERSONNEL Bachelor degree. Bachelor degree. Nieto González, José Valladares Millan, Arroyo Saborido, Luis. Ismael. Alejandro. PhD. Bachelor degree. Technician.

ABSTRACT Our research project is focused on the investigation of the molecular mechanisms involved in the maintenance of the structure and function of synaptic terminals which integrity can be compromised in neurodegenerative diseases. Most of our studies are based on the generation and/or phenotypic analysis of genetically modified mice with modifications in genes that encode for co-chaperones (CSPα, CSPγ), proteins of the synaptic vesicle cycle or express proteins with pathogenic mutations in humans such as mutations L115R and L116Del in Dnajc5 (that encodes for CSPa) which cause adult neuronal ceroidal lipofuscinosis or the G2019S mutation in LRRK2 associated with Parkinson’s disease. Our molecular, structural and functional studies are normally carried out in primary cultures of different cell types and slices of mouse brain. An important part of our recent investigations are based on a new line of mice in which Dnajc5 is floxed. This novel mouse line is allowing us to study the function of CSPα in specific neuronal and cell types by using different mouse lines that express cre-recombinase protein under specific promoters. One of these lines (Sox2CreERT2) has allowed us to show in vivo that the absence of CSPα in neural stem cells of the hippocampus interferes with the quiescence of these cells, increases the percentage of proliferating cells and compromises the size of the stem cell pool (submitted to Proc Natl Acad Sci USA).

222 CIBERNED 2018 ANNUAL REPORT

This line of research is being reinforced by our collaboration with Dr. Isabel Fariñas. Our observations in neurogenesis have led us to investigate the molecular mechanisms of a previously unsuspected relationship between CSPα and the mTOR pathway (mechanistic target of rapamycin), a key route in the regulation of cell growth and nutrient-dependent metabolism and growth factors and involved in autophagy and cell proliferation. Within our CIBERNED project, we have also considered the use of new viral vectors that cross the blood-brain barrier to investigate the possibilities of therapeutic intervention to rescue neurodegenerative phenotypes associated with the absence of CSPa. Part of this project has been selected by the Tatiana Perez de Guzman el Bueno Foundation in a proposal that also includes the use of two-photon microscopy (2P) to monitor functional changes in neural circuits in the context of the dysfunction / synaptic degeneration and recovery after rescue in lines in which CSPα has been eliminated in specific neurons. On the other hand, we have also designed new constructions based on fluorescent reporters to monitor brain synaptic activity with 2P in vivo. Importantly, this year our collaboration with the group of Dr. Lucas Lozano has advanced very well leading to the publication of findings of great interest related to the molecular basis of autistic spectrum disorder (ASD) (Parras et al., Na- ture, 560:441-446 (2018)).

KEYWORDS Synapse, synaptic vesicle cycle, hippocampus, neurogenesis, neuronal ceroid lipofuscinosis, single-cell sequencing, transcriptomic, skeletal muscle.

PUBLICATIONS 2018 Parras A, Anta H, Santos-Galindo M, Swarup V, Elorza A, Nieto-Gonzalez J.L. et al. Autism-like phenotype and risk gene mRNA deadenylation by CPEB4 mis-splicing. Nature. 2018;560(7719):441- 446. PMID: 30111840. Valenzuela-Villatoro M, Garcia-Junco-Clemente P, Nieto-Gonzalez J.L, Fernandez-Chacon R. Presyn- aptic neurodegeneration: CSP-α/DNAJC5 at the synaptic vesicle cycle and beyond. Current Opinion in Physiology. 2018;4:65-69. Ortega-de San Luis C, Sanchez-Garcia M.A, Nieto-Gonzalez J.L, Garcia-Junco-Clemente P, Monte- ro-Sanchez A, Fernandez-Chacon R. et al. Substantia nigra dopaminergic neurons and striatal interneurons are engaged in three parallel but interdependent postnatal neurotrophic circuits. Aging Cell. 2018;17(5). PMID: 30058223. Servian-Morilla E, Robles-Lanuza E, Sanchez-Hidalgo A.C, Camacho-Garcia R.J, Paez-Gomez J.A, Ma- villard F. et al. Proteolytic processing of neurexins by presenilins sustains synaptic vesicle release. Journal of Neuroscience. 2018;38(4):901-917. PMID: 29229705.

223 RESEARCH PROJECTS 2018 Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: Yes. CIBERNED groups: G604 ; G102; G606; G111; G306. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350000. Duration: 2016-2018.

Code: BFU2016-76050-P. Title: Mecanismos Moleculares del Mantenimiento a Largo Plazo de las Sinapsis Glutama- tergicas in Vivo. Principal investigator: Rafael Fernandez-Chacon. CIBERNED’s collaboration: No. CIBERNED groups: G606. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 280000. Duration: 2016-2019.

PHD DISSERTATIONS 2018 Author: José Antonio Martínez López. Title: Presynaptic calcium dynamics, neuronal excitability and synaptic vesicle cycle in central synapses lacking Cysteine String Protein-alpha (CSP-alpha). Date: 22/9/2018. Supervisor: Rafael Fernández Chacón.

224 CIBERNED 2018 ANNUAL REPORT

601 | Jon Infante Ceberio

SERVICIO DE NEUROLOGÍA HOSPITAL UNIVERSITARIO MARQUÉS DE VALDECILLA UNIVERSIDAD DE CANTABRIA

39008 Santander E-mail: [email protected]

PRINCIPAL De La Fuente García, Pelayo Negro, Ana Lara. INVESTIGATOR Marta Gema. PhD. Auxiliar Administrativo. Pozueta Cantudo, Ana. Infante Ceberio, Jon Gallardo Agromayor, Bachelor degree. Elena. Riancho Zarrabeitia, LIST OF PhD. Javier. PERSONNEL García García, Antonio. Bachelor degree. PhD. Berciano Blanco, José Rodríguez Rodríguez, Ángel. Gonzalez Aramburu, Eloy. PhD. Isabel. PhD. Bachelor degree. Berciano Blanco, María Sánchez Juan, Pascual. Teresa. González Mandly, PhD. PhD. Andrés. Sánchez Quintana, Bachelor degree. Bravo González, María. María del Coro. Diplomado. González suarez, Technician. Andrea. Casafont Parra, Iñigo. Sedano Tous, M ª Jóse. Bachelor degree. PhD. PhD. Kazimierczak, Martha. Castilla Castrillon, Sierra Peña, María. Diplomado. Joaquin. PhD. Bachelor degree. Lafarga Coscojuela, Tapia Martínez, Olga. Miguel Ángel. Bachelor Castillo Iglesias, María PhD. degree. Soledad. Vázquez Higuera, José Bachelor degree. Lage Martínez, Carmen. Luis. Bachelor degree. PhD. Narcis Majos, Oriol. Bachelor degree.

225 ABSTRACT Peripheral Neuropathies Our investigation has continued been focused on Charcot-Marie-Tooth disease (CMT) and Guillain-Barré syndrome (GBS), which will be analyzed separately. In a collaborative paper, we have defined the phenotype associated with homozygous MME mutations, which is characteristically included in AR-CMT2 of adult onset. Our contributions to intermediate CMT have resulted in a significant revision of the syndrome in the OMIM catalogue (cf. Cassandra L. Kniffin: 02/20/2018), and in the phenotypic re-definition of CMT associated to heterozygous GNB4 mutations causing not an intermediate but an axonal syndrome. We have reported the potential pathogenic role of endoneurial inflammatory oedema in CMT1A. We have established that the physiological background of normorreflexia in CAN- VAS (Cerebellar ataxia, neuropathy, vestibular areflexia syndrome) is preservation of fibres Ia in the hallmark of a severe somatic sensitive deficit. Likewise, selective involvement of Ia fibres may be of help for detecting reversible conduction failure in early GBS. We have updated the pathophysiology of axonal damage and the role of proximal nerve trunk lesions in early GBS; this allowed us to revisit the great legacy by Haymaker and Kernohan (Medicine 1949; 28: 50-141). We have proposed a new mechanism of nerve trunk pain that might be an inaugural manifestation in any GBS subtype. Our interpretation of initial semeiology of Franklin Delano Roosevelt’s acute paralytic disease, August 1921, has been helpful in establishing that the future US President suffered from not paralytic polio but GBS. We have proposed that in acute flaccid myelitis (AFM) there may be distal motor axonopathy, a fact suggesting the pathogenic relationship between AFM and GBS. We have continued collaborating in the IGOS study (International Guillain-Barré Outcome Study). Parkinson’s disease We have replicated in a multicentric study, together with the Hospital Clinic in Bar- celona, the association of a SNP in SNCA gene with the penetrance of the LRRK2 G2019S mutation in Spanish population. This polymorphism has an influence in the age of onset of this form of genetic Parkinson´s disease (PD), unlike the SNP rs2421947 of DMN3 gene, as it had been reported previously by other authors. In collaboration with the International Parkinson’s Disease Genomic Consortium an exome sequencing of 2835 PD patients, 5343 controls and 111 pathologically confirmed cases of Lewy body dementia (LBD) has been carried out. This study doesn´t support a pathogenic role of genetic variants in LRP10 in PD or LBD. We are undertaking an ambitious project searching for multimodal biomarkers in a large cohort of carriers of the LRRK2 G2019S mutation to deepen knowledge in prodromal Parkinson´s disease. Ataxias The results of survival at 10 years from the EUROSCA longitudinal cohort, a multicentric international study that includes 525 patients with the most frequent forms of dominant ataxias (SCA1, 2, 3 and 6) have been recently published, yielding the following figures: 57%, 74%, 72% y 87%, respectively. Within the same study, after a follow-up of 8 years, patients´ reported outcome measures have been evaluated, showing that they provide relevant complementary information to that obtained from neurological scales. The results from both studies have a direct application for the design of coming clinical trials. Alzheimer’s disease We have started the project “Valdecilla Cohort for Memory and Brain Aging” whose purpose is to set up a cohort with a large sample of volunteers cognitively healthy volunteers.

226 CIBERNED 2018 ANNUAL REPORT

Neuropsychology: During 2017 we carried on the neuropsychological description of one of the largest series of right semantic dementia patient. Three manuscripts have been written based on these data, as part of Ana Pozueta’s doctoral thesis. Genetics: Within the multicenter project PI16/01652. “Study of rare variants in genes associated with Alzheimer’s disease in the Spanish population” we have gathered a population of 772 DNA samples from Spanish patients with early-onset Alzheimer’s, which is one of the main samples studied to date. The study, funded by the Instituto de Salud Carlos III and whose PI and coordinator is Pascual Sánchez-Juan, is the first large project of DEGESCO (DEmentia GEnetics Spanish COnsortium) for a large-scale genomic study of dementias in the Spanish population. Within the international study of Alzheimer’s genomics IGAP, it is worth mentioning our participation in its la- test publication in Nature Genetics: “Genetic meta-analysis of diagnosed Alzheimer’s disease identifies new risk loci and implicates Aβ, tau, immunity and lipid processing. Biomarkers: In 2018, a manuscript was published in the Journal of Alzheimer’s Di- sease in which we evaluated the extensive experience of the Cognitive Impairment Unit in the clinical use of amyloid PET. Also during this year it is worth mentioning an editorial in the journal N Engl J Med in connection with our project “MicroRNA Profile in Patients with Alzheimer’s Disease” funded by CIBERNED, for the study of the diagnostic value of the determination of these structures in CSF both free and exosomes and whose results were published in 2017 in the Journal of Alzheimer’s Disease. Amyotrophic lateral sclerosis (ALS) Regarding ALS epidemiology, we have published two reviews discussing the role of environmental factors (Riancho et al, Int J Biomet 2018) and infectious agents (Cas- tanedo-Vazquez et al, J Neurol 2018) in the disease. We have continued with our scientific collaborations: i) Professor Ana Maria Cuervo (Albert Einstein College of Me- dicine, New York) focused on the role of autophagy in ALS; ii) Biopharma-University of Santiago de Compostela devoted to the identification of new Retinoid X Receptor agonists as a new potential therapy for ALS (Riancho et al, JAMA Neurol 2018) and iii) Professor Adolfo López de Munain (Biodonostia Research Institute-CIBERNED, San Sebastian) focused in the investigation of new biomarkers for ALS. In this line we have identified several lipidomic disturbances in ALS patients (Fernandez de Eulate G et al, Neurology, submitted). Our group has recently collaborated in the creation of the first Spanish ALS consortium (ALSGESCO) Cell neurobiology Our work has focused in two research topics: i) dysfunction of RNA metabolism in motor neurons (MNs) of the murine SMNΔ7 model of Type I spinal muscular atrophy (SMA), and ii) DNA damage processing in neurons and its relevance in neurodegeneration. Regarding the first point, survival motor neuron (SMN) deficiency in MNs from the SMA mice interferes with mRNA processing, particualrly pre-mRNA splicing. As a result, intron-containing and incorrectly processed polyadenylated mRNAs are retained in poly(A) RNA granules that also concentrate the splicing regulator Sam68. As a consequence, MNs undergo an important déficit in mRNA processing and translation that severely impacts on SMA pathogenesis. Concerning the second point, we have demonstrated in rat cortical neurons exposed to 4Gy of ionizing radiation that DNA damage processing takes place in a specific chromatin compartment and identified DNA sequences more vulnerable to DNA damage or refractory to DNA repair. Interestingly, some of these sequences are specifically involved in human neurodegenerative disorders.

227 KEYWORDS CMT, Guillain Barré syndrome, LRRK2, ataxia, Spinal muscular atrophy, Alzheimer´s disease, Parkinson´s disease, ALS, Cajal bodies, nucleolus.

PUBLICATIONS 2018 Kia DA, Sabir MS, Ahmed S, Trinh J, Bandres-Ciga S, International Parkinson’s Disease Genomics Consortium. et al. LRP10 in α-synucleinopathies.The Lancet. Neurology. 2018;17(12). PMID: 30507383. Castanedo-Vazquez D, Bosque-Varela P, Sainz-Pelayo A, Riancho J. Infectious agents and amyotrophic lateral sclerosis: another piece of the puzzle of motor neuron degeneration. Journal of Neurology. 2019;266(1):27-36. epub2018. PMID: 29845377. Cantero JL, Atienza M, Sanchez-Juan P, Rodriguez-Rodriguez E, Vazquez-Higuera JL, Pozueta A et al. Cerebral changes and disrupted gray matter cortical networks in asymptomatic older adults at risk for Alzheimer’s disease.Neurobiology of aging. 2018;64. PMID: 29331877. Garcia-Cerro S, Vidal V, Lantigua S, Berciano M.T, Lafarga M, Ramos-Cabrer P. et al. Cerebellar alterations in a model of Down syndrome: The role of the Dyrk1A gene. Neurobiology of Disease. 2018;110:206-217. PMID: 29221819. Borm C.D.J.M, Krismer F, Wenning G.K, Seppi K, Poewe W, Pellecchia M.T. et al. Axial motor clues to identify atypical parkinsonism: A multicentre European cohort study. Parkinsonism and Related Disorders. 2018;56:33-40. PMID: 29910157. Bergeron D, Gorno-Tempini M.L, Rabinovici G.D, Santos-Santos M.A, Seeley W, Miller B.L. et al. Prevalence of amyloid-β pathology in distinct variants of primary progressive aphasia. Annals of Neurology. 2018;84(5):729-740. PMID: 30255971. Mattsson N, Groot C, Jansen W.J, Landau S.M, Villemagne V.L, Engelborghs S. et al. Prevalence of the apolipoprotein E ε4 allele in amyloid β positive subjects across the spectrum of Alzheimer’s disease. Alzheimer’s and Dementia. 2018;14(7):913-924. PMID: 29601787. Fernandez-Santiago R, Garrido A, Infante J, Gonzalez-Aramburu I, Sierra M, Fernandez M. et al. α-synuclein (SNCA) but not dynamin 3 (DNM3) influences age at onset of leucine-rich repeat kinase 2 (LRRK2) Parkinson’s disease in Spain. Movement Disorders. 2018;33(4):637-641. Narcis J.O, Tapia O, Tarabal O, Piedrafita L, Caldero J, Berciano M.T. et al. Accumulation of poly(A) RNA in nuclear granules enriched in Sam68 in motor neurons from the SMNΔ7 mouse model of SMA. Scientific Reports. 2018;8(1). PMID: 29941967. Natera-de Benito D, Berciano J, Garcia A, M de Lucas E, Ortez C, Nascimento A. Acute Flaccid Myelitis With Early, Severe Compound Muscle Action Potential Amplitude Reduction: A 3-Year Follow-up of a Child Patient. Journal of clinical neuromuscular disease. 2018;20(2):100-101. PMID: 30439757. Berciano J. Additional arguments supporting that Franklin Delano Roosevelt’s paralytic illness was related to Guillain-Barré syndrome. Journal of Medical Biography. 2018;26(2):142-143. PMID: 29461141. Berciano J. Axonal pathology in early stages of Guillain-Barré syndrome.Neurologia (Barcelona, Spain). 2018. PMID: 30057217. Suner-Soler R, Grau-Martin A, Terceno M, Silva Y, Davalos A, Sanchez J.M. et al. Biological and psy- chological factors associated with smoking abstinence six years post-stroke. Nicotine and Tobacco

228 CIBERNED 2018 ANNUAL REPORT

Research. 2018;20(10):1182-1188. PMID: 29106659. Infante J, Garcia A, Serrano-Cardenas K.M, Gonzalez-Aguado R, Gazulla J, de Lucas E.M. et al. Cer- ebellar ataxia, neuropathy, vestibular areflexia syndrome (CANVAS) with chronic cough and pre- served muscle stretch reflexes: evidence for selective sparing of afferent Ia fibres. Journal of Neu- rology. 2018;265(6):1454-1462. PMID: 29696497. Lupo V, Frasquet M, Sanchez-Monteagudo A, Pelayo-Negro A.L, Garcia-Sobrino T, Sedano M.J. et al. Characterising the phenotype and mode of inheritance of patients with inherited peripheral neuropathies carrying MME mutations. Journal of Medical Genetics. 2018. PMID: 30415211. Riancho J, Sanchez-Juan P. Circulating Extracellular Vesicles in Human Disease. The New England journal of medicine. 2018;379(22):2180-. PMID: 30499643. Sconfienza LM, Albano D, Allen G, Bazzocchi A, Bignotti B, Chianca V et al. Clinical indications for musculoskeletal ultrasound updated in 2017 by European Society of Musculoskeletal Radiology (ESSR) consensus.European radiology. 2018. PMID: 29876703. Cortes-Donate V.E, Perez-Lorensu P.J, Garcia-Garcia A. Intraoperative neurophysiological monitoring in Spain: Its beginnings, current situation and future prospects Monitorización neurofisiológica intraoperatoria en España: Inicios, situación actual y perspectivas de futuro. Revista de Neurologia. 2018;66(9):315-320. PMID: 29696619. Jacobi H, du Montcel S.T, Bauer P, Giunti P, Cook A, Labrum R. et al. Long-term evolution of patient-reported outcome measures in spinocerebellar ataxias. Journal of Neurology. 2018;265(9):2040- 2051. PMID: 29959555. Gerace L, Tapia O. Messages from the voices within: regulation of signaling by proteins of the nuclear lamina. Current Opinion in Cell Biology. 2018;52:14-21. PMID: 29306725. Peran P, Barbagallo G, Nemmi F, Sierra M, Galitzky M, Traon A.P.-L. et al. MRI supervised and unsu- pervised classification of Parkinson’s disease and multiple system atrophy. Movement Disorders. 2018;33(4):600-608. PMID: 29473662. Berciano J. Neuropathic pain in early guillain-barré syndrome. Pain Physician. 2018;21(3):E279-E280. PMID: 29871382. Mata-Garrido J, Tapia O, Casafont I, Berciano MT, Cuadrado A, Lafarga M. Persistent accumulation of unrepaired DNA damage in rat cortical neurons: nuclear organization and ChIP-seq analysis of damaged DNA.Acta neuropathologica communications. 2018;6(1). PMID: 30049290. Garcia A, Sedano M.J, Alvarez-Paradelo S, Berciano J. Reversible conduction failure on the deep ten- don reflex response recording in early Guillain-Barré syndrome. Clinical Neurophysiology Practice. 2018;3:159-163. PMID: 30560219. Berciano J, Teran-Villagra N. Sneddon syndrome and non-bacterial thrombotic endocarditis: a clin- icopathological study. Journal of Neurology. 2018;265(9):2143-2145. PMID: 29980854. Diallo A, Jacobi H, Cook A, Labrum R, Durr A, Brice A. et al. Survival in patients with spinocerebellar ataxia types 1, 2, 3, and 6 (EUROSCA): a longitudinal cohort study. The Lancet Neurology. 2018;17(4):327-334. PMID: 29553382. Martinez-Dubarbie F, Herrera-Calvo G, Drake-Perez M, Riancho J. Unilateral isolated hypoglossal neuropathy associated to jugular paraganglioma. Neurological Sciences. 2018;39(5):971-972. PMID: 29353352. Lage C, Suarez A.G, Pozueta A, Riancho J, Kazimierczak M, Bravo M. et al. Utility of Amyloid and FDG-PET in Clinical Practice: Differences between Secondary and Tertiary Care Memory Units. Jour- nal of Alzheimer’s Disease. 2018;63(3):1025-1033. PMID: 29710706.

229 Riancho J, Gil-Bea F, Santurtun A, Lopez De Munain A. Amyotrophic lateral sclerosis: A complex syndrome that needs an integrated research approach. Neural Regeneration Research. 2019;14(2):193-196. epub2018. PMID: 30530996. Kishore A, Ashok Kumar Sreelatha A, Sturm M, von-Zweydorf F, Pihlstrom L, Raimondi F. et al. Understanding the role of genetic variability in LRRK2 in Indian population. Movement Disorders. 2018. PMID: 30485545. Peloso G.M, van der Lee S.J, Sims R, van der Lee S.J, Naj A.C, Bellenguez C. et al. Genetically elevated high-density lipoprotein cholesterol through the cholesteryl ester transfer protein gene does not associate with risk of Alzheimer’s disease. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2018;10:595-598. Riancho J, Bosque-Varela P, Perez-Pereda S, Povedano M, de Munain A.L, Santurtun A. The increasing importance of environmental conditions in amyotrophic lateral sclerosis. International Journal of Biometeorology. 2018;62(8):1361-1374. PMID: 29713861. Rueda N, Vidal V, Garcia-Cerro S, Narcis J.O, Llorens-Martin M, Corrales A. et al. Anti-IL17 treatment ameliorates Down syndrome phenotypes in mice. Brain, Behavior, and Immunity. 2018;73:235- 251. PMID: 29758264. Riancho J, Gil-Bea F.J, Castanedo-Vazquez D, Sedano M.J, Zufiria M, de Eulate G.F.G. et al. Clinical evidences supporting the Src/c-Abl pathway as potential therapeutic target in amyotrophic lateral sclerosis. Journal of the Neurological Sciences. 2018;393:80-82. PMID: 30125805. Alonso-Jimenez A, Kroon R.H.M.J.M, Alejaldre-Monforte A, Nunez-Peralta C, Horlings C.G.C, Van Engelen B.G.M. et al. Muscle MRI in a large cohort of patients with oculopharyngeal muscular dystrophy. Journal of Neurology, Neurosurgery and Psychiatry. 2018. PMID: 30530568.

Code: PREVAL 17/03 Carmen Lage Martinez. Title: Contrato Predoctoral IDIVAL-UC. Principal investigator: Sanchez Juan, Pascual Jesus. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: IDIVAL. Funding: 65688. Duration: 2017-2021.

230 CIBERNED 2018 ANNUAL REPORT

Code: INNVAL17/20. Title: Diseno de estructuras biopolimericas funcionalizadas con grafeno para el desarrollo de cultivos neuronales en modelos celulares de patologia de la motoneurona. Principal investigator: Olga Tapia Martinez. CIBERNED’s collaboration: No. CIBERNED groups: G601. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: IDIVAL. Funding: 12000. Duration: 2017-2019.

Code: NVAL16/21. Title: Diseno de nuevas moleculas para el tratamiento de la esclerosis lateral amiotrofica-Innopharma. Principal investigator: Javier Riancho. CIBERNED’s collaboration: No. CIBERNED groups: G601. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: IDIVAL. Funding: 15000. Duration: 2017-2018.

Code: NVAL17/22. Title: Efecto del deficit del factor de supervivencia de las neuironas motoras (SMN) sobre la organizacion estructural y molecular de lkos compartimentos nucleares implicados en la biogenesis de RNPs espliceosomales y en el procesamiento de RNAs: estudio experim. Principal investigator: Olga Tapia Martinez. CIBERNED’s collaboration: No. CIBERNED groups: G601. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: IDIVAL. Funding: 8000. Duration: 2017-2019.

Code: NVAL 16/21. Title: Enfermedad de Alzheimer y sindrome de Down. Una aproximacion clinica y experimental. Principal investigator: Javier Riancho. CIBERNED’s collaboration: No. CIBERNED groups: G601. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: IDIVAL. Funding: 15000. Duration: 2017-2018.

Code: PI17/00936. Title: Estudio de biomarcadores motores, geneticos y de imagen en la enfermedad de Parkinson prodromica asociada a la mutacion G2019S de LRRK2. Principal investigator: Jon Infante. CIBERNED’s collaboration: No. CIBERNED groups: G601. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 53240. Duration: 2018-2020.

231 Code: PI16/01652. Title: Estudio de variantes raras en genes asociados a Enfermedad de Alzheimer en poblacion espanola. Principal investigator: Pascual Sanchez Juan. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 139150. Duration: 2017-2019.

Code: ESMI. Title: European Spinocerebellar Ataxia Type 3/Machado Joseph disease initiative. Principal investigator: Thomas Klockgether. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: JPND. Funding: 3000. Duration: 2016-2019.

Code: IGOS. Title: International Guillain-Barre syndrome outcome study (IGOS). Principal investigator: Bart Jacobs. CIBERNED’s collaboration: No. CIBERNED groups: G601. Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Erasmus Medical Center. Funding: ND. Duration: 2012-2019.

Code: BFU2017-84046P. Title: Papel del dano en el DNA y de la reorganizacion epigenetica del DNA y de las his- tonas como determinantes del destino de las celulas progenitoras del esqueleto de las extremidades. Principal investigator: Juan Antonio Montero Simon y Juan Hurle Gonzalez. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 199650. Duration: 2018-2020.

Code: COPPADIS. Title: Proyecto COPPADIS (Cohort of patients with Parkinson’s disease in Spain, 2015). Principal investigator: Diego Santos. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: ABBVIE, ACEBRE, ALTER, ASOCIACIÓN PARKINSON ARAGON, ASOCIACIÓN PARKINSON PALENCIA, ASOCIACIÓN PARKINSON VALENCIA, AYUNTAMIENTO DE A CORUÑA, CLIMANOSA, CONSTRUDECO NORTE, FEDERACION GALLEGA ATLETISMO, ESTEVE, LUND- BECK, QUALIGEN, UCB PHARMA, VALENCIA CONCIE. Funding: 2557,38. Duration: 2016-2021.

232 CIBERNED 2018 ANNUAL REPORT

Code: SMA Europe Call 8 grant application. Title: Regulation of the survival motor neuron (SMN) protein by acetylation and its importance in snRNP biogenesis and molecular assembly of Cajal bodies. Principal investigator: Olga Tapia Martinez. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: SMA Europe. Funding: 40000. Duration: 2017-2018.

Code: Tau Consortium (UCSF, USA) 2016. Title: Tau Consortium. Principal investigator: Suzee Lee. CIBERNED’s collaboration: No. CIBERNED groups: G601 . Other CIBER’s collaboration: No. Type: Internacional. Funding agency: UCSF. Funding: 98000. Duration: 2014-2018.

233 609 | Adolfo López de Munain Arregui

BIODONOSTIA - INSTITUTO DE INVESTIGACIÓN SANITARIA. ÁREA DE NEUROCIENCIAS.

Pº Dr Begiristain s/n - Planta 1ª 20014 San Sebastián Guipúzcoa - España Tel. +34 943 006 295 Fax: +34 943 006 250 E-mail: mirenkarmele.arnaizperez@ osakidetza.eus

PRINCIPAL Fernandez Torron, Moreno Izco, Fermín. INVESTIGATOR Roberto. Bachelor degree. Bachelor degree. Naldaiz Gastesi, Neia. López de Munain Garcia-Bragado Acín, Bachelor degree. Arregui, Adolfo Federico. Navalpotro Gómez, PhD. Irene. LIST OF Gereñu Lopetegui, Bachelor degree. PERSONNEL Gorka. Poza Aldea, Juan Jose. PhD. Aiastui Pujana, Ana. Bachelor degree. PhD. Gil Bea, Francisco Javier. Rico Castro, Anabel. PhD. Arnaiz Perez, Miren Bachelor degree. Karmele. Gorostidi Pagola, Ana. Rodríguez Oroz, María Oficial Administrativo. PhD. Cruz. Barandiaran Amillano, Guembelzu Iraola, Bachelor degree. Myriam. Garazi. Ruiz Martínez, Javier. Bachelor degree. Bachelor degree. Bachelor degree. Belloso Iguerategui, Holt, Ian James. Sáenz Peña, Amets. Arantzazu. PhD. Bachelor degree. Bachelor degree. Jimenez Urbieta, Haritz. Sistiaga Berrondo, Bergareche Yarza, Jesus Bachelor degree. Andone. Alberto. Lasa Elgarresta, Jaione. Bachelor degree. Bachelor degree. Bachelor degree. Vallejo Illarramendi, Camaño González, María Lasa Fernández, Ainara. del Pilar. Haizpea. Bachelor degree. Bachelor degree. Bachelor degree. Zufiria García, Mónica. Dehesa Etxebeste, Martinez - Lage alvarez, Bachelor degree. Martxel Pedro. Pablo. Zulaica Ijurco, Miren. Bachelor degree. Bachelor degree. Bachelor degree. Merino Galán, Leyre. Bachelor degree.

234 CIBERNED 2018 ANNUAL REPORT

ABSTRACT PARKINSON DISEASE Main research lines: 1. Genetics and pre-motor markers of PD. The studies are focused c patients with the LRRK2 R1441G mutation (frequent in the Basque Country) and asymptomatic carriers, which allows to know pre-motor biomarkers of PD: clinical, molecular and imaging (DATSCAN and RM) as well as its evolutional behavior. 2. Design of compounds with kinase inhibitory activity and drug testing models 3. Clinical trials 4. Frontotemporal dementia. The group continues working on the cohort of patients with DT secondary to mutations in progranulin as well as on carriers, focusing their study on the search for progression biomarkers as well as the study of lysosomal function whose alteration seems to be the cause of the clinical manifestations. The group studies various known targets and works in the design of their modulators using as models immortalized cells of patients and Drosophila.

NEUROMUSCULAR DISORDERS The group is composed by neurologists, neuropsychologists and biologists who develop a comprehensive approach to neuromuscular pathology from a clinical and basic aspect (including ELA and hereditary-based muscular diseases), as well as on the role of the muscle in the metabolic control and its influence on the development of central phenomena associated with aging (brain proteinopathies and aging). Main research lines: 1. Myotonic dystrophy. The group continues studying aging mechanisms linked to the disease and its clinical consequences. There is an established collaboration to develop chemical compounds to act on the disease and to design phase I clinical trials and test them along with other groups. 2. Duchenne dystrophy. Study of alterations in calcium homeostasis and its rescue with compounds targeting ryanodine receptor stability. The study has resulted in a patent for a serial of compounds called generically AHULKEN. A company has been created for its preclinical development: MIRAMOON PHARMA (www.mira- moonpharma.com). This study has been extended to test the therapeutic potential on the cardiac manifestations of the disease. 3. Facioscapulohumeral dystrophy. The group continues collaborating with other groups in the clinical characterization of cases with scapuloperoneal syndrome and methylation patterns. A researcher based in London will soon join the group to promote this line using CRISPR / Cas13 editing techniques in this disease. 4. Limb Girdle Dystrophy type 2 A. Study of the influence of calpain 3 in proliferation, differentiation and replacement of muscle satellite cells mechanisms, as well as the relationship with the extracellular matrix proteins. The group continues working on the study of calcium homeostasis and RER and also on the potential of CRISPR mediated gene editing, for both using corrected cells as a therapy and creating a pig animal model in collaboration with the University of Murcia. The group is also developing strategies for searching targets related to the functioning of myogenesis in the body as well as in the regulation of the Wnt pathway. 5. Amyotrophic Lateral Sclerosis. Recent incorporations have strengthened the current intraciber network. The group has already completed an intraciber research

235 project and will present another one in the next call. The results of this line will be published throughout 2019 and have opened multiple collaboration possibilities with other groups of CIBERER, as well as the design of therapeutic strategies whose development will be done together with the startup MIRAMOON Pharma. The group has joined ENCALS and takes part actively in other international ALS consortiums.

KEYWORDS Parkinson Disease, frontotemporal dementia, cognitive impairment, LRRK2, progranulin, neuroinflammation, muscular dystrophies, calpain, biomarkers, neurogenetics, ALS, drug design.

PUBLICATIONS 2018 Contreras Muruaga MM, Reig G, Vivancos J, González A, Cardona P, Ramírez-Moreno JM et al. Factors associated with poor anticoagulation control with vitaminK antagonists among outpatients attended in Internal Medicine and Neurology. The ALADIN study.Revista clinica espanola. 2018;218(7). PMID: 29983190. Poza JJ, Pujol M, Ortega-Albás JJ, Romero O, en representación del Grupo de estudio de insomnio de la Sociedad Española de Sueño (SES). Melatonin in sleep disorders.Neurologia (Barcelona, Spain). 2018. PMID: 30466801. Riancho J, Gil-Bea F, Santurtun A, Lopez De Munain A. Amyotrophic lateral sclerosis: A complex syndrome that needs an integrated research approach. Neural Regeneration Research. 2019;14(2):193-196. epub2018. PMID: 30530996. Jimenez-Urbieta H, Gago B, Quiroga-Varela A, Rodriguez-Chinchilla T, Merino-Galan L, Oregi A. et al. Pramipexole-induced impulsivity in mildparkinsonian rats: a model of impulse control disorders in Parkinson’s disease. Neurobiology of Aging. 2019;75:126-135. epub2018. PMID: 30572183. Toral-Ojeda I, Aldanondo G, Lasa-Elgarresta J, Lasa-Fernandez H, Vesga-Castro C, Mouly V, de Munain AL, Vallejo-Illarramendi A. A Novel Functional In Vitro Model that Recapitulates Human Muscle Dis- orders. Muscle Cell and Tissue-Current Status of Research Field: IntechOpen; 2018. Riancho J, Bosque-Varela P, Perez-Pereda S, Povedano M, de Munain A.L, Santurtun A. The increasing importance of environmental conditions in amyotrophic lateral sclerosis. International Journal of Biometeorology. 2018;62(8):1361-1374. PMID: 29713861. Yakhine-Diop S.M.S, Rodriguez-Arribas M, Martinez-Chacon G, Uribe-Carretero E, Gomez-Sanchez R, Aiastui A. et al. Acetylome in human fibroblasts from Parkinson’s disease patients. Frontiers in Cellular Neuroscience. 2018;12. PMID: 29719501. Riancho J, Gil-Bea F.J, Castanedo-Vazquez D, Sedano M.J, Zufiria M, de Eulate G.F.G. et al. Clinical evidences supporting the Src/c-Abl pathway as potential therapeutic target in amyotrophic lateral sclerosis. Journal of the Neurological Sciences. 2018;393:80-82. PMID: 30125805. Hoogland J, van Wanrooij L.L, Boel J.A, Goldman J.G, Stebbins G.T, Dalrymple-Alford J.C. et al. Detect- ing Mild Cognitive Deficits in Parkinson’s Disease: Comparison of Neuropsychological Tests. Move- ment Disorders. 2018;33(11):1750-1759. PMID: 30216541.

236 CIBERNED 2018 ANNUAL REPORT

Yakhine-Diop S.M.S, Niso-Santano M, Rodriguez-Arribas M, Gomez-Sanchez R, Martinez-Chacon G, Uribe-Carretero E. et al. Impaired Mitophagy and Protein Acetylation Levels in Fibroblasts from Parkinson’s Disease Patients. Molecular Neurobiology. 2018. PMID: 30032424. Alonso-Jimenez A, Kroon R.H.M.J.M, Alejaldre-Monforte A, Nunez-Peralta C, Horlings C.G.C, Van Engelen B.G.M. et al. Muscle MRI in a large cohort of patients with oculopharyngeal muscular dystrophy. Journal of Neurology, Neurosurgery and Psychiatry. 2018. PMID: 30530568. Diez-Fairen M, Benitez B.A, Ortega-Cubero S, Lorenzo-Betancor O, Cruchaga C, Lorenzo E. et al. Pooled-DNA target sequencing of Parkinson genes reveals novel phenotypic associations in Spanish population. Neurobiology of Aging. 2018;70:325.e1-325.e5. PMID: 29887346. Skorvanek M, Martinez-Martin P, Kovacs N, Zezula I, Rodriguez-Violante M, Corvol J.-C. et al. Rela- tionship between the MDS-UPDRS and Quality of Life: A large multicenter study of 3206 patients. Parkinsonism and Related Disorders. 2018;52:83-89. PMID: 29625875. Lostal W, Urtizberea J.A, Richard I, Alonso-Jimenez A, Carlier R.-Y, Carson V. et al. 233rd ENMC Inter- national Workshop:: Clinical Trial Readiness for Calpainopathies, Naarden, The Netherlands, 15–17 September 2017. Neuromuscular Disorders. 2018;28(6):540-549. PMID: 29655529. Ezquerra-Inchausti M, Anasagasti A, Barandika O, Garai-Aramburu G, Galdos M, Lopez de Munain A. et al. A new approach based on targeted pooled DNA sequencing identifies novel mutations in patients with Inherited Retinal Dystrophies. Scientific reports. 2018;8(1):15457-. PMID: 30337596. Lucente G, Almendrote M, Ramos-Fransi A, Martinez-Pineiro A, Camano P, Ballester-Lopez A. et al. A novel mutation in the valosin-containing-protein gene found in a Spanish family. Journal of the Neurological Sciences. 2018;391:112-113. PMID: 30103957. Ferrero H, Larrayoz I.M, Gil-Bea F.J, Martinez A, Ramirez M.J. Adrenomedullin, a Novel Target for Neu- rodegenerative Diseases. Molecular Neurobiology. 2018;55(12):8799-8814. PMID: 29600350. Fernández-Eulate G, Alberro A, Muñoz-Culla M, Zulaica M, Zufiría M, Barandiarán M et al. Blood Markers in Healthy-Aged Nonagenarians: A Combination of High Telomere Length and Low Amy- loidβ Are Strongly Associated With Healthy Aging in the Oldest Old.Frontiers in aging neuroscience. 10. PMID: 30546303. Emparanza J.I, Lopez de Munain A, Greene M.H, Matheu A, Fernandez-Torron R, Gadalla S.M. Can- cer phenotype in myotonic dystrophy patients: Results from a meta-analysis. Muscle and Nerve. 2018;58(4):517-522. PMID: 30028904. Weinberg I, Giri J, Kolluri R, Arcelus J.I, Falga C, Soler S. et al. Characteristics, treatment patterns and outcomes of patients presenting with venous thromboembolic events after knee arthroscopy in the RIETE Registry. Journal of Thrombosis and Thrombolysis. 2018;46(4):551-558. PMID: 30196344. Petek B, Villa-Lopez M, Loera-Valencia R, Gerenu G, Winblad B, Kramberger M.G. et al. Connecting the brain cholesterol and renin–angiotensin systems: potential role of statins and RAS-modifying medications in dementia. Journal of Internal Medicine. 2018;284(6):620-642. PMID: 30264910. Johnson K, Bertoli M, Phillips L, Topf A, Van den Bergh P, Vissing J. et al. Detection of variants in dystroglycanopathy-associated genes through the application of targeted whole-exome sequencing analysis to a large cohort of patients with unexplained limb-girdle muscle weakness. Skeletal Muscle. 2018;8(1). PMID: 30060766. Meca-Lallana JE, Prieto-González JM, Jimenez-Veiga J, Carreón-Guarnizo E, Jiménez-Martín I, Hernán- dez-Clares R et al. Development and validation of a brief electronic screening test for cognitive impairment in multiple sclerosis (SCI-MS Test).Multiple sclerosis and related disorders. 2018;28. PMID: 30553169. Etxeberria A, Iribar J, Rotaeche R, Vrotsou K, Barral I, Barral I. et al. Evaluation of an educational inter-

237 vention and a structured review of polypharmacy in elderly patients in Primary Care Evaluación de una intervención formativa con revisión estructurada de la medicación en pacientes mayores poli- medicados en Atención Primaria. Revista Espanola de Geriatria y Gerontologia. 2018;53(6):319- 325. PMID: 30097319. Anasagasti A, Ezquerra-Inchausti M, Barandika O, Culla M.M, Caffarel M.M, Otaegui D. et al. Ex- pression profiling analysis reveals key Microrna-mRNA interactions in early retinal degeneration in retinitis pigmentosa. Investigative Ophthalmology and Visual Science. 2018;59(6):2381-2392. PMID: 29847644. Acuna G, Goma M, Temprana-Salvador J, Garcia-Bragado F, Alos L, Ordi J. et al. Human papillomavi- rus in laryngeal and hypopharyngeal lymphoepithelial carcinoma. Modern Pathology. 2018. PMID: 30552415. Munoz-Lopetegi A, Lopez-Gonzalez F.J, Rodriguez-Osorio X, Pato Pato A, Bellas Lamas P, Abella-Cor- ral J. et al. LACONORTE study: Efficacy and security of lacosamide as first add-on therapy for fo- cal-onset epilepsy in real-life setting. Epilepsy Research. 2018;145:51-54. PMID: 29890341. Durigon R, Mitchell A.L, Jones A.W.E, Manole A, Mennuni M, Hirst E.M.A. et al. LETM1 couples mitochondrial DNA metabolism and nutrient preference. EMBO Molecular Medicine. 2018;10(9). PMID: 30012579. Østergaard ST, Johnson K, Stojkovic T, Krag T, De Ridder W, De Jonghe P et al. Limb girdle muscular dystrophy due to mutations in POMT2.Journal of neurology, neurosurgery, and psychiatry. 2018;89(5). PMID: 29175898. Barandiaran M, Moreno F, de Arriba M, Indakoetxea B, Boda I, Gabilondo A et al. Longitudinal Neu- ropsychological Study of Presymptomatic c.709-1G>A Progranulin Mutation Carriers.Journal of the International Neuropsychological Society : JINS. 2018. PMID: 30369339. Ghadery C, Valli M, Mihaescu A, Strafella R, Navalpotro I, Kim J. et al. Molecular Imaging of Addictive Behavior in Idiopathic Parkinson’s Disease. International Review of Neurobiology. 2018;141:365- 404. PMID: 30314604. Garcia Martinez E, Ruiz Martinez J, Hernandez Gomez T.A, Hernandez-Gil Sanchez J, Brufau Redondo C. More than a cyst in the head Más allá de un quiste en la cabeza. Medicina Clinica. 2018;151(8):338-. PMID: 29398007. Thomalla G, Simonsen C.Z, Boutitie F, Andersen G, Berthezene Y, Cheng B. et al. MRI-Guided thrombolysis for stroke with unknown time of onset. New England Journal of Medicine. 2018;379(7):611- 622. PMID: 29766770. GoMez-Andres D, Diaz-Manera J, Alejaldre A, Pulido-Valdeolivas I, GonzaLez-mera L, Olive M. et al. Muscle imaging in laminopathies: Synthesis study identifies meaningful muscles for follow-up. Muscle and Nerve. 2018;58(6):812-817. PMID: 30066418. Diaz-Manera J, Fernandez-Torron R, Llauger J, James M.K, Mayhew A, Smith F.E. et al. Muscle MRI in patients with dysferlinopathy: Pattern recognition and implications for clinical trials. Journal of Neurology, Neurosurgery and Psychiatry. 2018;89(10):1071-1081. PMID: 29735511. Bueno G.E, Ruiz-Castaneda D, Martinez J.R, Munoz M.R, Alascio P.C. Natural history and clinical characteristics of 50 patients with Wolfram syndrome. Endocrine. 2018;61(3):440-446. PMID: 29728875. Garcia Martinez E, Ruiz Martinez J, Hernandez-Gil Sanchez J, Brufau Redondo C, Poblet Martinez E. Not always is an infection No siempre es una infección. Medicina Clinica. 2018;151(11):e67-. PMID: 29523339. Madero-Perez J, Fdez E, Fernandez B, Lara Ordonez A.J, Blanca Ramirez M, Gomez-Suaga P. et al.

238 CIBERNED 2018 ANNUAL REPORT

Parkinson disease-associated mutations in LRRK2 cause centrosomal defects via Rab8a phosphorylation. Molecular Neurodegeneration. 2018;13(1). PMID: 29357897. Pottier C, Zhou X, Perkerson R.B, Baker M, Jenkins G.D, Serie D.J. et al. Potential genetic modifiers of disease risk and age at onset in patients with frontotemporal lobar degeneration and GRN mutations: a genome-wide association study. The Lancet Neurology. 2018;17(6):548-558. PMID: 29724592. Figueroa-Bonaparte S, Llauger J, Segovia S, Belmonte I, Pedrosa I, Montiel E. et al. Quantitative muscle MRI to follow up late onset Pompe patients: A prospective study. Scientific Reports. 2018;8(1). PMID: 30022036. Delgado-Alvarado M, Dacosta-Aguayo R, Navalpotro-Gomez I, Gago B, Gorostidi A, Jimenez-Urbieta H. et al. Ratios of proteins in cerebrospinal fluid in Parkinson’s disease cognitive decline: prospective study. Movement Disorders. 2018;33(11):1809-1813. PMID: 30423201. Ferrero H, Larrayoz I.M, Solas M, Martinez A, Ramirez M.J, Gil-Bea F.J. Reduced Adrenomedullin Par- allels Microtubule Dismantlement in Frontotemporal Lobar Degeneration. Molecular Neurobiology. 2018;55(12):9328-9333. PMID: 29671277. Labayru G, Arenzana I, Aliri J, Zulaica M, Lopez De Munain A, Andone Sistiaga A. Social cognition in myotonic dystrophy type 1: Specific or secondary impairment?. PLoS ONE. 2018;13(9). PMID: 30248121. Moore U.R, Jacobs M, Fernandez-Torron R, Jang J, James M.K, Mayhew A. et al. Teenage exercise is associated with earlier symptom onset in dysferlinopathy: A retrospective cohort study. Journal of Neurology, Neurosurgery and Psychiatry. 2018. PMID: 29378789. Naldaiz-Gastesi N, Bahri O.A, Lopez de Munain A, McCullagh K.J.A, Izeta A. The panniculus carnosus muscle: an evolutionary enigma at the intersection of distinct research fields. Journal of Anatomy. 2018;233(3):275-288. PMID: 29893024. Cluett T.J, Akman G, Reyes A, Kazak L, Mitchell A, Wood S.R. et al. Transcript availability dictates the balance between strand-asynchronous and strand-coupled mitochondrial DNA replication. Nucleic acids research. 2018;46(20):10771-10781. PMID: 30239839. Valderrama-Carvajal A, Irizar H, Gago B, Jiménez-Urbieta H, Fuxe K, Rodríguez-Oroz MC et al. Tran- scriptomic integration of D4R and MOR signaling in the rat caudate putamen.Scientific reports. 2018;8(1). PMID: 29743514. Munoz-Lopetegi A, Fernandez Garcia de Eulate G, Rodriguez-Antiguedad Munoz J, Bergareche A, Jose Poza J. Two cases of opercular myoclonic-anarthric status epilepticus. Epileptic Disorders. 2018;20(3):189-194. PMID: 29905156. Dranca L, de Abetxuko Ruiz de Mendarozketa L, Goni A, Illarramendi A, Navalpotro Gomez I, Delga- do Alvarado M. et al. Using Kinect to classify Parkinson’s disease stages related to severity of gait impairment. BMC Bioinformatics. 2018;19(1). PMID: 30526473. dos Santos T.W, Miranda J, Teixeira L, Aiastui A, Matheu A, Gambero A. et al. Yerba Mate Stimulates Mitochondrial Biogenesis and Thermogenesis in High-Fat-Diet-Induced Obese Mice. Molecular Nu- trition and Food Research. 2018;62(15). PMID: 29851217. Kia DA, Sabir MS, Ahmed S, Trinh J, Bandres-Ciga S, International Parkinson’s Disease Genomics Consortium. et al. LRP10 in α-synucleinopathies.The Lancet. Neurology. 2018;17(12). PMID: 30507383. Del Rey N.L.-G, Quiroga-Varela A, Garbayo E, Carballo-Carbajal I, Fernandez-Santiago R, Monje M.H.G. et al. Advances in parkinson’s disease: 200 years later. Frontiers in Neuroanatomy. 2018;12. PMID: 30618654.

239 Estevez-Fraga C, Lopez-Sendon Moreno J.L, Martinez-Castrillo J.C, Perez-Perez J, Matarazzo M, Es- piga P.G.-R. et al. Phenomenology and disease progression of chorea-acanthocytosis patients in Spain. Parkinsonism and Related Disorders. 2018;49:17-21. PMID: 29317187. Toledo M, Carnero-Pardo C, Carreno-Martinez M, Escudero-Torrella J, Gaig C, Garcia-Ribas G et al. Apuntes en Neurologia» (Notes in Neurology): a synthesis of the evidence on common paroxysmal neurological disorders and on neurodegenerative disorders].Revista de neurologia. 2018;67(s01). PMID: 30779106. Garcia-Ruiz P.J, Sanz-Cartagena P, Martinez-Castrillo J.C, Ares-Pensado B, Aviles-Olmos I, Blazquez-Estrada M. et al. Myths and evidence on the use of botulinum toxin: Neuropharmacol- ogy and dystonia Mitos y evidencias en el empleo de la toxina botulínica: Neurofarmacología y distonías. Revista de Neurologia. 2018;66(5):163-172. Zamarbide M, Gil-Bea F.J, Bannenberg P, Martinez-Pinilla E, Sandoval J, Franco R. et al. Maternal imprinting on cognition markers of wild type and transgenic Alzheimer’s disease model mice. Sci- entific Reports. 2018;8(1). PMID: 29691440.

Code: RTC-2015-3750-1. Title: Desarrollo de agentes terapeuticos basados en acidos nucleicos para el tratamiento de enfermedades neuromotoras y neuromusculares. Principal investigator: Ruben Lopez Vales. CIBERNED’s collaboration: Yes. CIBERNED groups: G607 ; G609. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 204315. Duration: 2015-2018.

Code: ASAP. Title: Aligned and Standardized Neuroimaging in Atypical Parkinsonism. Principal investigator: Thilo Van Eimeren.

240 CIBERNED 2018 ANNUAL REPORT

CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: ND. Duration: 2017-2019.

Code: 2018222031. Title: Alteraciones metabolicas y perturbaciones de organulos celulares en la fisiopatolo- gia de la esclerosis lateral amiotrofica. Principal investigator: Ian Holt. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 36052. Duration: 2018-2018.

Code: BIO/ER/022. Title: Analisis del papel de la Calpaina 3 en la regulacion de las celulas satelite musculares. Principal investigator: Adolfo Lopez De Munain. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Eitb Maratoia. Funding: 40541,34. Duration: 2017-2019.

Code: FPU/04677. Title: Beca Formacion de Profesorado Universitario (FPU) 2015. Principal investigator: Irene Navalpotro. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MECD. Funding: 16422. Duration: 2016-2018.

Code: DPPE17/001. Title: Compuestos Ahulken: Nuevos moduladores de calcio como tratamiento para la distrofia muscular de Duchenne. Principal investigator: Ainara Vallejo. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Duchenne Parent Project. Funding: 25000. Duration: 2018-2019.

Code: CM16/00033. Title: Contrato Rio Hortega. Principal investigator: Irene Navalpotro Gomez. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 53732. Duration: 2017-2018. Code: 2018222021.

241 Title: Desarrollo preclinico de una terapia combinada frente a la distrofia miotonica con rycals y compuestos anti-envejecimiento. Principal investigator: Adolfo Lopez de Munain. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 35006. Duration: 2018-2018.

Code: PI17/01231. Title: Envejecimiento en la distrofia miotonica tipo 1: Analisis multifactorial desde una perspectiva biologica, neuropsicologica y neurorradiologica. Principal investigator: Andone Sistiaga Berrondo. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 105270. Duration: 2018-2020.

Code: PI14/00436. Title: Estudio de la capacidad regenerativa de los progenitores musculares derivados del ipss del pacientes con distrofia de cinturas tipo 2ª. Estudio in vitro y en un modelo murino de dano tisular. Principal investigator: Adolfo Lopez De Munain. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 131000. Duration: 2015-2018.

Code: PI17/001841. Title: Funcion de la calpaina 3 en las celulas satelite durante la regeneracion muscular y su modulacion farmacologica como posible tratamiento de la LGMD2A. Principal investigator: Adolfo Lopez de Munain. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 129470. Duration: 2018-2020.

Code: SAF2017-92696-EXPLORA. Title: Generation of “Lab-on-chip” systems to investigate neuromuscular disorders. Principal investigator: Francisco Javier Gil Bea. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 72600. Duration: 2018-2020.

Code: 2015111038. Title: Implicacion de la calpaina 3 en la senescencia prematura de las celulas satelite musculares: Estudio del ciclo celular en progenitores musculares derivados de IPSs de pacientes con LGMD2A.

242 CIBERNED 2018 ANNUAL REPORT

Principal investigator: Adolfo Lopez De Munain. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 53920. Duration: 2016-2018.

Code: BI017/ND/023-BD. Title: Metabolic distrubancer and organelle erturbations in the PAT. Principal investigator: Francisco Javier Gil Bea. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Eitb Maratoia. Funding: 81115. Duration: 2018-2020.

Code: 2018-000117-01-B. Title: Modulacion de SERCA como diana terapeutica para el tratamiento de la distrofia muscular LGMD2A. Principal investigator: Ainara Vallejo. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 67310. Duration: 2018-2019.

Code: PI16/01430. Title: Papel de SOX2 en la remodelacion dermica y en el deficit de cicatrizacion cutanea asociado al envejecimiento. Principal investigator: Ander Izeta. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 183315. Duration: 2017-2019.

Code: PI17/00676. Title: Pharmacological modulation of ryanodine receptor in Duchenne and Becker muscular dystrophies. Principal investigator: Ainara Vallejo. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 99220. Duration: 2018-2021.

Code: BECA FORMACION INVESTIGADOR-MODALIDAD PREDOCTORAL(A) ARANTZAZU BEL- LOSO. Title: Plasticidad sinaptica y actividad de los ganglios basales en un modelo de parkin- sonismo e impulsividad. Principal investigator: Maria Cruz Rodriguez Oroz. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No.

243 Type: CCAA. Funding agency: Gobierno Vasco. Funding: ND. Duration: 2018-2018.

Code: PI16/01325. Title: Regulacion de la expresion de FRZB in vitro e in vivo, para el restablecimiento de la homeostasis en la fibra muscular de pacientes con distrofia de cinturas tipo 2A (LGM- D2A). Principal investigator: Amets Saenz Pena. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 57475. Duration: 2016-2019.

Code: DFG17/003. Title: Terapia combinada con tioredoxinas ancestrales y nuevos compuestos rycals para tratar desequilibrios calcio-redox en enfermedades neurodegenerativas. Principal investigator: Adolfo Lopez De Munain. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: CCAA. Funding agency: Gobierno Vasco. Funding: 74000. Duration: 2017-2018.

Code: HR17-00268. Title: Therapeutic targeting of MBNL microRNAs as innovative treatments for myotonic dystrophy. Principal investigator: Ruben Artero. CIBERNED’s collaboration: No. CIBERNED groups: G609 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Caixa. Funding: 53000. Duration: 2018-2021.

PHD DISSERTATIONS 2018 Author: Neia Naldaiz Gastezi. Title: Caracterización celular y molecular de la miogénesis dérmica de ratón y su posible traslación al humano. Date: 12/3/2018. Supervisor: Adolfo López de Munain Arregui.

Author: Leire Casas Fraile. Title: doctoral defendida en 2018: Title: FRZB gene expression regulation in vitro to re- store muscle fibre homeostasis in limb-girdle muscular dystrophy type 2A (LGMD2A) and Frzb-/- murine model muscle analysis. Date: 26/9/2018. Supervisor: Amets Sáenz Peña.

244 CIBERNED 2018 ANNUAL REPORT

604 | Pura Muñoz-Cánoves

CELL BIOLOGY GROUP ICREA AND POMPEU FABRA UNIVERSITY (UPF) DEPARTMENT OF EXPERIMENTAL AND LIFE SCIENCES (DCEXS)

Tel.: +34 933 160 891 Fax: +34 933 160 901 E-mail: [email protected]

PRINCIPAL Jardí Ripoll, Mercé. Moiseeva, Victoria. INVESTIGATOR Technician. Bachelor degree. Lukesova, Vera. Perdiguero Santamaría, Muñoz Cánoves, Pura Technician. Eusebio. Martínez García, PhD. LIST OF Antonio. Serrano Sánchez, PERSONNEL Bachelor degree. Antonio L. Gerez Cruz, Rosa Maseres Javaloy, Pedro. PhD. Johanna. Bachelor degree. Technician.

ABSTRACT Muscle stem cells are subjected to circadian regulation, and this regulation is altered during aging. 1. Our group showed for the first time that muscle stem cells, despite being quiescent, are subject to a strict circadian control throughout the 24 hour day/night cycle, and this circadian regulation is altered with aging. Based on these findings from the group, we have addressed our research this year to answer two fundamental questions: 1.1. What are the long-term molecular and epigenetic consequences of muscle stem cell clock dampening during aging? These results will help us to identify which clock genes and their targets show aberrant epigenetic modifications that might affect their expression during aging. Since the enzymes responsible for regulating these chromatin marks are known, identifying changes in these may point toward strategies to rein- state normal clock function in aged muscle stem cells. 1.2. Does disruption of the clock machinery affect satellite cell quiescence and regenerative ability? The synchronization of physiological and metabolic processes to the appropriate time of the day is achieved by the regulation of systemic factors, and by the regulation of local tissue specific gene expression. We have characterized how both

245 clocks coordinate (systemic and tissue specific) and how each clock de-regulation affects muscular stem cells during aging. - Work under preparation (Kumar A, Vaca M and Muñoz-Cánoves P). 2. Studying the heterogeneity of quiescent muscular stem cells. Several studies based on transgenesis have probed that muscular stem cells is not a homogeneous population. Our group has developed a cellular sorting strategy by flux citometry (FACS) able to screen two different populations of satellite cells with different myogenic properties and we have demonstrated that the transcription factor FoxO3a is a regulator of both populations divergence. - Submitted work (García Prat L, Perdiguero E and Muñoz-Cánoves P). 3. Additional studies in stem cells show their autophagic activity loss during aging and the entrance in senescence. We have demonstrated that caloric restriction during three months in aged mice is able to restitute autophagy in their satellite cells and increase their regenerative capacity. -Work under preparation (García-Prat L, Perdiguero E, Serrano A and Muñoz-Cánoves P). 4. Studying Sestrins capacity to prevent muscular atrophy. We have demonstrated that Sestrins over-expression protects muscle from mass and strength loss caused by disuse, by 1) mTORC1 inhibition, which up-regulates autophagy, and 2) AKT activation which inhibits muscular proteolysis. These results have a potential implication in sarcopenia treatment. - Work under revision in Nature Communications (Segalés J, Perdiguero E, Serrano A, García-Prat and Muñoz-Cánoves P). Additional studies and reviews: - Muñoz-Cánoves P, Huch M. Definitions for adult stem cells debated. Nature 563(7731):328-329, 2018 - Sousa-Victor P, García-Prat L, Muñoz-Cánoves P. New mechanisms driving muscle stem cell regenerative decline with aging. Int J Dev Biol.;62:583-590, 2018 - Baar MP, Perdiguero E, Muñoz-Cánoves P, de Keizer PL. Musculoskeletal senescence: a moving target ready to be eliminated. Curr Opin Pharmacol. 40:147-155, 2018 - Chang NC, Sincennes MC, Chevalier FP, Brun CE, Lacaria M, Segalés J, Muñoz-Cánoves P, Ming H, Rudnicki MA. The Dystrophin Glycoprotein Complex Regulates the Epigenetic Activation of Muscle Stem Cell Commitment. Cell Stem Cell. 5:755-768, 2018.

KEYWORDS Skeletal muscle, regeneration, muscle stem cells, sarcopenia, aging, inflammation, fibrosis.

246 CIBERNED 2018 ANNUAL REPORT

PUBLICATIONS 2018 Munoz-Canoves P, Huch M. Definitions for adult stem cells debated. Nature. 2018;563(7731):328-329. PMID: 30378594. Baar M.P, Perdiguero E, Munoz-Canoves P, de Keizer P.L. Musculoskeletal senescence: a moving target ready to be eliminated. Current Opinion in Pharmacology. 2018;40:147-155. PMID: 29883814. Sousa-Victor P, Garcia-Prat L, Munoz-Canoves P. New mechanisms driving muscle stem cell regenerative decline with aging. International Journal of Developmental Biology. 2018;62(6- 8):583-590. PMID: 29938769. Chang N.C, Sincennes M.-C, Chevalier F.P, Brun C.E, Lacaria M, Segales J. et al. The Dystrophin Glycoprotein Complex Regulates the Epigenetic Activation of Muscle Stem Cell Commitment. Cell Stem Cell. 2018;22(5):755-768.e6. PMID: 29681515.

RESEARCH PROJECTS 2018 Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: Yes. CIBERNED groups: G604 ; G102; G606; G111; G306. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350000. Duration: 2016-2018.

Code: TV3. Title: New stem cell therapies for Duchenne Muscular Dystrophy. Principal investigator: Antonio Serrano Sanchez. CIBERNED’s collaboration: No. CIBERNED groups: G604 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 390000. Duration: 2018-2018.

Code: ERC. Title: Tissue regeneration and aging: the decisive quiescent stem-cell state. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: No. CIBERNED groups: G604 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 1250000. Duration: 2017-2022.

Code: MDA 2016. Title: Understanding and reversing muscle stem cell regenerative decline in DMD. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: No. CIBERNED groups: G604 . Other CIBER’s collaboration: No.

247 Type: Internacional. Funding agency: Muscular Dystrophy Association (MDA)-USA. Funding: 100000. Duration: 2016-2019.

Code: La Caixa. Title: Understanding muscle regenerative decline with aging. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: No. CIBERNED groups: G604 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Caixa. Funding: 400000. Duration: 2018-2021.

Code: SAF2015-67369-R. Title: Understanding skeletal muscle regenerative decline with aging. Principal investigator: Pura Munoz Canoves. CIBERNED’s collaboration: No. CIBERNED groups: G604 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 150000. Duration: 2016-2018.

PHD DISSERTATIONS 2018 Author: Pedro Maseres Javaloy. Title: Regulation of cell proliferaton and differentiation by p38MAPK in distinct phisyol- pathological processes. Date: 12/3/2018. Supervisor: Pura Muñoz Cánoves.

248 CIBERNED 2018 ANNUAL REPORT

607 | Xavier Navarro Acebes

DEPT. CELL BIOLOGY, PHYSIOLOGY AND IMMUNOLOGY, INSTITUTE OF NEUROSCIENCES UNIVERSITAT AUTONOMA DE BARCELONA.

Av. Can Domènech, Edif. M, Campus UAB 08193 Bellaterra Tel. +34 93 581 19 66 Fax. +34 93 581 29 86 E-mail: [email protected]

PRINCIPAL del Valle Maciá, Martínez Muriana, INVESTIGATOR Jaume. Anna. PhD. Bachelor degree. Navarro Acebes, Xavier Gaja Capdevilla, Nuria. Módol Caballero, Bachelor degree. Guillem. LIST OF García Alias, Guillermo. Bachelor degree. PERSONNEL PhD. Penas Pérez, Clara. PhD. Amo Aparicio, Jesús. Hernández Martín, Puigdoménech Poch, Bachelor degree. Joaquin. María. Badía Casahuja, Jordi. PhD. Bachelor degree. Bachelor degree. Hernández Solanes, Rodríguez Cañón, María. Baylo Marín, Olaia. Neus. Bachelor degree. Bachelor degree. Technician. Romeo Guitart, David. Bosch Merino, Herrando Grabulosa, Bachelor degree. Assumpció. Mireia. Rubio Pérez, Miguel PhD. PhD. Angel. Bruna Escuer, Jordi. Jaramillo Rodríguez, Bachelor degree. Bachelor degree. Jesica. Sánchez Fernández, Calls Cobos, Aina. Technician. Alba. Bachelor degree. Leiva Rodriguez, Bachelor degree. Casas Louzao, Caty. PhD. Tatiana. Udina Bonet, Esther. De la Oliva Muñoz, Bachelor degree. PhD. Natalia. López Serrano, Clara. Velasco Fargas, Roser. Bachelor degree. Bachelor degree. Bachelor degree. López Vales, Rubén. PhD.

249 ABSTRACT The Group of Neuroplasticity and Regeneration of the UAB is a multidisciplinary research group working on repair, regeneration and functional recovery after peripheral nerve and spinal cord lesions and in neurodegenerative diseases. The research activities of the Group of Neuroplasticity and Regeneration have focused on the study of physiopathological mechanisms of neural lesions, neuropathic pain and neurodegeneration, and on the application of novel therapeutic strategies for promoting neuroprotection and regeneration in traumatic and degenerative lesions of the nervous system. The active research lines include: • Cellular and molecular mechanisms implicated in motoneuron degenerative diseases in experimental models. Search for biomarkers. Neuroprotective strategies based on gene and pharmacological therapy. • Cell therapy by transplantation of mesenchymal stem cells and neural stem cells for the repair of spinal cord injuries and motoneuron degenerative diseases. • Repair of spinal root avulsion injuries by surgical reimplantation and pharmacological neuroprotection. • Etiopathogenic role of lipid mediators and modulators of the neuroinflammatory response in neurodegeneration induced by central nervous system lesions. • Activity-dependent therapies for enhancing axonal regeneration and functional recovery after peripheral nerve lesions and for preventing neuropathic pain. • Physiopathologic mechanisms in neuropathic pain induced by nerve and spinal cord lesions. Study of the relationship between neuroinflammation and hyperexcitability. • Study of etiopathogenic mechanisms and potential neuroprotective treatments in peripheral neuropathies induced by diabetes and by antitumoral agents. • Gene therapy using viral vectors for diseases affecting both the central (mucopolysaccharidosis type VII, megalencephalic leukodystrophy, amyotrophic lateral sclerosis) and peripheral nervous system (diabetic neuropathy, nerve regeneration). • Neuromodulation of neural plasticity for promoting functional restitution in spinal cord injuries. • Design and evaluation of neural interfaces for the development of neuroprostheses applied to neurorehabilitation. Study of new intraneural electrodes for the selective stimulation and recording of neural activity.

KEYWORDS Neurodegeneración, regeneración nerviosa, lesión de médula espinal, dolor neuropático, enfermedades de motoneuronas, neuropatías periféricas, neuroplasticidad, interfases neurales, terapia génica

250 CIBERNED 2018 ANNUAL REPORT

PUBLICATIONS 2018 Lopez-Serrano C, Santos-Nogueira E, Francos-Quijorna I, Coll-Miro M, Chun J, Lopez-Vales R. Lysophosphatidic acid receptor type 2 activation contributes to secondary damage after spinal cord injury in mice. Brain, Behavior, and Immunity. 2019;76:258-267. epub2018. PMID: 30550929. Romeo-Guitart D, Casas C. Network-centric medicine for peripheral nerve injury: Treating the whole to boost endogenous mechanisms of neuroprotection and regeneration. Neural Regen- eration Research. 2019;14(7):1122-1128. epub2018. PMID: 30804234. Lopez-Alvarez V.M, Puigdomenech M, Navarro X, Cobianchi S. Monoaminergic descending pathways contribute to modulation of neuropathic pain by increasing-intensity treadmill exercise after peripheral nerve injury. Experimental Neurology. 2018;299:42-55. PMID: 28993250. Mòdol-Caballero G, Santos D, Navarro X, Herrando-Grabulosa M. Neuregulin 1 Reduces Moto- neuron Cell Death and Promotes Neurite Growth in an in Vitro Model of Motoneuron Degen- eration.Frontiers in cellular neuroscience. 11. PMID: 29375317. Gonzalez-Perez F, Hernandez J, Heimann C, Phillips J.B, Udina E, Navarro X. Schwann cells and mesenchymal stem cells in laminin-or fbronectin-aligned matrices and regeneration across a critical size defect of 15 mm in the rat sciatic nerve. Journal of Neurosurgery: Spine. 2018;28(1):109-118. del Valle J, Santos D, Delgado-Martinez I, de la Oliva N, Giudetti G, Micera S. et al. Segregation of motor and sensory axons regenerating through bicompartmental tubes by combining extracellular matrix components with neurotrophic factors. Journal of Tissue Engineering and Regenerative Medicine. 2018. PMID: 29266822. Cortes M, Alias G.G, Tansey K.E. A “Snapshot” of the Advances in SCI Therapeutics. Neurother- apeutics. 2018;15(3):527-528. PMID: 30083985. Leiva-Rodriguez T, Romeo-Guitart D, Marmolejo-Martinez-Artesero S, Herrando-Grabulosa M, Bosch A, Fores J. et al. ATG5 overexpression is neuroprotective and attenuates cytoskeletal and vesicle-Trafficking alterations in axotomized motoneurons article. Cell Death and Dis- ease. 2018;9(6). Lago N, Pannunzio B, Amo-Aparicio J, Lopez-Vales R, Peluffo H. CD200 modulates spinal cord injury neuroinflammation and outcome through CD200R1. Brain, Behavior, and Immunity. 2018;73:416-426. PMID: 29870752. Cayuela N, Jaramillo-Jiménez E, Majós C, Velasco R, Boget T, Graus F et al. Corrigendum.Neu- ro-oncology. 2018. PMID: 30462315. De la Oliva N, Navarro X, del Valle J. Dexamethasone Reduces the Foreign Body Reaction to Intraneural Electrode Implants in the Peripheral Nerve of the Rat. Anatomical Record. 2018;301(10):1722-1733. PMID: 30353712. Penas C, Navarro X. Epigenetic modifications associated to neuroinflammation and neuropathic pain after neural trauma. Frontiers in Cellular Neuroscience. 2018;12. PMID: 29930500. Cervero C, Blasco A, Tarabal O, Casanovas A, Piedrafita L, Navarro X. et al. Glial activation and central synapse loss, but not motoneuron degeneration, are prevented by the sigma-1 receptor agonist pre-084 in the SMN2B/ mouse model of spinal muscular atrophy. Journal of Neuropathology and Experimental Neurology. 2018;77(7):577-597. Argyriou AA, Anastopoulou GG, Bruna J. Inconclusive evidence to support the use of minimal- ly-invasive radiofrequency denervation against chronic low back pain.Annals of translational

251 medicine. 2018;6(7). PMID: 29955587. Taphoorn M.J.B, Dirven L, Kanner A.A, Lavy-Shahaf G, Weinberg U, Taillibert S. et al. Influence of treatment with tumor-treating fields on health-related quality of life of patients with newly diagnosed glioblastoma a secondary analysis of a randomized clinical trial. JAMA Oncology. 2018;4(4):495-504. PMID: 29392280. Rando A, Pastor D, Viso-Leon M.C, Martinez A, Manzano R, Navarro X. et al. Intramuscular transplantation of bone marrow cells prolongs the lifespan of SOD1G93A mice and modulates expression of prognosis biomarkers of the disease. Stem Cell Research and Therapy. 2018;9(1). PMID: 29625589. Navarro X, Geuna S, Grothe C, Haastert-Talini K. Introduction: Thematic Papers Issue on Pe- ripheral Nerve Regeneration and Repair. Anatomical Record. 2018;301(10):1614-1617. PMID: 30299596. Izquierdo C, Alentorn A, Idbaih A, Simo M, Kaloshi G, Ricard D. et al. Long-term impact of temozolomide on 1p/19q-codeleted low-grade glioma growth kinetics. Journal of Neuro-On- cology. 2018;136(3):533-539. PMID: 29143276. Bayo-Puxan N, Terrasso A.P, Creyssels S, Simao D, Begon-Pescia C, Lavigne M. et al. Lysosomal and network alterations in human mucopolysaccharidosis type VII iPSC-derived neurons. Sci- entific Reports. 2018;8(1). PMID: 30413728. Sanchez-Brualla I, Calls-Cobos A, Udina E. Minocycline Does Not Reduce the Regenerative Ca- pacity of Peripheral Motor and Sensory Neurons after a Conditioning Injury in Mice. Anatom- ical Record. 2018;301(10):1638-1645. PMID: 29710422. David S, Kroner A, Greenhalgh A.D, Zarruk J.G, Lopez-Vales R. Myeloid cell responses after spinal cord injury. Journal of Neuroimmunology. 2018;321:97-108. PMID: 29957394. Amo-Aparicio J, Martinez-Muriana A, Sanchez-Fernandez A, Lopez-Vales R. Neuroinflamma- tion Quantification for Spinal Cord Injury. Current Protocols in Immunology. 2018;123(1). PMID: 30253064. Romeo-Guitart D, Fores J, Herrando-Grabulosa M, Valls R, Leiva-Rodriguez T, Galea E. et al. Neuroprotective Drug for Nerve Trauma Revealed Using Artificial Intelligence. Scientific Re- ports. 2018;8(1). PMID: 29382857. Pinol-Jurado P, Suarez-Calvet X, Fernandez-Simon E, Gallardo E, De La Oliva N, Martinez-Muri- ana A. et al. Nintedanib decreases muscle fibrosis and improves muscle function in a murine model of dystrophinopathy. Cell Death and Disease. 2018;9(7). PMID: 29991677. De La Oliva N, Mueller M, Stieglitz T, Navarro X, Del Valle J. On the use of Parylene C polymer as substrate for peripheral nerve electrodes. Scientific Reports. 2018;8(1). PMID: 29654317. Besora S, Santos C, Izquierdo C, Martinez-Villacampa M.M, Bruna J, Velasco R. Rechallenge with oxaliplatin and peripheral neuropathy in colorectal cancer patients. Journal of Cancer Research and Clinical Oncology. 2018;144(9):1793-1801. PMID: 29955956. Alberti P, Rossi E, Argyriou A.A, Kalofonos H.P, Briani C, Cacciavillani M. et al. Risk stratification of oxaliplatin induced peripheral neurotoxicity applying electrophysiological testing of dorsal sural nerve. Supportive Care in Cancer. 2018;26(9):3143-3151. PMID: 29594485. Bruna J, Velasco R. Sigma-1 receptor: A new player in neuroprotection against chemotherapy-induced peripheral neuropathy. Neural Regeneration Research. 2018;13(5):775-778. PMID: 29862996. Romeo-Guitart D, Leiva-Rodriguez T, Espinosa-Alcantud M, Sima N, Vaquero A, Dominguez-Mar- tin H. et al. SIRT1 activation with neuroheal is neuroprotective but SIRT2 inhibition with

252 CIBERNED 2018 ANNUAL REPORT

AK7 is detrimental for disconnected motoneurons. Cell Death and Disease. 2018;9(5). PMID: 29748539. Alé A, Argyriou AA, Bruna J. Sphingolipid metabolism products: potential new players in the pathogenesis of bortezomib-induced neuropathic pain.Annals of translational medicine. 2018;6(Suppl 1). PMID: 30613653. Lopez-Font I, Sogorb-Esteve A, Javier-Torrent M, Brinkmalm G, Herrando-Grabulosa M, Garcia-Lareu B. et al. Decreased circulating ErbB4 ectodomain fragments as a read-out of impaired signaling function in amyotrophic lateral sclerosis. Neurobiology of Disease. 2019;124:428-438. epub2018. PMID: 30594809. Simó M, Navarro X, Yuste VJ, Bruna J. Autonomic nervous system and cancer.Clinical autonomic research : official journal of the Clinical Autonomic Research Society. 2018;28(3). PMID: 29594605. Moreno-Galarza N, Mendieta L, Palafox-Sanchez V, Herrando-Grabulosa M, Gil C, Limon D.I. et al. Peripheral Administration of Tetanus Toxin Hc Fragment Prevents MPP+ Toxicity In Vivo. Neurotoxicity Research. 2018;34(1):47-61. PMID: 29460114. Graus F, Escudero D, Oleaga L, Bruna J, Villarejo-Galende A, Ballabriga J. et al. Syndrome and outcome of antibody-negative limbic encephalitis. European Journal of Neurology. 2018;25(8):1011-1016. PMID: 29667271.

RESEARCH PROJECTS 2018 Code: TV32016-GG. Title: Recuperacion de los movimientos del brazo y de la mano en pacientes con lesion de la medula espinal cervical mediante la neuromodulacion electrica espinal asistida con un exoesqueleto de brazo. Principal investigator: Guillermo Garcia-Alias. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: CIBER-BBN. Type: Privado. Funding agency: Fundacio La Marato de TV3. Funding: 399945. Duration: 2018-2021.

Code: E10668. Title: eBIONERVE: personalized nerve grafts for treating human nerve injurie. Principal investigator: Xavier Navarro. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Europeo.

253 Funding agency: Comision Europea. Funding: 800000. Duration: 2016-2019.

Code: SAF2016-79774-R. Title: Estudio de los mecanismos de accion de la interleucina-37 en el sistema nervioso central lesionado. Principal investigator: Ruben Lopez-Vales. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 160000. Duration: 2017-2020.

Code: PI15/01303. Title: Estudio exploratorio sobre la etiopatogenia, biomarcadores de riesgo y mecanismos implicados en la regeneracion de la neuropatia inducida por platinos. Principal investigator: Jordi Bruna. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 86515. Duration: 2015-2018.

Code: PI15/01271. Title: Estudio molecular y tratamiento de la Mucopolisacaridosis tipo VII. Principal investigator: Assumpcio Bosch. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 147015. Duration: 2016-2018.

Code: CI17-00019. Title: Immunoresolvent lipids for multiple sclerosis and amyotrophic lateral sclerosis. Principal investigator: Ruben Lopez-Vales. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Privado. Funding agency: Fundacio La Caixa. Funding: 52500. Duration: 2017-2019.

Code: WFL-ES-14-17. Title: Interleukin-37: a novel therapeutic approach for the treatment of spinal cord injury. Principal investigator: Ruben Lopez-Vales. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Internacional. Funding agency: Wings for Life Foundation. Funding: 193000. Duration: 2017-2020.

Code: SAF2016-79279-R. Title: Neuromodulacion electrica del conectoma cortico-medular para facilitar la recuperacion de la destreza manual tras una lesion medular.

254 CIBERNED 2018 ANNUAL REPORT

Principal investigator: Guillermo Garcia-Alias. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 193600. Duration: 2016-2020.

Code: AC16/00050. Title: Non-invasive electrical stimulation of the cervical spinal cord to facilitate arm and hand functional recovery in incomplete traumatic cervical spinal cord injured patients (CERMOD). Principal investigator: Guillermo Garcia-Alias. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 750000. Duration: 2017-2020.

Code: RYC-2014-16529. Title: Promoting plasticity of reticulospinal axons to recover skilled hand function after spinal cord injury. Principal investigator: Guillermo Garcia-Alias. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 20000. Duration: 2016-2021.

Code: SAF 2014-59701. Title: Reprogramacion neuronal para promover los mecanismos endogenos de neuropro- teccion usando biologia sintetica en un modelo de degeneracion retrograda de motoneuronas. Principal investigator: Caty Casas. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Nacional. Funding agency: MICINN. Funding: 108900. Duration: 2015-2018.

Code: CA15138. Title: TRANSAUTOPHAGY: European network for multidisciplinar research and translation of autophagy knowledge. Principal investigator: Caty Casas. CIBERNED’s collaboration: No. CIBERNED groups: G607 . Other CIBER’s collaboration: No. Type: Europeo. Funding agency: Comision Europea. Funding: 520000. Duration: 2016-2020.

255 PHD DISSERTATIONS 2018 Author: Author: Natalia de la Oliva Muñoz. Title: Biological response to implanted intraneural electrodes. Date: 9/2/2018. Supervisor: Xavier Navarro Acebes.

Author: Anna Martínez Muriana. Title: Modulation of the inflammatory response in amyotrophic lateral sclerosis.. Date: 26/7/2018. Supervisor: Rubén López Vales.

Author: Tatiana Leiva Rodríguez. Title: Investigation of the role and modulation of autophagy for neuroprotection and nerve regeneration using models of peripheral nerve injury.. Date: 9/11/2018. Supervisor: Caty Casas Louzao.

256 CIBERNED 2018 ANNUAL REPORT

COOPERATIVE RESEARCH

257 258 CIBERNED 2018 ANNUAL REPORT

COOPERATIVE RESEARCH

The cooperative research program continues to be viewed as an essential scientific tool for planning CIBERNED future actions. This program is one of the cornerstones of the Center’s activities, and aims to encourage collaborative research between the various research groups, joining forces and exploiting synergies and complementary skills. It is intended to identify collaborative research projects with a marked innovative and translational nature where the cooperative effort significantly increase the added value of the research activity. Proposals, although subject to final approval by the Steering Committee, are externally reviewed by the National Evaluation Agency (ANEP), so that the allocation of funds is independent and transparent. The cooperative research program has so far launched nine calls for projects overall. The first four (2010, 2011, 2013 and 2014) has already included seventeen completed projects and a total accumulated budget allocation of 5,422,174 €. In 2015, the fifth and sixth calls were launched. The fifth call included three collaborative projects with a total allocation of 690,000€ and was finished by the end of 2017. The sixth call included two collaborative projects with a total allocation of 700,000€ which, due to an extension that did not involve additional funding, ended during 2018. In 2016, the seventh call for cooperative projects was approved with a total allocation of 1,016,000€. This seventh call included five projects that ended in late 2018. In 2017, the eighth call was launched with the concession of three new projects with a total allocation of 650,000€. Finally, during the last semester of 2018, the ninth call was launched with the concession of three additional projects with a budget allocation of 600,000€.

259 2010 CALL

CODE TITLE COORDINATOR Generating a dopaminergic neuronal model from induced PI2010/05 pluripotent stem cells from patients with Parkinson's disease Tolosa Sarró, Eduardo associated to mutations in the LRRK2 gene

García de Yébenes PI2010/06 Neuroprotection in Huntington's disease Prous, Justo

PI2010/07 Reelin and GSK3 as therapeutic targets in Alzheimer's disease Ávila de Grado, Jesús

Glial activation during the neuroinflammatory process: a potential Vitorica Ferrández, PI2010/08 therapeutic target for Alzheimer's disease Francisco Javier

BESAD-P: Biomarkers of early stages of Alzheimer disease - PI2010/09 Ferrer Abizanda, Isidro prevention

Consortium to generate a common database aimed at PI2010/11 implementing clinical and basic research on neuromuscular Illa Sendra, Isabel diseases

2011 CALL

CODE TITLE COORDINATOR

The Nrf2 transcription factor as a new therapeutic target for PI2011/01 Cuadrado Pastor, Antonio Parkinson's disease

Onset and progression of Parkinson's disease. Vulnerability Obeso Inchausti, José PI2011/02 of the nigrostriatal pathway, events at origin and destination Ángel

Generation of dopaminergic neurons from somatic cells of PI2011/03 Moratalla Villalba, Rosario parkinsonian patients with cognitive failure

Multicenter study on LCR and neuroimaging biomarkers in PI2011/04 the continuum preclinical- prodromal Alzheimer's disease Lleó Bisa, Alberto (SIGNAL Study)

2013 CALL

CODE TITLE COORDINATOR Emergent properties of the neuron-glia relationship PI2013/01 underlying neurodegeneration and dementia in Alzheimer's Torres Alemán, Ignacio disease

Identification and molecular characterization of cannabinoid PI2013/05 Guzmán Pastor, Manuel receptors subpopulations in poliglutaminopatías

Role of GSK-3β in the cortical circuits alterations occurring in PI2013/07 Iglesias Vacas, Teresa Alzheimer's disease

Mitochondrial dynamics and mitophagy as therapeutic PI2013/08 Soriano García, Eduardo targets in Parkinson's and Huntington's diseases

Degeneración sináptica y desregulación de la neurogénesis PI2013/09 Fernández Chacón, Rafael del adulto en modelos murinos de neurodegeneración

260 CIBERNED 2018 ANNUAL REPORT

2014 CALL

CODE TITLE COORDINATOR Epigenetic mechanisms involved in the etiology and PI2014/02 progression of rapidly progressive neurodegenerative Calero Lara, Miguel dementias

Onset and progression of Parkinson's disease: role of glial PI2014/06 Rodríguez Oroz, Mª Cruz activation

2015-I CALL

CODE TITLE COORDINATOR Neuregulin gene therapy aimed at the treatment of motor neuron Navarro Acebes, PI2015/01 degeneration in ALS Xavier

Validation of new therapeutic targets and biomarkers in Labandeira García, PI2015/02 Parkinson's disease José Luis

Fuentes Rodríguez, PI2015/03 Differential metabolic profiles in Parkinson's disease José Manuel

2015-II CALL

CODE TITLE COORDINATOR Pathological potential of astrocytes: a new perspective on Comella Carnicé, Joan PI2015-2/02 Alzheimer's disease Xavier

Molecular mechanisms of brain and muscle stem cell function PI2015-2/06 Muñoz Cánoves, Pura in aging and neurodegeneration

2016 CALL

CODE TITLE COORDINATOR Alterations of gluco-lipid metabolism and development of PI2016/01 Torres Alemán, Ignacio Alzheimer's dementia

Monitoring the Onset and Evolution of Neuronal Dysfunctions Del Río Fernández, PI2016/02 in Propagative Neural Disorders using Microfluidic Devices and José Antonio Translational approaches

López de Munain PI2016/04 The ALS CIBERNED Challenge: Accelerating New Drug Discovery Arregui, Adolfo

Naranjo Orovio, José PI2016/05 Dream inhibitors and Alzheimer´s Disease Ramón

Identification of pathophysiological pathways and candidate PI2016/06 Vila Bover, Miquel biomarkers in the prediagnostic phase of Parkinson’s disease

261 These twenty-seven projects, which have been already finished, have actively involved up to 57 CIBERNED research groups, as well as other external ones, representing 90.4% of the existing groups. During the year 2018, the sixth and seventh calls for CIBERNED cooperative projects were completed, and the groups involved presented the results obtained in the corresponding final reports that were delivered to the CIBERNED scientific management. Below is a brief description of the results obtained by each of the projects during these calls:

Project PI2015-2/02 Pathological potential of astrocytes: a new perspective on Alzheimer’s disease

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Vall d'Hebron University Hospital, Comella Carnicé, Joan Xavier Barcelona Vitorica Ferrández, Francisco Javier CIBERNED, University of Seville Gutiérrez Pérez, Antonia CIBERNED, University of Malaga Vicario Abejón, Carlos CIBERNED, Cajal Institute CSIC, Madrid Moratalla Villalba, Rosario CIBERNED, Cajal Institute CSIC, Madrid

Although astrocytes have well characterized roles related with trophic support, homeostasis, elimination of toxic molecules, production of cytokines, vascularization, extracellular matrix reshaping and control of synaptic activity, it is unknown their specific role in the development of AD. It has been characterized that astrocytes activate and accumulate around b-amyloid plaques, but it is unknown the detailed changes in their genetic expression patterns, or how their secreted factors contribute to the development of the disease. In this project, we have tried to go deeper in the knowledge of the role of astrocytes in the development of Alzheimer’s disease. To achieve this goal, we have started different experimental approaches to study the role of astrocytes in the development of the disease, such as the generation of iPS cells from fibroblasts and differentiated into astrocytes. We have also generated a model that should allow modifying in vivo the levels of neuronal FAIM-L, a protein that could play an interesting role in the protection of neurons against the disease, and also in the cross-talk neuron/glia. Our results show that in murine models, as well as in Alzheimer’s patients, reactive astrocytes associated to the amyloid plaques, but nor activated microglia, have phagocytic activity, selectively eliminating axonal and presynaptic distrophies. In addition, when analyzing samples from different stages of Alzheimer’s disease, we observed that this phagocytic component is altered in the development of the disease, thus constituting a likely therapeutic target for the disease, since the functional impairment of astrocytes seems a key component in the progression of Alzheimer’s disease. Regarding the in vivo models of FAIM, we have characterized that the FAIM KO mouse has an epileptic phenotype, with recurrent attacks induced by manipulation, which are age-dependent, and characterized by electrophysiological alterations, locomotor hyperexcitability, deficits in social interaction, and an altered behavior in nest building, as well as some general deficits in learning and memory. We have additionally

262 CIBERNED 2018 ANNUAL REPORT generated adeno-associated viruses which will allow us to explore if neuronal altered expression levels of FAIM may induce changes in the development of the disease in different murine models. We have also explored the effects of ApoE polymorphisms, which clearly induce alterations in cholesterol homeostasis and autophagy, alterations that are systemic, because do not only affect neurons, but also peripheral cells, pointing to higher levels of cholesterol esters in patients fibroblasts as an useful metabolic trait to further explore in cellular models of the disease. In addition, fibroblasts from AD patients show dramatic alterations in lysosomes and mitochondria.

Project PI2015-2/06: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Pompeu Fabra University, ICREA, Muñoz Cánoves, Pura Barcelona Fariñas Gómez, Isabel CIBERNED, University of Valencia Center for Molecular Biology "Severo Ochoa" Lucas Lozano, José Javier CSIC-UAM, Madrid Fernández Chacón, Rafael CIBERNED, University of Seville CIBERNED, Institute of Biomedical Research Iglesias Vacas, Teresa CSIC-UAM, Madrid

The functional decline of the Nervous and the musculoskeletal systems is a major factor determining the quality of life in elderly populations. Neural and muscular stem cells must remain active throughout life for the functional maintenance of both tissues. However, these cells are sensitive to aging and, possibly to the pathogenic factors that cause neurodegeneration as well, thus progressively losing their regenerative potential by unknown molecular mechanisms. Our consortium has identified possible mechanisms that would affect both neural and muscular stem cells as a result of both aging and neurodegeneration. These include: (1) the aging-dependent inhibition of autophagy In stem cells mediated by target-of-rapamycin (mTOR) signaling pathway; (2) the premature exhaustion of stem cell pools due to cell cycle and cell division mode dysregulation; and (3) stem cell dysfunctions due to alterations in alternative splicing such as those occurring in neurodegenerative diseases associated to polyglutamine repeats. The coordinator of the consortium Dr. Munoz-Cánoves has recently described key molecular pathways underlying the functional decline and senescence of satellite cells that impair muscle rejuvenation (Sousa-Victor, 2014). The transcriptomic analysis included in this study also points to the mTOR signaling pathway as an additional mechanism as well as to three key genes that are the central subject of study in the consortium laboratories: (a) Cysteine String Protein-alpha (CSP-alpha), a synaptic co-chaperone that prevents presynaptic degeneration (Garcia-Junco et. al, 2010) that, unexpectedly, maintains the postnatal quiescence of neural stem cells through inhibition of mTOR (unpublished work from Dr. Fernandez-Chacon’s group); (b) Kinase D interacting substrate (Kidins220), a major effector of multiple receptor signaling pathways regulating cellular responses in nervous, vascular and skeletal muscle systems (Iglesias, 2000; Neubrand, 2012) present in neural stem cells of the subventricular

263 zone (SVZ) and regulates them (unpublished data from the collaboration between the groups Dr. Teresa Iglesias and Dr. Isabel Fariñas); (c) Rbfox1, a specific splicing factor from neurons and skeletal muscle previously linked to myotonic dystrophy DM1 and that is decreased in Huntington’s disease, resulting in a pathogenic splicing of CPEB4 (unpublished work by Dr. José Lucas’s group ). Although satellite cells are the main focus of the study, the SVZ neural stem cells will be studied in parallel, easier to approach methodologically and better known thanks to the pioneering work of Dr. Isabel Fariñas. Our consortium brings together groups with ample expertise in the biology of stem cells and neural development, the role of satellite cells in muscle fiber maintenance and the molecular basis of neurodegenerative diseases and synaptic dysfunction. Our laboratories will synergistically interact to maximize multidisciplinary methodological resources and expertise which include a highly specialized knowledge in the cell biology of neural stem and satellite cells, yet-unpublished biological resources, i.e. mouse models, and first-hand understanding of specific signaling molecules and pathways. We expect that our project will throw light on key mechanisms involved in the functional maintenance of adult stem cells, an essential point for the potential regeneration of muscle fibers and specific neuronal types.

Project PI2016/01: Alterations of gluco-lipid metabolism and development of Alzheimer’s dementia

PRINCIPAL INVESTIGATOR INSTITUTION

Torres Alemán, Ignacio CIBERNED, Cajal Institute CSIC, Madrid CIBERNED, Center for Molecular Biology Wandosell Jurado, Francisco "Severo Ochoa" CSIC-UAM, Madrid Camins Espuny, Antonio CIBERNED, University of Barcelona CIBERNED, Doce de Octubre University Carro Díaz, Eva Hospital, Madrid Cantero Lorente, José Luis CIBERNED, Pablo de Olavide University, Sevilla

The aim of the Project was to determine along time, possible metabolic disturbances associated to progressive cognitive alterations in Alzheimer´s disease (AD) patients. We carried out a parallel study in AD patients and animal models. We first analyzed the serum glyco-lipid pattern in an AD cohort encompassing the entire natural history of the disease. Brain imaging analysis in the same patients was conducted to obtain structural/functional data potentially associated with peripheral metabolism that could ideally characterize each of the stages of the disease. The results show no relationship between changes in glyco-lipid metabolism and structural/functional alterations seen in the brain along the different stages of AD, at least in our patient population. However, we observed a correlation between serum homocysteine and oxidative stress, seemingly mediated by sleep quality, in mild cognitive impairment (MCI) patients. Thus, we propose a multi-factorial model to predict cognitive deterioration based in the relationship between adiposity and quality of sleep. Further, and emphasizing the known sexual dimorphism of the disease, we found that cholinergic dennervation in temporal cortex from the basal nuclei of Meynert is more pronounced in MCI women than in men. In asymptomatic subjects we observed changes in diverse miRNAs that predict cerebral atrophy together with a decrease in brain metabolism, suggesting that affected miRNAs may serve as biomarkers of brain vulnerability dur-

264 CIBERNED 2018 ANNUAL REPORT ing aging. We have also determined, in a parallel study carried out in an additional AD cohort, that serum lactoferrin, an immunomodulator regulated by metabolic status, may constitute a new peripheral biomarker. In AD animal models, we examined how metabolic disturbances associated to high fat diet or to brain glucose alterations (typically found in AD patients) impact in the development of the disease. We determined amyloidosis, tauopathy, lipid disturbances and insulin resistance. We found a profound cognitive loss in obese JNK2-/- . In addition, we also found a relationship between obesity, the JNK1 isoform, cognitive improvement, and increased synapse numbers. Our results indicate that this isoform, together with inhibitory drugs, would potentiate cognitive processes. We also obtained the lipid profile of APP/PS1 mice and their controls after submitting them to a high fat diet. We used diets with n-6/n-3 fatty acids in intact, ovariectomized, and ovariectomized plus estradiol female mice. In these groups we have determined diverse neural markers. In megalin-deficient mice showing cognitive deterioration, obesity, and neuroinflammation, we analyzed the relationship between altered metabolism, amyloid levels and cognitive loss. Specifically, we determined whether metabolic disturbances are sufficient to trigger cognitive loss or whether the underlying amyloidosis is essential. In a fourth type of experiments, we analyzed the influence of altered glucose metabolism in the development of AD (using APP/PS1 mice lacking either insulin or IGF-I receptors in astrocytes) and found in both cases changes in disease progress. Furthermore, we observed dysfunctional brain perfusion and abnormal angiogenesis by merely deleting the insulin receptor in astrocytes, which leads to brain insulin resistance. Collectively, our results indicate that glyco-lipid metabolism plays a role in development of the pathology in animal models. However, the results obtained in AD patients make us to be cautious, even though a potential relationship could be envisaged. Indeed, more human studies are required.

Project PI2016/02: Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Catalonian Institute de Bioengi- Del Río Fernández, José Antonio neering, Barcelona CIBERNED, Center for Molecular Biology "Seve- Ávila de Grado, Jesús ro Ochoa" CSIC-UAM, Madrid Canela Campos, Enric Isidre CIBERNED, University of Barcelona

Numerous aged-related neurodegenerative diseases (END) are characterized by the slow but inexorable advance of neuronal dysfunctions in brain patients that progress though anatomically linked regions. This evolution of the disease shared with the progressive accumulation of abnormally folded proteins in affected neurons that stem for gradual neuronal dysfunction, which involves synaptic transmission abnormalities, axonal damage and cell death. However, in some cases the observed spread though brain parenchyma failed to follow the “classical” progression illustrated in seminar studies, e.g., in Alzheimer´s (AD) or Parkinson´s (PD) diseases. For example, some elderly individ-

265 uals without diagnostics showed Lewy bodies in the olfactory bulb, amygdala or cortex which should seem to violate the theory of progression and perhaps fit better with a multicentric disease process. In addition, anatomically connected regions of the brain (e.g., entorhino-hippocampal formation) showed different p-tau burden in AD and other tauphaties. In conclusion, these data suggested both an area-specific and a differential neuronal vulnerability during disease progression. Factor/s modulating this differential neuronal vulnerability are unknown. The goal of the present project is to recreate in vitro using microfluidic devices (MFDs) normal and diseased neuronal networks of selected neural pathways susceptible to suffer neurodegeneration by using mice disease models or differentiated human pluripotent stem cells. We would like to focus on α-synuclein and Tau. In particular, MFDs in our project will allows us for i) the recreation of specific neuronal networks of neuroanatomical relevance in neurodegenerative diseases in vitro; ii) to investigate factors mediating the propagation of the neurodegenerative process in models of ND and iii) to monitor early/late physiological changes mediated by amyloid proteins. Due that α-synuclein and Tau behave as “prion-like” elements we will study in our culture platforms whether crucial neural functions (e.g., neuronal networks dynamics) or the onset and progression of neuronal dysfunctions appeared during the spreading of deleterious signals triggered by these “prion-like” elements in order to understand their propagation and neuron specific effects in vivo in parallel experiments.

Project PI2016/04: The ALS CIBERNED Challenge: Accelerating New Drug Discovery

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Biodonostia Research Institute, López de Munain Arregui, Adolfo San Sebastian CIBERNED, Bellvitge Institute of Biomedical Ferrer Abizanda, Isidro Research, Barcelona Fernández Ruiz, Javier CIBERNED, Complutense University of Madrid Soriano García, Eduardo CIBERNED, University of Barcelona

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive weakness and atrophy of skeletal muscles until paralysis and death. Mul- ti-factorial and multi-systemic neurotoxic mechanisms are the cause of a common pathological entity: motor neuron (MN) degeneration in central nervous system (CNS). The disease may spread to frontal cortex and other regions leading to frontotemporal lobar degeneration (FTLD) and dementia. The enormous heterogeneity in the patterns of clinical symptoms, anatomical onset, disease progression and survival among patients suggests that pathogenic pathways for MN degeneration are variable at topo- graphical and pathophysiological levels. Moreover, involvement of the frontotemporal cortex in ALS and the motor system in FTLD-TDP-43 links ALS and FTLD-TDP-43 within the spectrum of TDP-43 proteinopathies. This complex scenario for ALS would explain the unfavorable outcomes collected in clinical trials.

Future research must consider:

1) Identification of clinico-pathological markers in brain, CSF and blood specific of ALS subtypes, which will allow the design of individual intervention treatments. 2) Identification of new altered molecular pathways in different CNS regions to unveil

266 CIBERNED 2018 ANNUAL REPORT

common and specific profiles of regional vulnerability. 3) Identification of converging pathways of MN and frontal cortex degeneration, which will allow the generation of neuroprotective strategies to halt MN degeneration. The present collaborative project integrates 4 CIBERNED clinical/basic research groups (Adolfo López-de-Munain, Isidro Ferrer, Eduardo Soriano and Javier Fernández-Ruiz), with the purpose to address these 3 hot points in ALS/FTLD-TDP43 complex. Collabora- tive efforts will be extended to other CIBERNED groups. The first part of the project will create a unique Spanish ALS cohort by unifying several Spanish individual registries with standardization of clinical data and methods of biological sample collection. Large ALS cohort provides necessary potency to achieve the following milestones: a) Identification of novel clinical and molecular signatures categorizing different ALS subtypes. b) Identification of new altered molecular pathways in the motor system and frontal cortex in post-mortem brains of ALS (in comparison with FTLD-TDP-43). c) Discovery of novel biomarkers in brain, CSF, blood and striate muscle for diagnosis and prognosis. d) Drawing a pathophysiological map of the disease to detect potential targets for treatment. These objectives will be accomplished by carrying out combined ‘omics and computational integrative analyses in samples from alive ALS patients and post-mortem tissues. The second part of the project is aimed at exploring neuroprotective targets following the hypothesis of a crosstalk between TDP-43 proteinopathy and mitochondrial defects as a common pathogenic pathway to MN degeneration (and probably frontal/temporal cortex degeneration. Three main points will be analyzed: a) Efficacy of patented compounds as novel inhibitors of RyR Ca2+ channel as a way to protect neuromuscular system from cytosolic Ca2+ overload induced by pathological TDP-43-mediated mitochondria defects. b) Study of pharmacological targets within the endocannabinoid system against TDP-43-mediated mitochondria defects. c) The role of Eutherian Armcx proteins on mitochondrial dynamics in relation to TDP-43 pathology as a promising target for MN neuroprotection. Both parts of the project are interconnected and will be carried out by the collaborative work of the four group members of the proposal.

267 Project PI2016/05: DREAM inhibitors and Alzheimer’s disease

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, National Center of Biotechnology, Naranjo Orovio, José Ramón CSIC, Madrid CIBERNED, Cajal Institute CSIC; Technical Uni- De Felipe Oroquieta, Javier versity, Madrid CIBERNED, University of Castilla La Mancha, Ceña Callejo, Valentín Albacete

Deregulated protein and Ca2+ homeostasis underlie synaptic dysfunction and neuronal death in neurodegenerative diseases including Huntington’s (HD) and Alzheimer’s disease (AD). Early symptoms of synaptic dysfunction in AD are characterized by a reduced response to external stimuli and a progressive loss of post-synaptic dendritic spines in excitatory synapses, which correlates strongly with AD symptoms and memory loss. Previous to spine loss, atrophy of the spine apparatus, a synaptopodin-positive ER-associated membranous structure, is particularly relevant since the spine apparatus has an essential role in regulating the fate of calcium entering the spine upon synaptic activation and could be considered the structural base for synaptic plasticity. Dendritic failure in AD is also largely related to the accumulation of pathogenic protein aggregates that have been related to a defective unfolded protein response (UPR) and to changes in actin polymerization as a result of increased cofilin de-phosphorylation in AD. DREAM, also known as calsenilin or KChIP3, is a Ca2+ binding protein that regulates Ca2+ homeostasis and neuronal survival through transcriptional control of target genes and through protein-protein interactions. DREAM was originally associated with AD because of its interaction with presenilins. Preliminary results from our group show changes in the expression of DREAM in cerebral cortex and hippocampus in J20 and Tg2576 mice, two murine models of AD, as well as in frontal cortex from AD patients. Genetic manipulation of DREAM levels or chronic repaglinide treatment modified cognition in Tg2576 and J20 mice. These data support the idea that, like in HD, an early down regulation of DREAM level in neurons during the pre-symptomatic phase of the AD, is part of a neuroprotective mechanism and suggest that DREAM could be a novel and wide spectrum target for therapeutic intervention in AD. The aim of this collaborative research program is to investigate whether cognition improvement in AD mouse models after repaglinide restores the early response to neuronal activation (induction of immediate-early genes Npas4, Nr4a1 and Arc) and is related to changes in synaptopodin-positive dendritic spines, in cofilin dephosphorylation and/or in increased ATF6 processing and UPR activation in hippocampal neurons.

268 CIBERNED 2018 ANNUAL REPORT

Project PI2016/06: Identification of pathophysiological pathways and candidate biomarkers in the prediagnostic phase of Parkinson’s disease

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Vall d'Hebron University Hospital, Vila Bover, Miquel Barcelona Tolosa Sarró, Eduardo CIBERNED, Hospital Clinic of Barcelona CIBERNED, Institute of Biomedical Research Trullas Oliva, Ramón IDIBAPS-CSIC , Barcelona CIBERNED, Marqués de Valdecilla Research Infante Ceberio, Jon Institute, Santander

Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common genetic cause of Parkinson’s disease (PD). Like idiopathic PD (iPD), it is believed that the onset of PD associated with mutations in LRRK2 (LRRK2-PD) occurs before the onset of motor symptoms, but information on the prodromal phase is scarce. The subjects with mutations in LRRK2 who do not show motor symptoms (LRRK2-NMC), but who present a higher risk of developing LRRK2-PD, represent a unique opportunity to investigate the early pathophysiology of LRRK2-PD and to identify molecular profiles that could be candidate biomarkers associated with the prodromal and / or manifest phases of LRRK2-PD. To address these fundamental issues, we have used four different and complementary approaches to analyze the biological samples of selected patients. In the first year of this collaborative project we carried out the first discovery phase, in which we studied LRRK2- NMC subjects (subdivided according to whether they present normal / abnormal DAT- SPECT), LRRK2-PD patients, iPD patients and the healthy controls. In these five groups, we have studied the metabolic profiles in serum and we have characterized the epigenome in peripheral blood to determine the metabolic and epigenomic changes that occur in the prodromal and manifest phases of LRRK2-PD. In addition, we have studied the number of copies of mitochondrial DNA (mtDNA) and its release in fibroblasts derived from patients. Our data show a significant alteration in the lipid metabolism of subjects with mutations in LRRK2 and point to certain metabolites as potential biomarkers of PD status. We have also identified significant epigenetic changes in the peripheral blood of patients with overt PD in incipient stages of motor disease. Finally, the results obtained up to now in fibroblasts indicate that the alteration of mtDNA replication and transcription is a common factor underlying idiopathic and familial PD, supporting the hypothesis that transcription and replication are alternative processes and that its regulation is altered in PD. To conclude, up to now we have identified some candidate biomarkers of different nature that may be useful as biomarkers of the prediagnostic phase of PD. Throughout the second year of this collaborative project we expect to complement the studies of the discovery phase, as well as to use an integrative approach using systems biology for the identification of the physiopathological pathways and the molecular profiles associated with the prodromal and manifest phases of LRRK2-PD. We will also carry out the validation phase of those markers of greatest interest identified in the first phase of discovery. Finally, during the second semester of 2017, the eighth call was launched, which was again evaluated by the ANEP and resolved at the end of that same year with the granting of three new projects:

269 2017 CALL

CODE TITLE COORDINATOR Study of the microRNA in the cerebrospinal fluid exosomal Clarimón Echavarria, PI2017/01 compartment as a biomarker of frontotemporal dementia and a Jordi tool for understanding the biological basis of the disease.

Lanciego Pérez, José PI2017/02 Glucocerebrosidase and Neurodegenerative Proteinopathies Luis

Glial dysfunction in Alzheimer's disease: pathogenic implications Vitoríca Fernández, PI2017/04 and clinical potential Francisco Javier

The funding approved for this eighth call was 325,000€ per year for 2 years, which meant a total allocation of 650,000€ to the eleven participating CIBERNED research groups. Be- low is a brief description of the scientific activity carried out by each of the projects during their first year of execution:

Project PI2017/1: Study of the microRNA in the cerebrospinal fluid exosomal compartment as a biomarker of frontotemporal dementia and a tool for understanding the biological basis of the disease.

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Santa Creu i Sant Pau Hospital, Clarimón Echavarria, Jordi Barcelona CIBERNED, Autonomous University of Barce- Rodríguez Álvarez, José lona CIBERNED, Autonomous Universidad of Bullido Gómez-Heras, Mª Jesús Madrid

Frontotemporal degeneration (FTD) is a clinical, genetic and pathologically heterogeneous neurodegenerative disease, characterized by atrophy of the frontal and temporal lobes, resulting in a progressive alteration of behavior and/or language. Neuropatholog- ical studies indicate that 50% of cases present brain aggregates of the TDP43 protein (FTD-TDP), which has a pivotal function in the RNA metabolism. There is no specific biomarker for FTD, nor any that could distinguish between patients with DFT-TDP from the rest (typically with Tau: FTD-Tau). Exosomes are vesicles of 50-100nm in diameter that act as intermediaries in intercellular communication and can alter the transcriptional metabolism of the recipient cells through the microRNAs (miRNA) contained within. Due to their presence in most of the body fluids, exosomes have become a novel and attractive source for the study of biomarkers in peripheral tissues. Preliminary results obtained by the PI’S group have allowed us to elucidate for the first time part of the miRNA content that is found in the exosomal compartment of the cerebrospinal fluid (CSF; see Annex). From a large panel containing 752 different miRNAs we were able to identify 130 that are stably expressed in the CSF. Our analysis has elucidated differences in some of these microRNAs among patients with the semantic variant of DFT (characterized by presenting aggregates of TDP43) and healthy controls. Objective: The present project intends to examine (i) the impact of candidate miRNAs on global gene expression pattern, (ii) the consequences of these miRNAs on synaptic function and neuronal survival and (iii) their sensitivity and specificity as a diagnostic biomarker for FTD and / or the FTD-TDP

270 CIBERNED 2018 ANNUAL REPORT subtype. Design: To analyze the sensitivity and specificity of these miRNAs as biomarkers of DFT, as well as their ability to distinguish the DFT-TDP variant of DFT-Tau, a total of 144 subjects have been included (30 controls, 20 with Alzheimer’s disease, 28 with FTD (behavioral variant), 32 with semantic dementia and 34 with PSP / DCB). Results: The study has shown 4 species of miRNAs that are differentially expressed in the subgroup of patients with PSP / DCB. These are miR-146, miR-15, miR-361 and miR-708. Conclusions: The conclusions of this first phase of the study are that there are expression profiles of miRNAs in the exosomal CSF compartment that could be used as possible biomarkers for the in vivo diagnosis of the pathological subtype of the PSP / DCB group, within the spectrum of the DFT. During the second half of the study (second year) the functional / mechanistic studies of these 4 miRNAs will be carried out, from neuronal cultures in which the levels of the miRNAs of interest will be modulated, analyzing the expression pattern by means of high density microarrays, neuronal viability and functionality and synaptic density, as well as its role in different autophagy functions.

Proyecto PI2017/2: Glucocerebrosidase and Neurodegenerative Proteinopathies.

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Center of Applied Medical Re- Lanciego Pérez, José Luis search, Univ. Navarra, Pamplona CIBERNED, University of Santiago de Com- Labandeira García, José Luis postela CIBERNED, Santa Creu i Sant Pau Hospital, Kulisevsky Bojarski, Jaime Barcelona

It has only been recently uncovered a link between GBA 1 mutations (the gene coding for glucocerebrosidase) and the development of synucleinopathies such as Parkinson’s disease (PD) and dementia with Lewy bodies (DLB). Studies performed on patients suffering from Gaucher disease (a lysosomal storage disorder caused by GBA1 mutations) revealed that GBA1 mutations are the most common genetic risk for PD and DLB. PD patients carrying GBA1 mutations develop PD ealier than non-carriers and are more frequently affected by cognitive decline and neuropsychiatric symptoms, as shown in studies performed by clinical researchers from the applicant team. Furthermore, it is worth noting that the aggregation of alpha-synuclein (SYN) per se induces a GCase loss-of-function. In other words, both types of PD patients (e.g., with and without GBA1 mutations) are indeed susceptible of being treated with GCase. This also applies to patients suffering from dementia with Lewy bodies. Moreover, how exactly the aggregation, degradation and subcellular processing of SYN and GCase activity are coupled together –and vice versa– still is an open question with limited experimental evidence to date. Preliminary studies carried out in our laboratory showed that neurons from the nucleus basalis of Meynert, substantia nigra pars compacta and locus ceruleus are the ones showing the highest GCase baseline levels of expression than any other brain area. Finally, it is also well known that the earliest neurodegenerative phenomena in Alzheimer’s disease (AD) were typically found in cholinergic neurons from the basal forebrain. Accordingly, the main aims of this project are summarized as follows: 1. To evaluate whether a viral vector-mediated increase of GCase activity in the substantia nigra could slow down/stop the progressive SYN-induced dopaminergic neurodegeneration.

271 2. To evaluate whether a viral vector-mediated increase of GCase activity in the nucleus basalis of Meynert could slow down/stop the progressive Tau-induced cholinergic neurodegeneration. 3. To disclose whether patients with mild cognitive impairment (MCI) carrying GBA1 mutations are progressing towards AD quicker than non-carriers. Neurospecific adenoassociated viral vectors (AAVs) will be used to overexpress either mutated SYN or Tau proteins into the substantia nigra pars compacta or in the nucleus basalis of Meyert, respectively. Experiments will be carried our in rats (AAV9-SynA53T) and in NHPs (AAV9 SynA53T and AAV9-Tau301L). Next, a GBA1-coding AAV5 will be used to increase GCase activity to further induce SYN and Tau clearance. In parallel, genetic studies will be conducted looking for a potential association between GBA1 mutations and the incidence of AD in patients. Besides the strong translational focus of this project, it is expected that the experiments described here will provide new clues elucidating the crossroads between GCase and SYN homeostasis as well as in tau-related neurodegenerative proteinopathies.

Project PI2017/4: Glial dysfunction in Alzheimer’s disease: pathogenic implications and clinical potential.

PRINCIPAL INVESTIGATOR INSTITUTION

Vitoríca Ferrández, Francisco Javier CIBERNED, Universidad of Seville Gutiérrez Pérez, Antonia CIBERNED, Universidad of Malaga Cuadrado Pastor, Antonio CIBERNED, Autonomous Universidad, Madrid Sáez Valero, Javier CIBERNED, Miguel Hernández University, Elche CIBERNED, Vall d'Hebron University Hospital, Comella Carnice, Joan Xavier Barcelona

In the present project, we propose a new pathogenic scenario in Alzheimer’s disease, where microglial dysfunction would suppose an astroglial hyperactivation. This hyperactivation could be involved in the neurodegenerative process and, therefore, it would be necessary to reproduce this dysregulation of the innate brain immune system, in animal models, to increase its predictive value and ensure greater success of human therapies. To emulate this microglial dysfunctionality, we are developing genetic models that allow us to reduce the number of microglia in Abeta (APPsl) or Tau (TauP301S) models. In the same way, we are also developing a genetic model that allows the elimination of active microglia. With these models, we hope to determine the microglial role on the pathology and, in parallel, their impact on astroglial activation. The study of the possible microglial and / or astroglial “signature” in cerebrospinal fluid (CSF) collections of subjects with amnestic-type mild cognitive impairment (prodromal stages), to Alzheimer’s subjects characterized with classic CSF markers has also stated during this year.

Finalmente, durante el último semestre de 2018, se lanzó la novena convocatoria que fue de nuevo evaluada por la ANEP y resuelta a finales de ese mismo año con la concesión de 3 nuevos proyectos:

272 CIBERNED 2018 ANNUAL REPORT

2018 CALL

CODE TITLE COORDINATOR CB1R-GRP78 interaction: a new mechanism regulating the Guzman Pastor, PI2018/01 neuroprotective activity of cannabinoids ? Manuel

Cellular and molecular analysis of seeding and progression of tau PI2018/02 Ferrer Abizanda, Isidro in animal and cell models of different human tauopathies

Targeting CPEB-dependent impaired mitochondrial metabolism Lucas Lozano, José PI2018/06 and synaptic and stem cell function in Huntingtons disease Javier

The funding approved for this eighth call was 300,000€ per year for 2 years, which meant a total allocation of 600,000€ to the twelve participating CIBERNED research groups. Be- low is a brief description of the scientific activity carried out by each of the projects during their first year of execution:

Project PI2018/01: CB1R-GRP78 interaction: a new mechanism regulating the neuroprotective activity of cannabinoids ?.

PRINCIPAL INVESTIGATOR INSTITUTION

Guzman Pastor, Manuel CIBERNED, Complutense University of Madrid Ginés Padrós, Silvia CIBERNED, University of Barcelona Fernández Ruiz, Javier CIBERNED, Complutense University of Madrid CIBERNED, Institute of Biomedical Research Mengod Los Arcos, Guadalupe IDIBAPS-CSIC , Barcelona

Understanding the processes of neuron survival/death is a pivotal issue for characterizing the aetiology and progression of neurodegenerative diseases and, therefore, for de- signing rational therapies for their treatment. In this context, since CIBERNED was funded, groups in the present consortium have been collaborating in the study of how type-1 cannabinoid receptor (CB1R), the main molecular target of endocannabinoids and cannabis active components, confers neuroprotection in preclinical models of neurodegeneration. Of note, two cannabinoid-based medicines have already been approved by EMA to palliate symptoms of neurodegenerative diseases as spasticity in multiple sclerosis (Sa- tivex®) and seizures in refractory epilepsies (Epidiolex®). However, the assessment of the physiopathological relevance and therapeutic potential of CB1R-evoked neuroprotection is still hampered, at least in part, by the lack of knowledge on the neuron-subpopulation selectivity of CB1R action. CB1R action may be modulated in different manners, being conceivably one of them its association to intracellular proteins through its large cytoplasmic C-terminal domain. Recently, we have purified the CB1R C-terminal domain and have conducted wide-scale proteomic analyses and yeast two-hybrid experiments aimed at finding receptor interactors. The combination of these two approaches has rendered a list of potential CB1R-interacting proteins, among which there is one that clearly stands out: glucose-regulated protein 78 (GRP78/BiP/Hspa5). Hence, the general objective of this project is to characterize in detail the neurobiological and neuropathological role of GRP78 as a potential CB1R interactor. Our initial hypothesis is that CB1R, through its C-terminal domain, is capable of binding GRP78 in a spatiotemporally-selective manner; this modulates CB1R-mediated signalling on specific neuron subpopulations; and this, in

273 turn, tunes CB1R neuroprotective action. We will appraise this issue through the following specific objectives: 1. Characterizing CB1R-GRP78 interaction in cultured cells. 2. Characterizing CB1R-GRP78 interaction in mouse brain. 3. Defining the functional relevance of CB1R-GRP78 interaction in mouse models of neurodegeneration. 4. Mapping CB1R-GRP78 interaction in post-mortem brain samples from human neurodegenerative diseases. These studies, which will put together the experience and complementary abilities of the 4 groups in the consortium, will provide a better understanding of the contextual features of CB1R-coupled neuronal signalling, which may thus contribute to (i) developing more selective neuroprotective strategies targeting the endocannabinoid system, and (ii) gaining deeper insight into unwanted effects of cannabinoid-based medicines used in the management of neurodegerative diseases.

274 CIBERNED 2018 ANNUAL REPORT

Project PI2018/02: Cellular and molecular analysis of seeding and progression of tau in animal and cell models of different human tauopathies.

PRINCIPAL INVESTIGATOR INSTITUCIÓN

CIBERNED, Bellvitge Institute of Biomedical Ferrer Abizanda, Isidro Research, Barcelona CIBERNED, Catalonian Institute de Bioengi- Del Río Fernández, José Antonio neering, Barcelona CIBERNED, Center for Molecular Biology "Seve- Ávila de Grado, Jesús ro Ochoa" CSIC-UAM, Madrid

Tauopathies are clinically, biochemically and anatomically heterogeneous neurodegenerative diseases, identified by the deposit of excessively phosphorylated tau protein, abnormally folded and forming aggregates in neurons, astrocytes and oligodendrocytes with particular characteristics for each disease. Neurons and glial cells have an active and interactive role in the pathogenesis. These diseases have a progressive character with selective affectation that extends over time to the entire brain and manifests itself with certain neurological deficits once a threshold of damage has been reached. The present project has the following objectives: Objective 1: To determine specific disease differences in the seeding and spreading of tau species using microfluidic devices with primary cultures of neurons, astrocytes and mixed, with different species of tau derived from human taupathies, as well as organotypic cultures with barriers. In a second phase, neural or mixed cultures derived from iPSc will be used. The neurons or glial cells cultured from I) wild mice (C57BL6 / J), II) mutant mice (P301S) or iii) iPSc, will be treated with sarkosil-insoluble fractions from human brain homogenates or with recombinant tau; Objective 2: To determine the role of microglia cells in the propagation of pathological tau. Specifically, to study potential receptors and mechanisms involved in the propagation of tau in these cells; Objective 3: To determine the specific differences in seeding and spreading of tau pathology after intracerebral inoculations (hippocampus) in wild-type mice of tau-enriched homogenates from pure and combined neuronal and glial human taupathies that accumulate 3Rtau or 4Rtau or 4Rtau + 3Rtau: Prima- ry age-related tauopathy (PART), Aging related tau astrogliopathy (ARTAG), globular glial tauopathy (GGT), Pick disease (PiD), Argyrophilic grain disease (AGD) and Gerst- mann-Straüssler-Scheinker syndrome (GSS). Assessment of the involvement of neurons and glial cells as effectors and as targets of taupathy; Objective 4: To analyze in vitro, possible changes in the activity of neural networks between the interconnected areas subjected to tau seeding, and to quantify the variations of the individual neuronal activity and the general alterations of the network. We will use genetically-encoded calcium indicators to monitor the evolution of the network over several days. In addition, we will modulate the neuronal activity of the network by using in vitro optogenetic techniques. The data of spontaneous activity at cellular resolution will be analyzed using advanced and customized software NETCAL or FLUOSNNAP. Ca2+ waves, spontaneous activity frequency, activity patterns that cover the network (network bursts) and their alteration during the process of seeding and spreading will be investigated. Then, theo- retical information algorithms will be used among all the pairs of neurons registered to identify functional alterations of the network; and Objective 5: To determine the effect on seeding and spreading after combined inoculation in wild mice of taupathy homogenates (PART, ARTAG) and α-synucleinopathy (Lewy body disease: LBD and multisystem atrophy: MSA); and use of inoculates in P301S and snca transgenic mice, for analysis of co-morbidity which is common in human pathology above sixty-five years.

275 Project PI2018/06: Targeting CPEB-dependent impaired mitochondrial metabolism and synaptic and stem cell function in Huntingtons disease.

PRINCIPAL INVESTIGATOR INSTITUTION

CIBERNED, Center for Molecular Biology "Seve- Lucas Lozano, José Javier ro Ochoa" CSIC-UAM, Madrid CIBERNED, Institute of Biomedical Research Iglesias Vacas, Teresa CSIC-UAM, Madrid CIBERNED, Pompeu Fabra University, ICREA, Muñoz Cánoves, Pura Barcelona Fernández Chacón, Rafael CIBERNED, University of Seville Fariñas Gómez, Isabel CIBERNED, University of Valencia CIBERNED, Virgen del Rocío University Hospi- Mir Rivera, Pablo tal, University of Seville

LHuntington’s disease (HD) and multiple inherited spinocerebellar ataxias are caused by polyQ-encoding expanded CAG repeats in different genes. Cytoplasmic Polyadeny- lation Element Binding Proteins (CPEB1-4) regulate translation of specific mRNAs by modulating the length of their poly(A)-tail. Interestingly, the CPEB Drosophila ortho- logue Orb2 was identified in an enhancer/suppressor screening as one of the few genes that modify both CAG mRNA- and polyQ-induced toxicity. However, a role of CPEBs in neurodegenerative disorders had not been fully explored until we recently analyzed the status of CPEBs in brain of HD patients and mouse models. We found increased CPEB1 and decreased CPEB4 levels (Parras et al., manuscript in preparation). Since neuronal mitochondrial function depends on proper CPEB1 mediated translation and CPEB1/CPEB4 crosstalk plays a key role in mitotic cell cycle progression, here we hypothesize that imbalanced CPEBs in HD leads to altered mitochondrial function and defective tissue homeostasis due to impaired stem cell function. In support of this hypothesis, we have recently reported that 9% of the transcripts in HD mouse brains are aberrantly deadeny- lated (Parras et al. Nature in press) and these include genes that are essential for mitochondrial function such as SLC19A3, several NDUFVs, and PRKD1, as well as CPEB4 itself -which controls FoxO3/4 translation- and is therefore expected to affect proper renewal of both neural and muscle stem cells. In this collaborative proposal we take advantage of the unique expertise of the participants to tackle specific aims that require multidisciplinary approaches. In the first objective, we aim to target mitochondrial deficit by: a) characterizing the role of the thiamine transporter SLC19A3 (hTHTR2) in HD pathogenesis and b) promoting a clinical trial of vitamin supplementation in collaboration with Dr. Pablo Mir at Hosp. Virgen del Rocío. In the second objective we will further characterize the effect of decreased expression of PRKD1, a kinase that preserves mitochondrial function and potentiates the elimination of mitochondrial reactive oxygen species (ROS) and neuronal survival in an excitotoxic environment as we have recently reported (Pose-Utrilla et al. Nat. Commun. 2017). We will also perform rescue experiments with AAVs for the neurospecific expression of a neuroprotective form of the kinase in HD models where synaptic activity will also be evaluated. Finally, in the third objective we will explore whether CPEB4 modulation may be used to enhance adult neural and muscle stem cell function as we have evidence to believe that the role of CPEBs in the division of adult neural stem cells (NSCs) from subventricular or subependymal zone (SVZ/SEZ) can be key to the maintenance and potential of these cells in normal and HD brains. On the other hand, regarding muscle stem cells (MuSCs), FoxO3/4 polyadenylation changes

276 CIBERNED 2018 ANNUAL REPORT in CPEB4 modified mice (Parras et al. Nature in press) and we have preliminary evidence of FoxO transcription factors are potential regulators of both basal autophagic activity required for renewal of stem cells and for defining the cellular heterogeneity of satellite cells. We will therefore also analyze whether CPEB4 modification attenuates the sarcopenia of HD-mice and their MuSC functional abnormalities. The cooperative activity program of CIBERNED has so far involved a global budget of ap- proximately 9,078,174€ distributed in the thirty-three cooperative projects mentioned above. A total of 59 CIBERNED research groups have actively participated at least in one cooperative project, which represents 93.6% of all the groups belonging to CIBERNED during this period, in addition to ten associated external groups. In addition, 51 groups (80.9%) have participated in more than one cooperative project and projects from the 2017 and 2018 calls are still active, with the involvement of 23 CIBERNED research groups (46%). By analyzing previous calls, and considering that some are already closed and others still ongoing, an estimation of the budget invested can be made by individual project and participating group. Thus:

Average budget Average budget Duration Number of Number of Call Budget for project and for project and in years groups projects year group

2010 2.322.100 € 3 40 6 129.005,56 € 19.350,83 €

2011 1.192.000 € 2 26 4 149.000,00 € 22.923,08 €

2013 1.376.000 € 2 23 5 137.600,00 € 29.913,04 €

2014 532.074 € 2 8 2 133.018,50 € 33.254,63 €

2015-I 690.000 € 2 11 3 115.000,00 € 31.363,64 €

2015-II 700.000 € 2 10 2 175.000,00 € 35.000,00 €

2016 1.016.000 € 2 19 5 101.600,00 € 26.736,84 €

2017 650.000 € 2 11 3 108.333,33 € 29.545,45 €

2018 600.000 € 2 12 3 100.000,00 € 33.333,33,€

277

INTERNATIONAL RELATIONS

CIBERNED 2018 ANNUAL REPORT

INTERNATIONAL RELATIONS

International scientific collaboration increases more and more, not only because of the availability of international funding and the drive of modern communication technologies, but also because science itself has become a truly international collaborative activity. In particular, the scope and scale of the problem of neurodegenerative diseases in today’s society require a global response to confront this great challenge and thus has been recognized by various international institutions such as the European Union (EU), the Organization for Economic Cooperation and Development (OECD), the World Health Organization (WHO), etc.), and the industrialized countries that constitute the G8. This global concern has led to the creation of the World Dementia Council (WDC) with the aim of collectively spur action against dementia worldwide in the areas of research, clinical care and social awareness. The leaders of governments, businesses and academia also recognize the need for a coordinated strategy to address this major global challenge for health systems. There is consensus among all stakeholders on the need to build capacities, infrastructures and R&D resources in the field of neurodegenerative diseases. As a result, WHO has decided to establish a global observatory on dementia to monitor the prevalence of the condition and resources to care for patients in Member States as well as to track the establishment of national plans and policies against dementia. There is also a pressing need for global participation and a commitment to a significant increase in investment in skills and resources to reduce the duration of these chronic brain pathologies and/or the number of people at risk. This budgetary effort should be accompanied by sound policies and legislative initiatives to encourage public-private partner- ships. History has shown that collaboration between academic researchers, government agencies and pharmaceutical and biotechnology companies is an essential ingredient in promoting this type of ambitious initiatives, especially when resources are limited. In this context, in recent years CIBERNED, has given a boost to its relations with international organizations in the area of research in neurodegenerative diseases such as the EU Joint Programme for Research in Neurodegenerative Diseases (JPND) and the Network of Centres of Excellence in Neurodegeneration (COEN), among other initiatives.

281 EUROPEAN UNION JOINT PROGRAMMING ON NEURODEGENERATIVE DISEASE RESEARCH (JPND) The EU Joint Programming for Research in Neurodegenerative Diseases (JPND) is an innovative collaborative research initiative created to address the growing challenges posed by these disorders. The JPND is a pioneering example of joint programming for the promotion of research within the European Union aimed at scientific challenges requiring a response that exceeds the capacity of a single country, based on the alignment of national research programs devoted to these challenges. Its objective is to enhance the impact of research by aligning existing national research programs and the identification of common objectives whose scope would benefit from joint action. The JPND Scientific Advisory Committee has significant participation of two CIBERNED researchers, Drs. Jesús Avila and Jesús de Pedro, as well as Dr. Angel Cedazo-Mínguez, from the Karolinska Ins- titute in Stockholm and member of the CIBERNED Scientific External Advisory Committee. The Research Strategy designed by JPND provides a framework for future investments and shows that the research effort within the European Union can be leveraged to improve care on prevention, diagnosis and treatment of patients suffering from these diseases. To achieve impact there is a need to encourage novel as well as multidisciplinary approaches, and to strengthen and extend existing capabilities across the full spectrum of basic, clinical, health and social care, and translational research. To that end, a number of priority areas for future research have been identified: The origins of neurodegenerative diseases; Disease mechanisms and models; Disease definition and diagnosis; Treatment and prevention; Health and social care. This Research Strategy also provides a framework of opportunities for countries involved in JPND and willing to participate in joint actions, which will be imple- mented through co-operative activities that realign or link national investments to achieve increased impact, and the provision of new funding. A guiding prin- ciple for its delivery will be that the research to be supported is of the highest scientific quality. In this regard, during 2011 took place the first call for European research projects JPND. Under the theme “Optimization of biomarkers and harmonization of their use in the clinic”, four transnational projects were awarded for the period 2012- 2015. Subsequently, in late 2012 a new call under the topic “Identifying protective factors and genetic and epigenetic risk in neurodegenerative diseases” was launched, resulting in five approved projects for the period 2014-2017. During 2018, there have been no CIBERNED research groups involved in active projects within the joint European Union program for research on neurodegenerative diseases (JPND). Unfortunately, despite repeated attempts from CIBERNED to modify this situation, the decrease in the budget dedicated to the JPND calls and the restrictions on the participation of the CIBERNED groups (especially the strict requirement of doing so only as a coordinating group) imposed in recent years by the ISCIII as the funding agency have ended up discouraging the participation of the CIBERNED groups in the JPND calls, with a gradual but very significant decrease in the number of applications submitted to these calls in recent years. From CIBERNED, as

282 CIBERNED 2018 ANNUAL REPORT the only Research Center specifically focused on the study of neurodegenerative diseases, we will continue to demand the possibility of participating in equal conditions with other institutions, so that it is the evaluation of the scientific merits of each application that determines the approval of the corresponding projects on a competitive basis.

CENTERS OF EXCELLENCE IN NEURODEGENERATION (COEN) A major obstacle to the advancement of research on neurodegenerative diseases is the relative lack of common standards and mechanisms for validation of potentially relevant results in preclinical studies, and clinical studies based on population. One approach to deal with these challenges on a large scale is through a more effective use of large centers and institutes, where there is already the necessary critical mass of resources and expertise. Increased collaboration between national centers of excellence should also provide the opportunity to accelerate progress in understanding the basic mechanisms of disease, and the identification of new therapeutic approaches. To this end, on June 10, 2010, the Canadian Institutes of Health Research (CIHR), the Ger- man Centre for Neurodegenerative Diseases (DZNE, Germany) and the Medical Research Council (MRC, UK) launched a funding initiative to establish a collaborative approach to research in neurodegenerative diseases, called “Centers of Excellence in Neurodegene- ration (COEN)”. These founding members were later joined by other European institutions and thus, in December 2011 the COEN membership application by CIBERNED-CIEN Foundation was approved, recognizing the scientific excellence in both basic and clinical science of the institution which became part of the COEN Oversight Group. In 2012, CIBERNED and CIEN Foundation joined this Committee to participate actively in the design of the future COEN scientific strategy. Both institutions are represented by Dr. Miguel Medina, CIBERNED Deputy Scientific Director and member of the CIEN Founda- tion Scientific Advisory Board. During 2015 the French Agence Nationale de la Recherche (ANR) has also been acknowledged as a new COEN member. Current COEN members are: • Canadian Institutes of Health Research (CIHR) • Deutsche Zentrum für Neurodegenerative Erkrankungen (DZNE, Germany) • Medical Research Council (MRC, United Kingdom) • Flanders Institute of Biotechnology (VIB Flanders, Belgium) • Health Research Board (HRB) / Science Foundation Ireland (SFI), Ireland • Ministero della Salute (MDS, Italy) Centre of Excellence for Brain Research (MES- RS), Slovakia • CIBERNED-Fundación CIEN, Spain • Agence Nationale de la Recherche (ANR), France The overlapping of the COEN group members with those of the JPND will ensure that their complementary objectives progress in close cooperation with each other. This is accomplished through a two-step process, involving expert workshops for the analysis of needs, followed by a call for proposals for collaborative teams between PIs within the participating national Centers of Excellence. . The first phase of the COEN initiative began at the end of 2010 and was intended to esta-

283 blish common resources and methodological approaches to support future studies. Some of the key issues addressed have been: the refinement and validation of cellular and animal disease models; the development of new measures to define patient subgroups for clinical trials; the identification of new biomarkers to support translational research; the development and harmonization of cognitive test batteries for diagnosis and follow-up of disease progression; and the establishment of common computer platforms to improve data analysis and exchange. Phase II of the initiative was launched during the year 2013, with the aim of catalyzing collaborative research between centers with a critical mass of resources and expertise to thus promote a radical change in research on neurodegeneration. To do this, the countries participating in COENs contributed a total amount of 5.5 million € (of which Spain has provided 600,000 €) in this call to establish an innovative and progressive research program to address the major challenges in this field. The call was intended to encourage the community to think outside the pre-established frameworks and stimulate new and creative approaches and solutions to the challenges of research in neurodegeneration. This call of Pathfinder projects intends to combine the strengths of research groups through Centers of Excellence in at least two partner countries to provide a truly collaborative effort and value that will advance our approach to research neurodegeneration. The projects would address issues that are not easily financed through standard grant mechanisms from COEN partners, and is expected to further collaboration between Cen- ters of Excellence, the projects would also serve to provide a platform for future collaboration with industry. The approved projects with the participation of CIBERNED that have been active during 2018 is described below:

COEN4016: Focused ultrasound modulation of neuromelanin accumulation in a humanized rat model of Parkinson’s disease.

Project partners: Miquel Vila (Spain), Stéphane Lehericy (France). Parkinson’s disease (PD) is a common neurodegenerative disorder which incidence is increasing due to the progressive aging of the world’s population. Despite the availability of symptomatic treatments, PD remains incurable, as current treatments do not halt nor slow the progressive loss of neurons in these patients. In PD there is a selective degeneration of neurons containing the pigment neuromelanin (NM), especially dopamine-producing neurons of the substantia nigra, which leads to the classical motor symptoms of the disease. In humans, NM appears in early childhood and accumulates progressively with age, since neurons cannot degrade or eliminate this pigment. We have recently demonstrated, using a novel rat model genetically engineered to produce human-like NM up to levels reached in elder humans, that age-dependent intracellular NM accumulation ultimately compromises neuronal function and triggers PD pathology when reaching a certain threshold. Here we will determine whether transcranial focused ultrasound (tFUS), an emerging non-invasive technology, is able to lower NM levels below their pathogenic threshold in NM-producing PD rats and prevent, halt or delay neuronal dysfunction and degeneration. If successful, this proposal will lay the ground- work for the development of a novel disease-modifying therapy for PD based on the modulation of NM levels with tFUS.

284 CIBERNED 2018 ANNUAL REPORT

COEN4017: Developing preclinical and clinical biomarkers of NRF2 pathway activation for therapeutic application in neurodegenerative diseases.

Project partners: Pamela Shaw (UK), Antonio Cuadrado (Spain) Neurodegenerative diseases such as Alzheimer’s (AD) and motor neuron disease (ALS/ MND) cause cell death of different populations of nerve cells. These conditions are very distressing for sufferers and their families. There is a severe lack of treatments available to slow disease progression and clinical trials have had high failure rates partly because there is no way to demonstrate that a drug is reaching the nervous system in the right amounts to protect the nerve cells from injury. Although the underlying causes that trigger these diseases are complex (multiple genes and environmental factors), there is substantial overlap in the cell pathways that lead to neurodegeneration. This project is focused on a master cellular pathway (sometimes called the programmed cell-life pathway) controlled by a molecule NRF2 that promotes cell survival in the face of stresses such as oxidative stress, inflammation, and failure of the energy-generating and protein quality-control pathways within neurons which are known to contribute to neurodegeneration. Our aim is to develop MRI imaging and body fluid markers to show NRF2-activating drugs working in the body. These results will be applied later in clinical trials to test the effectiveness of NRF2 activators for patients with MND and AD.

INTERNATIONAL CONGRESS FOR RESEARCH AND INNOVATION IN NEURODEGENERATIVE DISEASES (CIIIEN) During 19th to 21st September 2018, it was held in Alicante, Spain the VI International Congress on Research and Innovation in Neurodegenerative Diseases (CIIIEN), promoted by the Queen Sofia Foundation in collaboration with CIEN Foundation and CIBERNED. The main objective of CIIIEN is providing a forum in which to share progress and information of interest on neurodegenerative diseases among the scientific community. The CIIIEN, created in 2013, definitely consolidates the two major scientific conferences on neurodegenerative diseases organized in Spain: the International Symposium on Ad- vances in Alzheimer’s Disease, promoted annually by the Queen Sofia Foundation and CIEN Foundation, and the CIBERNED Scientific Forum, which brought together every year the research groups constituting the CIBER in Neurodegenerative Diseases. Unifying both congresses was a first step in creating a new operating structure in the two main institutions devoted to research on neurological and neurodegenerative diseases in Spain: the CIEN Foundation and CIBERNED, both dependent on the Ministry of Science, Innovation and Universities through the Carlos III Institute of Health. This new structure seeks greater effectiveness and efficiency in research, favoring an interaction between the different research groups. This sixth edition of CIIIEN was held at the Faculty of Medicine of the University of Santia- go de Compostela and during three intense days of presentations and sharing of knowledge, gathered well over a hundred international experts. Organized by the Queen Sofia Foundation, CIEN Foundation (Foundation Center for Research in Neurological Diseases) and CIBERNED (Network Center for Biomedical Research in Neurodegenerative Diseases), the VI Congress CIIIEN is a forum for exchange on the main advances in research and treatment of Alzheimer’s, Parkinson’s, Huntington’s, and other neurodegenerative diseases. The invited speakers included world leaders such as Harald-Jürgen Hampel (University of the Sorbonne, France), who spoke about the developments in precision medicine for

285 Alzheimer’s disease in his inaugural lecture; Michael T. Heneka (Medical Research Center of the University of Bonn, Germany), who focused on the relationship between the innate aspects of the immune system and Alzheimer’s disease; and Adriano Chiò (University of Turin, Italy), who delved into the role of cognitive phenotypes. In addition to the intervention of the scientific director of CIBERNED, Jesús Ávila, and the deputy scientific director, Miguel Medina, the Congress included internationally recognized speakers, including Isa- bel Fariñas (University of Valencia), whose presentation focused on the effects of certain extracellular proteins in the stem cells; Ángel Carracedo (University of Santiago de Com- postela), with a work on the search of genes involved in neurodegenerative diseases, and José Luis Labandeira-García. Likewise, and responding to the vocation to promote the training of young researchers of CIBERNED, the Young Researcher Award was awarded during the congress to Julia Pose Utrilla, who made presented the study which has been granted such recognition. In short, this event is consolidated in its sixth edition as a meeting point for the best national and international leading experts in neurodegenerative diseases, enabling sharing of knowledge, working methods, new advances and discoveries, in a field in which international cooperation between institutions is decisive for obtaining optimal results in research.

286 CIBERNED 2018 ANNUAL REPORT

OTHER INTERNATIONAL ACTIVITIES

H2020 MARIE SKŁODOWSKA-CURIE ACTIONS: INNOVATIVE TRAINING NETWORKS The Innovative Training Networks (ITN) are actions created by the European Union (within the Horizon 2020 program framework) to support research in the European Re- search Area and are aimed to form, through an international network of public and private centers, a new generation of creative and innovative researchers, capable of transforming knowledge and ideas into products and services for the economic and social benefit of the European Union. During 2018, the CIBERNED groups have participated in two of these actions: 1. Blood Biomarker-based Diagnostic Tools for Early-stage Alzheimer’s Disease – BBDiag The main objective of this project carried out within the international network “In- novative Training Networks” of the EU (ITN-BBDiag research project) is the development of a new non-invasive methodology aimed at the identification and validation of blood biomarkers with diagnostic value, in preclinical models of Alzheimer’s disease (AD). To do this, we will analyze AD biomarker levels in blood, related to the main changes that appear in the brain in animals with AD and in different disease stages (starting from the prodromal state). For this we have established 7 experimental groups (2, 3, 4, 6, 9, 12 and 15 months-old animals) according to the presymptomatic and postsymptomatic characterization of AD in these animals. Additionally, we have standardized a non-invasive method of taking plasma samples from the different age groups of AD transgenic mice. This non-invasive blood collection protocol was optimi- zed and cross-validated with other researchers in the field. We have also carried out several immunocytochemical and immunohistochemical analyzes to examine the specific load of beta-amyloid (Aβ) plaques in correlation with neuroinflammation and neurogenesis during the progression of pathology both in cortical and the hippocampal areas. Next, we have pre-validated, in collaboration with other laboratories members of the consortium, possible biomarkers that will finally be validated using technology based on the use of biosensors.

287 As an added value, we have analyzed in vitro neurogenesis in 2D cultures, and also in vivo. We have managed to establish primary cultures from our prodromal animal model and also in late stages. One of the main challenges in this regard is to find in vitro and in vivo the relationship between neurogenesis and the appearance of biomarkers. In addition, we are using new neuroprotective agents developed in the laboratory to discover how to improve the progression of the disease in preclinical models of Alzhei- mer’s disease. We are currently performing an in vitro analysis of factors involved in such neuroprotection for once the in vitro results have been obtained, to carry out a complete study in vivo. The development of a new model 2-3D in vitro to study neurodegenerative pathology, especially in AD, is mandatory in the understanding of the pathophysiological pathways involved in the disease and could lead to an advance in drug development and subsequent treatment of this disorder. 2. Interdisciplinary training network on the purinergic P2X7 receptor to control neuroinflammation and hyperexcitability in brain diseases - PurinesDX PurinesDX encompasses global leaders in translational research on purinergic signaling, clinical specialists in a wide range of brain disorders and industry partners specia- lizing in the drug development and biomarkers. Throughout this first year of the Puri- nesDX Project, we have focused on the study of the P2X7 receptor status in patients with Huntington’s disease in relation to its messenger RNA isoforms and protein levels. Regarding the activities related to interdisciplinary training, our Early Stage Researchers (ESR) have participated in several meetings and symposia in which they improve the collaborations with the other participants of the consortium. Starting in April, we attended the Introduction Program and the Mini-Symposium on Nervous Diseases: New Approaches in Diagnosis and Therapeutics. Our ESR also participated in the First Transferable Skills Course where they were able to learn about statistics, the importance of social networking in research and scientific writing skills, among others. In October, we attended the PurinesDX Project Follow up Meeting where the EU commission reviewed our work during the first months of project execution. Our ESR also participated in the Second Transferable Skills Course where they learned about 3D imaging, business plan organization and resource management, among others.

ALZHEIMER’S ASSOCIATION The Alzheimer’s Association is a non-for-profit organization that focuses on the care and support for patients with Alzheimer’s disease, and also funds research through competitive calls for research projects on Alzheimer’s disease. During the year 2018, CIBERNED researchers have received funding from the Alzheimer’s Association through 2 research projects: 1. A multicenter, randomized, double-blind, placebo-controlled, 4-arm, 26 week parallel-group study to evaluate the safety, tolerability and anti-inflammatory effect of three oromucosal doses of Sativex® in patients with mild cognitive impairment of Alzheimer type or early Alzheimer dementia (Sat-CIEN-02) During 2018 we continued with the activities of this clinical trial, included in an open and competitive call of the Alzheimer’s Association that was approved and financed by it to be developed in Spain during the period of Sept-2016 to Oct-2018. The primary end-point of the trial is to prove the safety and tolerability of the cannabinoids in these patients; in addition some hints about their potential therapeutic

288 CIBERNED 2018 ANNUAL REPORT

effect are expected and will be useful for the design of future efficacy studies. The selected doses in accordance with previous experimental animal studies are low and without psychoactive effects. The indication of these drugs in the Alzheimer’s disease is based on their modulatory action on the synaptic activity and their potent anti-inflammatory and neuroprotection effect. 2. AARG-17-528125: Novel Methods to Interrogate the Subcellular Machinery of AD Models in Vivo The hippocampus is one of the few brain regions that experience the existence of neurogenesis throughout life (1). In this brain area, a population of neurons that plays an important role in the acquisition of new memories, granular neurons, are severely damaged in patients suffering from Alzheimer’s disease (2). These alterations are also reflected in an animal model of the disease, the mouse that over-expresses the protein glycogen synthase kinase 3β (GSK-3β). This mouse shows a reduction in the maturation of the new neurons in the hippocampus, which could be related to the loss of hippocampal-dependent memory that is observed in these mice. The main objective of this project is to analyze in depth the causes of the malfunction that these cells undergo in the mouse that over-expresses GSK-3β, in order to be able to test two novel therapeutic strategies in these mice. To date, we have found important alterations in several of the cellular components that are part of these neurons (we are currently preparing two research articles summarizing these findings). Throughout the duration of this project, we intend to continue analyzing these alterations, as well as propose some of the strategies we are testing to slow down the progress of the pathology observed in mice. − Adult hippocampal neurogenesis is abundant in neurologically healthy subjects and drops sharply in patients with Alzheimer’s disease. Moreno-Jiménez EP, Flor-García M, Terreros-Roncal J, Rábano A, Cafini F, Pallas-Bazarra N, Ávila J, Llo- rens-Martín M. Nat Med. 2019 Apr;25(4):554-560. − GSK-3β overexpression causes reversible alterations on postsynaptic densities and dendritic morphology of hippocampal granule neurons in vivo. Llorens-Martín M, Fuster-Matanzo A, Teixeira CM, Jurado-Arjona J, Ulloa F, Defelipe J, Rábano A, Hernández F, Soriano E, Avila J. Mol Psychiatry. 2013 Apr;18(4):451-60.

289 290 CIBERNED 2018 ANNUAL REPORT

SCIENTIFIC PRODUCTIVITY AND OTHER ACTIVITIES

291 292 CIBERNED 2018 ANNUAL REPORT

ANALYSIS OF CIBERNED SCIENTIFIC PRODUCTIVITY IN 2018 The table below summarize the bibliometric indicators related to the scientific activity from years of 2017 and 2018. A quick analysis shows a consolidation of the improved scientific production in 2018 (in line with the ongoing historical improvement observed in the last years since the creation of the Centre), not only in quantity but particularly in quality (number of publications overall, within the first quartile, number of publications led by CIBERNED groups within quartiles 1 and 2; number of international publications in Q1 and Q2; or ).

CHANGE INDICATOR 2017 2018 %

(A) Total number of publications1 571,00 613,00 7,36

(B) Number of publications in quartiles (Q) 1 y 2 470,00 497,00 5,74

(E) Number of publications in 1st decile 189,00 167,00 -11,64

(F) Number of publications (1st decile) as percentage of total (A) 33,10 27,24 -17,69

(G) Number of publications in Q1 (includes 1st decile) 372,00 402,00 8,06

(H) Number of publications in Q1 as percentage of total (A) 65,15 65,58 0,66

(I) Number of publications in Q1 as percentage of total Q1 and Q2 79,15 80,89 2,19 (B)

(J) Number of publications in Q2 98,00 95,00 -3,06

(K) Number of publications in Q2 as percentage of total (A) 17,16 15,50 -9,70

(L) Number of publications (Q1+Q2) led by CIBERNED groups (1st 267,00 273,00 2,25 author or corresponding author)

(M) Number of publications in Q1 and Q2 led by CIBERNED groups 56,81 54,93 -3,31 as percentage of total (B)

(N) Number of publications (Q1 and Q2) not led by CIBERNED 203,00 224,00 10,34 groups (1st author or corresponding author)

293 CHANGE INDICATOR 2017 2018 %

(O) Number of publications in Q1 and Q2 not led by CIBERNED 43,19 45,07 4,35 groups as percentage of total (B)

(P) Number of publications in Q1 and Q2 in which “CIBERNED” 81,70 84,71 3,68 appears in the affiliation (as percentage of total (B))

(Q) Number of publications in Q1 and Q2 led by CIBERNED groups in which “CIBERNED” appears in the affiliation (as percentage of 90,26 90,11 -0,17 total (L))

(R) Number of publications in Q1 and Q2 not led by CIBERNED groups in which “CIBERNED” appears in the affiliation (as 70,44 78,13 10,90 percentage of total (N))

(S) Number of publications in Q1 and Q2 with international 260,00 282,00 8,46 groups

(T) Number of publications in Q1 and Q2 with international 55,32 56,74 2,57 groups (S) as percentage of total number of publications (B)

(U) Number of publications in Q1 and Q2 with international 123,00 118,00 -4,07 groups, and led by CIBERNED groups

(V) Number of publications in Q1 and Q2 with international groups, and led by CIBERNED groups as percentage of all 47,31 41,84 -11,55 international publications (S)

(W) Number of publications in Q1 and Q2 with other CIBERS 18,00 19,00 5,56

(X) Percentage of publications in Q1 and Q2 with other CIBERS led 27,78 31,58 13,68 by CIBERNED groups

(Y) Number Percentage of published in Q1 y Q2 5,00 - -

(Z) Percentage of Percentage of led by CIBERNED groups 40,00 50,00 25,00

(A2) Addition of impact factors of journals (Q1 y Q2) in which 2859,70 3345,81 17,00 articles have been published

(B2) Average of impact factors of journals (Q1 y Q2) in which 6,08 6,18 1,59 articles have been published

(C2) Number of publications in journals with impact factor >15 44,00 61,00 38,64

Number of publications in Q1 and Q2 with 2 CIBERNED groups 57,00 48,00 -15,79

Number of publications in Q1 and Q2 with 3 CIBERNED groups 20,00 12,00 -40,00

Number of publications in Q1 and Q2 with 4 CIBERNED groups 1,00 7,00 600,00

Number of publications in Q1 and Q2 with 5 CIBERNED groups - 1,00 -

Number of publications in Q1 and Q2 with 6 CIBERNED groups 2,00 - -

Number of publications in Q1 and Q2 with 7 CIBERNED groups - - -

Number of publications in Q1 and Q2 with 8 CIBERNED groups - - -

Cooperative projects – Program 1 68,00 96,00 41,18

294 CIBERNED 2018 ANNUAL REPORT

CHANGE INDICATOR 2017 2018 %

Cooperative projects – Program 2 157,00 147,00 -6,37

Cooperative projects – Program 3 - 62,00 -

Projects with other CIBERs/RETICs/CIEN networks 36,00 15,00 -58,33

Basic Research-type projects 79,00 74,00 -6,33

Translational research-type projects 146,00 175,00 19,86

Projects with other national research groups 121,00 163,00 34,71

Projects with international research groups 45,00 50,00 11,11

Projects with industry 82,00 92,00 12,20

The following are a few figures that summarize some of the main bibliometric indicators in relation to the scientific production of CIBERNED, including the total production (indexed and not indexed in PubMed and Scopus), during the year 2018 and its evolution since 2011:

No IF

FIGURE 1. Percentage of indexed CIBERNED 2018 publications distributed by quartiles. Q1 stands for publications in the 1st quartile; Q2 stands for publications in the 2nd quartile; Q3 stands for publications in the 3rd quartile; Q4 stands for publications in the last quartile. No IF stands for publications for which no impact factor is available.

295 Nr. Publications

FIGURE 2. CIBERNED 2011-2018 production by quartiles/1st decile. Q1+Q2 stands for the sum of publications in 1st and 2nd quartiles; D1 stands for publications in the 1st decile.

FIGURE 3. Percentage of publications of excellence (1st decile) CIBERNED 2011-2018.

296 CIBERNED 2018 ANNUAL REPORT

FIGURE 4. Number of CIBERNED 2011-2018 publications within 1st and 2nd quartiles and 1st decile.

Cumulative IF Mean IF

FIGURE 5. Evolution of the impact factor from 2011 to 2018. IF stands for Impact factor.

297 FIGURE 7. Scientific journals with 5 or more CIBERNED publications in 2018.

298 CIBERNED 2018 ANNUAL REPORT

FIGURE 8. Most frequent SUBJECT categories (accprding to WoS-JCR) in CIBERNED 2018 publications

299 CROSS-NETWORK PROGRAMS NEUROLOGICAL TISSUE BANKS 1. Neurological Tissue Bank and Biological Samples of the IDIBELL Institute of Neu- ropathology, Bellvitge Hospital (Head: Dr. Isidro Ferrer). 2. CIEN Foundation Tissue Bank for Neurological Research (BT-CIEIN) (Head: Dr. Alberto Rábano). 3. Neurological Tissue Bank (BTN) at the University of Barcelona (Head: Dr. Eduardo Tolosa)

OTHER PLATFORMS • DNA Analysis Service (Head: Dr. Jordi Pérez Tur). • Electron Microscopy Service (Head: Dr. José Manuel García Verdugo). • Neuroimaging Service (Head: Dr. Jose Luis Cantero Llorente) • Dementia Genetics Spanish Consortium (DEGESCO) (Coordinator: Dr. Jordi Clarimón)

300 CIBERNED 2018 ANNUAL REPORT

EVENTS AND OTHER ACTIVITIES CIBERNED has held several meetings throughout the year, including the VI Conference on Research and Innovation in Neurodegenerative Diseases (CIIIEN) which took place in Santiago de Compostela on September 19-21 and the Second DEGESCO Symposium on Genetics of Dementia (held on May 23 in the CosmoCaixa auditorium in Barcelona), in addition to other scientific, dissemination and international cooperative events described below.

VI INTERNATIONAL CONFERENCE ON RESEARCH AND INNOVATION IN NEURODEGENERATIVE DISEASES (CIIIEN) In 2018 CIBERNED held its annual Scientific Forum. This event, which has been held year after year since 2007, is essential for the proper functioning of CIBERNED, as it allows the principal investigators, the members of their groups, as well as all the attendees, to meet to discuss the findings of their research, present new data and establish collaborations. In 2013, a new format as an International Conference on Research and Innovation in Neurodegenerative Diseases (CIIIEN) was established with great success, which has been consolidated in the successive editions (2014, 2015, 2016, 2017 and 2018). Coinciding with the week in which World Alzheimer’s Day is celebrated, on September 19- 21, the VI Conference on Research and Innovation in Neurodegenerative Diseases - CIIIEN was held in Santiago de Compostela, which, during three intense days of presentations and knowledge exchanges, brought together well over a hundred international experts. Organized by the Queen Sofia Foundation, CIEN (Neurological Diseases Research Center Foundation) Foundation and CIBERNED (Network Center for Biomedical Research in Neu- rodegenerative Diseases), the VI CIIIEN Conference is an exchange forum on the main advances in research and treatment of Alzheimer’s, Parkinson’s, Huntington’s, and other neurodegenerative diseases. In short, this event is consolidated in its fifth edition as a meeting point for the greatest national and international experts in neurodegenerative diseases, allowing to share knowledge, work methods, new advances and discoveries, in a field in which international cooperation and between institutions is decisive for obtaining optimal results in research.

301 SECOND DEGESCO SYMPOSIUM ON GENETICS OF DEMENTIA DEGESCO (Dementia Genetics Spanish Consortium) is a nation-wide consortium of scientific and technical nature jointly promoted by eleven founding research centers under the institutional umbrella of CIBERNED. DEGESCO was constituted in 2013 with the general aim of promoting and strengthen the development of genetic studies in order to understand the genetic architecture of neurodegenerative dementias in the Spanish population through the implementation of collaborative projects and actions among its members. DEGESCO philosophy is inclusive, that is, it is open to the incorporation of new research groups, whether public or private, that show their capacity and interest to conduct research on dementias. DEGESCO is a coordination structure for sample repositories that share information and processing protocols. The samples selected by the participants for DEGESCO must be characterized by both scientific-technical and ethical quality, comply- ing with the Biomedical Research Law (Law 14/2007) and the Royal Decree of Biobanks (RD 1716/2011). The transfer of samples between the members of the consortium will be carried out in accordance with the requirements of the Royal Decree of Biobanks. At the end of 2018, DEGESCO is composed of 24 fully-fledged research groups with the institutional coverage of CIBERNED (Center for Biomedical Research in Neurodegenerative Diseases Network). Those member groups are the following:

DEGESCO (DEmentia GEnetics Spanish COnsortium)

San Sebastián: Santander: • Hospital Donostia • Hospital Marqués de Valdecilla • Fundación CITA-Alzheimer Vitoria: Oviedo: • UPI/EHU • Hospital Central de Asturias Pamplona: • Navarrabiomed Barcelona: • Hospital de Sant Pau Asturias Cantabria País Vasco • Hospital Clínic Galicia Navarra • Fundació ACE La Rioja • Fundació Pascual Maragall Terrassa: Cataluña Castilla y León Aragón • Hospital Mútua de Terrassa Madrid: • Hospital la Paz Lleida: • Hospital 12 de Octubre • IRB-Lleida • Hospital Ramón y Cajal Madrid • CIBERNED Islas Baleares • Fundación CIEN Comunidad Castilla - Valenciana • CBM Extremadura La Mancha P. Mallorca: • Hospital de la Princesa • Hospital Son Espases

Murcia València: • Institut de Biomedicina de València Andalucía (CSIC)

Murcia: • Hospital Clínico Universitario Virgen Málaga: de la Arrixaca • Instituto Andaluz de Neurociencia Sevilla: • Instituto de Biomedicina de Sevilla

302 CIBERNED 2018 ANNUAL REPORT

Continuing with the consortium dissemination activities, the Second DEGESCO (Dementia Genetics Spanish Consortium) Symposium on Genetics of Dementia was held at the Cos- moCaixa Auditorium in Barcelona on May 23, 2018. This symposium, which was organized for the first time as part of the XI edition of the Barcelona-Pittsburgh Conference, which Fundació ACE has organized during the last 20 years for its commitment to training and research, and aimed to bring together the best national experts on genetics of neurodegenerative diseases to address a key research area by thoroughly reviewing the genetic aspects of the different types of dementia and their implications in the Spanish population.

EXCELLENCE IN NEURODEGENERATION SEMINAR SERIES During 2018 CIBERNED has continued to carry out the series of lectures “Excellence in Neurodegeneration Seminar Series”, offering presentations by leading international experts in their areas of research on all aspects of frontier research in neurodegenerative diseases (molecular, cellular, genetic, cognitive, clinical, animal models, biomarkers, imaging, etc.) and related areas. Below there is a list of the seminars held during 2017 within this series: Speaker: Harald-Jürgen Hampel, Sorbonne University, France. Title: Development of Precision Medicine for Alzheimer Disease. Speaker: Michael T. Heneka, University of Bonn Medical Center, Germany. Title: Does innate immune activation drives Alzheimer disease?. Speaker: Adriano Chiò, University of Turin, Italy. Title: The determinants of motor and cognitive phenotypes in ALS. Speaker: Werner Poewe, Innsbruck Medical University, Austria. Title: Novel therapeutic targets for Parkinson’s Disease. Speaker: Ángel Carracedo, University of Santiago de Compostela, Spain. Title: The search for genes involved in neurodegenerative diseases: Challenges and new strategies

YOUNG INVESTIGATOR OF THE YEAR AWARDS To promote research excellence, CIBERNED annually awards a prize to an emerging scientific involved in clinical or basic research, and on a biennial basis, a prize to an emerging scientist involved in clinical research in neurodegenerative diseases. The candidates for the 2018 Young Investigator of the Year Award must be under 35 years old, members of a CIBERNED research group and main authors of an article published in an indexed scientific journal during 2017. The call remit was announced by email addressed to all personnel belonging to CIBERNED as well as through the Center’s website. Once the deadline for submitting applications had elapsed, a total of 8 applications were received that met the specifications of both calls in an appropriate and timely manner. For both calls, the selection committee was made up of members of the Steering Commit- tee and of Senior Researchers from outside the Committee, who assessed the scientific quality and impact of the publications, as well as the candidate’s specific contribution.

303 Once the evaluation process for the submitted applications was completed, the CIBERNED Scientific Directorate agreed to award the Young Investigator Award to Julia Pose Utrilla for her work “Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1”, published in Nature Communications (2017); 8 (1): 2275.

OTHER RELEVANT EVENTS UNIVERSITY OF EXTREMADURA (UEX) SUMMER-AUTUMN COURSE

Between 20th and 22nd June, 2018 the Course “Advances in neurobiology and neurodegenerative diseases” was held in Cáceres, as part of the XIX edition of the Interna- tional Summer Courses of the University of Extremadura, led by Dr. José Manuel Fuentes Rodríguez, Principal Investigator of CIBERNED and member of the Department of Bio- chemistry and Molecular Biology and Genetics of the Faculty of Nursing and Occupational Therapy of the University of Extremadura. This Course was focused on knowing the most current advances at the molecular and cellular level of the Nervous System, with special emphasis on what refers to neurodegenerative disorders, given its growing importance at the social, health and scientific level. It was also intended to integrate this knowledge to address aspects such as its understanding, its prevention, or its treatment; to allow an improvement in the acquisition of skills for future professional training, especially if it has to do with scientific research or clinical practice in relation to these or other diseases of the Nervous System; to obtain scientific training in the biomedical field that bridges basic and clinical research, and ac- quire cross-cutting knowledge to ensure a multidisciplinary vision of the neuroscientist acitivities. Among other specialists, the following CIBERNED Principal Investagtors participated in the Course: Drs. José Ramón Naranjo, Isidre Ferrer, Rosario Moratalla, José Luis Labandeira, Rafael Fernández Chacón, Adolfo López de Munain and Jordi Pérez Tur.

PURINESDX MINI-SYMPOSIUM – BRAIN DISEASES: NEW APPROACHES ON DIAGNOSTICS AND THERAPEUTICS

On April 24, a symposium of the European project PurinesDX entitled “Brain Diseases: New Approaches on Diagnostics and Therapeutics” was held at the Royal College of Sur- geons in Ireland (RCSI) in Dublin, where the main objectives of the project and the specific aims of each of the partners of the consortium were addressed. The PurinesDX project is part of the European Union’s Horizon 2020 research and innovation program, collaborates with leading research leaders in purinergic signaling, brain pathologies and in the development of biomarkers and therapies. The symposium was attended by Dr. Jose J. Lucas, Principal Investigator of CIBERNED, and Dr. Miguel Medina, Deputy Scientific Director of CIBERNED.

304 CIBERNED 2018 ANNUAL REPORT

II EUROTAU MEETING 2018

On April 26 and 27, 2018, the second edition of the Eurotau Meeting took place in Lil- le. This event, hosted by Dr. Luc Bueé, brought together the main European specialists involved in Tau research and constitutes a reference forum to share new ideas and hypotheses about the physiological and pathological roles of Tau proteins and the so-called tauopathies. Like last year, Drs. Jesús Ávila and Miguel Medina, who also participated as a speaker, were part of the scientific committee of the event. In addition, the program had the intervention of Drs. Lastres-Becker and Sánchez-Juan, both members of CIBERNED. The organizers and the scientific committee of the Eurotau Meeting aim to make the annual meeting one of the reference events for scientists and researchers around the world, especially as a forum for discussing alternative approaches to tauopathies in a context of disparate results in drug development.

I MOLECULAR NEUROPATHOLOGY DAY

On February 16, 2018, the First Molecular Neuropathology Day was held at the Center of Molecular Biology Severo Ochoa (CBMSO, for its acronym in Spanish) in Madrid. This event was organized jointly between the Department of Molecular Neuropathology of the Center of Molecular Biology Severo Ochoa and CIBERNED, along with other institutions. The event aimed to share the results of projects related to neurological pathologies and researchers from other research institutions and centers with research lines in this field were invited to participate. In addition, the participation of young researchers the field was promoted and encouraged.

XI NEUROBIOLOGY SYMPOSIUM: FUTURE TECHNICAL ADVANCES

This Symposium is a biennial scientific event organized by the Experimental Neurobiology section of the Catalan Society of Biology and on this occasion was held on November 12- 13, 2018 in Barcelona. The objective of the Symposium was to present recent scientific advances in the field of neuroscience from the research groups of our community. In this event, topics such as development, neurogenesis and stem cells, glial cells and neuroinflammation, neurotransmission, neuropharmacology, receptors and cell signaling or death mechanisms and neuronal regeneration, among others, were discussed. The organizing committee was formed by two members who belong to the CIBERNED network since its inception, Drs. Carles Saura and Silvia Ginés.

305 306 FINANTIAL REPORT 308 CIBERNED 2018 ANNUAL REPORT

FINANTIAL REPORT The Network Center for Biomedical Research in Neurodegenerative Diseases (CI- BERNED) is a research organization with its own legal personality, according to Article 6.5 of Law 30/1992 of November 26, on the Legal Regime of Public Administrations and the Common Administrative Procedure, whose mission is the broadly defined monographic research on neurodegenerative diseases. It is composed of research groups, without physical contiguity, belonging to different Administrations, Institu- tions and Regions, from the public and private sector with research lines and goals focused on the specific common area of neurodegenerative diseases and coordinating for the achievement of objectives, which could hardly be envisaged in a more restricted context of implementation. CIBERNED became part of the State Public Administrative Sector on May 14, 2010. Since January 1, 2011, it has been subject to the Public Accounting General Plan, approved by Order EHA/1037/2010, as an accounting standard, taking into account the Adaptation of the PGCP for Public Entities whose expenditure budget are estimated. Its public procurement system is governed by Law 9/2017, of November 8, on Public Sector Contracts which transposes the Directives of the European Parliament and of the Council 2014/23/EU and 2014/24/EU of February 26, 2014 to the Spanish legal system. CIBERNED is a virtual Research Center consisting of research groups belonging to different Administrations and Institutions in partnership: the member Principal Inves- tigators work in the institutions to which they belong by participating actively and simultaneously in CIBERNED’s own cooperative research agenda. It is therefore the result of the collaboration and association of entities as well as the sum of research groups. In 2018, it consisted of 50 groups, an average of 120.32 hired staff that add up to 119 by December 31, 2018.

309 Average staffing for the year 2018:

CATEGORY WOMEN MEN TOTAL Administrative 4,93 - 4.93 assistant Undergraduate 2,42 - 2,42 Doctor (PhD) 26,17 13,89 40,06 Graduate (BSc, MSc) 24,82 16,28 41,10 Technician 26,88 4,93 31,81 Total 85,22 35,32 120,32

Staff personnel at year end:

CATEGORY WOMEN MEN TOTAL Administrative 6 - 6 assistant Undergraduate 3 - 3 Doctor (PhD) 26 16 42 Graduate (BSc, MSc) 26 18 44 Technician 19 4 23 Total 80 38 118

The Scientific Head Office is located at the Center for Molecular Biology “Severo Ochoa” in Madrid. The headquarters and the Manager Office are located at Valderrebollo 5 in Madrid within the premises of the Alzheimer Center of the Queen Sofia Foundation.

FUNDING The Resolution of March 30, 2006 from the Carlos III Health Institute, which calls for grants to fund stable cooperative research structures in the area of biomedicine and health sciences within the framework of the initiative Ingenio 2010, Consolider program, and CIBER actions, establishes the conditions of the grants under point 13. In that point, it is indicated that the ISCIII assistance may reach a maximum of 80% of the budget of the Consortium’s activities. This budget will be used to cover expenses directly related to the development and execution of CIBERNED activities. In determining the total budget the corresponding expenditure to fund salaries for researchers that are part of the CIBERNED and do not have a working contract with the consortium, but rather with the associated institutions is considered included, but not eligible and therefore allocated in the 20% not funded by the grant. In this financial year, the contribution of the Associated Institutions accounted for 2,185,041.12€. During 2018, the Consortium counts on the funding from the Carlos III Institute of Health as a promoter organization for the execution of the activities and general objectives already described.

310 CIBERNED 2018 ANNUAL REPORT

On March 2, 2018, the Resolution of the Carlos III Institute of Health was approved, which regulates the conditions for the license to CIBER in the thematic area of Neurodegener- ative Diseases of the nominative assignments provided for in the Statement of Expend- iture of the 2017 Expanded Budget for 2018 of the Carlos III Institute of Health. That resolution was issued respecting the limits imposed by the seventh paragraph of the Ministerial Agreement of December 29, 2017, which decided: To assign to CIBERNED the following nominative contributions foreseen in the Statement of Expenditure of the Carlos III Institute of Health Budget for the year 2017: • 1.961.620 euros from budget heading 27.107.465A.44605 • 81.690 euros from budget heading 27.107.465A.74605 Corresponding to 50% of the total credit existing in the aforementioned budgetary head- ings, leaving the remaining 50% of the credit subject to the approval of the General State Budget Law for 2018. Once the General State 2018 Budget Law 6/2018 of July 3 was approved, the amounts initially allocated were adjusted and by resolution of July 31, 2018, the amounts granted were increased: • 1.826.620 euros from budget heading 27.107.465A.44605 • 81.690 euros from budget heading 27.107.465A.74605 Other sources of income for the development of the activity and general objectives of the Consortium are: • Contributions that may be obtained as a result of research, technical assistance or advice that is performed. • Income from private law and any other income or consideration authorized by current legislation.

BASIS OF PRESENTATION OF THE ACCOUNTS True and fair view

The financial statements have been prepared from the accounting records of the Entity following the accounting principles and standards established in the General Public Ac- counting Plan (PGCP, for its acronym in Spanish), showing the true and fair view of the net worth, changes in the same, financial situation, budget execution, statement of cash flows and results. The accounting principles and valuation criteria are applied in accordance with the regis- tration and valuation rules of the PGCP, Order EHA / 1037/2010 of April 13 and the adaptation of the PGCP for public bodies whose expenditure budget is estimated, approved by Resolution of July 28, 2011 of the General Intervention of the State Administration and other legal provisions in force regarding accounting matters. These financial statements have been prepared by the Managing Director of the Institu- tion and will be submitted to approval by the Governing Council.

311 Balance sheet year 2018

ASSETS A) Non-current assets 1.320.631,82 I. Intangible assets 40.239,20 1. Investment in research and development - 2. Industrial and intellectual property - 3. IT applications 40.239,20 4 Investments in assets used under lease or assignment - 5. Other intangible assets - II. Material assets 1.279.789,92 1. Lands - 2. Buildings - 3. Infrastructures - 4. Historic heritage assets - 5. Other material assets 1.279.789,92 6. Fixed assets under construction and advances - III. Real state investemnts - IV. Long-term financial investments in group companies, joint - ventures and associates V. Long-term financial investments 602,70 1. Financial investments in equity - 2. Credit and debt securities - 3. Financial derivatives - 4. Other financial investments 602,7 VI. Debtors and other long-term accounts receivables - B) Current assets 3.404.044,99 I. Assets assigned for sale - II. Inventory - III. Debtors and other accounts receivable 932.560,35 1. Debtors from management operations 534.764,17 2. Other accounts receivable - 3. Public Administrations 397.796,18 IV. Short-term financial investments in group companies, joint - ventures and associates V. Short-term financial investments 4.285,76 1. Financial investments in equity - 2. Credits and debt securities 4.285,76 3. Financial derivatives - 4. Other financial investments - VI. Accrual 14.342,99 VII. Cash and cash equivalents 2.452.855,89 1. Treasury 2.452.855,89 TOTAL ASSETS (A+B) 4.724.676,81

312 CIBERNED 2018 ANNUAL REPORT

EQUITY AND LIABILITIES A) Equity 2.414.470,19 I. Contributed equity - II. Generated equity 1.111.824,39 1. Results from previous years 1.436.086,28 2. Results in the year -324.261,89 3. Reserves - III. Valuation adjustments - IV. Other assets increases pending allocation to results 1302645,8 B) Non-current liabilities 417.360,38 I. Long-term provisions 60.466,86 II. Long term debts 356.893,52 1. Debentures and other marketable securities - 2. Debts with credit institutions - 3. Financial derivatives - 4. Other debts 356.893,52 5. Debtors for long-term capital leasing agreements - III. Debts with group companies, joint ventures and associates - IV. Creditors and other long-term payables - V. adjustments for long-term accrual - C) Current liabilities 1.892.846,24 I. Short-term provisions 38.240,05 II. Short-term debts 1.346.782,32 1. Debentures and other marketable securities - 2. Debts with credit institutions - 3. Financial derivatives - 4. Other debts 1.346.782,32 5. Debtors for short-term capital leasing agreements - III. Debts with group companies, joint ventures and associates - IV. Accrual 507.823,87 1. Creditors from management operations 305.676,47 2. Other accounts payaable 16,13 3. Public Administrations 202.131,27 V. Adjustments for accrual - TOTAL EQUITY AND LIABILITIES (A+B+C) 4.724.676,81

313 Economic assets income statement

2018 ECONOMIC ASSETS INCOME STATEMENT

1. Income tax and social contributions -

2. Transfers and subsidies received 6.564.336,41

a) Del ejercicio 6.416.950,54

a.1) subsidies received to finance expenditures for the year 442.169,42

a.2) transfers 5.974.781,12

b) Allocation of subsidies for non-financial assets 147.385,87

3. Net sales and services 240.131,19

4. Changes in inventory of finished and in progress of manufacturing - goods and impairment loss

5. Work by the company for its fixed assets -

6. Other income from ordinary management -

7. Provisions surpluses 4.781,36

A) TOTAL REVENUES FROM ORDINARY MANAGEMENT (1+2+3+4+5+6+7) 6.809.248,96

8. Personnel costs -3.502.828,43

a) Wages and salaries and similar expenses -2.642.824,19

b) Social security costs -860.004,24

9. Transfers and grants awarded -2.257.329,11

10. Procurements -728.435,40

a) Consumption of goods and other supplies -728.435,40

11. Other ordinary management expenses -474.724,70

a) Supplies and other external services -474.622,69

b) Taxes -102,01

c) Others -

12. Depreciation of fixed assets -149.906,44

314 CIBERNED 2018 ANNUAL REPORT

B) TOTAL COSTS OF ORDINARY MANAGEMENT (8+9+10+11+12) -7.113.224,08

I Results (surplus or deficit) of ordinary management (A+B) -303.975,12

13. Impairment loss and results on disposals of non-finacial assets and -5.420,57 selling state assets

14. Other non-ordinary items 827,67

a) Income 827,99

b) Expenditure -0,32

II results of non-dinancial operations (I +13+14) -308.568,02

15. Financial income 713,41

a) participations in equity instruments -

b) marketable securities and loans of fixed assets 713,41

16. Financial expenses -

17. Financial expenses charged to assets

18. Change of fair value of financial assets and liabilities -

19. Exchange differences -16.407,28

20. Impairment loss, retirements and disposals of financial assets and - liabilities

21. Grants to fund financial operations -

III Result of financial operations (15+16+17+18+19+20+21) -15.693,87

IV Net surplus (or deficit) in the year (II + III) -324.261,89

(+-) Adjustments to the account of the outcome of the prior year -

Adjusted profit for the prior year -

315 IDENTIFICATION OF PROGRAMS Programs and other activities have the following breakdown of financial resources for their implementation, the description of each, as well as data and information is detailed in the scientific section of this report:

IMPLEMEN- PERSONNEL AMORTIZATION TOTAL TATION COSTS

Administration, management and 122.210,10 138.170,29 11.763,62 272.144,01 coordination

Program 1 "Alzheimer's disease and other 795.168,85 141.776,13 58.464,60 995.409,58 degenerative dementias"

Program 2 "Parkinson's disease and other 1.307.412,44 258.920,21 65.923,03 1.632.255,68 degenerative motor disorders""

Deputy Scientific Director / Cross- 234.352,42 36.273,37 278,53 270.904,32 network plataforms

Training / Young Investigator Award / - 26.383,18 - 26.383,18 Mobility

Media service, internationalization, 14.265,49 56.937,77 - 71.203,26 and transfer of knowledge

COOPERATIVE RESEARCH

Cooperative projects, 0,00 431,00 1.219,31 1.650,31 4th call (PI2014/2013)

Cooperative projects,54th call 156.327,46 49.312,62 - 205.640,08 (PI2015)

Cooperative projects, - 936,85 - 936,85 6th call (PI2015-2)

Cooperative projects, 356.218,49 211.838,94 855,21 568.912,64 tth call (PI2016)

Cooperative projects, 184.587,31 20.953,04 - 205.540,35 8th call (PI2017)

Coordination and management of call for 12.187,97 - - 12.187,97 pojects

316 CIBERNED 2018 ANNUAL REPORT

IMPLEMEN- PERSONNEL AMORTIZATION TOTAL TATION COSTS

COMPETITIVE PROJECTS

PI11/03028 - - 1.952,47 1.952,47

COEN Projects - 2013 - - 68,16 68,16

COEN Projects - 2017 15882,42 36.173,59 - 52.056,01

BBVA Research Projects - 2.198,28 - 2.198,28

RTC-2014-1877-1 - - - -

InDiWIP/ Ramon Areces 15.600,70 4125,43 19.726,13 Foundation

PIE14/00061 - - 827,79 827,79

COHORTE BBN-CIBERNED - 40.987,75 - 40.987,75

ALZ.ASSOC/SATIVEX 4.876,72 -18.533,71 - -13.656,99

BBDiag 69.566,66 24.879,02 - 94.445,68

AC16-00050 30.108,84 3540,25 384,95 34.034,04

AFTD 2016 - 2.686,15 116,84 2.802,99

SAF2016-786 14790,03 8.128,03 - 22.918,06

MS16/00041. Miguel 40.500,04 - - 40.500,04 Servet program

CP16/00041. 20.552,45 16.335,60 12,01 36.900,06 Associated project

PurinesDX 52866,76 4007,84 - 56.874,60

AARG 2017 16351,97 17.199,10 - 33.551,07

OTHER PROJECTS

GW PHARMA 2.852,39 - - 2.852,39 VIVACELL 36148,92 6.237,28 - 42.386,20 EHP - 13.751,02 - 13.751,02 HEMP SOLUTION - 13.039,31 - 13.039,31 FGCSIC - 36,18 - 36,18 ASSOCIATED INSTITUTIONS - 69.287,99 - 69.287,99 AGREEMENTS ANNUAL FORUM - 89.435,58 - 89.435,58 AMORTIZATION - - 8039,92 8.039,92 CONTRIBUTIONS ASSOCIATED - 2.185.041,12 - 2.185.041,12 INSTITUTIONS 3.502.828,43 3.460.489,21 149.906,44 7.113.224,08

317 The data in the table below show that CIBERNED has allocated 2,627,655.26€ for implementing research programs, which represents 53% of the ordinary operating expenses, excluding taxes and without taking into account the contribution of Associated Institu- tions.

2017 2018 VARIACIÓN % Research programs expenditure 2.534.372,53 2.627.655,26 +3,68%

The amount allocated in 2018 to cover the costs of the internal calls for cooperative projects adds to 994,868.20€, representing 20% of the overall budget. Without considering the contribution from the Associated Institutions.

2017 2018 VARIACIÓN % Cooperative research expenditure 1.073.770,34 994.868,20 -7,35%

The total expenditure allocated to fund internal research activities, either through the CI- BERNED research groups or through the calls for cooperative projects, amounts to a total of 3.622.533,46€, which represents 73.5% of the year’s expenditure, without taking into account the contribution of the associated institutions.

2017 2018 VARIACIÓN % Research programs/Cooperative research 3.608.142,87 3.622.533,46 +0,40% expenditure

The expenditure on personnel dedicated to stable research structures (groups and cooperative projects) represents 56.64% of total expenditure, excluding the contribution of the associated institutions.

2017 2018 VARIACIÓN % Personal costs Research groups/ 2.791.727,22 2.811.902,52 +0,72% Cooperative

The resources allocated to the Administration department decrease compared to previous years, representing 5.52% of the total of ordinary expenses, excluding the valuation of the contribution of the associated institutions.

2017 2018 VARIACIÓN % Administration 346.822,60 272.144,01 -21,50%

318 CIBERNED 2018 ANNUAL REPORT

ACQUISITION OF FIXED ASSETS The acquisition of assets made during the year are as follows:

PURCHASE EQUIPMENT DESCRIPTION DATE COST ORDER 0012171885 LAPTOP HP 250 G6 12/01/2018 598,04 0012171886 NETGEAR CABIN CAPACITY 48 TB 12/01/2018 11.107,80 0012171887 HP PROBOOK 640 G3 11/01/2018 1.344,74 0012171888 HP PROBOOK 640 G3 12/01/2018 1.344,73 4102141889 QX200 DROPLET PCR READER 24/01/2018 88.198,11 4102141890 C1000 TOUCH CYCLER THERM CYCLER BASE 24/01/2018 8.788,23 4102141891 PX1 PCR PLATE SEALER 24/01/2018 3.699,22 2042171892 LAPTOP ASUS SENBOOK UX4900UA 19/01/2018 1.784,08 4112141893 SHAKER GRANT BIO PS-3D 05/02/2018 389,62 COMPUTER HP BUNDLE22_260-P10979 + 5032171894 22/01/2018 929,92 MONITOR 4092171895 COMPUTER HP PAVILION 12/02/2018 1.754,77 5042141896 ULTRAMICROTOME 27/02/2018 49.610,00 4092141897 CYTOMETER 23/02/2018 59.834,50 5112141898 VERTICAL FREEZER - 80 MODEL EXF32086 29/01/2018 9.196,00 MICROSCOPE WITHPHOTOMIRCOGRAPHY 6072141899 13/02/2018 36.166,90 SYSTEM 4012171900 APPLE AIRPORT TIME CAPSULE-2TB 06/03/2018 349,69 ADJUSTABLE ARMCHAIR ANTHEA RED 0012161901 09/04/2018 195,90 CHROME ASINCRO ADJUSTABLE ARMCHAIR ANTHEA RED 0012161902 09/04/2018 195,90 CHROME ASINCRO ADJUSTABLE ARMCHAIR ANTHEA RED 0012161903 09/04/2018 195,90 CHROME ASINCRO ADJUSTABLE ARMCHAIR ANTHEA RED 0012161904 09/04/2018 195,90 CHROME ASINCRO ADJUSTABLE ARMCHAIR ANTHEA RED 0012161905 09/04/2018 195,90 CHROME ASINCRO 2092141906 4D NUCLEOFECTOR CORE UNIT 30/01/2018 37.510,00 3062171907 DESKTOP PC 18/04/2018 1.292,47 3062171908 DESKTOP PC 18/04/2018 1.292,47 3062171909 DESKTOP PC 18/04/2018 1.292,48 1032141910 MOLECULAR INVERTED MICROSCOPE 22/02/2018 78.625,84 4112141911 FREEZER LIEBHERR 30/01/2018 1.832,52 5092171912 MONITOR 28" SAMSUG MODEL CODE 23/04/2018 358,16 4012141913 FREEZER - 86 RLE00086V 09/05/2018 11.495,00

319 PURCHASE EQUIPMENT DESCRIPTION DATE COST ORDER 3062171914 MONITOR LCD LG 24 MP58VQ-P 25/05/2018 159,72 3062171915 MONITOR LCD LG 24 MP58VQ-P 25/05/2018 159,72 0012171916 SSD SAMSUNG 2.5 11/04/2018 138,19 1012141917 ROCKER 25-D WITH DOUBLE PLATAFORM 19/07/2018 798,60 1012141918 ROCKER 25-D WITH DOUBLE PLATAFORM 19/07/2018 798,60 6042141919 STEREOMICROSCOPE LEICA S9I 29/06/2018 7.782,94 5112141920 ANALYTICAL BALANCE 210G/0,1MG 18/07/2018 1.129,84 5112141921 EPPENDORF XPLORER, 8 CHANNELS 01/07/2018 888,62 PHOTOMICROGRAPHY CAMERA DS-RI2 DE 2032141922 31/08/2018 8.499,04 NIKON + COMPONENTS PE-300 WHITE STANDARD FLUORESCENCE 1052141923 01/10/2018 4.987,84 ILLUMINATION 3032141924 CRYOSTAT MICROTOME 10/09/2018 34.076,79 AREX-6 DIGITAL WITH PT100 PROBE 6042141925 17/08/2019 550,25 230V/50 6072141926 INFRARED CAMERA FOR REACHING 31/05/2018 3.136,32 6072141927 REGISTRY VIDEO CAMERA FOR REACHING 25/10/2018 1.005,39 6072141928 REGISTRY VIDEO CAMERA FOR REACHING 25/10/2018 1.005,39 5032141929 EPPENDORFRESEARCH PLUS PIPETTE 19/09/2018 714,38 5032141930 FINNPIPETTE F2 MULTICHANNEL 23/10/2018 739,55 2012141931 ELECTROPHORESIS CUVETTE 10/11/2018 912,10 1032141932 INCUBATOR FOR MICROSCOPE 18/10/2018 11.562,69 VARIABLE SPEED NUTATION SHAKER, 100- 1032141933 28/12/2018 554,91 240 V 1032141934 CS-C-DBCP-VR DB CORE 31/10/2018 3.870,86 4122141935 ROCKER VARIABLE SPEED ROCKER II 230V 14/12/2018 653,91

320 CIBERNED 2018 ANNUAL REPORT

HUMAN RESOURCES During 2018, the number of people who have been part of the consortium’s research groups has totaled 458, of which 310 have been personnel assigned to the research groups and 148 have been staff hired by CIBERNED at some point of the year. Below we detail the personnel segmented by each research group.

PRINCIPAL GROUP INSTITUTION ASSIGNED HIRED INVESTIGATOR

Cuadrado Pastor, Autonomous University of 101 3 2 Antonio Madrid 102 Fariñas Gómez, Isabel University of Valencia 9 4 Fuentes Rodríguez, 103 University of Extremadura 2 1 José Manuel Andalusian Public Foundation 105 López Barneo, José for Health Research 12 2 Management in Seville 106 Ceña Callejo, Valentín University ofCastilla La Mancha 2 3 Higher Council of Scientific 108 Vicario Abejón, Carlos 2 1 Research Fundación Instituto de Recerca 109 Vila Bover, Miquel del Hospital Universitario Valle 10 4 de Hebrón Pérez Castillo, Ana Higher Council of Scientific 110 1 5 María Research Higher Council of Scientific 111 Iglesias Vacas, Teresa 2 4 Research García Verdugo, José 113 University of Valencia 1 2 Manuel Del Río Fernández, Bioengineering Institute of 114 1 1 José Antonio Catalonia (IBEC) Canela Campos, Enric 201 University of Barcelona 4 3 Isidre Foundation Research Institute Kulisevsky Bojarski, 202 of the Hospital de la Santa 12 1 Jaime Creu and Sant Pau Lanciego Pérez, José Foundation for Applied Medical 203 4 2 Luis Research (FIMA) Moratalla Villalba, Higher Council of Scientific 204 3 2 Rosario Research Obeso Inchausti, José 205 HM – Hospitals of Madrid 4 3 Ángel Rodríguez Díaz, 206 University of La Laguna 4 1 Manuel Barcelona Clinical and 207 Tolosa Sarró, Eduardo 7 2 Provincial Hospital Labandeira García, University of Santiago de 208 11 2 José Luis Compostela Higher Council of Scientific 209 Pérez Tur, Jordi 1 1 Research 301 Alberch Vié, Jordi University of Barcelona 2 4

321 PRINCIPAL GROUP INSTITUTION ASSIGNED HIRED INVESTIGATOR Complutense University of 303 Fernández Ruiz, Javier 13 3 Madrid Guzmán Pastor, Complutense University of 305 8 2 Manuel Madrid Lucas Lozano, José Higher Council of Scientific 306 1 6 Javier Research Naranjo Orovio, José Higher Council of Scientific 307 0 3 Ramón Research Higher Council of Scientific 401 Ávila de Grado, Jesús 3 6 Research Camins Espuny, 402 University of Barcelona 6 1 Antonio De Felipe Oroquieta, Higher Council of Scientific 403 10 4 Javier Research University of the Basque 404 Matute Almau, Carlos 14 3 Country Rodríguez Álvarez, Autonomous University of 406 5 2 José Barcelona Miguel Hernández University 407 Sáez Valero, Javier 0 2 of Elche Soriano García, 408 University of Barcelona 3 4 Eduardo Torres Alemán, Higher Council of Scientific 409 4 2 Ignacio Research Higher Council of Scientific 410 Trullas Oliva, Ramón 1 3 Research Vitorica Ferrández, 411 University of Sevilla 3 2 Francisco Javier Wandosell Jurado, Higher Council of Scientific 412 2 2 Francisco Research Foundation Research Institute Comella Carnice, Joan 413 of the University Hospital Valle 0 4 Xavier de Hebron Gutiérrez Pérez, 415 University of Malaga 8 2 Antonia Foundation for Biomedical 502 Carro Díaz, Eva María Research of the 12 de Octubre 7 3 University Hospital Ferrer Abizanda, Bellvitge Biomedical Research 503 3 7 Isidro Institute (IDIBELL) Foundation Research Institute 504 Lleó Bisa, Alberto of the Hospital de la Santa 25 1 Creu and Sant Pau Mengod Los Arcos, Higher Council of Scientific 508 2 2 Guadalupe Research De Pedro Cuesta, 509 Carlos III Institute of Health 9 3 Jesús Bullido Gómez-Heras, Autonomous University of 510 2 2 María Jesús Madrid Cantero Lorente, José 511 Pablo de Olavide University 4 3 Luis

322 CIBERNED 2018 ANNUAL REPORT

PRINCIPAL GROUP INSTITUTION ASSIGNED HIRED INVESTIGATOR Marqués de Valdecilla 601 Infante Ceberio, Jon 13 4 Foundation 604 Muñoz Cánoves, Pura Pompeu Fabra University 6 2 Fernández Chacón, 606 University of Seville 6 3 Rafael Navarro Acebes, Autonomous University of 607 26 3 Xavier Barcelona López de Munain Biodonostia Health Research 609 29 2 Arregui, Adolfo Institute Administration 12 Total 310 148

Comparing these data with those collected in 2017, we can observe a stabilization of personnel, producing small variations compared to the immediately previous year. Spe- cifically, there has been an increase of 12 assigned researchers and a decrease of 10 hired workers. The distribution of these personnel by the different Regions is as follows.

Distribution of personnel by Region

Assigned Hired

323 INCORPORATION OF NEW TALENT Maintaining our quest to attract the greatest amount of talent to CIBERNED’s research, during the year 2018 a total of 12 call for personnel have been published, in which 33 job positions have been posted. This year we have increased the dissemination of our job calls through different social networks in order to reach the largest number of potential candidates, whether they are located in the national territory or not.

CALL YEAR NUMBER OF CALLS AVAILABLE POSITIONS 2015 8 47 2016 8 37 2017 10 36 2018 12 33

All jobs posted are defined with a specific profile, required qualifications, job requirements and roles to be developed space as well as the working hours and location of the workplace.

Out of the 33 positions filled-in during 2018, thirteen people had a PhD degree, fourteen were graduates with Higher Degree, one graduate with Middle Degree, and five were advanced technicians.

CATEGORY 2017 2018 Doctor (PhD) 10 13 Graduate (Higher Degree) 22 14 Graduate (Middle Degree) - 1 Advanced technician 4 5 36 33

Another interesting fact that we can highlight from these results is that of the 33 job profiles that have been posted in 2018, 25 of them have been filled in by women and 8 have been filled in by men, which means that 76% of the job postings called had a woman as the best candidate.

New incoporations by gender Personnel by gender

Female Male Female Male

324 CIBERNED 2018 ANNUAL REPORT

INCORPORATION AND ORIENTATION OF NEW PERSONNEL CIBERNED is a consortium for research, with its own legal personality, without physical contiguity, and this can be a stumbling block for the incorporation and orientation of new staff in research groups. To facilitate this quickly and efficiently, a Welcome Hand- book was written that is provided to workers from day one. This guide contains all the information that will be useful in providing the new personnel with an overview of our organization, as well as some basic guidelines that facilitate their working relationship. Its contents includes a brief presentation of CIBERNED, its origin and its objectives, job classification, organization of work, leaves and vacations, a summary of Occupational Risk Prevention (ORP) and templates to handles vacations and ORP. Likewise, they are provided with a Safety and Health Handbook with the information related to their job position. In order to guarantee the health and safety of all its researchers in all activities undertaken at the workplace, CIBERNED provides a health-friendly work environment in accordance with the highest standards of reliability, hygiene and safety. The Human Resources department ensures the safety of each worker and compliance with the Occupational Health and Safety protocols, seeing it as an opportunity to detect room for improvement in those areas most closely linked to work procedures, professional development, compe- tences, training and communication with researchers. To improve all this type of information and facilitate the procedures that CIBERNED workers may need, this 2018 we have been working on improving our intranet so that from the first day our workers can obtain all the information related to the Consortium, they can already access and perform all work procedures in a simple and efficient way.

Distribution of personnel in the research groups during 2018

During 2018 the distribution of personnel by type of contract was 27 permanent hirings and 121 part-time ones.

Types of work contracts

Part-time Permanent

325 During 2018 the distribution of personnel according to their degree was 53 Doctors (PhD), 51 graduates Higher Degree), 4 graduates (Middle Degree), 34 advanced technicians, and 6 administrative assistants.

Professional category

Doctors (PhD)

Higher Degree

Middle Degree Technicians

Administrative assistants

326 CIBERNED 2018 ANNUAL REPORT

TRAINING PROGRAM During 2018, the activities included in the CIBERNED‘s Training Program have continued to be promoted and semi-annual grants have been called for the realization of short secondments in other research centers, as well as for attending courses or training activities that contribute to the training of our researchers in the field of neurodegenerative diseases. In 2018, two semi-annual calls were launched that included the following actions: • Grants for the realization of short stays in other research centers. • Grants for carrying out training activities Below are both the number of applications received, and those granted throughout this year:

Applications 2018

1st. Call 2nd. Call

Mobility applications Training applications Budget requested

327 Grants awarded 2018

1st. Call 2nd. Call

Mobility grants awarded Training grants awarded Budget approved

328 CIBERNED 2018 ANNUAL REPORT

PREVENTION OF OCCUPATIONAL RISKS During the management period analyzed, the following preventive activities have been carried out: • Coordination of the visits to the research centers with the External Prevention Service and the different associated Institutions, for data collection related to personnel risk assessment. • Management of the training of employees in Prevention of Occupational Hazards, requesting the Prevention Service the specific training schedule for existing job positions. • Delivery of laboratory, offices and health sector handbooks to inform workers of the risks of their jobs and preventive measures to be observed for safe work. • The monitoring of individual health has been carried out for more than 73% of the workforce. The medical examinations carried out were 93, subject to specific protocols according to the risks to which the employees are exposed. Finally, the objectives of improving the safety and health conditions of employees and reducing accident rates taken as a reference by the Mutual Insurance Company for Occupa- tional Accidents and Disease have been met, keeping those indexes to zero and therefore below the performance benchmarks of the Research and Development sector:

CNAE REFERENCE CIBERNED

LIMIT VALUE VALUE ACCIDENT RATE OF ACCIDENT RATE INDEX 1 5,98 0 0 INDEX 2 0,54 0 0 INDEX 3 0,33 0 0

QUALITY In 2018, the transition to the new version of ISO 9001:2015 standard was made; as well as the renewal of the certificate of said quality standard. For this purpose, we worked on the evaluation, administration, elimination and/or mini- mization of risks and opportunities and measures were taken to ensure the reduction of the effects of said risks; opportunities were also effectively addressed and monitored. The understanding of needs and expectations established the way in which the -CIBERNED responded to these measures. CIBERNED Management Office of the is open to any improvement proposal, where it will be the organization’s own leaders who promote and develop a quality culture and adaptation to the organization’s change.

329 COLLABORATION AGREEMENTS In 2018, the following agreements have been kept in force by the institution:

SIGNING PARTNER INSTITUTIONS SUBJECT

Establishing a cooperation framework to perform joint IRTA (PERSONNEL IDIBELL)- activities in the filed of neurodegenerative diseases, CIBERNED including prion disorders or transmissible spongiform encephalopathies

STATE GENERAL ADMINISTRATION (MINISTRY OF JUSTICE, STATE’S ATTORNEY, DIRECTORATE OF THE Legal assistance LEGAL SERVICE OF THE STATE)- CIBERNED RAMÓN ARECES FOUNDATION XVIII National call for research grants on life and -CIBERNED materials sciences Collaboration agreement for the implementation of BIOCROSS- CIEN FOUNDATION public-private cooperation actions within the call RIS3 -CIBERNED 2016 Collaboration agreement for the implementation of BIOCROSS- CIEN FOUNDATION public-private cooperation actions within the call retos -CIBERNED colaboración 2017 Regulate the participation of the research groups of CIBER ISCIII-CIBERNED ciberned in the Vallecas project (Cohortes p. Vallecas) UNIVERSITY OF BARCELONA Assignment of use to the university of barcelona CLÍNIC FOUNDATION FOR Co-ownership agreement related to patent “Method BIOMEDICAL RESEARCH, CLINICAL for the subclassification for patients suffering from HOSPITAL OF BARCELONA, UNIV. OF parkinson disease" BARCELONA, CSIC AND CIBERNED Co-ownership agreement related to patent “A fast and BIOCROSS-ISCIII-CIBERNED cost-effective method for apolipoprotein e isotyping as an alternative to apoe genotyping” NATIONAL EVALUATION AND Scientific-technical evaluation of ciberned cooperative PROSPECTIVE AGENCY (ANEP)- projects CIBERNED Co-ownership agreement related to patent nº CSIC-AUTONOMOUS UNIVERSITY OF 201431898 "Compuestos modulares del sensor neuronal MADRID-CIBERNED de calcio dream y sus usos terapéuticos" Co-ownership agreement related to patent CSIC-UAM-CIBERNED nº201431898 “Compuestos moduladores del sensor neuronal de calcio dream y sus usos terapéuticos" Management of training and mobility grants from the SECRETARIAT OF STATE OF promotion of talent program and its employability of UNIVERSITIES MINISTERY OF the state plan of scientific and technical research and of EDUCATION, CULTURE AND SPORTS innovation 2013-2016 and other actions of grants calls -CIBERNED for the mobility of students and college teachers UNIVERSITY OF EXTREMADURA- Curricular and extracurricular practices of students CIBERNED Co-ownership agreement related to patent "Regulación COMPLUTENSE UNIVERSITY OF de receptores CB1 de cannabinoides para la generación MADRID-CIBERNED de neuronas corticoespinales a partir de células troncales embrionarias murinas"

330 CIBERNED 2018 ANNUAL REPORT

SIGNING PARTNER INSTITUTIONS SUBJECT UNIVERSITY OFEXTREMADURA- Curricular and extracurricular practices of students CIBERNED Establish a framework for the development of relations CIBERNED ASSOCIATED INSTITUTIONS with institutions that are integrated in the consortium HOSPITALES DE MADRID Establish a framework for the development of relations FOUNDATION -CIBERNED with institutions that are integrated in the consortium MIGUEL HERNÁNDEZ UNIVERSITY OF ELCHE-FOUNDATION FOR THE Co-ownership agreement related to patent P201130290 PROMOTION OF HEALTH AND "Método de diagnóstico y/o pronóstico de la enfermedad BIOMEDICAL RESEARCH OF THE de alzheimer" REGION OF VALENCIA -CIBERNED Co-ownership agreement related to patent pct/ FIBIO 12 DE OCTUBRE HOSPITAL es2013/070693 "Modelo animal de déficit cognitivo, -CIBERNED procedimiento de obtención y aplicaciones" Co-ownership agreement related to patent CSIC - CIEN- FOUNDATION CIBERNED ep13382108.2 “Methods for the prognosis and diagnosis of neurodegenerative diseases” Non-disclosure agreement related to patent BIOCROSS – CIEN FOUNDATION ep13382108.2 “Methods for the prognosis and -CIBERNED diagnosis of neurodegenerative diseases” Non-disclosure agreement related to patent P201231654 “Proteína viral recombinante SGG2/y/o CSIC - NEURON BIO-CIBERNED complejos binarios SGG2-FNS para su uso en crecimiento y/o regeneración axonal” Co-ownership agreement related to patent p201231654 “Proteína viral recombinante SGG2/y/o complejos CSIC-CIBERNED binarios SGG2-fns para su uso en crecimiento y/o regeneración axonal” FUNDACIÓN ACE-CIBERNED Collaboration framework agreement Collaboration framework agreement to promote and TEOFILO HERNANDO INSTITUTE strengthen study, research, and teaching activities, (UAM)-CIBERNED as well as the dissemination and holding of events of scientific, academic and cultural interest Co-ownership agreement related to patent "ADN CSIC-CIBERNED mitocondrial como marcador para el diagnóstico de enfermedades neurodegenerativas" FIBIO RAMÓN Y CAJAL HOSPITAL Co-ownership agreement related to patent "Medio -CIBERNED condicionado de glía como agente neuroprotector" Co-ownership agreement related to patent "método MIGUEL HERNÁNDEZ UNIVERSITY OF para determinar la enfermedad de alzheimer mediante la ELCHE-CIBERNED detección de glicoproteínas portadoras del glicoepitodo HNK-1" Co-ownership agreement related to patent p201130033 CIEN FOUNDATION - CSIC - CRG - "Compuestos para el tratamiento de enfermedades CIBERNED neurodegenerativas" Establish an association agreement in the framework of CIEN FOUNDATION (UIPA. ALZHEIMER art. 29 of the ciberned statutes for the development of PROJECT RESEARCH UNIT)-CIBERNED scientific-technological research on neurodegenerative diseases Collaboration agreement for for the creation of the CIBER ISCIII-ASSOCIATED INSTITUTIONS consortium for the thematic area of neurodegenerative diseases

331

INVESTIGATORS INDEX ALPHABETICAL INDEX

112 | Alberch Vié, J / Program 2 - Group 301 023 | Ávila de Grado, J - Program 1 - Group 401 026 | Bullido Gómez-Heras, MJ - Program 1 - Group 510 029 | Calero Lara, M - Program 1 - Group 509 034 | Camins Espuny, A - Program 1 - Group 402 040 | Cantero Lorente, JL - Program 1 - Group 511 043 | Carro Díaz, E - Program 1 - Group 502 119 | Ceña Cañejo, V - Program 2 - Group 106 048 | Comella Carnicé, J - Program 1 - Group 413 121 | Cuadrado Pastor, A - Program 2 - Group 101 051 | de Felipe Oroquieta, J - Program 1 - Group 403 194 | del Río Fernández, JA - Program 2 - Group 114 125 | Fariñas Gómez, I - Program 2 - Group 102 222 | Fernández-Chacón, R - Program 3 - Group 606 129 | Fernández Ruiz, J - Program 2 - Group 303 055 | Ferrer Abizanda, I - Program 1 - Group 503 134 | Franco Fernández, R - Program 2 - Group 201 140 | Fuentes Rodríguez, JM - Program 2 - Group 103 144 | García Verdugo, JM - Program 2 - Group 113 061 | Gutiérrez Pérez, A - Program 1 - Group 415 148 | Guzmán Pastor, M - Program 2 - Group 305 152 | Iglesias vacas, T - Program 2 - Group 111 225 | Infante Ceberio, J - Program 3 - Group 601 155 | Kulisevsky Bojarsky, J - Program 2 - Group 202 161 | Labandeira García, JL - Program 2 - Group 208

334 CIBERNED 2018 ANNUAL REPORT

166 | Lanciego Pérez, J - Program 2 - Group 203 065 | Lleó Bisa, A - Program 1 - Group 504 169 | López Barneo, J - Program 2 - Group 105 234 | López de Munain Arregui, A - Program 3 - Group 609 174 | Lucas Lozano, JJ - Program 2 - Group 306 075 | Matute Almau, C - Program 1 - Group 404 080 | Mengod Los Arcos, G - Program 1 - Group 508 177 | Moratalla Villalba, R - Program 2 - Group 204 245 | Muñoz-Cánoves, P - Program 3 - Group 604 181 | Naranjo Orovio, JR - Program 2 - Group 307 249 | Navarro Acebes, X - Program 3 - Group 607 184 | Obeso Inchausti, J - Program 2 - Group 205 190 | Pérez Castillo, AM - Program 2 - Group 110 192 | Pérez Tur, J - Program 2 - Group 209 082 | Rodríguez Álvarez, J - Program 1 - Group 406 198 | Rodríguez Díaz, M - Program 2 - Group 206 085 | Sáez Valero, J - Program 1 - Group 407 089 | Soriano García, E - Program 1 - Group 408 201 | Tolosa Sarró, E - Program 2 - Group 207 093 | Torres Alemán, I - Program 1 - Group 409 096 | Trullás Oliva, R - Program 1 - Group 410 211 | Vicario Abejón, C - Program 2 - Group 108 214 | Vila Bover, M - Program 2 - Group 109 099 | Vitorica Ferrández, JV - Program 1 - Group 411 102 | Wandosell Jurado, F - Program 1 - Group 412

335

C/ Valderrebollo, 52 8031 Madrid Tel.: + 34 913 852 200 Fax: +34 913 852 118 www.ciberned.es