Report Annual

2017 ANNUAL REPORT

MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD

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: Duomo Comunicación INDEX

• Letter from the Scientific Director...... 7

• Overview...... 8

• Aims...... 10

• Directory of research groups and associated institutions...... 11

• Geographic distribution of CIBERNED research groups...... 13

• Organizational structure...... 14

• Organization chart...... 15

• External scientific advisory committee...... 16

• Consortium members ...... 17

• Scientific report...... 19

• Research programs...... 21

• Program 1: Alzheimer’s disease and other degenerative dementias...... 23

• Program 2: Parkinson’s disease and other neurodegenerative movement disorders ...... 107

• Cooperative research...... 237

• International relations...... 257

• Scientific productivity and other activities...... 271

• Financial report...... 287

• Principal investigator index...... 313

LETTER FROM THE SCIENTIFIC DIRECTOR

JESÚS ÁVILA DE GRADO SCIENTIFIC DIRECTOR

During the year 2017, the 52 research groups that make up our Network Center have carried out a fruitful work, which allows us to continue leading the Spanish Centers with publications with the highest impact factor in the field of Biomedicine. This accomplishment is a consequence of the rigorous, efficient and quality activities carried out by the members of CIBERNED. The level of productivity of the different research groups is gradually increasing.

In addition to recognizing the effort of our researchers, I would also like to thank the funding of our activities through the financial support of the Carlos III Institute of Health and the collaboration we have with the associated institutions. Furthermore, with regard to the management of the Center, I would like to thank the efforts of my colleagues on the Steering Committee and the Center’s Management Office, as well as all the support staff.

As a brief summary it can be highlighted that this year more than 40 articles have been published in journals with an impact factor (IF) greater than 10. Twelve of those research papers are in journals with an IF higher than 23, such as Science, Cell, Nature Medicine or Lancet Neurology. Moreover, considering the translational research nature of our Center, new patents have been requested, more than 75 clinical trials have been kept active and new clinical guidelines have been published. Various training courses, congresses or seminars have also been organized. On the other hand, the Training Plan has continued to be developed. Additionally, I would like to congratulate those colleagues who have received international or national awards or have been granted relevant European and international projects.

Finally, I would like to mention the high and valuable participation in our Scientific and Social Forums, specially highlighting the Global Summit held in Lisbon in partnership with the CIEN, Champalimaud and Queen Sofia Foundations.

In short, this has been a good year. Thank you all.

Jesús Ávila de Grado Scientific Director

2017 CIBERNED Annual Report / 7 OVERVIEW

The Network Center for Biomedical Research of cutting research in neurodegenerative in Neurodegenerative Diseases (CIBERNED) disorders through a Network Research emerges in 2006 from the Center for Research Structure, thus contributing to scientifically in Neurological Diseases (CIEN), one of the four substantiate the programs and policies of the Centers together with the CNIO, CNIC or CNMVIS National Health System in the priority areas of that were created with the mission of fighting the National R+D+I. the most prevalent health problems of Spanish society: cancer, cardiovascular diseases, infectious diseases and neurodegenerative disorders. As for the other Centers, a supporting CIBERNED emerges foundation named CIEN Foundation was created in 2003 as the CIEN’s management in 2006 to fight body, although unlike the other Centers, it was neurodegenerative created as the first Network Center in Spain, eventually being renamed as CIBERNED. Since disorders 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 CIBERNED is a research organism, endowed with located in Vallecas (Madrid). legal status under the Article 6.5 of Law 30/1992 of November 26 on the Legal Regime of Public CIBERNED was founded under the auspices Administrations and Common Administrative of the Carlos III Institute of Health, under Procedure. It is formed by the association of a cooperation agreement signed by the research groups, with no physical contiguity, Government and other participating institutions belonging to different administrations, with the idea of creating a Research Center institutions and regional governments, from in which basic, clinical and population-based public and private sectors with research lines studies are integrated in order to develop and goals focused on the specific common a single joint research program, focusing area of neurodegenerative diseases and being on certain pathologies of great importance coordinated to achieve scientific objectives for the National Health System either for its that could hardly be considered in a more prevalence or because of its social impact. It restricted execution context. CIBERNED is has a multidisciplinary and multi-institutional governed in the internal operating rules, by character and it is aimed at boosting the impact way of a Regulation.

2017 CIBERNED Annual Report / 8 Currently, the CIBERNED-CIEN Foundation participating on CIBERNED’s own cooperative partnership is the only research center in Spain research agenda. (and one of the few in the world) integrated into the international network of Centres of Thus, it is the result of a collaborative partnership Excellence in Neurodegeneration (COEN), an between different institutions and the sum of 52 initiative arising from the European Union Joint research groups. Depending on the results of Programme for Research in Neurodegenerative the scientific evaluation of the various research Diseases (JPND). This joint program constitutes groups, there is the possibility to agree on the an innovative collaborative research initiative discontinuation of some groups. Depending on created to address the growing challenges budget availability, it may also consider adding posed by this group of diseases. Its goal is to some new research group currently outside boost the impact of research by aligning existing CIBERNED that satisfy the requirements of national research programs and identifying scientific quality and translational activity and the common objectives whose scope would aligns with CIBERNED priorities. benefit through joint action.

Its own statutes govern CIBERNED and since the end of 2014 its activity is structured around two CIBERNED is the result Scientific Programs: of a collaborative ► Program 1: Alzheimer’s disease and other degenerative dementias partnership between different institutions ► Program 2: Parkinson’s disease and other neurodegenerative motor disorders and research groups During 2017 CIBERNED has consisted of 52 research groups supported by different universities, hospitals and the Research Council (CSIC), each led by a principal investigator or The Scientific Director Office has been set since responsible. CIBERNED is a network research October 2011 at the Center for Molecular center, composed of research groups Biology “Severo Ochoa” (CSIC) in Madrid. The belonging to different Administrations and headquarters, including the General Manager affiliated Institutions: researcher members Office is located in the Alzheimer Center of the physically work in their parent institutions they Queen Sofia Foundation, 5 Valderrebollo Street belong to while simultaneously and actively in Madrid.

2006 2 52 Foundation of Scientific Research CIBERNED programs groups

2017 CIBERNED Annual Report / 9 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-platforms (biobanks, brain imaging ...).

► To involve society in the enormous medical and socioeconomic impact of neurodegenerative diseases and facilitate their involvement in the fight against neurodegeneration.

José Antonio del Río Fernández, Group 114, Molecular and Cellular Neurobiotechnology

2017 CIBERNED Annual Report / 10 DIRECTORY OF RESEARCH GROUPS AND ASSOCIATED INSTITUTIONS

Group Principal investigator Institution

301 Alberch Vié, Jordi University of Barcelona 401 Ávila de Grado, Jesús Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid 510 Bullido Gómez-Heras, Mª Jesús Autonomous University, Madrid 509 Calero Lara, Miguel1 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 Díaz, Eva Doce de Octubre University Hospital, Madrid 106 Ceña Callejo, Valentín University of Castilla La Mancha, Albacete 413 Comella Carnicé, Joan Xavier Vall d’Hebron University Hospital, Barcelona 101 Cuadrado Pastor, Antonio Autonomous University of Madrid 403 de Felipe Oroquieta, Javier Cajal Institute CSIC; Technical University, 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 University of Extremadura, Caceres 113 García Verdugo, José Manuel Cavanilles Institute, University of Valencia 104 González Castaño, José Autonomous University of Madrid 415 Gutiérrez Pérez, Antonia University of Malaga 305 Guzmán Pastor, Manuel Complutense University of Madrid 111 Iglesias Vacas, Teresa Instituto de Investigaciones Biomédicas CSIC-UAM, Madrid 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 University of Santiago de Compostela 203 Lanciego Pérez, José Luis Center of Applied Medical Research, Univ. Navarra, Pamplona 504 Lleó Bisa, Alberto Santa Creu i Sant Pau Hospital, Barcelona 105 López Barneo, José Virgen del Rocio University Hospital, University of Sevilla 609 López de Munain Arregui, Adolfo Biodonostia Research Institute, San Sebastian 306 Lucas Lozano, José Javier Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid 404 Matute Almau, Carlos University of the Basque Country, Bilbao 508 Mengod Los Arcos, Guadalupe Institute of Biomedical Research IDIBAPS-CSIC , Barcelona 204 Moratalla Villalba, Rosario Cajal Institute CSIC, Madrid

1 Since March 2017

2017 CIBERNED Annual Report / 11

Group Principal investigator Institution

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 University of Barcelona 205 Obeso Inchausti, José Ángel Hospitales de Madrid Foundation 507 Pastor Muñoz, María Asunción Center of Applied Medical Research, Univ. Navarra, Pamplona 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 of Barcelona 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 Hospital Clinic of Barcelona 409 Torres Alemán, Ignacio Cajal Institute CSIC, Madrid 410 Trullás Oliva, Ramón Institute of Biomedical Research IDIBAPS-CSIC , Barcelona 108 Vicario Abejón, Carlos Cajal Institute CSIC, Madrid 109 Vila Bover, Miquel Vall d’Hebron University Hospital, Barcelona 411 Vitorica Ferrández, Fco. Javier University of Seville 412 Wandosell Jurado, Francisco Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid

52 RESEARCH GROUPS

PROGRAM 1: 21 GROUPS

PROGRAM 2: 31 GROUPS

2017 CIBERNED Annual Report / 12 GEOGRAPHIC DISTRIBUTION OF CIBERNED RESEARCH GROUPS

1 2 1 2 16

1 1 4

2017 CIBERNED Annual Report / 13 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 for Research in 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 Governing 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 the organs of government:

● The Steering Committee ● The External Scientific Advisory Committee

The Steering Committee consists of:

● Dr. Jesús Ávila de Grado – Scientific Director ● Ms. María Ángeles Pérez Muñoz – Manager ● Dr. Miguel Medina Padilla – Deputy Scientific Director

And the Program coordinators:

● Dr. Eduardo Tolosa Sarró ● Dr. Albert Lleó Bisa ● Dr. José Javier Lucas Lozano ● Dr. Adolfo López de Munaín ● Dra. Teresa Iglesias Vacas ● Dr. Rafael Fernández Chacón ● Dra. Isabel Fariñas Gómez1

1 Since December 2017

2017 CIBERNED Annual Report / 14 ORGANIZATION CHART

INSTITUTE OF HEALTH CARLOS III

GOVERNING COUNCIL

ASSOCIATED INSTITUTIONS

STANDING COMMITTEE

STEERING COMMITTEE SCIENTIFIC DIRECTOR MANAGER

DEPUTY SCIENTIFIC DIRECTOR RESEARCH PROGRAM COORDINATOR

RESEARCH GROUPS

2017 CIBERNED Annual Report / 15 EXTERNAL SCIENTIFIC ADVISORY COMMITTEE

Dr. George Perry University of Texas, San Antonio, USA (Chairman)

Dr. Vincenzo Bonifati Erasmus University, Rotterdam, The Netherlands

Dr. Ángel Cedazo-Mínguez Karolinska Institute, Stockholm, Sweden

Dra. Mary Reilly Institute of Neurology, London, United Kingdom

2017 CIBERNED Annual Report / 16 CONSORTIUM MEMBERS

The Central Administration, represented by:

- The Institute of Health Carlos III.

- 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 Foundation (IDIVAL).

- Autonomous Region of Castilla La Mancha, through the University of Castilla-La Mancha.

- Autonomous Region of Catalonia, through the Pompeu Fabra University, University Hospital- Research Institute Vall d’Hebron Foundation, Bellvitge Biomedical Research Institute Foundation (IDIBELL), Catalonian Institute of Bioengineering, University of Barcelona and Autonomous University of Barcelona.

- Autonomous Region of Valencia, through the University of Valencia and 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 Madrid Health Services (Doce de Octubre University Hospital), 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).

- Center for Applied Medical Research (CIMA, Navarra).

- Hospital Clinic of Barcelona (Catalonia).

- HM Hospitales Research Foundation (Madrid).

2017 CIBERNED Annual Report / 17

Scientific Report

RESEARCH PROGRAMS

From an organizational point of view, CIBERNED research groups are organized around two scientific programs, whose goals, composition and structure are maintained as in previous years, and whose budget allocation is based on the results of the internal scientific evaluation.

This organizational structure highlights CIBERNED commitment to encourage translational research and the generation of scientific knowledge resulting in improving the prevention, diagnosis and treatment of neurodegenerative diseases. All this, without undermining the development of transversal research projects of interest to different areas (see below). The objective of the structure described is to provide a coherent conceptual framework on which the actual cooperative scientific activity carried out by the different research groups materializes.

The Programs are described below:

2017 CIBERNED Annual Report / 21

PROGRAM 1 Alzheimer’s disease and other degenerative dementias

Ávila de Grado, Jesús...... 27 Matute Almau, Carlos...... 75 Bullido Gómez-Heras, Mª Jesús...... 30 Mengod Los Arcos, Guadalupe...... 79 Calero Lara, Miguel...... 33 Pastor Muñoz, María Asunción...... 81 Camins Espuny, Antonio...... 38 Rodríguez Álvarez, José...... 84 Cantero Lorente, José Luis...... 43 Sáez Valero, Javier...... 87 Carro Díaz, Eva...... 47 Soriano García, Eduardo...... 90 Comella Carnicé, Joan Xavier...... 52 Torres Alemán, Ignacio...... 93 de Felipe Oroquieta, Javier...... 55 Trullás Oliva, Ramón...... 96 Ferrer Abizanda, Isidro...... 59 Vitorica Ferrández, Francisco Javier...... 99 Gutiérrez Pérez, Antonia...... 65 Wandosell Jurado, Francisco...... 102 Lleó Bisa, Alberto...... 68

PROGRAM 1: ALZHEIMER’S DISEASE AND OTHER DEGENERATIVE DEMENTIAS

Aging is one of the major risk factors for some of the most prevalent diseases such as cancer, cardiovascular disorders or neurodegenerative diseases, but while the number of cases of the formers tends to stabilize, the number of patients with neurodegenerative pathologies, especially dementia, continues to grow exponentially. According to EUROSTAT, Spain is among the oldest countries in Europe and the world and the demographic estimates from the National Statistics Institute indicate that there are 8.7 million people aged 65 or older live in Spain in 2017 (18.7% of the population) whereas it is anticipated that in 2050 about 15 million Spaniards (approximately one third of our population) will be over 65 years.

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 almost one million 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 reproduced in cellular or animal models.

Laboratories around the world work with great interest in identifying new causal and risk genes involved in this pathology that could help clarify its pathophysiological basis and lead to the identification of new therapeutic targets. One of the main problems in the EA 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.

In this program, 21 groups of clinical and basic researchers are committed both to the diagnosis and care of patients suffering EA. Thus, research in the laboratory will continue over the next year 2016 combining experience 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.

2017 CIBERNED Annual Report / 25 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, Miguel1 Institute of Health Carlos III, Madrid

Camins Espuny, Antonio University de Barcelona

Cantero Lorente, José Luis Pablo de Olavide University, Sevilla

Carro Díaz, Eva Hospital Universitario Doce de Octubre, Madrid

Comella Carnice, Joan Xavier Foundation Vall d’Hebron University Hospital, Barcelona

De Felipe Oroquieta, Javier Cajal Institute CSIC, Center of Biomedical Technology, 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

Pastor Muñoz, María Asunción Center of Applied Medical Research, University of, Pamplona

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, CSIC , 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).

1 PI of the group since March 2017 due to retirement of Dr. Jesus de Pedro Cuesta

2017 CIBERNED Annual Report / 26 Jesús Ávila de Grado 401

Principal Investigator Jesús Ávila de Grado

Research team

Félix Hernández Pérez Noemí Pallas Bazarra Miguel de la Flor UAM Professor – Co-IP Researcher PhD student

Marta Bolos Jurado Jesús Merchán Robira Elena Moreno Jiménez Researcher PhD student PhD student

María Llorens-Martín Alberto García Rodríguez Raquel Cuadros Catalán Researcher PhD student Technician

Vega García-Escudero Juan Ramón Perea Esther García García Researcher PhD student Technician

Julia Terreros Roncal Nuria de la Torre Alonso PhD student Administrative Assistant

Centro de Biología Molecular “Severo Ochoa” C/ Nicolás Cabrera, 1 (Laboratorio 208) Campus de Cantoblanco 28049 Madrid (Spain) Tel.: +34 91 196 45 64 Fax: +34 91 196 44 20 [email protected]

2017 CIBERNED Annual Report / 27 SUMMARY

During the last year, among other studies, that fractalkine is expressed in neurons and it we have found the devastating effect of could be involved in signaling mechanisms extracellular tau on the structural plasticity in microglia. In this way, we have found that (morphology and connectivity) of newborn overexpression of GSK3 in neurons results in hippocampal granule neurons. On the other changes in fractalkine expression, changes that hand, we have also analyzed the effect of could affect fractalkine signaling in microglia. extracellular tau on microglia cells. Our results have indicated, that fractalkine receptor, Our working hypothesis is that in tauopathies, CX3CR1, is acting as microglia receptor like Alzheimer disease, could take place for extracellular tau. In addition, we have changes in fractalkine expression that could observed that absence of microglial CX3CR1 affect to microglia function and tau clearance impairs the synaptic integration of adult-born (or uptake), by these glia cells and that it may hippocampal granule neurons. It is known result in the spreading of tau pathology.

KEYWORDS

Tau, GSK3, Alzheimer disease, Neurogenesis PUBLICATIONS

2017 CIBERNED Annual Report / 28 Program 1 Group: Jesús Ávila de Grado

• Tau has devastating effects on the structural plasticity of hippocampal granule neurons. Translational Psychiatry. 2017;7(12). • Martín-Maestro P, Gargini R, García E, Perry G, Avila J, García-Escudero V. Slower Dynamics and Aged Mitochondria in Sporadic Alzheimer’s Disease.Oxidative medicine and cellular longevity. ;2017. • Quinlan S, Kenny A, Medina M, Engel T, Jimenez-Mateos EM, Ávila de Grado Jesús. MicroRNAs in Neurodegenerative Diseases.International review of cell and molecular biology. ;334. • Avila J. Our Working Point of View of Tau Protein.Journal of Alzheimer’s disease : JAD. 2017. • Anton-Fernandez A., Merchan-Rubira J., Avila J., Hernandez F., Defelipe J., Munoz A. Phospho- Tau Accumulation and Structural Alterations of the Golgi Apparatus of Cortical Pyramidal Neurons in the P301S Tauopathy Mouse Model. Journal of Alzheimer’s Disease. 2017;60(2):651-661. • Avila J., Llorens-Martin M., Pallas-Bazarra N., Bolos M., Perea J.R., Rodriguez-Matellan A. et al. Cognitive decline in neuronal aging and Alzheimer’s disease: Role of NMDA receptors and associated proteins. Frontiers in Neuroscience. 2017;11(NOV). • Garcia-Escudero V., Gargini R., Martin-Maestro P., Garcia E., Garcia-Escudero R., Avila J. Tau mRNA 3′UTR-to-CDS ratio is increased in Alzheimer disease. Neuroscience Letters. 2017;655:101-108. • Sotiropoulos I, Galas MC, Silva JM, Skoulakis E, Wegmann S, Maina MB et al. Atypical, non- standard functions of the microtubule associated Tau protein.Acta neuropathologica communications. 2017;5(1). • Mudher A, Brion JP, Avila J, Medina M, Buée L. EuroTau: towing scientists to tau without tautology. Acta neuropathologica communications. 2017;5(1). Bolos M., Perea J.R., Avila J. Alzheimer’s disease as an inflammatory disease. Biomolecular Concepts. 2017;8(1):37-43. • Medina M., Khachaturian Z.S., Rossor M., Avila J., Cedazo-Minguez A. Toward common mechanisms for risk factors in Alzheimer’s syndrome. Alzheimer’s and Dementia: Translational Research and Clinical Interventions. 2017;3(4):571-578. • Gomez-Ramos A., Picher A.J., Garcia E., Garrido P., Hernandez F., Soriano E. et al. Validation of Suspected Somatic Single Nucleotide Variations in the Brain of Alzheimer’s Disease Patients. Journal of Alzheimer’s Disease. 2017;56(3):977-990.

RESEARCH PROJECTS

• Code: 2016-COHORTE BBN-NED. • Code: I2016/02. Title: COHORTE-BBN-NED. Title: Monitoring the onset and evolution of Búsqueda de biomarcadores para la neuronal dysfunctions in propagative neural detección temprana de la enfermedad disorders using microfluidic devices and de Alzheimer en la cohorte del “Proyecto translational approaches. Vallecas”. Principal Investigator: José Antonio del Río. Principal Investigator: Miguel Medina. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G114, G401, G201. CIBERNED groups: G209, G401. Other CIBER’s collaboration: No. Other CIBER’s collaboration: CIBER-BBN. Type: Intramurales. Type: National. Funding agency: CIBERNED. Funding agency: Instituto de Salud Carlos III. Funding: 210.000 €. Funding: 150.000€. Duration: 2016-2018 Duration: 2016-2018

2017 CIBERNED Annual Report / 29 María Jesús Bullido Gómez-Heras 510

Principal Investigator María Jesús Bullido

Research team

Ana Frank-García Ángel Martín Montes Patricia Llorente Ginés Associated professor Researcher PhD student

Jesús Aldudo Soto Isabel Sastre Merlín Researcher Researcher

María Recuero Vicente Henrike Kristen Researcher PhD student

Centro de Biología Molecular “Severo Ochoa”(CSIC-UAM) Universidad Autónoma de Madrid Edificio CBM Tel.: +34 911 964 45 67 Fax: +34 911 96 44 20 [email protected]

2017 CIBERNED Annual Report / 30 SUMMARY

To identify genes and mechanisms involved in like neurodegenerative events induced by the neurodegeneration process characteristic HSV-1 and OS. To do this, we are characterizing of Alzheimer’s disease (AD), we developed several LAMP2-deficient cell lines (SK-N-MC, cellular models mimicking different aspects of N2a and MEFs). Preliminary results showed the AD pathogenesis intended to identify novel infected LAMP2-deficient cells to produce genes/processes involved in AD pathogenesis. less viral DNA, proteins and mature infectious. We perform gene expression and functional These results suggest that LAMP2 might affect studies in the models, and then validate the most HSV-1 viral cycle, which in turn could affect interesting candidates using genetic association the neurodegenerative events induced by the and biomarkers studies in AD cases and control. virus. To test if the alterations observed in our cellular models do also occur in AD patients, we Currently, we work on models of oxidative stress have started the analysis of biomarkers related (OS) and herpes simplex virus type 1 (HSV-1) to HSV-1 infection and lysosomal function in infection, which present markers characteristic AD patients at different prodromal and clinical of AD, like the alterations in amyloid precursor phases compared with control individuals. protein (APP) traffic, metabolism and These biomarkers include genetic variants proteolysis and in tau protein phosphorylation. (association studies), HSV antibodies in serum The genomic analysis of these models revealed and cerebrospinal fluid, and lysosomal markers that the main pathway altered was the in leukocytes. The data obtained to date suggest lysosomal system, and the functional study of the genetic association of LAMP2 variants with the autophagy lysosome shown HSV-1 infection AD in a gender-dependent manner, as well as and OS to profoundly affect the final steps of the alterations in the anti-HSV IgM levels in serum and route, causing an increased cellular burden of the lysosomal content in leukocytes of patients. lysosomes, a significant reduction in the activity of different lysosomal hydrolases, a defect in In summary, the results obtained from the the cathepsin B maturation, and an impaired functional, gene expression and biomarker trafficking and decreased degradation studies reinforce the hypothesis that a failure of lysosomal substrates as EGF receptor. of the lysosomal function could be a relevant neurodegeneration mechanism, both in cellular In addition, recent results indicate the models and AD patients. We are therefore involvement of cathepsin B in the metabolism currently focused on the analysis of this functional and proteolysis of APP induced by OS, revealing pathway, extending the study to new models a direct link between lysosomal dysfunction including iPSCs derived from patients and and amyloidogenesis. The gene encoding for controls skin fibroblasts. We also participate in the lysosomal membrane protein LAMP2 was, the investigation of the genetic architecture of among potential mediators of the autophagic- AD, in the framework of the Dementia Genetics lysosomal dysfunction, the most modulated Spanish Consortium (DEGESCO), and in one at the transcriptional level. Consequently, international projects (EADI EADB, IGAP, etc ...) we have initiated the study of the role of focused on the search of factors involved in the LAMP2 in the lysosomal dysfunction and AD- AD pathogenesis using “omics” approaches.

KEYWORDS Alzheimer, Herpesvirus, oxidative stress, neurodegeneration, lysosomal function, functional genomics, genetic association PUBLICATIONS • Sims R., Van Der Lee S.J., Naj A.C., Bellenguez C., Badarinarayan N., Jakobsdottir J. et al. Rare coding variants in PLCG2, ABI3, and TREM2 implicate microglial-mediated innate immunity in Alzheimer’s disease. Nature Genetics. 2017;49(9):1373-1384.

2017 CIBERNED Annual Report / 31 RESEARCH PROJECTS

• Code: - . 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: María J. Bullido Gómez-Heras. CIBERNED’s collaboration: No. CIBERNED groups: G510. Other CIBER’s collaboration: No. Type: Private. Funding agency: Fundación Ramón Areces. Funding: 108.107,5€. Duration: 2017-2019

PhD DISSERTATIONS

• Author: Diana Oviedo • Author: Meritxell Valentí Soler. Title: Análisis de las capacidades cognitivas, Title: Robots sociales y animales en la terapia biomarcadores vasculares y APOE4 en de personas con demencia avanzada. deterioro cognitivo leve y la enfermedad de Date: 20th April 2017. Alzheimer en panameños adultos. Supervisor: Ana Frank García. Date: 3rd July 2017. Supervisor: Ana Frank García. • Author: : Henrike S. Kristen. Title: Analysis of a cell model of neurodegeneration induced by herpes simplex virus 1 and oxidative stress. implication of the lysosomal pathway. Date: 1st September 2017. Supervisor: María Jesús Bullido Gómez-Heras.

2017 CIBERNED Annual Report / 32 Miguel Calero Lara 509

Pincipal Investigator Miguel Calero Lara

Research team

Jesús de Pedro Alexandra Moreno García Fernando José García López Consultant PhD Student Postdoctoral researcher

Pablo Martínez Juan Manuel Castellote Olivito María del Carmen Rodríguez Postdoctoral researcher Graduate Blázquez Postdoctoral researcher Olga Calero Rueda Alejandra Elizabeth Kun Postdoctoral researcher González María Ruiz Tovar Visiting researcher Postdoctoral researcher Javier Damián Moreno Postdoctoral researcher Javier Almazán Isla María Belén Frades Payo Technician Graduate Enrique Alcalde Cabrero Postdoctoral researcher Herminia Jiménez Muñoz Technician

Unidad de Encefalopatías Espongiformes Unidad Funcional de Investigación de Enfermedades Crónicas-CROSADIS Instituto de Salud Carlos III Tel.: +34 918 223 709 [email protected] [email protected]

2017 CIBERNED Annual Report / 33 SUMMARY

Public Health/aging Subgroup disability in Parkinson’s disease and behavioral disorders in Huntington’s disease have been In the field of epidemiology and etiology carried out. Several “recommended” scales of neurodegenerative diseases, the most were identified for predicted use in disability for important contribution with the participation Parkinson’s disease and behavioral disorders in of the three subgroups is the publication of Huntington, but the development of a specific the model of “drivers”. This model combines scale for gait and balance in Parkinson’s molecular elements, clinical and general disease was recommended. We have also epidemiology and animal models, providing a carried-out several studies on quality of life in transversal view of sporadic neurodegenerative different populations, transcultural adaptation pathologies, suggesting interpretations that of questionnaires, global assessment scales for encompass other endocrine and vascular Parkinson’s disease, analysis of properties and conformational pathologies, and the idea of performance of different evaluation scales for continuity between certain neurodegenerations neuropsychological disorders, and analysis of from clinical onset to advanced age. the effects of stereotactic surgery on non-motor symptoms in Parkinson’s disease. Regarding aging and disability, several articles deal with the analysis of confinement related Molecular Biology Subgroup to disability and with the loss of key functions, mainly mental and secondly musculoskeletal. The current objectives of this subgroup are Finally, a consensus article among Spanish (i) molecular diagnosis, (ii) basic research on professionals points out the characteristics of the genetic susceptibility factors in Alzheimer’s relationship between mental health and public disease and prion diseases, and (iii) molecular health, as well as the priorities in prevention basis of conformational diseases, including and surveillance in this field. According to data AD and CJD. The subgroup maintains a from the surveillance of human transmissible close collaboration with different CIBERNED spongiform encephalopathies, no new groups in two collaborative projects: the cases of variant Creutzfeldt-Jakob disease DEGESCO consortium, and the National Prion (vCJD) have been diagnosed. A system of Network (AGL2015-71764-REDT Network of pharmacovigilance in Multiple Sclerosis is Excellence). In the context of the DEGESCO designed at the request of the AEMPS. consortium, a number of shared or differential genetic risk factors have been studied in Results subgroup different neurodegenerative diseases. In collaboration with Dr. M. Medina (PI1 of the During the past decade, since the development project and CIBERNED deputy director), we and validation of instruments to evaluate non- are carrying out the project entitled “miRNA motor manifestations of Parkinson’s Disease, and lipid metabolism markers as potential links which have provided information on the between vascular dysfunction and Alzheimer’s importance of these symptoms, it has begun to pathophysiology” (MINECO). The group also consider this disease as a complex multisystem collaborates with the company AMO Pharma disorder. However, a critical review of Non-Motor Ltd, in the exploration of treatment response Symptoms Scale showed shortcomings and markers (tideglusib) for juvenile-onset myotonic implementation problems that have prompted a dystrophy. On the other hand, the group new version, whose validation study (sponsored collaborates with other institutions that provide by the International Parkinson and Movement a fundamental support, highlighting the Disorder Society (IPMDS) began in 2017). collaboration with the CIEN Foundation for the early diagnosis of Alzheimer’s disease (Vallecas In this year and with the support of the IPMDS, project, and the University of the República de several systematic reviews of scales for Uruguay in the study of peripheral expression of assessment of posture, gait and balance, and CNS neurodegenerative diseases. KEYWORDS

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

2017 CIBERNED Annual Report / 34 Program 1 Group: Miguel Calero Lara PUBLICATIONS

• Ayala A., Trivino-Juarez J.M., Forjaz M.J., Rodriguez-Blazquez C., Rojo-Abuin J.-M., Martinez- Martin P. et al. Parkinson’s disease severity at 3 years can be predicted from non-motor symptoms at baseline. Frontiers in Neurology. 2017;8(OCT). • Virues-Ortega J., Vega S., Seijo-Martinez M., Saz P., Rodriguez F., Rodriguez-Laso A. et al. A protective personal factor against disability and dependence in the elderly: an ordinal regression analysis with nine geographically-defined samples from Spain. BMC Geriatrics. 2017;17(1):1-10. • Diack A.B., Boyle A., Ritchie D., Plinston C., Kisielewski D., De Pedro-Cuesta J. et al. Similarities of variant Creutzfeldt-Jakob disease strain in mother and son in Spain to UK reference case. Emerging Infectious Diseases. 2017;23(9):1593-1596. • Lopez F.J.G., Ruiz-Tovar M., Almazan-Isla J., Alcalde-Cabero E., Calero M., de Pedro-Cuesta J. Risk of transmission of sporadic Creutzfeldt-Jakob disease by surgical procedures: Systematic reviews and quality of evidence. Eurosurveillance. 2017;22(43). • Darling A., Tello C., Marti M.J., Garrido C., Aguilera-Albesa S., Tomas Vila M. et al. Clinical rating scale for pantothenate kinase-associated neurodegeneration: A pilot study. Movement Disorders. 2017;32(11):1620-1630. • Skorvanek M, Goldman JG, Jahanshahi M, Marras C, Rektorova I, Schmand B et al. Global scales for cognitive screening in Parkinson’s disease: Critique and recommendations.Movement disorders : official journal of the Movement Disorder Society. 2017. • Luquin MR, Kulisevsky J, Martinez-Martin P, Mir P, Tolosa ES. Consensus on the Definition of Advanced Parkinson’s Disease: A Neurologists-Based Delphi Study (CEPA Study).Parkinson’s disease. ;2017. • Dafsari H.S., Reker P., Stalinski L., Silverdale M., Rizos A., Ashkan K. et al. Quality of life outcomes after subthalamic stimulation in Parkinson’s disease depends on age. Movement Disorders. 2017. • Mestre T.A., Bachoud-Levi A.-C., Marinus J., Stout J.C., Paulsen J.S., Como P. et al. Rating scales for cognition in Huntington’s disease: Critique and recommendations. Movement Disorders. 2017. • Carro E., Bartolome F., Bermejo-Pareja F., Villarejo-Galende A., Molina J.A., Ortiz P. et al. Early diagnosis of mild cognitive impairment and Alzheimer’s disease based on salivary lactoferrin. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2017;8:131-138. • Mestre T.A., van Duijn E., Davis A.M., Bachoud-Levi A.-C., Busse M., Anderson K.E. et al. Response to letter by Saenz-Farret et al. on “Rating scales for behavioral symptoms in Huntington’s disease: Critique and recommendations”. Movement Disorders. 2017;32(3):482-. • Martino D., Pringsheim T.M., Cavanna A.E., Colosimo C., Hartmann A., Leckman J.F. et al. Systematic review of severity scales and screening instruments for tics: Critique and recommendations. Movement Disorders. 2017;32(3):467-473. • Martinez-Martin P. What is quality of life and how do we measure it? Relevance to Parkinson’s disease and movement disorders.Movement disorders : official journal of the Movement Disorder Society. 2017;32(3). • Buetow S.A., Martinez-Martin P., McCormack B. Ultrabilitation: beyond recovery-oriented rehabilitation. Disability and Rehabilitation. 2017;:1-6. • Martinez-Martin P., Ray Chaudhuri K. Comprehensive grading of Parkinson’s disease using motor and non-motor assessments: addressing a key unmet need. Expert Review of Neurotherapeutics. 2018;18(1):41-50. [ePub2017]. • Sauerbier A, Jitkritsadakul O, Titova N, Klingelhoefer L, Tsuboi Y, Carr H et al. Non-Motor Symptoms Assessed by Non-Motor Symptoms Questionnaire and Non-Motor Symptoms Scale in Parkinson’s Disease in Selected Asian Populations.Neuroepidemiology. ;49(1-2). • Padron-Monedero A., Damian J., Pilar Martin M., Fernandez-Cuenca R. Mortality trends for accidental falls in older people in Spain, 2000-2015. BMC Geriatrics. 2017;17(1). • Caron A., Ayala A., Damian J., Rodriguez-Blazquez C., Almazan J., Castellote J.M. et al. Physical activity, body functions and disability among middle-aged and older Spanish adults. BMC Geriatrics. 2017;17(1).

2017 CIBERNED Annual Report / 35 • Dafsari H.S., Reker P., Silverdale M., Reddy P., Pilleri M., Martinez-Martin P. et al. Subthalamic Stimulation Improves Quality of Life of Patients Aged 61 Years or Older With Short Duration of Parkinson’s Disease. Neuromodulation. 2017. • van den Berg MEL, Castellote JM, Mayordomo JI, Mahillo-Fernandez I, de Pedro-Cuesta J. Spinal Cord Injury due to Tumour or Metastasis in Aragón, Northeastern Spain (1991-2008): Incidence, Time Trends, and Neurological Function.BioMed research international. ;2017. • Almazan-Isla J., Comin-Comin M., Alcalde-Cabero E., Ruiz C., Franco E., Magallon R. et al. Disability, support and long-term social care of an elderly Spanish population, 2008-2009: An epidemiologic analysis. International Journal for Equity in Health. 2017;16(1). • Bhidayasiri R., Martinez-Martin P. Clinical Assessments in Parkinson’s Disease: Scales and Monitoring. International Review of Neurobiology. 2017;132:129-182. • Martinez-Martin P., Rodriguez-Blazquez C., Forjaz M.J., Kurtis M.M., Skorvanek M. Measurement of Nonmotor Symptoms in Clinical Practice. International Review of Neurobiology. 2017. • De Roos P., Bloem B.R., Kelley T.A., Antonini A., Dodel R., Hagell P. et al. A Consensus Set of Outcomes for Parkinson’s Disease from the International Consortium for Health Outcomes Measurement. Journal of Parkinson’s Disease. 2017;7(3):533-543. • Titova N., Martinez-Martin P., Katunina E., Chaudhuri K.R. Advanced Parkinson’s or “complex phase” Parkinson’s disease? Re-evaluation is needed. Journal of Neural Transmission. 2017;:1-9. Balestrino R., Martinez-Martin P. Neuropsychiatric symptoms, behavioural disorders, and quality • of life in Parkinson’s disease. Journal of the Neurological Sciences. 2017;373:173-178. Balestrino R., Martinez-Martin P. Reprint of Neuropsychiatric symptoms, behavioural disorders, • and quality of life in Parkinson’s disease. Journal of the neurological sciences. 2017;374:3-8. Martinez-Martin P, Manuel Rojo-Abuin J, Rizos A, Rodriguez-Blazquez C, Trenkwalder C, Perkins • L et al. Distribution and impact on quality of life of the pain modalities assessed by the King’s Parkinson’s disease pain scale.NPJ Parkinson’s disease. ;3. Buetow SA, Martínez-Martín P, McCormack B. Falling upward with Parkinson’s disease.NPJ • Parkinson’s disease. ;3. Radicati FG, Martinez Martin P, Fossati C, Chaudhuri KR, Torti M, Rodriguez Blazquez C et al. Non • motor symptoms in progressive supranuclear palsy: prevalence and severity.NPJ Parkinson’s disease. ;3. • Rodríguez-Blázquez C, Alvarez M, Arakaki T, Campos Arillo V, Chaná P, Fernández W, Garretto N, Martínez-Castrillo JC, Rodríguez-Violante M, Serrano-Dueñas M, Ballesteros D, et al. Self- Assessment of Disability in Parkinson’s Disease: The MDS-UPDRS Part II Versus Clinician-Based Ratings. Movement Disorders Clinical Practice. 2017 Jul 1;4(4):529-35. • Rodríguez-Violante M, Alvarado-Bolaños A, Cervantes-Arriaga A, Martinez-Martin P, Rizos A, Chaudhuri K. Clinical Determinants of Parkinson’s Disease-associated Pain Using the King’s Parkinson’s Disease Pain Scale. Movement Disorders Clinical Practice. 2017 Jul 1;4(4):545-51. RESEARCH PROJECTS

• Code: AGL2015-71764-REDT. Títle: Red nacional de priones. Principal Investigator: José Antonio Del Río. CIBERNED’s collaboration: Yes. CIBERNED groups: G114 ; G509; G503; G609; G207. Other CIBER’s collaboration: No. Type: National. Funding agency: MINECO. Funding: 39.000€. Duration: 2016-2017

2017 CIBERNED Annual Report / 36 Program 1 Group: Miguel Calero Lara

• Code: AMO-02-MD-2-001. • Code: PI15CIII/00037. Title: GSK3 en la distrofia miotónica congénita Títle: Mortalidad en población que vive en de aparcición juvenil. residencias de ancianos de Madrid. Principal Investigator: Miguel Calero. Principal Investigator: Javier Damián. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G509 CIBERNED groups: G509 Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: National. Funding agency: AMO PHARMA. Funding agency: Instituto de Salud Carlos III. Funding: 30.090€. Funding: 0€. Duration: 2016-2017 Duration: 2015-2019

2017 CIBERNED Annual Report / 37 Antonio Camins Espuny 402

Principal Investigator Antonio Camins Espuny

Research team

Mercè Pallàs Lliberia Anna María Canudas Teixidó Elena Sánchez López Full Professor Associated professor Assistant professor

Carme Auladell Costa Carme Pelegrí Gavalda Christian Griñán Farre Associated professor Associated professor Assistant professor

Francesc Sureda Batlle Jordi Vilaplana Hortensi Amanda Cano Fernández Associated professor Associated professor Fellow researcher

Jaume Folch López Ester Verdaguer Cardona Oriol Busquets Figueras Associated professor Lecturer PhD student

María Luisa García López Miren Ettcheto Ariola Dolors Puigoriol Illamola Associated professor CIBERNED researcher Fellow researcher

Foteini Vasilopoulou Fellow researcher

Facultad de Farmacia Universidad de Barcelona Av. Joan XXIII, s/n 08028 Barcelona (Spain) Tel.: +34 934 02 45 31 [email protected]

2017 CIBERNED Annual Report / 38 SUMMARY

Pre-clinical model of Alzheimer’s disease by natural antibodies that exist from birth in the same mice appear in the brain of mice. The results obtained during the last years with This same situation has been determined later APPSwe / PS1dE9 (APP / PS1) mice produce in the corpora amylacea of the human brain. higher levels of β-amyloid than wild type (WT) These facts indicate a certain relationship and mice, treated with a high-fat diet, develop possible interaction between the degenerative memory loss compared with mice fed with a and aging processes on the one hand and the normal diet. Although the reason is unknown, natural immune system on the other. there are many studies that point out that the inactivation of the insulin receptor, at the level General objective: of the hippocampus, area of the brain related to the process of memory and learning, is the • Study the relationship between degenerative main responsible of this cognitive loos. In this and aging processes on the one hand and line, different works suggest that cognitive the natural immune system on the other. loss associated with metabolic disorders (type II diabetes), is a key factor in the onset of Concrete objectives: Alzheimer’s disease, suggests that Alzheimer’s disease not affects only the brain, if not whole 1.- Characterize, in different neurodegenerative organism. diseases and in aging, brain structures that present neo-epitopes A key molecule involved in the regulation of the activity of the insulin receptor is JNK1, 2.- Determine the nature of these neo-epitopes which in turn responds to different types of stress signals and, in particular, glial activation 3.- Determine the natural immune response that and the release of pro-inflammatory mediators. occurs in relation to the formation of these Antonio Camins Espuny For this reason, we are working with this drug, epitopes during aging or neurodegeneration. Licochalcone A, an inhibitor of JNK1 kinase. Through this inhibition, the inactivation of the Development of drugs for the treatment of brain insulin receptor can be prevented being Alzheimer’s disease and neurodegenerative suitable for the treatment of Alzheimer’s disease. diseases in general General objective: Studying and clarifying the process of neuronal death after the stimulation of the ionotropic • To act specifically on the activity of JNK1, glutamate receptors is key to understanding as a key target, preventing phosphorylation neuronal death. We already have drugs and inhibition of the insulin receptor in synthesized to the Pharmaceutical Chemistry the hippocampus and preventing the Unit of the Faculty of Pharmacy that can development of AD. potentially be effective in treating Alzheimer’s disease. On the one hand it is a series of more Concrete objectives: than 400 compounds from 2008 until now (Campos et al., Bioorg Med Chem, 2008, 16, 1.- Assessment of cognitive loss in the process 9925-9936, Duque et al., Bioorg Med Chem, of obesity in relation to the activity of JNK, in 2009, 17, 3198- 3206 and 2010, 18, 46-57, Torres C57Bl6 mice of the wild strain exposed to a et al., Bioorg Med Chem, 2012, 20, 942-948, chronic intake during 8 months of a high-fat Valverde et al., Bioorg Med Chem, 2014, 22, diet (HFD). 2678-2683, Leiva et al., Tetrahedron. Lett, 2015, 56, 1272-1275). On the other hand, it is derived 2.- To evaluate and characterize the impact from huprins (Canudas et al., Exp. Neurol, of the chronic administration of a selective 2003, 180, 123-130), which may have multidian inhibitor of JNK1, licochalcone A, in APPsw / activity, both on NMDA receptors and on PS1E9 mice, subjected to a chronic intake of acetylcholinesterase. a high-fat diet (HFD), for 8 months. General objective: Neurodegeneration, brain aging and innate immunity • To determine the activity of drugs developed as NMDA receptor antagonists and In recent studies we have determined that neurotoxins of marine origin, as well as their during aging, degenerative granular structures action on other targets that provide them containing neo-epitopes that are recognized with neuroprotective capacity.

2017 CIBERNED Annual Report / 39 Concrete objectives: 3.- To evaluate the neuroprotective activity of the compounds against the induced 1.- To evaluate the activity of the new neurodegeneration in cultures of granular compounds against the increase of calcium calcium neurons and in animal models (APP induced by NMDA in cultures of cerebellar / PS1 mice) granule neurons

2.- Determine the activity on the monoamine oxidase (MAO) of both type A and B KEYWORDS

Alzheimer’s disease, type 3 diabetes, obesity, cognition, drug development, aging PUBLICATIONS

• Ettcheto M., Abad S., Petrov D., Pedros I., Busquets O., Sanchez-Lopez E. et al. Early Preclinical Changes in Hippocampal CREB-Binding Protein Expression in a Mouse Model of Familial Alzheimer’s Disease. Molecular Neurobiology. 2017; 1-11. • Sánchez-López E, Egea MA, Davis BM, Guo L, Espina M, Silva AM et al. Memantine-Loaded PEGylated Biodegradable Nanoparticles for the Treatment of Glaucoma.Small (Weinheim an der Bergstrasse, Germany). 2017. • de Lemos L., Junyent F., Camins A., Castro-Torres R.D., Folch J., Olloquequi J. et al. Neuroprotective Effects of the Absence of JNK1 or JNK3 Isoforms on Kainic Acid-Induced Temporal Lobe Epilepsy-Like Symptoms. Molecular Neurobiology. 2017; 1-16. • Busquets O, Ettcheto M, Verdaguer E, Dario R, Auladell C, Beas-Zarate C et al. JNK1 inhibition by Licochalcone A leads to neuronal protection against excitotoxic insults derived of kainic acid.Neuropharmacology. 2017. • Leiva R., Grinan-Ferre C., Seira C., Valverde E., McBride A., Binnie M. et al. Design, synthesis and in vivo study of novel pyrrolidine-based 11β-HSD1 inhibitors for age-related cognitive dysfunction. European Journal of Medicinal Chemistry. 2017;139:412-428. • Bahous R.H., Jadavji N.M., Deng L., Cosin-Tomas M., Lu J., Malysheva O. et al. High dietary folate in pregnant mice leads to pseudo-MTHFR deficiency and altered methyl metabolism, with embryonic growth delay and short-term memory impairment in offspring. Human Molecular Genetics. 2017;26(5):888-900. Ettcheto M, Sánchez-López E, Pons L, Busquets O, Olloquequi J, Beas-Zarate C et al. • Dexibuprofen prevents neurodegeneration and cognitive decline in APPswe/PS1dE9 through multiple signaling pathways.Redox biology. 2017;13.

• Auge E., Cabezon I., Pelegri C., Vilaplana J. New perspectives on corpora amylacea in the human brain. Scientific Reports. 2017;7.

• Cabezón I, Augé E, Bosch M, Beckett AJ, Prior IA, Pelegrí C et al. Serial block-face scanning electron microscopy applied to study the trafficking of 8D3-coated gold nanoparticles at the blood-brain barrier.Histochemistry and cell biology. 2017;148(1).

• Sanchez-Lopez E., Ettcheto M., Egea M.A., Espina M., Calpena A.C., Folch J. et al. New potential strategies for Alzheimer’s disease prevention: pegylated biodegradable dexibuprofen nanospheres administration to APPswe/PS1dE9. Nanomedicine: Nanotechnology, Biology, and Medicine. 2017;13(3):1171-1182.

2017 CIBERNED Annual Report / 40 Program 1 Group: Antonio Camins Espuny

• Folch J, Busquets O, Ettcheto M, Sánchez-López E, Castro-Torres RD, Verdaguer E et al. Memantine for the Treatment of Dementia: A Review on its Current and Future Applications. Journal of Alzheimer’s disease : JAD. 2017. • Folch J, Busquets O, Sánchez-López ME, Pallàs M, Beas-Zarate C, Marin M et al. Experimental models for aging and their potential for novel drug discovery.Current neuropharmacology. 2017. • 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.Current neuropharmacology. 2018. [ePub2017]. • 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. Experimental Gerontology. 2018;102:3-11. [ePub2017]. • Palomera-Avalos V., Grinan-Ferre C., Izquierdo V., Camins A., Sanfeliu C., Pallas M. Metabolic Stress Induces Cognitive Disturbances and Inflammation in Aged Mice: Protective Role of Resveratrol. Rejuvenation Research. 2017;20(3):202-217. • Busquets O., Ettcheto M., Pallas M., Beas-Zarate C., Verdaguer E., Auladell C. et al. Long-term exposition to a high fat diet favors the appearance of β-amyloid depositions in the brain of C57BL/6J mice. A potential model of sporadic Alzheimer’s disease. Mechanisms of Ageing and Development. 2017;162:38-45. • Bartolome F., Antequera D., Tavares E., Pascual C., Maldonado R., Camins A. et al. Obesity and neuroinflammatory phenotype in mice lacking endothelial megalin. Journal of Neuroinflammation. 2017;14(1). • Patraca I., Martinez N., Busquets O., Marti A., Pedros I., Beas-Zarate C. et al. Anti-inflammatory role of Leptin in glial cells through p38 MAPK pathway inhibition. Pharmacological Reports. 2017;69(3):409-418. • Hernandez-Ojeda M., Urena-Guerrero M.E., Gutierrez-Barajas P.E., Cardenas-Castillo J.A., Camins A., Beas-Zarate C. KB-R7943 reduces 4-aminopyridine-induced epileptiform activity in adult rats after neuronal damage induced by neonatal monosodium glutamate treatment. Journal of Biomedical Science. 2017;24(1). • Auladell C, de Lemos L, Verdaguer E, Ettcheto M, Busquets O, Lazarowski A et al. Role of JNK isoforms in the kainic acid experimental model of epilepsy and neurodegeneration.Frontiers in bioscience (Landmark edition). 2017;22. • Akiguchi I., Pallas M., Budka H., Akiyama H., Ueno M., Han J. et al. SAMP8 mice as a neuropathological model of accelerated brain aging and dementia: Toshio Takeda’s legacy and future directions. Neuropathology. 2017;37(4):293-305. • Viveros-Paredes JM, González-Castañeda RE, Gertsch J, Chaparro-Huerta V, López-Roa RI, Vázquez-Valls E et al. Neuroprotective Effects of β-Caryophyllene against Dopaminergic Neuron Injury in a Murine Model of Parkinson’s Disease Induced by MPTP.Pharmaceuticals (Basel, Switzerland). 2017;10(3). RESEARCH PROJECTS

• Code: SAF2016-77703-C2-1-R. CIBERNED groups: G402. Title: Estudio de la epoxido hidrolasa soluble Other CIBER’s collaboration: No. como una nueva diana farmacológica para Type: National. la enfermedad de Alzheimer: modulación Funding agency: MINECO. del estrés oxidativo y la función mitocondrial. Funding: 100.000 €. Principal Investigator: Merce Pallas. Duration: 2015-2019 CIBERNED’s collaboration: No.

2017 CIBERNED Annual Report / 41 • Code: BFU2016-78398-P. • Code: PI2016/01. Title: Estudio de la presencia de neo- Title: Alteraciones del metabolismo gluco- epitopos en estructuras degenerativas lipídico y desarrollo de la demencia de cerebrales y de la existencia en el plasma alzhéimer. de anticuerpos naturales dirigidos contra Principal Investigator: Ignacio Torres dichos neo-epitopos. Alemán. Principal Invetigator: Carme Pelegrí CIBERNED’s collaboration: Yes. Gabalda. CIBERNED groups: G409; G402; G511; G502; CIBERNED’s collaboration: No. G412. CIBERNED groups: G402. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: CCAA. Funding agency: CIBERNED. Funding agency: Generalitar de Catalunya. Funding: 200.000 €. Funding: 120.000 €. Duration: 2016-2018 Duration: 2016-2020 PhD DISSERTATIONS

• Author: Itsaso Cabezón Rodríguez. • Author: : Nohora Milena Martínez López. Title: Estudio del transporte de sustancias Title: Efecto de la melatonina, leptina a través de la barrera hematoencefálica y ácidos grasos: Docosahexaenoico, mediante el anticuerpo 8D3 dirigido contra eicosapentaenoico y palmítico, en la el receptor de transferrina. modulación de la inflamación en células de Date: 4th March 2017. glía. Supervisor: Antonio Camins Espuny. Date: 14th September 2017. Supervisor: Jaume Folch.

2017 CIBERNED Annual Report / 42 José Luis Cantero Lorente 511

Principal Investigator José Luis Cantero Lorente

Research team

Mercedes Atienza Ruiz Cynthia Ayala Rodríguez David García Solís Researcher Technician Collaborator

Sofía Rodríguez Peñuela Maité Crespo García Eulogio Gil Néciga Technician Collaborator Collaborator

Laboratorio de Neurociencia Funcional Servicios Centrales de Investigación (Edificio 21) Universidad Pablo de Olavide Ctra. de Utrera, Km. 1 41013-Sevilla (Spain) Tel.: +34 95 497 74 33 [email protected]

2017 CIBERNED Annual Report / 43 SUMMARY

In 2017, we have performed experiments aimed status, and on the other hand, the association at determining the damage of the cholinergic between higher HCY levels and volume loss system and their innervated regions in the of the temporal lobe depended on the serial prodromal stage of Alzheimer’s disease (AD). relationship between poorer sleep quality and lower antioxidant capacity. These findings Patients with mild cognitive impairment (MCI) provided novel insights into how sleep deficits showed bilateral atrophy of the nucleus basalis may contribute to maintain the relationship of Meynert (NBM/Ch4), and patterns of NBM between HCY and oxidative stress in prodromal atrophy were correlated with bilateral thinning AD, and offer empirical foundations to design of the temporal cortex and volume loss of the therapeutic interventions aimed to weaken this basolateral nucleus of amygdala, regions that link (Sánchez-Espinosa et al., 2017). receive massive cholinergic projections from the NBM. Therefore, these results revealed, for the In collaboration with Dr. Eva Carro’s group, first time in humans, that the entire cholinergic we have carried out experiments aimed at pathway is structurally affected years before assessing the diagnostic value of salivary the diagnosis of AD (Cantero et al., 2017). lactoferrin in MCI/AD patients. We showed that MCI/AD patients have significantly lower We have continued exploring the association lactoferrin levels than controls, and that lower between impaired sleep and biomarkers of AD lactoferrin paralleled cognitive impairment in MCI patients. These experiments have shown and pathological CSF levels of amyloid-beta that poorer sleep quality in MCI correlated and total tau (Carro et al., 2017). Given the with lower antioxidant capacity and increased non-invasive and inexpensive nature of salivary homocysteine (HCY) levels. We further found markers, these results are promising to be used that sleep deficits in MCI mediated the in screening of AD biomarkers in the general relationship between HCY and oxidative population. KEYWORDS

Trajectories of vulnerable aging, Biomarkers of preclinical and prodromal stages of Alzheimer’s disease, Structural and functional connectivity of human neocortex PUBLICATIONS

• 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. 2017;64. • Huertas I., Oldehinkel M., van Oort E.S.B., Garcia-Solis D., Mir P., Beckmann C.F. et al. A Bayesian spatial model for neuroimaging data based on biologically informed basis functions. NeuroImage. 2017;161:134-148. • Huertas I., Jesus S., Garcia-Gomez F.J., Lojo J.A., Bernal-Bernal I., Bonilla-Toribio M. et al. Genetic factors influencing frontostriatal dysfunction and the development of dementia in Parkinson’s disease. PLoS ONE. 2017;12(4). • Carro E., Bartolome F., Bermejo-Pareja F., Villarejo-Galende A., Molina J.A., Ortiz P. et al. Early diagnosis of mild cognitive impairment and Alzheimer’s disease based on salivary lactoferrin. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2017;8:131-138. • Cantero JL, Zaborszky L, Atienza M. Volume Loss of the Nucleus Basalis of Meynert is Associated with Atrophy of Innervated Regions in Mild Cognitive Impairment.Cerebral cortex (New York, N.Y. : 1991). 2017.

2017 CIBERNED Annual Report / 44 Program 1 Group: José Luis Cantero Lorente

• 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. 2017;:1-11. • Huertas I., Jesus S., Lojo J.A., Garcia-Gomez F.J., Caceres-Redondo M.T., Oropesa-Ruiz J.M. et al. Lower levels of uric acid and striatal dopamine in non-tremor dominant Parkinson’s disease subtype. PLoS ONE. 2017;12(3). • Sanchez-Espinosa MP, Atienza M, Cantero JL. Sleep mediates the association between homocysteine and oxidative status in mild cognitive impairment.Scientific reports. 2017;7(1). • Clausner T., Dalal S.S., Crespo-Garcia M. Photogrammetry-based head digitization for rapid and accurate localization of EEG electrodes and MEG fiducial markers using a single digital SLR camera. Frontiers in Neuroscience. 2017;11(MAY). • Balsa MA, Camacho V, Garrastachu P, García-Solís D, Gómez-Río M, Rubí S et al. A new era for Nuclear Medicine neuroimaging in Spain: Where do we start from in Spain?. Revista espanola de medicina nuclear e imagen molecular. ;36(4). RESEARCH PROJECTS

• Code: PI2016/01. CIBERNED’s collaboration: No. Title: Alteraciones del metabolismo gluco- CIBERNED groups: G511. lipídico y desarrollo de la demencia de Other CIBER’s collaboration: No. alzhéimer. Type: National. Principal Investigator: Ignacio Torres Alemán. Funding agency: MINECO. CIBERNED’s Collaboration: Yes. Funding: 117.975 €. CIBERNED groups: G409; G402; G511; G502; Duration: 2015-2017 G412. Other CIBER’s collaboration: No. • Code: PSI2016-81881-REDT. Type: Intramurales. Tile: Aplicaciones clínicas de la neuroimagen Funding agency: CIBERNED. funcional. Funding: 200.000 €. Principal Investigator: Fernando Maestu Duration: 2016-2018 Unturbe. CIBERNED’s collaboration: No. • Code: P12-CTS 2327. CIBERNED groups: G511. Title: Caracterización de las fases Other CIBER’s collaboration: No. presintomáticas y preclínicas de la Type: National. enfermedad de Alzheimer mediante Funding agency: MINECO. marcadores biológicos y de neuroimagen. Funding: 20.000 €. Principal Investigator: José Luis Cantero Duration: 2017-2019 Lorente. CIBERNED’s collaboration: No. • Code: SES_2017. CIBERNED groups: G511. Title: Efectos de la concentración de beta Other CIBER’s collaboration: No. amiloide cerebral sobre el sueno fisiológico Type: CC.AA. en personas mayores. Funding agency: Junta de Andalucía. Principal Investigator: José Luis Cantero Funding: 225.419 €. Lorente. Duration: 2014-2017 CIBERNED’s collaboration: No. CIBERNED groups: G511. • Code: PSI2014-55747-R. Other CIBER’s collaboration: No. Title: Estudio de las oscilaciones cerebrales Type: Private. EEG relacionadas con la memoria como Funding agency: Sociedad Española de potencial biomarcador de la enfermedad Sueño. de Alzheimer. Funding: 10.000 €. Principal Investigator: Mercedes Atienza Ruiz. Duration: 2017-2018

2017 CIBERNED Annual Report / 45 PhD DISSERTATIONS

• Author: Laura Prieto del Val. • Author: : Mayely Paola Sánchez Espinosa. Title: Correlatos neurales del déficit de Title: Relación entre las alteraciones del memoria asociativa en el deterioro cognitivo sueño, cambios de la estructura cortical leve: efecto del gen APOE y conversión a la y marcadores en sangre periférica en enfermedad de Alzheimer. personas mayores con deterioro cognitivo Date: 16th June 2017. leve. Supervisor: José Luis Cantero Lorente. Date: 30th June 2017. Supervisor: José Luis Cantero Lorente.

2017 CIBERNED Annual Report / 46 Eva María Carro Díaz 502

Principal Investigator Eva María Carro Díaz

Research team

Desiree Antequera Tienda Julián Benito León David Andrés Pérez Martínez Technician Research staff Research staff

Fernando Bartolomé Robledo Jesús Hernández Gallego Alejandro Herrero San Martín Postdoctoral fellow Research staff Research staff

Marta González Sánchez Rocío Trincado Soriano Sara Llamas Velasco Predoctoral fellow Technician Research staff

José Antonio Molina Arjona Alberto Villarejo Galende Research staff Research staff

Instituto de Investigación Hospital 12 de Octubre Avenida de Córdoba s/n 28041 Madrid Tel.: +34 913 90 87 65 Fax: +34 913 90 85 44 [email protected]

2017 CIBERNED Annual Report / 47 SUMMARY

Our scientific activity has been focused at important role in neurodegeneration. studying pathophysiological mechanisms On this regard, we also studied of neurodegenerative diseases, mainly alterations on mitochondrial dynamics Alzheimer’s disease and associated dementias, and bioenergetics. Our results found identifying new therapeutic targets, risk factors proteinopathies resulting in cellular and biomarkers. Specific activities are: alterations including dementias.

• Preclinical studies on new risk factors, • Epidemiological research on biomarkers of disease, mechanisms and new neurodegenerative diseases, particularly risk therapeutic strategies in Alzheimer’s disease factors. and other neurodegenerative disorders. a) Epidemiological analysis NEDICES-I a) Research on risk factors that highlight cohort. the relationship between metabolic disorders and neurodegenerative b) Implementation of NEDICES-II cohort: diseases, including Alzheimer’s, through harvesting, processing and biobank mechanisms and inflammatory responses storage of ~2000 subject samples at central and peripheral level. recruited in Primary Care. Classification and verification of cases with b) Preclinical research searching for new neurodegenerative pathology. therapeutic targets and design of potential drugs using cellular and animal • Clinical research in dementia. experimental models of Alzheimer’s disease and other degenerative a) Collaboration searching for genetic risk dementias. Particularly we analysed factors, and participation in genetic the therapeutic efficiency of GDNF consortia (DEGESCO). in Parkinson’s disease animal model. The intranasal administration of this b) Determination of clinical utility of compound allowed it to go through the molecular and imaging biomarkers BBB and avoided its quick degradation. recently introduced: new useful biomarkers for early detection of disease. c) Studies on molecular pathology in In collaboration with other CIBERNED neurodegenerative disorders analysing groups (M. Calero, A. Rábano and JL. new pathophysiological mechanisms, Cantero), we have proposed salivary in particular, mitochondrial alterations, lactoferrin levels as early AD biomarker as as mitochondrial disfunction plays an it is reduced with the disease progression. KEYWORDS Alzheimer’s disease, Parkinson’s disease, Amyotrophic Lateral Sclerosis, Frontotemporal Dementia, biomarkers, inflammation, bioenergetics, therapeutic drugs

PUBLICATIONS

• Carro E., Bartolome F., Bermejo-Pareja F., Villarejo-Galende A., Molina J.A., Ortiz P. et al. Early diagnosis of mild cognitive impairment and Alzheimer’s disease based on salivary lactoferrin. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2017;8:131-138. • Virues-Ortega J., Vega S., Seijo-Martinez M., Saz P., Rodriguez F., Rodriguez-Laso A. et al. A protective personal factor against disability and dependence in the elderly: an ordinal regression analysis with nine geographically-defined samples from Spain. BMC Geriatrics. 2017;17(1):1-10.

2017 CIBERNED Annual Report / 48 Program 1 Group: Eva María Carro Díaz

• Bartolome F., Antequera D., Tavares E., Pascual C., Maldonado R., Camins A. et al. Obesity and neuroinflammatory phenotype in mice lacking endothelial megalin. Journal of Neuroinflammation. 2017;14(1). • Garzo-Caldas N., Ruiz-Sainz E., Vila-Bedmar S., Llamas-Velasco S., Hernandez-Lain A., Ruiz- Morales J. et al. Enteroviral T-cell encephalitis related to immunosuppressive therapy including rituximab. Neurology. 2017;89(4):408-409. • Sanchez-Ferro A., Benito-Leon J., Louis E.D., Contador I., Hernandez-Gallego J., Puertas-Martin V. et al. Cognition in non-demented Parkinson’s disease vs essential tremor: A population- based study. Acta Neurologica Scandinavica. 2017. • Benito-Leon J. Viral hepatitis and the risk of Parkinson disease. Neurology. 2017;88(17):1596-1597. • Arroyo-Gallego T., Ledesma-Carbayo M.J., Sanchez-Ferro A., Butterworth I., Mendoza C.S., Matarazzo M. et al. Detection of Motor Impairment in Parkinson’s Disease Via Mobile Touchscreen Typing. IEEE Transactions on Biomedical Engineering. 2017;64(9):1994-2002. • Serrano J.I., Lambrecht S., del Castillo M.D., Romero J.P., Benito-Leon J., Rocon E. Identification of activities of daily living in tremorous patients using inertial sensors. Expert Systems with Applications. 2017;83:40-48. • Ettcheto M., Abad S., Petrov D., Pedros I., Busquets O., Sanchez-Lopez E. et al. Early Preclinical Changes in Hippocampal CREB-Binding Protein Expression in a Mouse Model of Familial Alzheimer’s Disease. Molecular Neurobiology. 2017;:1-11. • 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). [ePub2017]. • Labiano-Fontcuberta A., Benito-Leon J. Radiologically isolated syndrome should be treated with disease-modifying therapy – No. Multiple Sclerosis. 2017;23(14):1820-1821. • Lopez-Blanco R., Posada Rodriguez I.J., Benito-Leon J. Learning disability in a son and premature ovarian failure as clinical pointers to identify a premutation on the X chromosome in a female with long-standing tremor. Parkinsonism and Related Disorders. 2017;42:107-108. • Serrano J.I., Romero J.P., Castillo M.D.D., Rocon E., Louis E.D., Benito-Leon J. A data mining approach using cortical thickness for diagnosis and characterization of essential tremor /692/53/2421 /692/617/375/346 /129 /141 /123 article. Scientific Reports. 2017;7(1). • Bartolome F, Esteras N, Martin-Requero A, Boutoleau-Bretonniere C, Vercelletto M, Gabelle A et al. Pathogenic p62/SQSTM1 mutations impair energy metabolism through limitation of mitochondrial substrates.Scientific reports. 2017;7(1). • Benito-Leon J., Mato-Abad V., Louis E.D., Hernandez-Tamames J.A., Alvarez-Linera J., Bermejo- Pareja F. et al. White matter microstructural changes are related to cognitive dysfunction in essential tremor. Scientific Reports. 2017;7(1). • Hernando S., Herran E., Figueiro-Silva J., Pedraz J.L., Igartua M., Carro E. et al. Intranasal Administration of TAT-Conjugated Lipid Nanocarriers Loading GDNF for Parkinson’s Disease. Molecular Neurobiology. 2017;:1-11. • Ludtmann M.H.R., Arber C., Bartolome F., De Vicente M., Preza E., Carro E. et al. Mutations in valosin-containing protein (VCP) decrease ADP/ATP translocation across the mitochondrial membrane and impair energy metabolism in human neurons. Journal of Biological Chemistry. 2017;292(21):8907-8917. • Benito-Leon J., Laurence M. The role of fungi in the etiology of multiple sclerosis. Frontiers in Neurology. 2017;8(OCT). • Agundez J.A.G., Garcia-Martin E., Alonso-Navarro H., Ayuso P., Esguevillas G., Benito-Leon J. et al. Delta-amino-levulinic acid dehydratase gene and essential tremor. European Journal of Clinical Investigation. 2017;47(5):348-356.

2017 CIBERNED Annual Report / 49 • 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 International. 2018;35(2):270-279. [ePub2017]. • Villarejo-Galende A., Llamas-Velasco S., Gomez-Grande A., Puertas-Martin V., Contador I., Sarandeses P. et al. Amyloid pet in primary progressive aphasia: case series and systematic review of the literature. Journal of Neurology. 2017;264(1):121-130. • De Pablo-Fernandez E., Sierra-Hidalgo F., Benito-Leon J., Bermejo-Pareja F. Association between Parkinson’s disease and diabetes: Data from NEDICES study. Acta Neurologica Scandinavica. 2017. • Contador I, Fernández-Calvo B, Boycheva E, Rueda L, Bermejo-Pareja F. Normative Data of the Story and Six-Object Memory Recall Tests in Older Spanish Adults: NEDICES Population-Based Cohort.Archives of clinical neuropsychology : the official journal of the National Academy of Neuropsychologists. 2017. • Sardinero-Garcia C., Santiago-Saez A., Bravo-Llatas M.D.C., Perea-Perez B., Albarran-Juan M.E., Labajo-Gonzalez E. et al. Liability for loss of chance in neurological conditions in the Spanish public healthcare system Responsabilidad por pérdida de oportunidad asistencial en patologías neurológicas en la medicina pública española. Gaceta Sanitaria. 2017;31(1):30-34. • Laurence M, Benito-León J. Epstein-Barr virus and multiple sclerosis: Updating Pender’s hypothesis.Multiple sclerosis and related disorders. 2017;16. • Leon Ruiz M., Benito-Leon J., Garcia-Soldevilla M.A., Rubio-Perez L., Parra Santiago A., Lozano Garcia-Caro L.A. et al. Innovative and effective immunosuppressive bitherapy for an unusual paraneoplastic opsoclonus-myoclonus-ataxia syndrome of the adult Biterapia inmunosupresora efectiva e innovadora en un síndrome opsoclono-mioclono-ataxia paraneoplásico e inusual del adulto. Neurologia. 2017;32(2):122-125. • Villarejo Galende A., Eimil Ortiz M., Llamas Velasco S., Llanero Luque M., Lopez de Silanes de Miguel C., Prieto Jurczynska C. Report by the Spanish Foundation of the Brain on the social impact of Alzheimer disease and other types of dementia Informe de la Fundación del Cerebro. Impacto social de la enfermedad de Alzheimer y otras demencias. Neurologia. 2017. • Mendez-Guerrero A., Uriarte-Perez de Urabayen D., Llamas-Velasco S. Spinocerebellar ataxia type 3 presenting with writer’s cramp without ataxia. International Journal of Neuroscience. 2017;:1-2. • Contador I., Bermejo-Pareja F., Pablos D.L., Villarejo A., Benito-Leon J. High education accelerates cognitive decline in dementia: A brief report from the population-based NEDICES cohort Maior educação acelera o declínio cognitive em demência: Relato breve da coorte de base populacional NEDICES. Dementia e Neuropsychologia. 2017;11(3):297-300. • Lenka A., Benito-Leon J., Louis E.D. Is there a premotor phase of essential tremor?. Tremor and Other Hyperkinetic Movements. 2017;7. RESEARCH PROJECTS

• Code: PI2016/01. • Code: PI15-00780. Title: Alteraciones del metabolismo gluco- Title: Inflamación periférica como factor de lipídico y desarrollo de la demencia de riesgo en la patología de la enfermedad de alzhéimer. alzhéimer: nuevas herramientas diagnósticas Principal Investigator: Ignacio Torres Alemán. y dianas terapéuticas. CIBERNED’s collaboration: Yes. Principal Investigator: Eva Carro. CIBERNED groups: G409; G402; G511; G502; CIBERNED’s collaboration: No. G412. CIBERNED groups: G502. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: National. Funding agency: CIBERNED. Funding agency: Instituto de Salud Carlos III. Funding: 200.000 €. Funding: 100.000€. Duration: 2016-2018 Duration: 2016

2017 CIBERNED Annual Report / 50 Program 1 Group: Eva María Carro Díaz

• Code: RTC-2015-3967-1. • Code: DTS15/00141. Title: Plataforma para el seguimiento de Title: Evaluación del impacto de la imagen trastornos del movimiento (NETMD). PET de amiloide en el diagnóstico de Principal Investigator: Julián Benito León. los pacientes con deterioro cognitivo CIBERNED’s collaboration: No. evaluados por sospecha de alzhéimer. CIBERNED groups: G502. Principal Investigator: Dr. Javier Arbizu. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: National. CIBERNED groups: G504; G502; G609. Funding agency: MINECO. Other CIBER’s collaboration: No. Funding: N.D. Type: National. Duration: 2015-2017 Funding agency: Instituto de Salud Carlos III. Funding: 59.139 €. Duration: 2016

2017 CIBERNED Annual Report / 51 Joan Xavier Comella Carnicé 402

Principal Investigator Joan Xavier Comella Carnicé

Research team

Víctor Yuste Mateos Isabel Calleja Yagüe Raquel Badillos Rodríguez PhD Fellow PhD student PhD student

Joaquín López Soriano Elena Coccia Julia Soler Flores PhD Fellow PhD student Student

María José Pérez García Anna Martínez Sirés PhD Fellow PhD student

VHIR – Vall Hebron Institut de Recerca Passeig Vall d’Hebron 119-129 08035 Barcelona (Spain) Tel.: +34 93 489 38 07 Web: www.vhir.org [email protected]

2017 CIBERNED Annual Report / 52 SUMMARY

Our main interest is to characterize the Our lab has recently characterize new mechanisms controlling neuronal death induced unexpected physiological roles for FAIM-L, by a group of receptors collectively known as since we have observed that it is implicated in death receptors, and also the relevance of the control of some non-apoptotic effects of some of their intracellular antagonists. Among caspases, such as axonal degeneration and those, we are most interested in the antagonists long term depression (LTD), through modulation expressed in nervous system, such as FAIM-L, of XIAP levels (Martínez-Mármol et al., 2016). as well as their molecular partners. Our main These events of synaptic plasticity are also objective is to characterize these proteins modulated in neurodegenerative diseases, at the molecular level, their involvement in thus positioning FAIM-L as a good candidate neuronal differentiation and physiology, and for the treatment of the disease. At present we their possible role in different pathologies, are also characterizing the interaction among mainly neurodegenerative diseases. The FAIM-L and XIAP with other proapototic partners knowledge of the molecular basis of these such as Siva-1, which may help better explain receptor antagonists and the activation of the role of FAIM in neuron physiology. survival signaling pathways could be of high relevance to understand the etiology or We have also characterized two new to open new therapeutic strategies for the isoforms of FAIM, one of them specific from treatment of neurodegenerative diseases, such neurons (Coccia et al., 2017), although as Alzheimer’s disease. their physiological role is still not fully characterized. We have recently characterized, in collaboration with different CIBERNED groups, Our main research lines at present are: the role of FAIM-L in the development of Alzheimer’s disease, both in animal models 1) To study FAIM-L function in in vitro and in vivo Joan Xavier Comella Carnicé (APP/PS1) and patients (Carriba et al., 2015). At models of AD, and its relation with TNF (its present we are further characterizing this role duality as a pro-apoptotic molecule and at of FAIM-L, and with this aim we have started the same time as a survival promoter) and its the characterization of the Faim knockout signaling pathways, particularly NFkB, as well model, through immunohistochemistry, in the cross-talk neuron/glia electrophysiological and behavior studies. Surprisingly, this model shows an epileptic 2) To characterize FAIM-L functional partners, phenotype, which enlarges the relevance particularly XIAP and Siva-, and their of FAIM-L function in nervous system. We implications in neuron physiology (neuronal have also developed adenoassociated viral plasticity) and different neurodegenerative vectors, which will allow us to overexpress or diseases (neuronal apoptosis, synaptic downregulate FAIM-L levels in brain, at present degeneration). already in use in the murine models of Alzheimer’s disease, in order to study the role of FAIM-L in 3) To study microRNAs that coul modulate FAIM the development or progression of the disease, expression levels, both in physiological and or its potential protector effect in these models. pathologic states. KEYWORDS

Alzheimer, neuroinflammation, death receptors, FAIM, TNF, Fas PUBLICATIONS

• Coccia E., Calleja-Yague I., Planells-Ferrer L., Sanuy B., Sanz B., Lopez-Soriano J. et al. Identification and characterization of new isoforms of human fas apoptotic inhibitory molecule (FAIM). PLoS ONE. 2017;12(10).

2017 CIBERNED Annual Report / 53 • 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. [ePub2017]. RESEARCH PROJECTS

• Code: SAF2016-80236-R. Title: Relevancia del antagonista de receptores de muerte, FAIM-L, en la enfermedad de Alzheimer. Principal Investigator: Joan Xavier Comella Carnicé. CIBERNED’s collaboration: No. CIBERNED groups: G413. Other CIBER’s collaboration: No. Type: National. Funding agency: MINECO. Funding: 273.459 €. Duration: 2016-2019

2017 CIBERNED Annual Report / 54 Javier de Felipe Oroquieta 403

Principal Investigator Javier de Felipe Oroquieta

Research team

Lidia Alonso-Nanclares Marta Turégano Marta Domínguez PhD Fellow PhD student PhD student Ruth Benavides-Piccione PhD Fellow Débora Cano Laiseca Sandra Ostos Lab technician PhD student Rodrigo Rodríguez Sánchez Research staff Mari Carmen Álvarez Pérez María del Carmen Regalado Lab technician PhD student Lidia Blazquez-Lorca Marta Montero PhD Fellow Silvia Tapia González PhD Fellow PhD student Isabel Fernaud Espinosa Concepción Rojo PhD Fellow Gonzalo León Espinosa PhD Fellow Visiting researcher Asta Kastanauskaite Miguel Miguens Vázquez PhD Fellow Mirian Marín Horcajada Lab technician Visiting researcher Ángel Merchán-Pérez Juncal González PhD Fellow Alejandro Antón Fernández PhD student Visiting researcher Alberto Muñoz Céspedes Pilar Flores Romero PhD Fellow Liulia Diana Furcila PhD student Research and Project manager Lorena Valdés Lora Montserrat Fernández Bouzo Lab technician Andrea Santuy Muñoz Research and Project manager PhD student Ana Isabel García Ramírez Yago Rodríguez Cela Lab technician Guillermo Aparicio Torres Administrative officer PhD student Marta Barbado Amigo Laboratorio Cajal de Circuitos Corticales, Research and Project manager CTB, UPM. Campus de Montegancedo Laura Fernández García Pozuelo de Alcorcón 28223 Madrid (Spain) PhD student Tel.: +34 91 336 46 60 Diana Sánchez Ponce [email protected] PhD student

2017 CIBERNED Annual Report / 55 SUMMARY

Microorganization of the normal cerebral microscopy stacks of images of pyramidal cortex and alterations of cortical circuits in cells to analyze the distribution and brain pathologies morphology of dendritic spines in different cortical areas, layers and species. The CCCL principally focuses on the microorganization of the normal cerebral Intrinsic and extrinsic connectivity of the cortex (including hippocampus) in various cortical column: species (particularly humans) and on the alterations of cortical circuits in epilepsy and • Target identification of cortical synapses Alzheimer disease. These studies are performed (FIB/SEM) to determine the proportion through the use of anatomical tracers, high of synapses on dendritic spines and resolution immunocytochemistry and 3D light dendritic shafts. and electron microscopy. Another major • Analysis of synaptic sizes (FIB/SEM) and aim is to develop informatics technologies to development of computer models to examine the brain. In particular, the research explore the possible relationship between lines carried out at the CCCL are as follows: synaptic morphology and physiology. • Characterization of the afferent and Study of the components of the column: efferent connections of the hindlimb representation area of the primary • Development and validation of software somatosensory cortex in P14 rats in tract- tools for 3D segmentation of cells of tracing experiments. the complete cortical column in stacks obtained by confocal microscopy Quantification of cellular and subcellular in samples from the somatosensory alterations in Alzheimer’s Disease (AD) and neocortex. the possible influence of these alterations on • Spatial analysis distribution of the cognition: segmented cell to determine the distribution patterns of different cell types. • Quantification of specific synaptic • Analysis of the spatial distribution of alterations in the neuropil and in identified synapses in the six cortical layers by means 3D reconstructed cortical neurons from of spatial statistical tools. Calculation areas showing histopathological changes of the volume fraction occupied by in AD. mitochondria in the six cortical layers. • Analysis of the alterations of the Golgi- • Development of a new method that apparatus of neurons accumulating makes it possible to obtain high-resolution phospho-tau (in a mouse model scanning EM images of large areas of taupathy) to investigate the possible the brain in the millimeter to nanometer neuronal functional alterations during the range. AD. • Application of systematic methods to Pyramidal cells: human tissue from AD patients, using conventional three dimensional (3D) dual • Intracellular injections and beam electron microscopy (FIB/SEM) to 3D-reconstruction of pyramidal cells and which allows analyzing synaptic densities to analyze the microanatomy of these as well as their morphological properties. cells in different cortical areas, layers and • Correlation of quantitative species. microanatomical data with cognitive • Generation of high resolution confocal impairment. KEYWORDS

Microanatomy; Cortical Circuits; Cerebral cortex; Rat, Mouse and Human Brain

2017 CIBERNED Annual Report / 56 Program 1 Group: Javier de Felipe Oroquieta PUBLICATIONS

• Anton-Fernandez A., Merchan-Rubira J., Avila J., Hernandez F., Defelipe J., Munoz A. Phospho- Tau Accumulation and Structural Alterations of the Golgi Apparatus of Cortical Pyramidal Neurons in the P301S Tauopathy Mouse Model. Journal of Alzheimer’s Disease. 2017;60(2):651-661. • Pallas-Bazarra N., Kastanauskaite A., Avila J., Defelipe J., Llorens-Martin M. GSK-3β overexpression alters the dendritic spines of developmentally generated granule neurons in the mouse hippocampal dentate gyrus. Frontiers in Neuroanatomy. 2017;11. • Rodriguez-Moreno J, Rollenhagen A, Arlandis J, Santuy A, Merchan-Pérez A, DeFelipe J et al. Quantitative 3D Ultrastructure of Thalamocortical Synapses from the Lemniscal Ventral Posteromedial Nucleus in Mouse Barrel Cortex.Cerebral cortex (New York, N.Y. : 1991). 2017. • Leon-Espinosa G., Regalado-Reyes M., DeFelipe J., Munoz A. Changes in neocortical and hippocampal microglial cells during hibernation. Brain Structure and Function. 2017;:1-15. • Gonzalez-Riano C., Tapia-Gonzalez S., Garcia A., Munoz A., DeFelipe J., Barbas C. Metabolomics and neuroanatomical evaluation of post-mortem changes in the hippocampus. Brain Structure and Function. 2017;222(6):2831-2853. • Santuy A., Rodriguez J.R., DeFelipe J., Merchan-Perez A. Volume electron microscopy of the distribution of synapses in the neuropil of the juvenile rat somatosensory cortex. Brain Structure and Function. 2017;:1-14. • Anton-Fernandez A., Aparicio-Torres G., Tapia S., DeFelipe J., Munoz A. Morphometric alterations of Golgi apparatus in Alzheimer’s disease are related to tau hyperphosphorylation. Neurobiology of Disease. 2017;97:11-23. • Urrecha M., Romero I., DeFelipe J., Merchan-Perez A. Influence of cerebral blood vessel movements on the position of perivascular synapses. PLoS ONE. 2017;12(2). • Anton-Sanchez L., Larranaga P., Benavides-Piccione R., Fernaud-Espinosa I., Defelipe J., Bielza C. Three-dimensional spatial modeling of spines along dendritic networks in human cortical pyramidal neurons. PLoS ONE. 2017;12(6). • Blazquez-Llorca L., Valero-Freitag S., Rodrigues E.F., Merchan-Perez A., Rodriguez J.R., Dorostkar M.M. et al. High plasticity of axonal pathology in Alzheimer’s disease mouse models. Acta neuropathologica communications. 2017;5(1):14-. RESEARCH PROJECTS

• Code: Cajal Blue Brain Project. • Code: PI2016/05. Title: Cajal Blue Brain Project. International Title: Dream inhibitors and Alzheimer´s Disease. Blue Brain Proyect. Principal Investigator: José Ramón Naranjo Principal Investigator: Javier de Felipe. Orovio. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G204, G403. CIBERNED groups: G307; G106; G403. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: Intramurales. Funding agency: Ecole Polytechnique Funding agency: CIBERNED Federale de Lausanne (Suiza). Funding: 210.000 €. Funding: ND. Duration: 2016-2018 Duration: 2009-2019

2017 CIBERNED Annual Report / 57 • Code: ZEN-15-321663. • Code: SAF2015-66603-P. Title: The Pyramidal Neuron in Cognition and Title: Estudio de la microorganización de la Alzheimer's Disease. corteza cerebral en pacientes de azhéimer Principal Investigator: Javier de Felipe. y del hamster como modelo para estudiar la CIBERNED’s collaboration: No. fosforilación de tau. CIBERNED groups: G403. Principal Investigator: Javier de Felipe. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: International. CIBERNED groups: G403. Funding agency: Alzheimer's Association. Other CIBER’s collaboration: No. Funding: ND. Type: National. Duration: 2015-2018 Funding agency: MINECO. Funding: 196.000€. Duration: 2016-2018

2017 CIBERNED Annual Report / 58 Isidro Ferrer Abizanda 503

Principal Investigator Isidro Ferrer Abizanda

Research team

Marta Barrachina Castilla Margalida Frau Méndez Mayelín Domínguez González Research staff Student PhD Fellow

Franc Llorens Raissa Camila Alvear Irene López González PhD Fellow Contreras PhD Fellow Visiting student Paula García-Esparcia Ester Asó PhD Fellow Margarita Carmona Murillo Research staff Lab technician Pol Andrés Benito Marta Povedano PhD student Jesús Moreno Researcher Lab technician Daniela Díaz Lucena Belén Ansoleaga Ávila PhD Fellow PhD student

Departamento de Patología y Terapeútica Experimental Universidad de Barcelona C/ Feixa Llarga sn, 08907 Hospitalet de Llobregat Barcelona Tel.: +34 93 260 74 52 [email protected]

2017 CIBERNED Annual Report / 59 SUMMARY

Study is mainly focused on molecular globular glial tauopathy (GGT) which are now neuropathology using combined ‘omics’ of accepted as specific disorders by the scientific selected neurodegenerative diseases with community. An additional topic, not yet finished, abnormal protein aggregates in aging, in order has been the study of the locus ceruleus (LC) at to identify de-regulated pathways at progressive early stages of neurofibrillary tangle pathology. stages of disease and in different regions, both NFT pathology is very common in the LC in the related to disease-specific vulnerability. The elderly, as a possible substrate (together with principal achievements have been obtained alterations in the raphe nuclei) of depression in Creutzfeldt-Jakob disease and humanized in old age. We are exploring its relationship murine models of prion diseases, Alzheimer’s with primary aging tauopathy and Alzheimer’s disease, synucleinopathies, and amyotrophic disease. The work is being carried out in lateral sclerosis. Common alterations, albeit collaboration with experts in neuroimaging and with disease specificities, involve mitochondrial clinicians in a population of aged individuals function and energy metabolism, purine with depression and patients with mild cognitive metabolism, protein synthesis from the impairment, assessing in both groups a wide nucleolus to the ribosome, neurotransmission array of tests and following up on possible and synapses, and inflammatory responses. conversion to Alzheimer’s disease. Combined Collaborative studies have also been carried clinical and neuropathological observations out in X-linked adrenoleukodystrophy. on this particular topic are promising. Dr. Marta Barrachina, a PhD member of our group running This combined and comprehensive approach her particular line of research, has created has permitted us to uncover certain aspects a company geared to developing tests of of the molecular neuropathology of these biomarkers for the diagnosis of Alzheimer’s neurodegenerative diseases. Studies have been disease. performed in parallel to recognize biomarkers in the CSF in dementia, mainly those of use in the The incorporation of new members such as diagnosis of different types of rapid dementias. Dr. Franc Llorens will promote further research Different transgenic murine models have been on the identification of biomarkers in the CSF utilized to look at similarities and differences and blood, in parallel with their correlates with human diseases and to test combined with expression levels in brain tissue. Close drugs for therapeutic purposes; cannabinoids collaboration with Dr. Antonella Consiglio, have been proven to be useful potential drugs incorporated as adjunct professor, will prove for reducing certain symptoms in Alzheimer’s highly beneficial. The IP, Dr. I. Ferrer is now disease models, and we have moved forward retired from his position at the Bellvitge University to test their use in human clinical trials. Hospital, the Institute of Neuropathology does not exist anymore, and the laboratories An additional point of interest has been the of molecular neuropathology, including the study of astrogliopathy in the pathogenesis of biosecurity facility, are dismantled. Dr. Ferrer this group of disorders considering that these is now a senior teacher in the hospital and diseases are the result of the abnormal function working as full professor at the laboratories of of neurons and glial cells. This concept is now the Barcelona University and the neighboring being expanded to oligodendrocytes as key Hospital Duran Reynals. partners in the degenerative process. Certain studies have contributed to defining new glial Present situation and agreements with other tauopathies by international consent, such as centers will permit research continuity and age-related tau astrogliopathy (ARTAG) and collaborations with other groups of CIBERNED. KEYWORDS

Neurodegenerative diseases with abnormal protein aggregates, molecular neuropathology, Alzheimer, synucleionopathies, tauopathies, prion diseases, biomarkers

2017 CIBERNED Annual Report / 60 Program 1 Group: Isidro Ferrer Abizanda PUBLICATIONS

• Rodriguez-Rodriguez P, Sandebring-Matton A, Merino-Serrais P, Parrado-Fernandez C, Rabano A, Winblad B et al. Tau hyperphosphorylation induces oligomeric insulin accumulation and insulin resistance in neurons.Brain : a journal of neurology. 2017. • Garcia-Esparcia P., Lopez-Gonzalez I., Grau-Rivera O., Garcia-Garrido M.F., Konetti A., Llorens F. et al. Dementia with lewy bodies: Molecular pathology in the frontal cortex in typical and rapidly progressive forms. Frontiers in Neurology. 2017;8(MAR). • Urrea L, Ferrer I, Gavín R, Del Río JA. The cellular prion protein (PrPC) as neuronal receptor for α-synuclein.Prion. 2017;11(4). • Ozaita A., Aso E. The cannabis paradox: When age matters. Nature Medicine. 2017;23(6):661-662. • Launay N., Ruiz M., Grau L., Ortega F.J., Ilieva E.V., Martinez J.J. et al. Tauroursodeoxycholic bile acid arrests axonal degeneration by inhibiting the unfolded protein response in X-linked adrenoleukodystrophy. Acta Neuropathologica. 2017;133(2):283-301. • Llorens F., Thune K., Tahir W., Kanata E., Diaz-Lucena D., Xanthopoulos K. et al. YKL-40 in the brain and cerebrospinal fluid of neurodegenerative dementias. Molecular Neurodegeneration. 2017;12(1). • Fourcade S., Morato L., Parameswaran J., Ruiz M., Ruiz-Cortes T., Jove M. et al. Loss of SIRT2 leads to axonal degeneration and locomotor disability associated with redox and energy imbalance. Aging Cell. 2017;16(6):1404-1413. • Ferrer I. Diversity of astroglial responses across human neurodegenerative disorders and brain aging. Brain Pathology. 2017;27(5):645-674. • Santpere G., Garcia-Esparcia P., Andres-Benito P., Lorente-Galdos B., Navarro A., Ferrer I. Transcriptional network analysis in frontal cortex in Lewy body diseases with focus on dementia with Lewy bodies. Brain Pathology. 2017. • Zafar S., Younas N., Sheikh N., Tahir W., Shafiq M., Schmitz M. et al. Cytoskeleton-Associated Risk Modifiers Involved in Early and Rapid Progression of Sporadic Creutzfeldt-Jakob Disease. Molecular Neurobiology. 2017;:1-21. • 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). [ePub2017]. • Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, García-Aznar JM et al. Erratum to: Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons.Molecular neurobiology. 2017. • Cabré R, Naudí A, Dominguez-Gonzalez M, Jové M, Ayala V, Mota-Martorell N et al. Lipid Profile in Human Frontal Cortex is Sustained Throughout Healthy Adult Lifespan to Decay at Advanced Ages.The journals of gerontology. Series A, Biological sciences and medical sciences. 2017. • Andres-Benito P., Fernandez-Duenas V., Carmona M., Escobar L.A., Torrejon-Escribano B., Aso E. et al. Locus coeruleus at asymptomatic early and middle Braak stages of neurofibrillary tangle pathology. Neuropathology and Applied Neurobiology. 2017;43(5):373-392. • 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. [ePub2017]. • Jiménez-García MP, Lucena-Cacace A, Robles-Frías MJ, Ferrer I, Narlik-Grassow M, Blanco- Aparicio C et al. Inflammation and stem markers association to PIM1/PIM2 kinase-induced tumors in breast and uterus.Oncotarget. 2017;8(35). • Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Aznar JM et al. Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons.Molecular neurobiology. 2017. • Matamoros-Angles A., Gayosso L.M., Richaud-Patin Y., Di Domenico A., Vergara C., Hervera A. et al. iPS Cell Cultures from a Gerstmann-Sträussler-Scheinker Patient with the Y218N PRNP Mutation Recapitulate tau Pathology. Molecular Neurobiology. 2017;:1-16. • Gamir-Morralla A, Belbin O, Fortea J, Alcolea D, Ferrer I, Lleó A et al. Kidins220 Correlates with Tau in Alzheimer’s Disease Brain and Cerebrospinal Fluid.Journal of Alzheimer’s disease : JAD. 2017;55(4).

2017 CIBERNED Annual Report / 61 • Cabre R., Naudi A., Dominguez-Gonzalez M., Ayala V., Jove M., Mota-Martorell N. et al. Sixty years old is the breakpoint of human frontal cortex aging. Free Radical Biology and Medicine. 2017;103:14-22. • Ferrer I., Gil-Villar M.P., Ayala V., Jove M., Mota-Martorell N., Portero-Otin M. et al. Region-specific vulnerability to lipid peroxidation and evidence of neuronal mechanisms for polyunsaturated fatty acid biosynthesis in the healthy adult human central nervous system. Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids. 2017;1862(5):485-495. • Marin R., Fabelo N., Martin V., Garcia-Esparcia P., Ferrer I., Quinto-Alemany D. et al. Anomalies occurring in lipid profiles and protein distribution in frontal cortex lipid rafts in dementia with Lewy bodies disclose neurochemical traits partially shared by Alzheimer’s and Parkinson’s diseases. Neurobiology of Aging. 2017;49:52-59. • Llorens F., Schmitz M., Knipper T., Schmidt C., Lange P., Fischer A. et al. Cerebrospinal fluid biomarkers of Alzheimer’s disease show different but partially overlapping profile compared to vascular dementia. Frontiers in Aging Neuroscience. 2017;9(SEP). • Vanni S., Moda F., Zattoni M., Bistaffa E., De Cecco E., Rossi M. et al. Differential overexpression of SERPINA3 in human prion diseases. Scientific Reports. 2017;7(1). • Lachen-Montes M., Gonzalez-Morales A., Zelaya M.V., Perez-Valderrama E., Ausin K., Ferrer I. et al. Olfactory bulb neuroproteomics reveals a chronological perturbation of survival routes and a disruption of prohibitin complex during Alzheimer’s disease progression. Scientific Reports. 2017;7(1). • Andres-Benito P., Moreno J., Aso E., Povedano M., Ferrer I. Amyotrophic lateral sclerosis, gene deregulation in the anterior horn of the spinal cord and frontal cortex area 8: Implications in frontotemporal lobar degeneration. Aging. 2017;9(3):823-851. • Boada M., Anaya F., Ortiz P., Olazaran J., Shua-Haim J.R., Obisesan T.O. et al. Efficacy and safety of plasma exchange with 5% albumin to modify cerebrospinal fluid and plasma amyloid-β concentrations and cognition outcomes in Alzheimer’s disease patients: A multicenter, randomized, controlled clinical trial. Journal of Alzheimer’s Disease. 2017;56(1):129-143. • Zafar S., Shafiq M., Younas N., Schmitz M., Ferrer I., Zerr I. Prion Protein Interactome: Identifying Novel Targets in Slowly and Rapidly Progressive Forms of Alzheimer’s Disease. Journal of Alzheimer’s Disease. 2017;59(1):265-275. • Ferrer I. Sisyphus in Neverland.Journal of Alzheimer’s disease : JAD. 2017. • Cayuela N, Simó M, Majós C, Rifà-Ros X, Gállego Pérez-Larraya J, Ripollés P et al. Seizure-susceptible brain regions in glioblastoma: identification of patients at risk.European journal of neurology. 2017. • Nicolia V., Cavallaro R.A., Lopez-Gonzalez I., Maccarrone M., Scarpa S., Ferrer I. et al. DNA methylation profiles of selected pro-inflammatory cytokines in Alzheimer disease. Journal of Neuropathology and Experimental Neurology. 2017;76(1):27-31. • Gonzalez de San Roman E., Manuel I., Giralt M.T., Ferrer I., Rodriguez-Puertas R. Imaging mass spectrometry (IMS) of cortical lipids from preclinical to severe stages of Alzheimer’s disease. Biochimica et Biophysica Acta - Biomembranes. 2017;1859(9):1604-1614. • Andres-Benito P., Moreno J., Dominguez R., Aso E., Povedano M., Ferrer I. Inflammatory gene expression in whole peripheral blood at early stages of sporadic amyotrophic lateral sclerosis. Frontiers in Neurology. 2017;8(OCT). • Kovacs G.G., Xie S.X., Lee E.B., Robinson J.L., Caswell C., Irwin D.J. et al. Multisite assessment of aging-related tau astrogliopathy (ARTAG). Journal of Neuropathology and Experimental Neurology. 2017;76(7):605-619. • Lopez-Gonzalez I., Cordomi C.T., Ferrer I. Regional gene expression of inflammation and oxidative stress responses does not predict neurodegeneration in aging. Journal of Neuropathology and Experimental Neurology. 2017;76(2):135-150. • Candelise N., Schmitz M., Da Silva Correia S.M., Arora A.S., Villar-Pique A., Zafar S. et al. Applications of the real-time quaking-induced conversion assay in diagnosis, prion strain- typing, drug pre-screening and other amyloidopathies. Expert Review of Molecular Diagnostics. 2017;17(10):897-904.

2017 CIBERNED Annual Report / 62 Program 1 Group: Isidro Ferrer Abizanda

• Nicolia V., Ciraci V., Cavallaro R.A., Ferrer I., Scarpa S., Fuso A. GSK3β 5’-flanking DNA methylation and expression in Alzheimer’s disease patients. Current Alzheimer Research. 2017;14(7):753-759. • Arena A, Iyer AM, Milenkovic I, Kovacs GG, Ferrer I, Perluigi M et al. Developmental Expression and Dysregulation of miR-146a and miR-155 in Down’s Syndrome and Mouse Models of Down’s Syndrome and Alzheimer’s Disease.Current Alzheimer research. ;14(12). • Llorens F., Zarranz J.-J., Fischer A., Zerr I., Ferrer I. Fatal Familial Insomnia: Clinical Aspects and Molecular Alterations. Current Neurology and Neuroscience Reports. 2017;17(4). • Armand-Ugon M, Ansoleaga B, Berjaoui S, Ferrer I. Reduced Mitochondrial Activity is Early and Steady in the Entorhinal Cortex but it is Mainly Unmodified in the Frontal Cortex in Alzheimer’s Disease.Current Alzheimer research. ;14(12). • Gil L., Federico C., Pinedo F., Bruno F., Rebolledo A.B., Montoya J.J. et al. Aging dependent effect of nuclear tau. Brain Research. 2017;1677:129-137. • Reumann R, Vierk R, Zhou L, Gries F, Kraus V, Mienert J et al. The serine protease inhibitor neuroserpin is required for normal synaptic plasticity and regulates learning and social behavior. Learning & memory (Cold Spring Harbor, N.Y.). 2017;24(12). • Llorens F., Thune K., Andres-Benito P., Tahir W., Ansoleaga B., Hernandez-Ortega K. et al. MicroRNA Expression in the Locus Coeruleus, Entorhinal Cortex, and Hippocampus at Early and Middle Stages of Braak Neurofibrillary Tangle Pathology. Journal of Molecular Neuroscience. 2017;:1-10. • Llorens F, Thüne K, Sikorska B, Schmitz M, Tahir W, Fernández-Borges N et al. Altered Ca2+ homeostasis induces Calpain-Cathepsin axis activation in sporadic Creutzfeldt-Jakob disease. Acta neuropathologica communications. 2017;5(1). • Sanchez-Mut J.V., Heyn H., Vidal E., Delgado-Morales R., Moran S., Sayols S. et al. Whole genome grey and white matter DNA methylation profiles in dorsolateral prefrontal cortex. Synapse. 2017;71(6). • Garcia-Esparcia P., Sideris-Lampretsas G., Hernandez-Ortega K., Grau-Rivera O., Sklaviadis T., Gelpi E. et al. Altered mechanisms of protein synthesis in frontal cortex in Alzheimer disease and a mouse model. American Journal of Neurodegenerative Diseases. 2017;6(2):15-25. • Canerina-Amaro A., Hernandez-Abad L.G., Ferrer I., Quinto-Alemany D., Mesa-Herrera F., Ferri C. et al. Lipid raft ER signalosome malfunctions in menopause and Alzheimer’s disease. Frontiers in Bioscience - Scholar. 2017;9(1):111-126. • Fernandez B., Ferrer I., Gil F., Hilfiker S. Biomonitorization of iron accumulation in the substantia nigra from Lewy body disease patients. Toxicology Reports. 2017;4:188-193. RESEARCH PROJECTS

• Code: AGL2015-71764-REDT. • Code: Sat CIEN-02. Title: Red nacional de priones. Title: The part of the cloud challenge on Principal Investigator: José Antonio del Río. neuroinflammation. SATIVEX. Alzheimer’s CIBERNED’s collaboration: Yes. Association. CIBERNED groups: G114; G509; G503; G609; Principal Investigator: Isidro Ferrer Abizanda. G207. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G503. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: International. Funding: 39.000 €. Funding agency: Alzheimer’s Association. Duration: 2016-2017 Funding: 950.000 €. Duration: 2016-2017

2017 CIBERNED Annual Report / 63 • Code: PI14/00268. • Code: PI2016/04. Title: Establecimiento de un algoritmo Title: The ALS CIBERNED Challenge: mitocondrial para predecir el riesgo de Accelerating New Drug Discovery. progresión a la enfermedad de Alzheimer Principal Investigator: Adolfo López De en pacientes con deterioro cognitivo leve. Munain Arregui. Principal Investigator: Marta Barrachina. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G609, G303, G503, G408. CIBERNED groups: G503. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: National. Funding agency: CIBERNED. Funding agency: Instituto de Salud Carlos III. Funding: 200.000 €. Funding: ND. Duration: 2016-2018 Duration: 2015-2017

PhD DISSERTATIONS

• Author: Mayelin Domínguez González. Title: Daño oxidativo y regulación redox en el envejecimiento cerebral: vulnerabilidad regional. Date: 1st May 2017. Supervisor: Isidro Ferrer Abizanda.

2017 CIBERNED Annual Report / 64 Antonia Gutiérrez Pérez 415

Principal Investigator Antonia Gutiérrez Pérez

Research team

José Carlos Dávila Cansino Elisabeth Sánchez Mejías Ángela Gómez Arboledas Full Professor Postdoctoral researcher PhD student

Zafaruddin Khan Juan Antonio García León Cristina Núñez Díaz Associate professor Postdoctoral researcher PhD student

David Baglietto Vargas Laura Trujillo Estrada Juan José Fernández Assistant professor Postdoctoral researcher Valenzuela PhD student Raquel Sánchez Varo Inés Moreno González Postdoctoral researcher Postdoctoral researcher Mercedes Aneiros Ferrer Technician

Dpto. Biología Celular, Genética y Fisiología (Área de Biología Celular) Facultad de Ciencias, Universidad de Málaga Campus de Teatinos 29071, Málaga (Spain) Tel.: +34 95 213 33 44 [email protected]

2017 CIBERNED Annual Report / 65 SUMMARY

In 2017, we continued investigating neuronal phenotype although their exact role is not and glial pathology in Alzheimer’s disease known at present. The term reactive is insufficient animal models and human post-mortem to define whether astrocytes contribute to the samples. We analyzed the effect of Abeta on pathology by gain or loss of function, or if on neuronal and glial proliferation in the APP/PS1 the contrary they are protective, or even have hippocampus. Neurogenesis was early affected diverse functions depending on the disease with significant decrease of neuronal stem cells stage and its location. (type-1 cells) and intermediate progenitors (type-2 cells) numbers. We demonstrated that We have identified that plaque-associated soluble Aβ damages neuronal proliferation reactive astrocytes phagocytose axonal/ using neurosphere cultures. presynaptic dystrophies. These phagocytic astrocytes are present in models and patients, On the other hand, oligomeric Aβ stimulated however the ability of these astrocytes to microglial proliferation while not modifying degrade the phagocytic material seems to astroglial population. Therefore, Aβ has a dual be limited/defective and therefore they might effect by inhibiting neuronal proliferation and not be effective in restricting the pathology. promoting the formation of microglial cells. We Our translational objective is to propose novel have also analyzed the pathological potential therapeutic targets for the development of of astrocytes in models and patients under a effective treatments aimed at restoring glial CIBERNED collaborative project and Fundació function and modifying the disease course. La Marató grant. On contrary to microglial cells, Finally, we have collaborated with several astrocytes do not show signs of degeneration in national and international research groups in Alzheimer’s patients. Astrocytes have a reactive various scientific projects on Alzheimer’s disease. KEYWORDS

Alzheimer, neuroinflammation, neuropathology, microglia, astroglia, hippocampus, transgenic models, patients PUBLICATIONS

• Baglietto-Vargas D, Sánchez-Mejias E, Navarro V, Jimenez S, Trujillo-Estrada L, Gómez-Arboledas A et al. Dual roles of Aβ in proliferative processes in an amyloidogenic model of Alzheimer’s disease.Scientific reports. 2017;7(1). • 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. [ePub2017]. • Garcia-Sanz P., Orgaz L., Bueno-Gil G., Espadas I., Rodriguez-Traver E., Kulisevsky J. et al. N370S- GBA1 mutation causes lysosomal cholesterol accumulation in Parkinson’s disease. Movement Disorders. 2017;32(10):1409-1422. • Peñalver A, Campos-Sandoval JA, Blanco E, Cardona C, Castilla L, Martín-Rufián M et al. Glutaminase and MMP-9 Downregulation in Cortex and Hippocampus of LPA1 Receptor Null Mice Correlate with Altered Dendritic Spine Plasticity.Frontiers in molecular neuroscience. ;10.

2017 CIBERNED Annual Report / 66 Program 1 Group: Antonia Gutiérrez Pérez RESEARCH PROJECTS

• Code: PI2015-2/02. • Code: 20141431. Title: Potencial patológico de los astrocitos: Title: Deciphering the link between astrocyte una nueva perspectiva en la enfermedad reactivity and neuronal damage in de Alzheimer. Alzheimer´s disease. Principal Investigator: Joan X. Comella Principal Investigator: Elena Galea Carnice. Rodríguez de velasco. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G413, G415, G204, G108, CIBERNED groups: G411, G413, G415. G411. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Private. Type: Intramurales. Funding agency: La Maratò de TV3. Funding agency: CIBERNED. Funding: 382.250 €. Funding: 350.000 €. Duration: 2015-2017 Duration: 2016-2017 • Code: FIS PI15/00796. Title: Evaluando la disfunción microglial y astroglial como base del proceso neurodegenerativo y la demencia en la enfermedad de Alzheimer: nuevas aproximaciones terapéuticas. Principal Investigator: Antonia Gutiérrez Pérez. CIBERNED’s collaboration: Yes. CIBERNED groups: G415, G411. Other CIBER’s collaboration: No. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: 99.220 €. Duration: 2016-2018

2017 CIBERNED Annual Report / 67 Alberto Lleó Bisa 504

Principal Investigator Alberto Lleó Bisa

Research team

Rafael Blesa González Isabel Sala Marta Querol Neurologist Neuropsycologist PhD student

Juan Fortea M. Belén Sánchez-Saudinós Laura Cervera Neurologist Neuropsycologist PhD student

Jordi Clarimón Echavarría Andrea Subirana Eduard Vilaplana Biologist Neuropsycologist PhD student

Olivia Belbin Lidia García Victor Montal Biologist Nurse PhD student

Roser Ribosa Laia Muñoz Neurologist Lab technician

Servicio de Neurología Hospital Santa Cruz y San Pablo Sant Antoni María Claret 167 08025 Barcelona (España) Tel.: +34 93 556 59 86 Fax: +34 93 556 56 02 [email protected]

2017 CIBERNED Annual Report / 68 SUMMARY

Our group is composed of a multidisciplinary also involved in different imaging studies, using team of neurologists, neuropsychologists, magnetic resonance imaging and amyloid biologists, engineers and lab technicians. positron emission tomography (PET) to determine The activities of the group are focused the structural correlates of different biomarkers on translational and clinical aspects of along the AD continnum, Down syndrome neurodegenerative dementias. In 2017 the and other neurodegenerative dementias. group has been working on novel genetics factors and novel biomarkers of the most Finally, the group also has uncovered common neurodegenerative dementias. We important aspects of the molecular basis have investigated new genetic risk variants in of neurodegenerative in human brain. We Alzheimer’s disease (AD) and frontotemporal have used novel microscopy techniques dementia (FTD). (Array Tomography) to show the loss of synapses in human brain in patients with Lewy In parallel, the group has conducted different body dementia. The group offers a unique investigations in cerebrospinal fluid biomarkers environment for clinicians and investigators along the continuum of AD, dementia with Lewy to tackle basic and translational aspects of bodies, FTD and Down syndrome. The group is neurodegeneration. KEYWORDS

Alberto Lleó Bisa Alzheimer, dementia, cerebrospinal fluid, amyloid, imaging, biomarkers PUBLICATIONS

• Jansen WJ, Ossenkoppele R, Tijms BM, Fagan AM, Hansson O, Klunk WE et al. Association of Cerebral Amyloid-β Aggregation With Cognitive Functioning in Persons Without Dementia. JAMA psychiatry. 2017. • Montal V., Vilaplana E., Alcolea D., Pegueroles J., Pasternak O., Gonzalez-Ortiz S. et al. Cortical microstructural changes along the Alzheimer’s disease continuum. Alzheimer’s and Dementia. 2017. • Colom-Cadena M., Grau-Rivera O., Planellas L., Cerquera C., Morenas E., Helgueta S. et al. Regional overlap of pathologies in lewy body disorders. Journal of Neuropathology and Experimental Neurology. 2017;76(3):216-224. • 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. [ePub2017]. • Alcolea D., Vilaplana E., Suarez-Calvet M., Illan-Gala I., Blesa R., Clarimon J. et al. CSF sAPPβ, YKL- 40, and neurofilament light in frontotemporal lobar degeneration. Neurology. 2017;89(2):178-188. • 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. 2017;64. • Dols-Icardo O, García-Redondo A, Rojas-García R, Borrego-Hernández D, Illán-Gala I, Muñoz- Blanco JL et al. Analysis of known amyotrophic lateral sclerosis and frontotemporal dementia genes reveals a substantial genetic burden in patients manifesting both diseases not carrying the C9orf72 expansion mutation.Journal of neurology, neurosurgery, and psychiatry. 2017. • Spataro N., Roca-Umbert A., Cervera-Carles L., Valles M., Anglada R., Pagonabarraga J. et al. Detection of genomic rearrangements from targeted resequencing data in Parkinson’s disease patients. Movement Disorders. 2016.

2017 CIBERNED Annual Report / 69 • Verheijen J., van der Zee J., Gijselinck I., Van den Bossche T., Dillen L., Heeman B. et al. Common and rare TBK1 variants in early-onset Alzheimer disease in a European cohort. Neurobiology of Aging. 2017. • Tell-Marti G, Puig-Butille JA, Potrony M, Plana E, Badenas C, Antonell A et al. A Common Variant in the MC1R Gene (p.V92M) is associated with Alzheimer’s Disease Risk.Journal of Alzheimer’s disease : JAD. 2017. • De Roeck A., Van den Bossche T., van der Zee J., Verheijen J., De Coster W., Van Dongen J. et al. Deleterious ABCA7 mutations and transcript rescue mechanisms in early onset Alzheimer’s disease. Acta Neuropathologica. 2017;134(3):475-487. • Garcia-Ayllon M.-S., Lopez-Font I., Boix C.P., Fortea J., Sanchez-Valle R., Lleo A. et al. C-Terminal fragments of the amyloid precursor protein in cerebrospinal fluid as potential biomarkers for Alzheimer disease. Scientific Reports. 2017;7(1). • Colom-Cadena M., Pegueroles J., Herrmann A.G., Henstridge C.M., Munoz L., Querol- Vilaseca M. et al. Synaptic phosphorylated α-synuclein in dementia with Lewy bodies. Brain. 2017;140(12):3204-3214. • Irwin D.J., Lleo A., Xie S.X., McMillan C.T., Wolk D.A., Lee E.B. et al. Ante mortem cerebrospinal fluid tau levels correlate with postmortem tau pathology in frontotemporal lobar degeneration. Annals of Neurology. 2017;82(2):247-258. • Cortes-Vicente E., Pradas J., Marin-lahoz J., De Luna N., Clarimon J., Turon-Sans J. et al. Early diagnosis of amyotrophic lateral sclerosis mimic syndromes: pros and cons of current clinical diagnostic criteria. Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration. 2017;18(5-6):333-340. • Carmona-Iragui M., Balasa M., Benejam B., Alcolea D., Fernandez S., Videla L. et al. Cerebral amyloid angiopathy in Down syndrome and sporadic and autosomal-dominant Alzheimer’s disease. Alzheimer’s and Dementia. 2017. • Illan-Gala I., Vilaplana E., Pegueroles J., Montal V., Alcolea D., Blesa R. et al. The pitfalls of biomarker-based classification schemes. Alzheimer’s and Dementia. 2017;13(9):1072-1074. • Gelpi E., Carrato C., Grau-Lopez L., Becerra J.L., Garcia-Armengol R., Massuet A. et al. Incidental neuronal intermediate filament inclusion pathology: unexpected biopsy findings in a 37-year- old woman with epilepsy. Neuropathology and Applied Neurobiology. 2017;43(7):636-640. • Querol-Vilaseca M, Colom-Cadena M, Pegueroles J, San Martín-Paniello C, Clarimon J, Belbin O et al. YKL-40 (Chitinase 3-like I) is expressed in a subset of astrocytes in Alzheimer’s disease and other tauopathies.Journal of neuroinflammation. 2017;14(1). • Gamir-Morralla A, Belbin O, Fortea J, Alcolea D, Ferrer I, Lleó A et al. Kidins220 Correlates with Tau in Alzheimer’s Disease Brain and Cerebrospinal Fluid.Journal of Alzheimer’s disease : JAD. 2017;55(4). • van der Zee J., Gijselinck I., Van Mossevelde S., Perrone F., Dillen L., Heeman B. et al. TBK1 Mutation Spectrum in an Extended European Patient Cohort with Frontotemporal Dementia and Amyotrophic Lateral Sclerosis. Human Mutation. 2017;38(3):297-309. • Calabria M., Cattaneo G., Marne P., Hernandez M., Juncadella M., Gascon-Bayarri J. et al. Language deterioration in bilingual Alzheimer’s disease patients: A longitudinal study. Journal of Neurolinguistics. 2017;43:59-74. • Sims R., Van Der Lee S.J., Naj A.C., Bellenguez C., Badarinarayan N., Jakobsdottir J. et al. Rare coding variants in PLCG2, ABI3, and TREM2 implicate microglial-mediated innate immunity in Alzheimer’s disease. Nature Genetics. 2017;49(9):1373-1384. • Jimenez A., Pegueroles J., Carmona-Iragui M., Vilaplana E., Montal V., Alcolea D. et al. Weight loss in the healthy elderly might be a non-cognitive sign of preclinical Alzheimer’s disease. Oncotarget. 2017;8(62):104706-104716. • van Waalwijk van Doorn LJ, Gispert JD, Kuiperij HB, Claassen JA, Arighi A, Baldeiras I et al. Improved Cerebrospinal Fluid-Based Discrimination between Alzheimer’s Disease Patients and Controls after Correction for Ventricular Volumes.Journal of Alzheimer’s disease : JAD. 2017;56(2). • Carmona-Iragui M., Santos T., Videla S., Fernandez S., Benejam B., Videla L. et al. Feasibility of Lumbar Puncture in the Study of Cerebrospinal Fluid Biomarkers for Alzheimer’s Disease in Subjects with Down Syndrome. Journal of Alzheimer’s Disease. 2017;55(4):1489-1496.

2017 CIBERNED Annual Report / 70 Program 1 Group: Alberto lleó Bisa

• Lleó A, Blesa R. Clinical course of Alzheimer’s disease. In: Contribution towards Alzheimer’s disease. Waldemar G, editor. 2017. • Engelborghs S., Niemantsverdriet E., Struyfs H., Blennow K., Brouns R., Comabella M. et al. Consensus guidelines for lumbar puncture in patients with neurological diseases. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2017;8:111-126. • Youssif C, Flix B, Belbin O, Comalada M. Measuring NLR Oligomerization IV: Using FRET-FLIM to Determine the Close Proximity of Inflammasome Components. In: NLR Proteins: Methods and Protocols. Di Virgilio F, Pelegrín P, editors. 2017. RESEARCH PROJECTS

• Code: : 2014 SGR 235. Type: National. Title: Grup de Recerca en Demècies: Sant Funding agency: Instituto de Salud Carlos III. Pau. Funding: 134.915 €. Principal Investigator: Jordi Clarimón. Duration: 2015-2017 CIBERNED’s collaboration: No. CIBERNED groups: G504. • Code: PI15/00058. Othet CIBER’s collaboration: No. Title: Valoración y caracterización de Type: CC.AA. biomarcadores en la fase preclínica de la Funding agency: Generalitat Catalunya. enfermedad de Alzheimer. Funding: 18.000 €. Principal Investigator: Olivia Belbín. Duration: 2014-2017 CIBERNED’s collaboration: No. CIBERNED groups: G504. • Code: GBHI_ALZ-18-543740. Other CIBER’s collaboration No. Title: Domiciliary Alzheimer Visiting in Down Type: National. syndrome. Funding agency: Instuto de Salud Carlos III. Principal Investigator: María Carmona Funding: 68.365 €. Iragui. Duration: 2016-2018 CIBERNED’s collaboration: No. CIBERNED groups: G504. • Code: PI14/01126. Other CIBER’s collaboration: No. Title: La estructura cerebral y el metabolismo Type: International. en la enfermedad de Alzheimer preclínica. Funding agency: Alzheimer’s Association. Interacciones entre “nuevos y viejos” Funding: 2.0974,19€. biomarcadores. Duration: 2017-2019 Principal Investigator: Juan Fortea. CIBERNED’s collaboration: No. • Code: SLT002/16/00408. CIBERNED groups: G504. Title: Estudio de biomarcadores y desarrollo Other CIBER’s collaboration: No. de nuevas estrategias terapéuticas en Type: National. una cohorte multicéntrica de Demencia Funding agency: Instuto de Salud Carlos III. Frontotemporal. Funding: 122.815€. Principal Investigator: Alberto Lleó. Duration: 2015-2019 CIBERNED’s collaboration: No. CIBERNED groups: G504. • Code: PI13/01532. Other CIBER’s collaboration: No. Title: Enfermedad de Alzheimer y síndrome Type: CC.AA. de Down. Estudios multimodales de líquido Funding agency: Generalitat de Catalunya. cefalorraquideo, resonancia magnetica y Funding: 399.363€. PET de amiloide. Duration: 2017-2019 Principal Invesitigator: Rafael Blesa. Colaboración CIBERNED: No. • Code: PI14/01561. Grupos CIBERNED: G504. Title: Marcadores sinápticos en la Colaboración otros CIBERs/Centros: No. enfermedad de Alzheimer. Tipo: Nacional. Principal Investigator: Alberto Lleó. Agencia financiadora: Instuto de Salud CIBERNED’s collaboration: No. Carlos III. CIBERNED groups: G504. Financiación: 130.075€. Other CIBER’s collaboration: No. Duración: 2014-2018

2017 CIBERNED Annual Report / 71 • Code: 201412 10. Funding agency: FUNDELA. Title: La malaltia d’Alzheimer a la sindrome Funding: 30.000€. de Down. Estudis multimodals amb LCR, RM, Duration: 2017-2018 EEG i PET. Principal Investigator: Juan Fortea. • Code: EU-JG1. CIBERNED’s collaboration: No. Title: Horizon 21 Genetics Consortium CIBERNED groups: G504. (H21GC): Large-scale pooling/genotyping Other CIBER’s collaboration: No. of individuals with Down syndrome for insight Type: Private. into predictive pathways for Alzheimer’s Funding agency: Fundació La Marató de disease. TV3. Principal Investigator:Cornelia Van Duijn Funding: 199.000€. And Tonnie Coppus. Duration: 2015-2018 CIBERNED’s collaboration: No. CIBERNED groups: G504. • Code: 201437 10. Other CIBER’s collaboration: No. Title: Biomarkers profile in different Type: Private. phenotypes of Motor Neuron Disease. Funding agency: Fundación Jerome Principal Investigator: Ricard Rojas. Lejeune. CIBERNED’s collaboration: No. Funding: 40.000€. CIBERNED groups: G504. Duration: 2017-2018 Other CIBER’s collaboration: No. Type: Private. • Code: 201614 31. Funding agency: Fundació La Marató de Title: Pre-clinical Alzheimer’s disease and TV3. Type 2 diabetes and obesity. Effects of Funding: 150.625€. bariatric surgery: a multimodal study. Duration: 2015-2018 Principal Investigator:Rafael Blesa, Amanda Jiménez. • Code: 20142610. CIBERNED’s collaboration: No. Title: Synaptic markers in preclinical CIBERNED groups: G504. Alzheimer’s disease. Other CIBER’s collaboration: CIBEROBN. Principal Investigator: Alberto Lleó. Type: Private. CIBERNED’s collaboration: No. Funding agency: Fundació La Maratá de CIBERNED groups: G504. TV3. Other CIBER’s collaboration: No. Funding: ND. Type: Private. Duration: 2017-2020 Funding agency: Fundació La Marató de TV3. • Code: IR16-P7. Funding: 199.878,75€. Title: Unitat genètica malalties Duration: 2015-2018 neurodegeneratives. Principal Investigator:Jordi Clarimon. • Code: TAUDEM. CIBERNED’s collaboration No. Title: Study with the tau Positron Emission CIBERNED groups: G504. Tomography radiotracer [18F]THK-5351 in Other CIBER’s collaboration: No. patients with different Tauopathies. Type: Private. Principal Investigator:Alberto Lleó. Funding agency: Fundació Privada Hospital CIBERNED’s collaboration: No. de la Santa Creu i Sant Pau. CIBERNED groups: G504. Funding: 7.000€. Other CIBER’s collaboration: No. Duration: 2017 Type: Private. Funding agency: Fundacion BBVA. • Code: EMIF-AD. Funding: ND. Title: EMIF-AD. A biomarker platform for AD Duration: 2016-2019 biomarkers. Principal Investigator: Simon Lovestone, • Code: FUNDELA 2017. Pieter J. Visser. Title: Estudio del splicing alternativo en ARN de CIBERNED’s collaboration: No. muestras con diagnóstico neuropatológico CIBERNED groups: G504. de ELA asociada a TDP-43. Other CIBER’s collaboration: No. Principal Investigator: Jordi Clarimon. Type: National. CIBERNED’s collaboration: No. Funding agency: Instituto de Salud Carlos III. CIBERNED groups: G504. Funding: 15.000€. Other CIBER’s collaboration: No. Duration: 2013-2018 Type: Private.

2017 CIBERNED Annual Report / 72 Program 1 Group: Alberto lleó Bisa

• Code: CP13/00091. Principal Investigator: Jordi Clarimon. Title: Search for synaptic biomarkers for AD. CIBERNED’s collaboration: No. Principal Investigator: Olivia Belbin. CIBERNED groups: G504. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G504. Type: National. Other CIBER’s collaboration: No. Funding agency: RTVE. Type: National. Funding: 99.822€. Funding agency: Instituto de Salud Carlos III. Duration: 2015-2017 Funding: ND. Duration: 2014-2017 • Code: EPISTASIS. Title: Epistasis Project. • Code: PI15/00026. Principal Investigator: A. David Smith. Title: Estudio del perfil de expresión de CIBERNED’s collaboration: No. microRNA exosomal en biofluídos para la CIBERNED groups: G504. identificación de biomarcadores de uso Other CIBER’s collaboration: No. diagnóstico en la demencia frontotemporal. Type: International. Principal Investigator: Jordi Clarimón. Funding agency: Moulton Charitable CIBERNED’s collaboration: No. Foundation & Herbertpur Trust. CIBERNED groups: G504. Funding: 40.000€. Other CIBER’s collaboration: No. Duration: 2008-2017 Type: National. Funding agency: Instituto de Salud Carlos III. • Code: 1 R21 AG056974-01 Funding: 122.815€. Title: Biological Correlates of Alzheimer in Duration: 2016-2018 Down Syndrome. Principal Investigator: Ann-Charlotte • Code: DTS15/00141. Granholm-Bentley, D.A. Paredes, A. Ledreux, Title: Evaluación del impacto de la imagen M. Margittai. PET de amiloide en el diagnóstico de CIBERNED’s collaboration: No. los pacientes con deterioro cognitivo CIBERNED groups: G504. evaluados por sospecha de alzhéimer. Other CIBER’s collaboration: No. Principal Investigator: Dr. Javier Arbizu. Type: International. CIBERNED’s collaboration: Sí. Funding agency: National Institute of Aging. CIBERNED groups: G504, G502, G609. Funding: 414.425€. Other CIBER’s collaboration: No. Duration: 2017-2019 Type: National. Funding agency: Instituto de Salud Carlos III. • Code: 4560/6393 La caixa-Down Alzheimer. Funding: 59.139€. Title: Down Alzheimer Barcelona Duration: 2016-2018 Neuroimaging Initiative (DABNI). Principal Investigator: Rafael Blesa. • Code: PI16/01825. CIBERNED’s collaboration: No. Title: Estudio del trazador de tomografía CIBERNED groups: G504. por emisión de positrones (PET) para Tau Other CIBER’s collaboration: No. [18F]THK-5351 en pacientes con diferentes Type: Private. Taupatias. Funding agency: Fundació La Caixa. Principal Investigator: Rafael Blesa. Funding: 870.000€. CIBERNED’s collaboration: No. Duration: 2017-2020 CIBERNED groups: G504. Other CIBER’s collaboration: No. • Code: SLT002/16/00099. Type: National. Title: Evaluation and characterization of Funding agency: Instituto de Salud Carlos III. synaptic proteins as biomarkers for the Funding: 122.815€. synaptic damage related to AD. Duration: 2017-2019 Principal Investigator: Olivia Belbin. CIBERNED’s collaboration: No. • Code: MRTVE/P29. CIBERNED groups: G504. Title: A comprehensive genomic analysis Other CIBER’s collaboration: No. of patients with motor neuron disease and Type: CC.AA. frontotemporal dementia to disentangle the Funding agency: Generalitat de Catalunya. missing genetic architecture of amyotrophic Funding: 65.109€. lateral sclerosis. Duration: 2017-2020

2017 CIBERNED Annual Report / 73 PhD DISSERTATIONS

• Author: Eduard Vilaplana Martinez. • Author: Martí Colom Cadena. Title: A biphasic model for cortical structural Title: Neuropathology of Lewy body diseases: changes in preclinical AD: a multimodal MRI, Confluence of multiple pathologies and role CSF and PET study. of synaptic p-?-synuclein. Date: 31th October 2017. Date: 14th June 2017. Supervisor: Alberto Lleó Bisa. Supervisor: Alberto Lleó Bisa.

• Author: María Carmona-Iragui. • Author: Miguel Ángel Santos Santos. Title: Sporadic cerebral amyloid angiopathy, Title: Clinicopathologic correlations and beyond lobar intracerebral hemorrhage: neuroimaging biomarkers in primary multimodal biomarker studies of atypical progressive aphasia. presentations. Date: 29th September 2017. Date: 12th September 2017. Supervisor: Alberto Lleó Bisa. Supervisor: Alberto Lleó Bisa.

2017 CIBERNED Annual Report / 74 Carlos Matute Almau 404

Principal Investigator Carlos Matute Almau

Research team

Alberto Pérez Samartín Susana Mato Santos Alazne Zabala Olaizola Associate professor Ramón y Cajal researcher PhD student

Fernando Pérez Cerdá Fabio Cavaliere Carolina Ortiz Sanz Associate professor Research Associate PhD student

Elena Alberdi Alfonso Asier Ruiz Núñez Maripaz Serrano Regal Assistant professor PhD student PhD student

María Domercq García Tania Quintela López Ainara Martínez González Assistant professor PhD student Technician

María Victoria Sánchez Gómez Andrea Manterola Juaristi Saioa Marcos Ezquerro Assistant professor PhD student Technician

Juan Carlos Chara Technician

Department of Neurosciences Faculty of Medicine and Dentistry University of País Vasco 48940-Leioa (Spain) Tel.: +34 94 601 32 44 Fax: +34 601 50 55 [email protected]

2017 CIBERNED Annual Report / 75 SUMMARY

Our laboratory is focused on the study of Aβ1-42 promotes translocation of the NR2B the molecular and cellular mechanisms of subunit of NMDA receptors into the plasma glia contributing to the pathophysiology of membrane via its interaction with integrin β1 Alzheimer disease (AD) and Parkinson (PD). and subsequent activation of PKC. This feature results in increased NMDA receptor activity, We recently observed that Aβ1-42 favors and in structural changes in dendritic spines. oligodendrocyte maturation and myelin repair after demyelination via activation of Fyn kinase Lastly, we found that pathogenic human in experimental settings in vitro. In turn, we found α-synuclein in Lewy bodies from Parkinson´s a substantial hypermyelination in the APP/PS1/ disease brains is taken up by neurons and tau triple transgenic mice model of AD as well as astroctyes, and transported intracellularly, in AD postmortem brain. In addition, the myelin being astrocytes resistant to its toxicity. These sheath in these mice is thicker while compound findings point to a relevant role of astroglia action potentials have lower amplitude and synthitium in PD disease propagation. In the slower conduction velocity, a feature that may last year, we have started to analyze the underlie cognitive deficits in AD. mechanisms making astrocytes less sensitive to α-synuclein and to halt its intracellular In the last year, we have carried out a transport; preliminary results indicate that these prospective study in CSF in collaboration with cells degrade this toxic protein more readily Dr. Cedazo-Mínguez and detected an increase that neurons. To validate these findings in in myelin protein levels in patients with mild murine astrocytes we have generated human cognitive deficits, which suggests that these astrocytes differentiated from iPSCs derived proteins can serve as surrogate early markers from fibroblasts obtained from controls and of AD. On the other hand, we have discovered from patients with sporadic Parkinson disease that acute exposure of cortical neurons to and with LRRK2 mutations. KEYWORDS

β-amyloid toxicity, α-synuclein, astrocytes, oligodendrocytes, myelin, oxidative stress PUBLICATIONS

• Akwa Y, Gondard E, Mann A, Capetillo-Zarate E, Alberdi E, Matute C et al. Synaptic activity protects against AD and FTD-like pathology via autophagic-lysosomal degradation.Molecular psychiatry. 2017. • Verkhratsky A., Zorec R., Parpura V. Stratification of astrocytes in healthy and diseased brain. Brain Pathology. 2017;27(5):629-644. • Lissek T., Adams M., Adelman J., Ahissar E., Akaaboune M., Akil H. et al. Building Bridges through Science. Neuron. 2017;96(4):730-735. • Fattorini G., Melone M., Sanchez-Gomez M.V., Arellano R.O., Bassi S., Matute C. et al. GAT-1 mediated GABA uptake in rat oligodendrocytes. GLIA. 2017;65(3):514-522. • Zorec R., Parpura V., Verkhratsky A. Astroglial vesicular network: Evolutionary trends, physiology and pathophysiology. Acta Physiologica. 2017. • Cisneros-Mejorado A, Gottlieb M, Ruiz A, Chara JC, Pérez-Samartín A, Marambaud P et al. Blockade and knock-out of CALHM1 channels attenuate ischemic brain damage.Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism. 2017. • Cavaliere F, Cerf L, Dehay B, Ramos-Gonzalez P, De Giorgi F, Bourdenx M et al. In vitro α-synuclein neurotoxicity and spreading among neurons and astrocytes using Lewy body extracts from Parkinson disease brains.Neurobiology of disease. 2017;103.

2017 CIBERNED Annual Report / 76 Program 1 Group: Carlos Matute Almau

• Cipriani R., Chara J.C., Rodriguez-Antiguedad A., Matute C. Effects of FTY720 on brain neurogenic niches in vitro and after kainic acid-induced injury. Journal of Neuroinflammation. 2017;14(1). • Rebai O., Belkhir M., Sanchez-Gomez M.V., Matute C., Fattouch S., Amri M. Differential Molecular Targets for Neuroprotective Effect of Chlorogenic Acid and its Related Compounds Against Glutamate Induced Excitotoxicity and Oxidative Stress in Rat Cortical Neurons. Neurochemical Research. 2017;42(12):3559-3572. • Pérez-Samartín A, Garay E, Moctezuma JPH, Cisneros-Mejorado A, Sánchez-Gómez MV, Martel- Gallegos G et al. Inwardly Rectifying K+ Currents in Cultured Oligodendrocytes from Rat Optic Nerve are Insensitive to pH.Neurochemical research. 2017;42(9). • Matute C. Mechanisms of Glial Death and Protection. In: Primer on Cerebrovascular Diseases. Caplan L.R., Biller J., Leary M.C., Lo E.H., Thomas A.J., Yenari M., Zhang J.H. editors. 2017. • Verkhratsky A., Zorec R., Rodriguez J.J., Parpura V. Neuroglia: Functional paralysis and reactivity in alzheimer’s disease and other neurodegenerative pathologies. Advances in Neurobiology. 2017;15:427-449. • 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. [ePub2017]. RESEARCH PROJECTS

• Code: PI_2016_1_0016. Type: CC.AA. Title: Plasticidad de la mielina como Funding agency: Gobierno Vasco. estrategia reparadora. Funding: 444.000 €. Principal investigator: Maria Domercq. Duration: 2013-2018 CIBERNED’s collaboration: No. CIBERNED groups: G404. • Code: PI_2016_1_0009. Other CIBER’s collaboration: No. Title: : Modulación de la interacción glia- Type: National. sinapsis como diana terapéutica en la Funding agency: Instituto de Salud Carlos III. enfermedad de Alzheimer. Funding: 51.351,9967€. Principal investigator: Estibaliz Capetillo. Duration: 2016-2018 CIBERNED’s collaboration: No. CIBERNED groups: G404. • Code: Coen Pathfinder 2015. Other CIBER’s collaboration: No. Title: Protection of neurons in vitro and in Type: CC.AA. vivo from Synuclein toxicity by molecular Funding agency: Gobierno Vasco. tweezers. Funding: 63.224€. Principal investigator: Erwan Bezard. Duration: 2016-2018 CIBERNED’s collaboration: No. CIBERNED groups: G404. • Code: KK-2017/00067. Other CIBER’s collaboration: No. Title: Polifenoles bioactivos con probado Type: European. efecto neuroprotector: desarrollo de Funding agency: JPND. ingredientes innovadores para la industria Funding: 280.000€. alimentaria vasca. Duration: 2015-2017 Principal investigator: Estibaliz Capetillo. CIBERNED’s collaboration: No. • Code: IT702-13. CIBERNED groups: G404. Title: Bases moleculares y celulares de la Other CIBER’s collaboration: No. neurodegeneración. Type: CC.AA.. Principal investigator: Carlos Matute Almau. Funding agency: Gobierno Vasco. CIBERNED’s collaboration: No. Funding: 400.000€. CIBERNED groups: G404. Duration: 2017-2018 Other CIBER’s collaboration: No.

2017 CIBERNED Annual Report / 77 • Code: Merck KGaA. • Code: SAF2015-74332-JIN. Title: Could reeducating microglia by P2X4R Title: Role of lactate and monocarboxylate manipulation constitute an alternative transporters in oligodendrocyte progenitor therapy for MS? cell energy metabolism and remyelination. Principal Investigator: Maria Domercq Principal Investigator: Vanja Tepavcevic. García. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G404. CIBERNED groups: G404. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: International. Funding agency: MINECO. Funding agency: Merck. Funding: 160.000€. Funding: 250.000€. Duration: 2017-2019 Duration: 2015-2017 • 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: National. Funding agency: MINECO. Funding: 340.000€. Duration: 2017-2019

2017 CIBERNED Annual Report / 78 Guadalupe Mengod Los Arcos 508

Principal Investigator Guadalupe Mengod Los Arcos

Research team

Roser Cortés Colomer Silvia Serrano Acedo Natascha Schmidt Research staff Researcher Erasmus student

Mª Teresa Vilaró Comas Álex Moreno Giró Felipe Jiménez Herández Research staff Master student Researcher

Rocío Martín-Álvarez Genís Castellano Mayós Lab technician PhD Fellow

IIBB-CSIC, IDIBAPS, CIBERNED Tel.: +34 93 363 83 23 Fax: +34 93 363 83 01 [email protected]

2017 CIBERNED Annual Report / 79 SUMMARY

Our group is composed of two sub- chronically with a new PDE7 inhibitor (VP3.15) lines, whose main and common theme is that significantly reduces the clinical symptoms neurodegeneration. of these mice (Martin-Alvarez et al. , 2017). During 2017 we have continued to investigate Together with the group of Dr. de Gortari the relationship between cAMP and the cellular from the Molecular Physiology group of the and molecular aspects of neurodegenerative INPRFM of Mexico DF we have observed diseases with an inflammatory component. that the treatment of rats with a classic PDE7 We are carrying out a series of studies of the inhibitor (BRL 50481) induces anxiolytic and effect of the manipulation of cAMP levels with anorexigenic-like behavior, depending on the drugs on murine macrophage cell cultures (cell dose used (Valdes-Moreno et al., 2017). line and primary cultures) on their polarization toward pro-inflammatory or anti-inflammatory We are also analyzing in a transgenic triple phenotypes. mouse model of Alzheimer’s disease the effect of stimulation of serotonin 5-HT4 receptors We have shown the possible influence of cAMP on the formation of deposits of amyloid Aβ on the development of EAE (experimental peptide and on other neuropathological and autoimmune encephalomyelitis, model of behavioral alterations observed in this animal multiple sclerosis) in mice, by treating them model. KEYWORDS

Neurodegeneration, neuroinflammation, molecular neuropharmacology PUBLICATIONS

• Code: PI15/00148. CIBERNED goups: G508. Title: ¿Están implicados los cambios Other CIBER’s collaboration: No. fenotípicos de las microglia/macrofagos en Type: National. los efectos beneficiosos de los inhibidores de Funding agency: Instituto de Salud Carlos III. PDEs del AMPc en la EAE crónica? Funding: 101.500€. Principal Investigator: Guadalupe Mengod. Duration: 2015-2018 CIBERNED’s collaboration: No.

2017 CIBERNED Annual Report / 80 María Asunción Pastor Muñoz 507

Principal Investigator María Asunción Pastor Muñoz

Research team

Pau Pastor Muñoz Javier Bernacer María Juan Pablo Tartari Co-Principal Investigator PhD MD

Miquel Aguilar Barberá Gonzalo Arrondo Luis Eudave Ramos PhD PhD PhD student

Martín Martínez Villar María Asunción Fernández Silvia Zarraluqui López PhD Seara PhD student PhD Maite Aznarez Sanado Sara Ortega Cubero PhD Elisa Mengual Poza PhD student PhD Elkin Luis García PhD Silvia Romero Contreras Nurse

Laboratorio de Neuroimagen Center for Applied Medical Research Universidad de Navarra 31008 Pamplona, España Tel.: +34 94 825 54 00 Fax: +94 829 65 00 [email protected]

2017 CIBERNED Annual Report / 81 SUMMARY

The group of Maria Pastor/Pau Pastor parkinsonism throughout magnetic resonance (Neuroimaging and Neurogenetics Laboratories, and next generation sequencing. Currently, Center for Applied Medical Research, CIMA, the projects focus on the influence of genetic University of Navarra) is working on projects variants in the evolution of dementias, novel focused to the identification of diagnostic patterns of amyloid-PET and genetic and markers that would help to understand and cerebrospinal fluid markers. In addition, we are personalize neurodegenerative diseases. working on the identification of novel genetic The Group is working actively on genetic risk variants for Parkinson disease and Essentials and neuroimaging markers in dementia and tremor. KEYWORDS

Neuroimaging, Functional magnetic resonance imaging, ASL, genetics, SNPs, Gen PUBLICATIONS

• Giri A., Mok K.Y., Jansen I., Sharma M., Tesson C., Mangone G. et al. Lack of evidence for a role of genetic variation in TMEM230 in the risk for Parkinson’s disease in the Caucasian population. Neurobiology of Aging. 2017;50:167.e11-167.e13. • Fernández-Santiago R, Iranzo A, Gaig C, Serradell M, Fernández M, Pastor P et al. MAPT association with REM sleep behavior disorder.Neurology. Genetics. 2017;3(1). • Verheijen J., van der Zee J., Gijselinck I., Van den Bossche T., Dillen L., Heeman B. et al. Common and rare TBK1 variants in early-onset Alzheimer disease in a European cohort. Neurobiology of Aging. 2017. • Gendron T.F., Daughrity L.M., Heckman M.G., Diehl N.N., Wuu J., Miller T.M. et al. Phosphorylated neurofilament heavy chain: A biomarker of survival for C9ORF72-associated amyotrophic lateral sclerosis. Annals of Neurology. 2017;82(1):139-146. • Lecumberri A., Lopez-Janeiro A., Corral-Domenge C., Bernacer J. Neuronal density and proportion of interneurons in the associative, sensorimotor and limbic human striatum. Brain Structure and Function. 2017;:1-11. • Jimenez-Jimenez F.J., Gomez-Tabales J., Alonso-Navarro H., Zurdo M., Turpin-Fenoll L., Millan- Pascual J. et al. Association between the rs1229984 polymorphism in the alcohol dehydrogenase 1B gene and risk for restless legs syndrome. Sleep. 2017;40(12). • Alvarez I., Aguilar M., Gonzalez J.M., Ysamat M., Lorenzo-Bosquet C., Alonso A. et al. Clinic-Based Validation of Cerebrospinal Fluid Biomarkers with Florbetapir PET for Diagnosis of Dementia. Journal of Alzheimer’s Disease. 2017;61(1):135-143. • Jiménez-Jiménez FJ, García-Martín E, Alonso-Navarro H, Martínez C, Zurdo M, Turpín-Fenoll L et al. Thr105Ile (rs11558538) polymorphism in the histamine-1-methyl-transferase (HNMT) gene and risk for restless legs syndrome.Journal of neural transmission (Vienna, Austria : 1996). 2017;124(3).

2017 CIBERNED Annual Report / 82 Program 1 Group: María Asunción Pastor Muñoz RESEARCH PROJECTS

• Code: COH-PAK-2014-01. • Code: SAF 2016-81016-R. Title: COPADIS. Title: Marcadores de Neuroimagen de la Principal Investigator: Diago Santos García. enfermedad de Parkinson genéticamente CIBERNED’s collaboration: No. determinada y Parkinsonismos mediante CIBERNED groups: G507. secuencias de RM de neuromelanina y SWI Other CIBER’s collaboration: No. del tronco del encéfalo. Type: ND. Principal Investigator: María Asunción Pastor Funding agency: ND. Munoz. Funding: 2.000.000€. CIBERNED’s collaboration: No. Duration: 2017 CIBERNED groups: G507. Other CIBER’s collaboration: No. Type: National. Funding agency: MINECO. Funding: 100.000€. Duration: 2017

2017 CIBERNED Annual Report / 83 José Rodríguez Álvarez 406

Principal Investigator José Rodríguez Álvarez

Research team

José Aguilera Ávila Miriam Javier Torrent Carlos Saura Antolín Academic staff PhD student Academic staff

Judit Catala Solsona Alfredo Jesús Miñano Molina Dolores Siedlecki Wullich PhD student PhD Fellow PhD student

Lilian Enríquez Barreto Laura Ortega Hernández Sergio Marco Martín PhD Fellow PhD Fellow PhD Fellow

Instituto de Neurociencias Universidad Autónoma de Barcelona Edificio M Campus de Bellaterra 08193 Cerdanyola del Vallés, Barcelona (Spain) Tel.: +34 93 581 38 61 [email protected]

2017 CIBERNED Annual Report / 84 SUMMARY

Alzheimer’s disease (AD), the major cause of using viral-mediated CRTC1 gene therapy, we dementia in the elderly, affects 47 million people have provided the first evidence that targeting worldwide and represents nowadays a large CRTC1-Nr4a2 reverses spatial and contextual health, social and economic burden. In the memory deficits in AD and neurodegeneration last decades, AD research has mainly focused mouse models. Moreover, our findings show on the classical pathological hallmarks of the that CRTC1 regulates synapse morphology, disease. Unfortunately, several promising clinical which is crucial for hippocampus-dependent trials based on these hallmarks have failed, synaptic plasticity and long-term memory. including recent anti β-amyloid strategies. The current lack of effective therapies to prevent In addition, the CRTC1-Nr4a2 axis controls the or delay the progression of the disease urges expression of the AMPAR GluA1 and GluA2 for alternative treatments. In our laboratory subunits and is involved in the activity-dependent we are deeply involved in understanding the regulation of BDNF. Other mechanisms that early events involved in AD which is crucial to could be related to early synaptic impairment in develop novel therapeutic strategies for this AD are alteration of glutamate AMPA receptors devastating disease. Synaptic dysfunction regulation by auxiliary synaptic proteins and the and loss of synapses are major correlates of control of local synaptic synthesis by miRNAs. cognitive impairment in AD. In this context, we have identified a group of miRNAs (miRNAs signature; patent pending), In the past years our research efforts were related to synaptic proteins regulation, that focused on deciphering the molecular events change significantly their expression levels underlying synaptic dysfunction at early stages in human plasma samples of early AD (Braak of AD. We and others have shown that memory III/IV stages). The early alteration of synaptic deficits appear before the appearance of function in AD can also be associated with the classical AD pathological hallmarks in an alteration of the permeability of the blood- José Rodríguez Álvarez mouse models of AD. Transcriptomic analyses brain barrier that could generate an alteration revealed that these deficits are caused by the in the homeostasis in the cerebral parenchyma alteration of a gene transcriptional program of certain angioneurines. The studies carried dependent on the CREB-regulated transcription out last year in a cellular model of the blood- coactivator-1 (CRTC1). Indeed, activity and brain barrier indicate that the activity of certain levels of CRTC1 and its target gene Nr4a2, which endothelial enzymes causes changes in BDNF is involved in synaptic plasticity and memory, and pro-inflammatory factors levels, released are reduced in human brain at intermediate by the endothelium, causing changes in Braak III/IV pathological stages. Importantly, synaptic maturation. KEYWORD

Alzheimer disease, memory, intracelular signaling, gene regulation, glutamate receptors, CRTC1, early synaptic dysfunction PUBLICATIONS

• Parra-Damas A., Chen M., Enriquez-Barreto L., Ortega L., Acosta S., Perna J.C. et al. CRTC1 Function During Memory Encoding Is Disrupted in Neurodegeneration. Biological Psychiatry. 2017;81(2):111-123. • Saura C.A., Cardinaux J.-R. Emerging Roles of CREB-Regulated Transcription Coactivators in Brain Physiology and Pathology. Trends in Neurosciences. 2017. • Vargas-Estevez C., Duch M., Duque M., del Campo F.J., Enriquez-Barreto L., Murillo G. et al. Suspended Silicon Microphotodiodes for Electrochemical and Biological Applications. Small. 2017;13(41).

2017 CIBERNED Annual Report / 85 • 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. [ePub2017]. • 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. [ePub2017]. • Parra-Damas A, Rubió-Ferrarons L, Shen J, Saura CA. CRTC1 mediates preferential transcription at neuronal activity-regulated CRE/TATA promoters.Scientific reports. 2017;7(1). RESEARCH PROJECTS

• Code: SAF2014-59697-R. • Code: 2015/01-1. Title: Mecanismos moleculares Title: Terapia Génica dirigida a neuregulinas implicados en la disfunción de la sinapsis para el tratamiento de la degeneración glutamatergicas en fases tempranas de la de las motoneuronas en la esclerosis lateral enfermedad de Alzheimer. amiotrófica. Principal Investigator: José Rodríguez Álvarez. Principal Investigator: Xavier Navarro CIBERNED’s collaboration: No. Acebes. CIBERNED groups: G406. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G607, G113, G406, G407. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Funding: 181.500 €. Funding agency: CIBERNED. Duration: 2015-2017 Funding: 240.000 €. Duration: 2015-2017 • 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: José Rodríguez Álvarez. CIBERNED’s collaboration: No. CIBERNED groups: G406. Other’s collaboration: No. Type: Private. Funding agency: Fundació La Maraton TV3. Funding: 199.875 €. Duration: 2015-2018

2017 CIBERNED Annual Report / 86 Javier Sáez Valero 407

Principal investigator Javier Sáez Valero

Research team

Inmaculada Cuchillo Ibáñez María Salud García Ayllón Aitana Sogorb Esteve PhD Fellow PhD Fellow PhD student

Inmaculada López Font Jordi Alom Poveda Claudia P. Boix PhD Fellow Head of neurologist PhD student department Trinidad Mata Balaguer PhD student

Instituto de Neurociencias de Alicante Universidad Miguel Hernández-CSIC Avenida Ramón y Cajal, s/n 03550 Sant Joan d’Alacant (Spain) Tel.: +34 96 591 95 80 Fax: +34 96 591 95 61 [email protected]

2017 CIBERNED Annual Report / 87 SUMMARY

The aim of our research group is focus into the amyloid precursor protein (APP) in CSF and Alzheimer’s disease (AD) from a basic point of their potential as biomarkers for AD (García- view, but also regarding translational benefits Ayllón et al., 2017); as well a shift in the balance including therapy and diagnostic applications. of the soluble forms of APP (sAPP) in CSF that In our basic research line, we have extended probably reflect pathophysiological changes in our examination on reelin signaling impairment the brain (Lopez-Font et al., 2017). in AD. We previously demonstrated that the C-terminal fragments (CTFs) of the reelin/ApoE Furthermore, regarding therapy, since the receptor, ApoER2, generated by proteolytic clinical trials with the potential therapeutic processing after reelin binding, regulate reelin agents, γ-secretase inhibitors (GSIs) have expression. proven disappointing, we addressed the possibility that γ-secretase inhibition can Now, we found that the ApoER2-CTF levels are provoke a rebound effect, elevating the levels decreased, while reelin expression is increased in of the catalytic γ-secretase subunit, presenilin-1 AD brain at advanced Braak stages, and in cell (PS1). In vitro and in vivo models demonstrate cultures after Aβ treatment. Our data support that treatments with GSIs augments PS1 levels. the view that ApoER2-CTF fragments exert a Sustained γ-secretase inhibition did not exert a modulatory role on reelin expression and that long-term effect on PS1 activity, evident through the mechanism fails in AD (Mata-Balaguer et al., the decrease in the γ-secretase substrates, the 2018). We also extend our analysis on potential CTFs of APP and ApoER2 (Sogorb-Esteve et al., cerebrospinal fluid (CSF) biomarkers for AD. 2017). The rebound increase in PS1 in response to GSIs must be taken into consideration for We have demonstrated the presence of CTFs of future drug development. KEYWORDS

Alzheimer, Amyotrophic Lateral Sclerosis, APP, secretase, reelin, biomarker, therapy PUBLICATIONS

• Garcia-Ayllon M.-S., Lopez-Font I., Boix C.P., Fortea J., Sanchez-Valle R., Lleo A. et al. C-Terminal fragments of the amyloid precursor protein in cerebrospinal fluid as potential biomarkers for Alzheimer disease. Scientific Reports. 2017;7(1). • Sogorb-Esteve A., Garcia-Ayllon M.-S., Llansola M., Felipo V., Blennow K., Saez-Valero J. Inhibition of γ-Secretase Leads to an Increase in Presenilin-1. Molecular Neurobiology. 2017;:1-12. • Gimenez-Molina Y., Villanueva J., Nanclares C., Lopez-Font I., Viniegra S., Frances M.M. et al. The differential organization of F-actin alters the distribution of organelles in cultured when compared to native chromaffin cells. Frontiers in Cellular Neuroscience. 2017;11.

2017 CIBERNED Annual Report / 88 Program 1 Group: Javier Sáez Valero RESEARCH PROJECTS

• Code: PI15/00665. • Code: PI2015/01. Title: Interacciones de las vias Reelina/ApoE Title: Terapia Génica dirigida a neuregulinas y ligandos alternativos con el β-amiloide, su para el tratamiento de la degeneración disfunción en la enfermedad de Alzheimer. de las motoneuronas en la esclerosis lateral Principal Investigator: Javier Sáez Valero. amiotrofica. CIBERNED’s collaboration: No. Principal Investigator: Xavier Navarro CIBERNED groups: G407. Acebes. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Si. Type: National. CIBERNED groups: G607, G113, G406, G407. Funding agency: Instituto de Saud Carlos III. Other CIBER’s collaboration: No. Funding: 80.465 €. Type: Intramurales. Duration: 2016-2018 Funding agency: CIBERNED. Funding: 240.000 €. • Code: PI14/00566. Duration: 2015-2017 Title: Procesamiento por secretasas de la forma colinérgica de acetilcolinesterasa, su actividad transcripcional e implicación en la enfermedad de Alzheimer. Principal Investigator: María Salud García Ayllon. CIBERNED’s collaboration: No. CIBERNED groups: G407. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 90.144,99 €. Duration: 2015-2017

2017 CIBERNED Annual Report / 89 Eduardo Soriano García 408

Principal Investigator Eduardo Soriano García

Research team

Marta Pascual Sánchez Ashraf Muhaisen Antoni Parcerisas Mosqueda Researcher PhD Fellow PhD Fellow

Jesús Mariano Ureña Bares Serena Mirra Cristina Roselló Busquets Researcher PhD Fellow PhD student

Ferran Burgaya Márquez Daniela Rossi Marc Hernaiz Llorens Researcher PhD Fellow PhD student

Fausto Alexánder Ulloa Yasmina Manso Sanz Irene Lobón García Darquea PhD Fellow PhD student Researcher Mónica Pardo Muñoz Alba del Valle Vilchez Acosta Tiziana Cotrufo PhD Fellow PhD student PhD Fellow Jonatan Dorca Arévalo Marco Solís Benites Lluís Pujadas Puigdomènech PhD Fellow Administrative assistant PhD Fellow Núria Masachs Janoher Universidad de Barcelona, Facultad de Biología PhD student Departamento de Biología Celular, Fisiología e Inmunología Pablo Barrecheguren Manero Avenida Diagonal 643, Edificio Prevosti 1r piso PhD student Barcelona 08028 Tel.: +34 93 403 71 17 Fax: +34 93 403 71 16 Eva Dávila Bouziguet [email protected] PhD student

2017 CIBERNED Annual Report / 90 SUMMARY

In Alzheimer’s disease less than 1.5% of cases genetic and molecular Reelin-dependent are hereditary (FAD), while the causes of late- networks. onset sporadic Alzheimer’s disease (SAD) remain largely unknown. Several genetic 2) Some human diseases, including cancer, are factors (identified in GWAS) increase the risk of caused by somatic mutations that alter the developing this disease. Throughout this year function of specific cell populations. Through we have focused mainly on two hypotheses the complete sequencing of the exome regarding the pathogenesis of AD and potential we have recently tested the hypothesis therapeutic strategies: that variations and somatic mutations (referenced here as Single Nucleotide 1) Reelin is an extracellular protein crucial for Variations-SNVS) are present in the brain of brain development, which is also expressed SAD. We have found a remarkable number in the adult brain. Recent studies support of specific SNVS in SAD, which were not the relationship between Reelin and AD and detected in the blood of the same donors. we have hypothesized that by regulating SNVS specific loci with recurrent SNVS were metabolism Aß42, phosphorylation of Tau common to several patients and were not and synaptic plasticity in adults, the Reelin found in the brains of control donors. These cascade may be an important regulator of results reveal that the blood and brain brain functions in adults, whose deregulation have different genomic DNA variations and may be related to the pathology of AD, suggest that somatic genome remodeling suggesting that Reelin may be protective of the brain may contribute to the against AD. We recently tested this hypothesis pathogenesis of SAD. In this year we have by overexpression of Reelin (Reelin-OE mice) tried to understand more deeply the signing in a model of AD (J20). The Reelin-OE / J20 of somatic mutations in brain SAD: mice showed decreased plaque burden Eduardo Soriano García and the rescue of synaptic and cognitive • Determining somatic gene signature deficits, suggesting that Reelin protects from in SAD, including INFDELS, variations AD pathology. We have studied: on chromosome X and copy number variations (CNVs). • If the Reelin improves AD in mice, once the disease has developed, and in mice • Integrating these data in defined molecular that exhibit AD pathology related to Tau pathways through systems biology. (Tau-GSK3 WLW -OE and mice). • We have begun to understand how • Whether a downregulation of Reelin specific somatic genetic events SAD may specifically in the adult (using conditional contribute to the pathogenesis of the KO mice, FlReelin / AD mice) accelerates disease. AD pathology. We believe these data reveal important • Started to understand the physiological new molecular and genetic information to and molecular mechanisms by which understand the mechanisms that trigger the affects Reelin AD pathology, including SAD and to open new therapeutic avenues. KEYWORDS

Reelin, Synapses, Alzheimer Disease, Somatic Mutations PUBLICATIONS

• Lissek T., Adams M., Adelman J., Ahissar E., Akaaboune M., Akil H. et al. Building Bridges through Science. Neuron. 2017;96(4):730-735.

2017 CIBERNED Annual Report / 91 • Gomez-Ramos A., Picher A.J., Garcia E., Garrido P., Hernandez F., Soriano E. et al. Validation of Suspected Somatic Single Nucleotide Variations in the Brain of Alzheimer’s Disease Patients. Journal of Alzheimer’s Disease. 2017;56(3):977-990. • del Mar Masdeu M., Armendariz B.G., Torre A.L., Soriano E., Burgaya F., Urena J.M. Identification of novel Ack1-interacting proteins and Ack1 phosphorylated sites in mouse brain by mass spectrometry. Oncotarget. 2017;8(60):101146-101157. • Soler H., Dorca-Arevalo J., Gonzalez M., Rubio S.E., Avila J., Soriano E. et al. The GABAergic septohippocampal connection is impaired in a mouse model of tauopathy. Neurobiology of Aging. 2017;49:40-51. • Barrecheguren P.J., Ros O., Cotrufo T., Kunz B., Soriano E., Ulloa F. et al. SNARE proteins play a role in motor axon guidance in vertebrates and invertebrates. Developmental Neurobiology. 2017. RESEARCH PROJECTS

• Code: 2017SGR1280 (Solicitado). • Code: SAF2016-76340-R. Title: Ayudas para apoyar las actividades Title: Novel approaches to understand de los grupos de investigación de Cataluña Alzheimers Disease pathogenesis and SGR 2017-2019. therapeutics. Principal Investigator: Eduardo Soriano Principal Investigator: Eduardo Soriano García. Garcia. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G408. CIBERNED groups: G408. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: CC.AA.. Type: National. Funding agency: Generalitat de Catalunya. Funding agency: MINECO. Funding: 70.500€. Funding: 380.010€. Duration: 2017-2019 Duration: 2017-2019 • Code: 20143330-31. • Code: : PI2016/04. Title: The role of reelin at the crossroads of Title: The ALS CIBERNED Challenge: alzheimer’s disease mechanisms: Tauopathy, accelerating new drug discovery. amyloid toxicity and transmissibility. Principal Investigator: Adolfo López De Principal Investigator: Lluis Pujadas Munain Arregui. Puigdomenech. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G609, G303, G503, G408. CIBERNED groups: G408. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: Private. Funding agency: CIBERNED. Funding agency: Fundació la Marato de TV3. Funding: 200.000 €. Funding: 187.635,5 €. Duration: 2016-2018 Duration: 2016-2018

PhD DISSERTATIONS

• Autor: Antoni Parcerisas Mosqueda Fecha: 20 de julio de 2017. Titulo: Paper de la molècula d’adhesió Director: Eduardo Soriano García. neuronal Ncam2 en desenvolupament i plasticitat neuronal.

2017 CIBERNED Annual Report / 92 Ignacio Torres Alemán 409

Principal Investigator Ignacio Torres Alemán

Research team

Ana M. Fernández Raquel Herrero Roberto Cañadas Researcher Researcher Researcher

Miguel García Jaime Pignatelli Laura Martínez Researcher Researcher Researcher

Jonathan Zegarra Estrella Fernández de Sevilla Researcher Researcher

Instituto Cajal Avenida Dr Arce, 37 28002 Madrid (Spain) Tel.: +34 91 585 47 23 Fax: +34 91 585 47 54 [email protected]

2017 CIBERNED Annual Report / 93 SUMMARY

Our laboratory works in the physio-pathology the brain vasculature between the ovarian of insulin factors in the adult nervous system. hormone estradiol and IGF-I. In this vein, we During 2017 we finished studying the control of also concluded that circulating IGF-I may brain glucose metabolism by IGF-I and insulin, serve as a biomarker of stress vulnerability in and started to explore the relationship with humans (in collaboration with other group). We brain perfusion, another early pathological continue work on the protective role of IGF-I in perturbation in Alzheimer´s disease (AD). ischemic damage, identifuing its receptor in astrocytes as a central element in the progress We also completed the analysis of mechanism of the pathology. Finally, we have started a underlying changes in mood during the peri- new line of study on the relationship between menopause transition, as they may be related IGF-I and the hypothalamic orexinergic system to common co-morbidities in AD. We ascribed that connects metabolism, mood and sleep these changes to a loss of the interaction at patterns. All these traits are involved in AD. KEYWORDS

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

PUBLICATIONS

• Fernandez A.M., Hernandez-Garzon E., Perez-Domper P., Perez-Alvarez A., Mederos S., Matsui T. et al. Insulin regulates astrocytic glucose handling through cooperation with IGF-I. Diabetes. 2017;66(1):64-74. • Santi A, Genis L, Torres Aleman I. A Coordinated Action of Blood-Borne and Brain Insulin-Like Growth Factor I in the Response to Traumatic Brain Injury.Cerebral cortex (New York, N.Y. : 1991). 2017. • Fernandez A.M., Hernandez E., Guerrero-Gomez D., Miranda-Vizuete A., Torres Aleman I. A network of insulin peptides regulate glucose uptake by astrocytes: Potential new druggable targets for brain hypometabolism. Neuropharmacology. 2017. • Stein A.M., Munive V., Fernandez A.M., Nunez A., Aleman I.T. Acute exercise does not modify brain activity and memory performance in APP/PS1 mice. PLoS ONE. 2017;12(5). • Franco C., Genis L., Navarro J.A., Perez-Domper P., Fernandez A.M., Schneuwly S. et al. A role for astrocytes in cerebellar deficits in frataxin deficiency: Protection by insulin-like growth factor I. Molecular and Cellular Neuroscience. 2017;80:100-110.

2017 CIBERNED Annual Report / 94 Program 1 Group: Ignacio Torres Alemán RESEARCH PROJECTS

• Code: PI2016/01. • Code: SAF2016-76462-C2-1-P. Title: Alteraciones del metabolismo gluco- Title: El sistema IGF-I/orexina/acetilcolina lipidico y desarrollo de la demencia de como nexo de union entre co-morbilidad. alzhéimer. Principal Investigator: Ignacio Torres Principal Investigator: Ignacio Torres Alemán. Alemán. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: G409. CIBERNED groups: G409 ; G402; G511; G502; Other CIBER’s collaboration: No. G412. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Fundig: 260.000€. Funding agency: CIBERNED. Duration: 2017-2020 Funding: 200.000€. Duration: 2016-2018 PhD DISSERTATIONS

• Author: Víctor Munive Herrera. Title: Papel de la interacción de IGF-I y estradiol en las diferencias sexuales en regulación del estado de ánimo mediante ejercicio físico. Date: 12th April 2017. Supervisor: Ignacio Torres Alemán.

2017 CIBERNED Annual Report / 95 Ramón Trullás Oliva 410

Principal Investigator Ramón Trullás Oliva

Research team

Anna Colell Riera Manuel Rodríguez Allué Margalida Puigròs Serra Research Scientist Assistant professor Predoctoral Student

Petar Podlesniy Nuria Serra Barea Research Scientist Technician

Unidad de Neurobiología IIBB-CSIC IDIBAPS Calle Rosselló 161, 7ª Planta, Laboratorio 711 08036 Barcelona (Spain) Tel.: +34 659 69 61 87 Fax: +34 93 363 83 01 [email protected]

2017 CIBERNED Annual Report / 96 SUMMARY

During the year 2017, we have completed the from patients with familial Parkinson’s have development of a new method to measure a significant increase in the transcription, but absolute values of mitochondrial DNA (mtDNA) not in the replication, of mtDNA. These results transcription and replication, based on the support the hypothesis that transcription and digital analysis of the polymerase chain reaction. replication are alternative processes and that The new method has been called Selfie-digital their regulation is altered in in both idiopathic PCR (Selfie-dPCR), because it allows to measure and familial forms of Parkinson’s disease. in a specific way the number of transcripts of the heavy (H) and light (L) chains in relation to On the other hand, we have completed the its own coding mtDNA. Using this method, we studies investigating the content of mtDNA in have investigated the replication, transcription the cerebrospinal fluid of patients at different and release of mtDNA in fibroblasts of patients stages of multiple sclerosis disease. The results with idiopathic Parkinson’s disease and with obtained indicate that patients with progressive Parkinson’s disease related to the G2019S multiple sclerosis show a significant increase in mutation in the LRRK2 gene. the content of mtDNA in the cerebrospinal fluid and that treatment with Fingolimod lowers the We have also started the analysis of the increase in the concentration of mtDNA at the transcription and replication of mtDNA control levels. These results indicate that the in neuroblastoma lines that express gene concentration of mtDNA in the cerebrospinal mutations that cause familial Alzheimer’s fluid responds to pharmacological treatment disease. The results obtained so far demonstrate and that it can be used as a biological indicator that both the fibroblasts of patients with of the therapeutic effect of novel multiple idiopathic Parkinson’s disease and those sclerosis treatments. KEYWORDS

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

• Hernando S., Herran E., Figueiro-Silva J., Pedraz J.L., Igartua M., Carro E. et al. Intranasal Administration of TAT-Conjugated Lipid Nanocarriers Loading GDNF for Parkinson’s Disease. Molecular Neurobiology. 2017;:1-11. • Leurs CE, Podlesniy P, Trullas R, Balk L, Steenwijk MD, Malekzadeh A et al. Cerebrospinal fluid mtDNA concentration is elevated in multiple sclerosis disease and responds to treatment. Multiple sclerosis (Houndmills, Basingstoke, England). 2017. • Podlesniy P., Trullas R. Absolute measurement of gene transcripts with Selfie-digital PCR. Scientific Reports. 2017;7(1). • Jorba I., Menal M.J., Torres M., Gozal D., Pinol-Ripoll G., Colell A. et al. Ageing and chronic intermittent hypoxia mimicking sleep apnea do not modify local brain tissue stiffness in healthy mice. Journal of the Mechanical Behavior of Biomedical Materials. 2017;71:106-113.

2017 CIBERNED Annual Report / 97 RESEARCH PROJECTS

• Code: PI2016/06. • Code: SAF2014-56644-R. Title: Identificación de vías fisiopatológicas y Title: Mecanismos de regulación del número biomarcadores candidatos en la fase pre- de copias de ADN mitocondrial para el diagnóstica de la enfermedad de Parkinson. tratamiento del alzhéimer. Principal Investigator: Miquel Vila Bover. Principal Investigator: Ramón Trullás. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G109, G601, G207, G410. CIBERNED groups: G410. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: National. Funding agency: CIBERNED. Funding agency: MINECO. Funding: 196.000 €. Funding: 160.000 €. Duration: 2016-2018 Duration: 2015-2017 • Code: PI042977. Title: Mitochondrial DNA copy number in LRRK2 fibroblasts. Principal Investigator: Eduardo Tolosa. CIBERNED’s collaboration: Si. CIBERNED groups: G410, G207. Other CIBER’s collaboration: No. Type: International. Funding agency: Michael J. Fox Foundation. Funding: 48.000 €. Duration: 2016-2017

PhD DISSERTATIONS

• Author: Mónica Mendoza Blanco. Title: Expanding Insights of mitophagy and mitochondrial morphology in a Parkinson’s Disease Genetic Model. Date: 18th July 2017. Supervisor: Ramón Trullás Oliva.

2017 CIBERNED Annual Report / 98 Javier Vitorica Ferrández 411

Principal Investigator Javier Vitorica Ferrández

Research team

Marisa Vizuete Chacón MªVirtudes Sánchez Mica Carmen Romero Molina Full professor PhD student PhD student

Victoria Navarro Garrido Clara Muñoz Castro Researcher PhD student Sebastián Jiménez Muñoz Technician

Mª Luisa García-Calvo Technician

Dpto. Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad de Sevilla Instituto de Biomedicina de Sevilla, Hospital Universitario Virgen del Rocío Sevilla (Spain) Tel.: +34 95 455 67 70 Fax: +34 95 592 30 53 [email protected]

2017 CIBERNED Annual Report / 99 SUMMARY

During this year, 2017, we have continued with that, unlike the microglial cells, astrocytes in the characterization of the microglial response human samples presented a clear activation. in both murine models of Alzheimer’s disease However, the phenotype and / or function of and in patient samples. As part of this project, the astrocytic activation in the pathology are we are now developing new models to better not yet established. In this regard, and taking mimic the microglial dysfunction observed into account the phagocytic capacity of in Alzheimer’s patients. Specifically, we are active astrocytes, observed in both, models developing Abeta and Tau models, conditional and human samples (this objective of our knockouts for the Csf1R gene specifically in current work is included in an intramural project microglial cells. The main goal is to resemble CIBERNED (IP Joan Comella) and is also done the microglial pathology, decreasing either in collaboration with Dr. E. Galea (Marató de their proliferative capacity and/or function, as TV3), we have also started in vitro “pulse and observed in Braak V-VI samples. These models chase” experiments to determine the effect will allow us to establish the function of microglial of astrocytic stimulation on their degradative cells in the development of Abeta and / or tau capacity of neuronal dystrophies (unpublished pathology. data). On the other hand, they will also allow us to Finally, we have established collaborations to study the astroglial response in the absence of determine, on the one hand, the role of hypoxia microglia. In this sense, also during this year, we in the microglial response in Alzheimer’s disease have studied the astrocyte response both in and, on the other hand, we are studying the vitro and in vivo in Abeta models and in human role of possible Trem2 ligands in microglial Alzheimer’s samples. The results have shown activation in this pathology. KEYWORDS

Alzheimer, degeneration, inflammation, Abeta, tau, oligomers, soluble, transgenic models, neuropathology PUBLICATIONS

• Serrano-Pozo A., Sanchez-Garci M.A., Heras-Garvin A., March-Diaz R., Navarro V., Vizuete M. et al. Acute and Chronic Sustained Hypoxia Do Not Substantially Regulate Amyloid-β Peptide Generation In Vivo. PLoS ONE. 2017;12(1). • Baglietto-Vargas D, Sánchez-Mejias E, Navarro V, Jimenez S, Trujillo-Estrada L, Gómez-Arboledas A et al. Dual roles of Aβ in proliferative processes in an amyloidogenic model of Alzheimer’s disease.Scientific reports. 2017;7(1). • 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. [ePub2017].

2017 CIBERNED Annual Report / 100 Program 1 Group: Javier Vitorica Ferrández RESEARCH PROJECTS

• Code: FIS PI15/00796. Title: Evaluando la disfunción microglial y astroglial como base del proceso neurodegenerativo y la demencia en la enfermedad de Alzheimer: nuevas aproximaciones terapéuticas. Principal Investigator: Antonia Gutiérrez Pérez. CIBERNED’s collaboration: Yes. CIBERNED groups: G415, G411. Other CIBER’s collaboration: No. Type: Nacional. Funding agency: Instituto de Salud Carlos III. Funding: 99.220 €. Duration: 2016-2018

2017 CIBERNED Annual Report / 101 Francisco Wandosell Jurado 412

Principal Investigator Francisco Wandosell Jurado

Research team

Inés M. Antón Lara Ordóñez Gutiérrez Carlota Gil Martín CSIC Staff Scientist-Researcher Postdoctoral Fellow Bachelor student

María José Benítez Moreno Irene Benito Alazne Arrazola Sastre Associate professor PhD student Master Thesis student

María José Pérez Álvarez Sergio Rivas Catalina Lapuente Chala Assist. Prof. UAM. PhD student Investigador visitante

Juan José Garrido Jurado Mario Villa Ángel Céspedes Staff Scientist-Researcher PhD student Visiting Phd and Postdoc.

Centro de Biología Molecular “Severo Ochoa” CSIC-UAM Universidad Autónoma de Madrid Calle Nicolás Cabrera nº1, Cantoblancon 28049 Madrid (Spain) Tel.: +34 91 196 45 61/45 91 Fax: +34 91 196 44 20 [email protected]

2017 CIBERNED Annual Report / 102 SUMMARY

Our group, “Molecular mechanisms of using pharmacological and shRNA approaches. Neurodegeneration“, is generated from an We reported that this via is modified after established line in the CBM over several years. At Ischemia and that the neuroprotective role of present, this CIBERNED line is formally composed neurosteroids, such as estradiol, is due to the by three sub-groups (IP´s-F Wandosell (CBM), I. activation of PI3K-Akt pathway, at least in part Anton (CNB) and J. J. Garrido, Cajal Inst.). (CoIPs: Dr. J.J.Garrido & F Wandosell). Our group is mostly devoted to the analysis of Second, we are analysing the role of the neurodegenerative disorders. Now is focusing Akt downstream element, mTORC1. This in a group of brain disorders associated with protein complex controls general aspects of aging, such as Alzheimer, disease, Stroke and protein synthesis and autophagy; whereas some type of brain tumours. We are interested its dysfunction has been considering a key in the analysis of which cellular signals are factor of beta amyloid generation and/or altered throughout the aging process and degradation in Alzheimer disease. how the modification of some of them triggers a pathological process. Therefore a relevant At the present we are analysing, in detail, the question is ... “are there the common molecular macroautophagy process in neurons from mechanisms in these age-dependent control and Alzheimer’s mice model (IPs:F. pathologies? Wandosell). Third, we are analysing some Akt- downstream elements (such as FoxO, Bim, Data from different animal models strongly β-catenin, YAP/TAZ), and our data indicated support that idea that longevity is controlled by that they are responsible of the cell division of several genetic pathways, such as: IGF1/Insulin- cancer-stem cells (CSC) and the maintenance PI3K-Akt-FoxO. And some reports indicate of its phenotype. that insulin and IGF-1 regulate longevity in a conserved manner. IGF-1/Insulin through their Our work is now focusing on the conversion tyrosine kinase receptors controls the activity of from astrocyte-astrocytoma-glioma trying to Class1 phosphatidylinositol 3-kinase (PI3K) and define the role Akt-downtream elements and several serine-threonine kinases such as Akt. how WIP regulates actin cytoskeleton and glioma proliferation. We just describe a new We are interested in the analysis of the pro-oncogenic pathway in which Akt and WIP molecular mechanisms of some brain disorders, are controlling the proliferation of CSC derived mainly focusing in the role of PI3K-Akt and some from glioma, via YAP/TAZ (CoIPs: Dr. I. Anton of the Akt substrates, trying to understand some & F. Wandosell). In summary, we are focusing key points of these signalling. Our initial studies on track PI3K-Akt signalling and elements that were focused on axonal differentiation and control some physiological process, from cell polarization, we defined the role of PI3K class division to differentiation, and how some of I, GSK3 and Akt in the neuronal morphogenesis these proteins are modified in pathology. KEYWORDS Neurodegeneration, Cellular Signaling, Neuroprotection, Estrogens, Alzheimer’s Disease, Neuritogenesis, Axonal Polarity, Glia Growth, Astrocyte / Glioma Conversion, Cytoskeleton, Actin PUBLICATIONS

• Pfajfer L., Seidel M.G., Houmadi R., Rey-Barroso J., Hirschmugl T., Salzer E. et al. WIP deficiency severely affects human lymphocyte architecture during migration and synapse assembly. Blood. 2017;130(17):1949-1953. • Ordonez-Gutierrez L., Posado-Fernandez A., Ahmadvand D., Lettiero B., Wu L., Anton M. et al. ImmunoPEGliposome-mediated reduction of blood and brain amyloid levels in a mouse model of Alzheimer’s disease is restricted to aged animals. Biomaterials. 2017;112:141-152. • Tapia M., Dominguez A., Zhang W., Puerto A.D., Ciorraga M., Benitez M.J. et al. Cannabinoid receptors modulate neuronal morphology and ankyring density at the axon initial segment. Frontiers in Cellular Neuroscience. 2017;11.

2017 CIBERNED Annual Report / 103 • Pose-Utrilla J., Garcia-Guerra L., Del Puerto A., Martin A., Jurado-Arjona J., De Leon-Reyes N.S. et al. Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1. Nature Communications. 2017;8(1). • Carradori D, Balducci C, Re F, Brambilla D, Le Droumaguet B, Flores O et al. Antibody- functionalized polymer nanoparticle leading to memory recovery in Alzheimer’s disease-like transgenic mouse model.Nanomedicine : nanotechnology, biology, and medicine. 2017;14(2). • Escoll M., Gargini R., Cuadrado A., Anton I.M., Wandosell F. Mutant p53 oncogenic functions in cancer stem cells are regulated by WIP through YAP/TAZ. Oncogene. 2017;36(25):3515-3527. Benito-Cuesta I, Diez H, Ordoñez L, Wandosell F. Assessment of Autophagy in Neurons and Brain Tissue.Cells. 2017;6(3). RESEARCH PROJECTS

• Code: PI2016/01. • Code: Fundacion RAMÓN ARECES. Title: Alteraciones del metabolismo gluco- Title: Interactoma diferencial de WIP: lipídico y desarrollo de la demencia de Actividad oncogénica versus supresora de Alzheimer. tumores. Principal Investigator: Ignacio Torres Principal Investigator: Francisco Wandosell, Alemán. Inés Antón. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G409 ; G402; G511; G502; CIBERNED groups: G412. G412. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Private. Type: Intramurales. Funding agency: Fundación Ramón Areces. Funding agency: CIBERNED Funding: 40.000€. Funding: 200.000€. Duration: 2017-2019 Duration: 2016-2018 • Code: PCIN-2016-108. • Code: SAF2015-70368-R. Title: Regulación del reflejo de micción Title: Análisis de la señalización mediada después de lesionmedular: abolición por por akt en neurodegeneración y en silenciamiento de los aferentes de las fibras proliferación, migración/invasión celulares. C de la vejiga hiper-excitados mediante Principal Investigator: Francisco Wandosell, terapia génica para restaurar la continencia Ines M. Antón Gutiérrez. y la micción”. CIBERNED’s collaboration: No. Principal Investigator: Francisco Wandosell. CIBERNED groups: G412. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G412. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: European. Funding: 220.000€. Funding agency: Comisión Europea. Duration: 2016-2018 Funding: 150.000€. Duration: 2017-2019 • 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: European. Funding agency: Comisión Europea. Funding: ND. Duration: 2016-2020

2017 CIBERNED Annual Report / 104 Program 1 Group: Francisco Wandosell Jurado PhD DISSERTATIONS

• Author: Irene Benito Cuesta. Title: Análisis de las vías de inducción de Autofagia en neuronas y su papel neuroprotector en un modelo transgénico de la enfermedad de Alzheimer. Date: 26th July 2017. Supervisor: Francisco Wandosell Jurado.

2017 CIBERNED Annual Report / 105

PROGRAM 2 Parkinson’s disease and other neurodegenerative movement disorders

Alberch Vie, Jordi...... 112 Lanciego Pérez, José Luis...... 174 Canela Campos, Enric Isidre...... 117 López Barneo, José...... 177 Ceña Callejo, Valentín...... 121 López de Munain Arregui, Adolfo...... 182 Cuadrado Pastor, Antonio...... 124 Lucas Lozano, José Javier...... 189 del Río Fernández, José Antonio...... 128 Moratalla Villalba, Rosario...... 192 Fariñas Gómez, Isabel...... 131 Muñoz Cánoves, Pura...... 195 Fernández Chacón, Rafael...... 135 Naranjo Orovio, José Ramón...... 199 Fernández Ruiz, Javier...... 138 Navarro Acebes, Xavier...... 202 Fuentes Rodríguez, José Manuel...... 142 Obeso Inchausti, José Ángel...... 208 García Verdugo, José Manuel...... 146 Pérez Castillo, Ana María...... 214 González Castaño, José...... 150 Pérez Tur, Jordi...... 217 Guzmán Pastor, Manuel...... 152 Rodríguez Díaz, Manuel...... 219 Iglesias Vacas, Teresa...... 156 Tolosa Sarró, Eduardo...... 222 Infante Ceberio, Jon...... 159 Vicario Abejón, Carlos...... 229 Kulisevsky Bojarski, Jaime...... 165 Vila Bover, Miquel...... 232 Labandeira García, José Luis...... 170

PROGRAMA 2: PARKINSON’S DISEASE AND OTHER NEURODEGENERATIVE MOVEMENT DISORDERS

This Program brings together 31 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 neuromodulation systems. It is estimated to affect around 70,000 people in Spain, a figure that is expected to be increasing due to the progressive aging of the population.

Although preventing and correcting DA deficit are still important goals, they cannot be considered the ultimate challenge in PD nowadays. Within this area, it is considered of vital importance to make progress on defining the following issues:

• Key aspects related to the ethiopathogenesis in PD. • Physiopatological mechanisms related to disease onset and progression. • 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. • 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.

2017 CIBERNED Annual Report / 109 Research Line 2: Huntington’s disease and ataxias

This program also focuses on research into other movement disorders such as Huntington’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-terminus. 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. The estimated prevalence of HD is 10 cases for every 100,000 persons, which means 4,500 patients in Spain and about 50,000 in the European 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.

There is a large number of studies with neuroprotective drugs which modify the supposed pathogenic mechanisms or that are used in other neurodegenerative diseases. These drugs include inhibitors of neuronal excitation, 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.

Research Line 3: Neurodegenerative motor disorders

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 muscles. Diagnosis involves using sophisticated methods covering: neurophysiological studies, muscle or nerve biopsy with the use of immunohistochemical techniques and some others, metabolic analysis and tests, RMN studies, quantitative muscle assessment, and in many cases, genetic studies.

Just a few epidemiological studies are available in Spain, from which it can be inferred that the prevalence of chronic neurodegenerative and/or hereditary pathology would account for around 60,000 patients in our country.

Members of the neuromuscular pathology program are committed to investigate into specific aspects including:

• Clinical characterization of neuromuscular 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.

2017 CIBERNED Annual Report / 110 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 University of Madrid Del Río Fernández, José Antonio Catalonian Institute of Bioengineering, Barcelona Fariñas Gómez, Isabel University of Valencia Fernández Chacón, Rafael University of Seville Fernández Ruiz, Javier Complutense University of Madrid Fuentes Rodríguez, José Manuel University of Extremadura, Cáceres García Verdugo, José Manuel Cavanilles Institute, University of Valencia González Castaño, José Autonomous University of Madrid Guzmán Pastor, Manuel Complutense University of Madrid Iglesias Vacas, Teresa Biomedical Research Institute CSIC-UAM, Madrid Infante Ceberio, Jon Instituto de Investigación Marqués de Valdecilla, Santander Kulisevsky Bojarski, Jaime Santa Creu i Sant Pau Hospital, Barcelona Labandeira García, José Luis University of Santiago de Compostela Lanciego Pérez, José Luis Center for Applied Medical Research, Univ. of Navarra, Pamplona López Barneo, José Virgen del Rocío University Hospital, Universidad de Sevilla López de Munain Arregui, Adolfo Biodonostia Research Institute, San Sebastian Lucas Lozano, José Javier Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid Moratalla Villalba, Rosario Cajal Institute CSIC, Madrid Muñoz Cánoves, Pura Pompeu Fabra University , ICREA, Barcelona Naranjo Orovio, José Ramón National center of Biotechnology CSIC, Madrid Navarro Acebes, Xavier Autonomous University of Barcelona Obeso Inchausti, José Ángel Hospitales de Madrid Foundation

Pérez Castillo, Ana María Biomedical Research Institutes CSIC-UAM, Madrid Pérez Tur, Jordi Biomedicine Institute CSIC, Valencia Rodríguez Díaz, Manuel University of La Laguna, Tenerife Tolosa Sarró, Eduardo Clinic Hospital, Barcelona Vicario Abejón, Carlos Cajal Institute CSIC, Madrid Vila Bover, Miquel Vall d’Hebron University Hospital, Barcelona

Program 2 is coordinated by Drs. Isabel Fariñas (University of Valencia), Rafael Fernández Chacón (University of Seville), Teresa Iglesias Vacas (Biomedical Research Institute CSIC-UAM, Madrid), Madrid), Adolfo López de Munain Arregui (Biodonostia Research Institute, San Sebastián), José J. Lucas (Center for Molecular Biology, CSIC, and Eduardo Tolosa (Clinic Hospital, Barcelona).

1 Since December 2017

2017 CIBERNED Annual Report / 111 Jordi Alberch Vié 301

Principal Investigator Jordi Alberch Vié

Research team

Josep M Canals Coll Raquel Martín Ibáñez Jordi Creus Muncunill Cristina Herranz Associate Professor Postdoctoral PhD researcher Sotoca researcher Technician Silvia Ginés Padrós Sara Fernández Assistant Professor Mercè Massana García Ana López Alonso Postdoctoral PhD researcher Technician Esther Pérez Navarro researcher Associate Professor Marta García Forn Mª Teresa Muñoz Alarcón Andrés Míguez PhD researcher Technician Cristina Malagelada González Grau Postdoctoral researcher Alfonso Gerardo Mireia Galofre Assistant Professor García Díaz Barriga Centelles Ana Cristina Saavedra PhD researcher Technician Albert Giralt Torroella Postdoctoral researcher Postdoctoral Laura López Molina Verónica Monforte researcher Phil Sanders PhD researcher Pizarro Postdoctoral Technician Xavier Xifró researcher Núria Suelves Collsamata Caballol Gemma Orpella Assistant Professor Unai Perpiñá Martín PhD researcher Aceret Postdoctoral Technician Glòria Blázquez researcher Laura Vidal Sancho Postdoctoral PhD researcher Felipe Chiape Rizzo researcher Elena Álvarez Periel Cristina Salado Technician PhD researcher Manzano Verónica Brito PhD researcher David Vanneste Postdoctoral Andrea Comella Bolla Project manager researcher PhD researcher Georgina Bombau Martínez Paola Lucero Technician Calabiug Project manager Departamento de Biomedicina. Facultat de Medicina, Universitat de Barcelona Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) Tel.: +34 93 403 52 58 / 93 403 72 52 Fax: +34 93 402 19 07 [email protected]

2017 CIBERNED Annual Report / 112 SUMMARY

During 2017, our research topics have neuroprotective effects of the TrkB agonists been focussed on the identification of new 7,8-dihydroxyflavone on motor and cognitive therapeutical targets in Huntington’s disease impairments in Huntington’s disease. Our results and in the study of intracellular mechanisms provide new insights into the mechanisms of activated by mutant huntingtin inducing action of 7,8-dihydroxyflavone through the neuronal dysfunction and neurodegeneration. activation of PLC1 pathway (García-Díaz We have also continued the studies about Barriga et al., 2017). the function of mutant huntingtin during development and differentiation of iPSCs to In the study of the pathogenic mechanisms, we striatal neuronal phenotypes. described that phosphorylation of huntingtin at residue T3 is decreased in Huntington’s disease To study the involvement of Cdk5 deficit in and modulates mutant huntingtin protein memory and learning in Huntington’s disease, conformation and aggregate properties we have developed a new animal model of (Cariulo et al., 2017; Daldin et al., 2017). About Huntington’s disease that expresses mutant our research line in regenerative medicine in huntingtin and low levels of Cdk5. The reduction Huntington’s disease, we have participated in of Cdk5 levels in these mice prevented the the HD iPSC Consortium in the study of the effects onset of hippocampal and corticostriatal of mutant huntingtin during development. Our cognitive impairment through the modulation results showed that mutant huntingtin impairs of the expression and presynaptic localization neurodevelopmental pathways that could of the NR2B subunit (Alvarez-Periel et al., 2017). disrupt synaptic homeostasis and increase Another protein that is involved in the alterations vulnerability to the pathologic consequence of neuronal plasticity in Huntington’s disease of expanded polyglutamine repeats over time is Pyk2. KO mice for Pyk2 showed a similar (HD iPSC Consortium, 2017). phenotype to R6/1 mice, which express mutant huntingtin. In mouse models and samples In collaboration with other CIBERNED group from patients with Huntington’s disease, we (Carlos Vicario-Abejón), we described that observed a decrease in Pyk2 levels. Normalizing the transcription factor Ikaros is involved in Pyk2 levels in the hippocampus of R6/1 mice, the development of a specific set of striatal after injection of AAV expressing Pyk2, rescued projection neurons, which are mainly affected memory deficits, spines pathology and PSD-95 in Huntington’s disease (Martin-Ibáñez et al., localization (Giralt et al., 2017). 2017). Furthermore, we have described a new We have also characterized human fetal mechanism for Chelerythrine. This drug was stem cells in different stages of development originally described as PKC inhibitor. We can analyzing the potentiality to differentiate observe that Chelerythrine promotes Ca2+- to several specific neuronal phenotypes in dependent calpain activation in neuronal different brain areas (Martin-Ibáñez et al., 2017). cells in a PKC-independent manner (Saavedra In another collaboration with a CIBENED group et al., 2017). Another mechanism that we (José López-Barneo) we studied the genetic characterized is the involvement of histone rescue of mitochondrial and skeletal muscle deacetylase 3 (HDAC3) in the cognitive deficit, impairment in an iPSC model of coenzyme Q10 suggesting that the inhibition of HDAC3 can be deficiency. The COQ4 mutation in iPSC was a good therapeutic approach in Huntington’s associated with CoQ10 deficiency, metabolic disease (Suelves et al., 2017). Searching dysfunction, and respiration defects (Romero- for new treatments, we characterized the Moya et al., 2017). KEYWORDS

BDNF, neuronal plasticity, biomarkers, kinases, phosphatases, neurogenesis, regenerative medicine, iPSCs

2017 CIBERNED Annnual Report / 113 PUBLICATIONS

• Martin-Ibanez R., Pardo M., Giralt A., Miguez A., Guardia I., Marion-Poll L. et al. Helios expression coordinates the development of a subset of striatopallidal medium spiny neurons. Development (Cambridge). 2017;144(8):1566-1577. • Illa M., Brito V., Pla L., Eixarch E., Arbat-Plana A., Batalle D. et al. Early Environmental Enrichment Enhances Abnormal Brain Connectivity in a Rabbit Model of Intrauterine Growth Restriction. Fetal Diagnosis and Therapy. 2017. • Lim R.G., Salazar L.L., Wilton D.K., King A.R., Stocksdale J.T., Sharifabad D. et al. Developmental alterations in Huntington’s disease neural cells and pharmacological rescue in cells and mice. Nature Neuroscience. 2017;20(5):648-660. • Giralt A., Brito V., Chevy Q., Simonnet C., Otsu Y., Cifuentes-DIaz C. et al. Pyk2 modulates hippocampal excitatory synapses and contributes to cognitive deficits in a Huntington’s disease model. Nature Communications. 2017;8. • 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. [ePub2017]. • Cariulo C., Azzollini L., Verani M., Martufi P., Boggio R., Chiki A. et al. Phosphorylation of huntingtin at residue T3 is decreased in Huntington’s disease and modulates mutant huntingtin protein conformation. Proceedings of the National Academy of Sciences of the United States of America. 2017;114(50):E10809-E10818. • Alvarez-Periel E., Puigdellivol M., Brito V., Plattner F., Bibb J.A., Alberch J. et al. Cdk5 Contributes to Huntington’s Disease Learning and Memory Deficits via Modulation of Brain Region-Specific Substrates. Molecular Neurobiology. 2017;:1-19. • Giralt A, Gómez-Climent MÁ, Alcalá R, Bretin S, Bertrand D, María Delgado-García J et al. The AMPA receptor positive allosteric modulator S 47445 rescues in vivo CA3-CA1 long-term potentiation and structural synaptic changes in old mice.Neuropharmacology. 2017;123. • Romero-Moya D, Santos-Ocaña C, Castaño J, Garrabou G, Rodríguez-Gómez JA, Ruiz-Bonilla V et al. Genetic Rescue of Mitochondrial and Skeletal Muscle Impairment in an Induced Pluripotent Stem Cells Model of Coenzyme Q10 Deficiency.Stem cells (Dayton, Ohio). 2017;35(7). • Barriga G.G., Giralt A., Anglada-Huguet M., Gaja-Capdevila N., Orlandi J.G., Soriano J. et al. 7,8-dihydroxyflavone ameliorates cognitive and motor deficits in a Huntington’s disease mouse model through specific activation of the PLCγ1 pathway. Human Molecular Genetics. 2017;26(16):3144-3160. • Saavedra A., Fernandez-Garcia S., Cases S., Puigdellivol M., Alcala-Vida R., Martin-Flores N. et al. Chelerythrine promotes Ca2+-dependent calpain activation in neuronal cells in a PKC- independent manner. Biochimica et Biophysica Acta - General Subjects. 2017;1861(4):922-935. • Suelves N., Kirkham-McCarthy L., Lahue R.S., Gines S. A selective inhibitor of histone deacetylase 3 prevents cognitive deficits and suppresses striatal CAG repeat expansions in Huntington’s disease mice. Scientific Reports. 2017;7(1). • Daldin M., Fodale V., Cariulo C., Azzollini L., Verani M., Martufi P. et al. Polyglutamine expansion affects huntingtin conformation in multiple Huntington’s disease models. Scientific Reports. 2017;7(1). • Martin-Ibanez R., Guardia I., Pardo M., Herranz C., Zietlow R., Vinh N.-N. et al. Insights in spatio-temporal characterization of human fetal neural stem cells. Experimental Neurology. 2017;291:20-35. • Moreno E, Chiarlone A, Medrano M, Puigdellívol M, Bibic L, Howell LA et al. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum.Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology. 2017. • Cases S., Saavedra A., Tyebji S., Giralt A., Alberch J., Perez-Navarro E. Age-related changes in STriatal-Enriched protein tyrosine Phosphatase levels: Regulation by BDNF. Molecular and Cellular Neuroscience. 2018;86:41-49. [ePub2017].

2017 CIBERNED Annual Report / 114 Program 2 Group: Jordi Alberch Vié RESEARCH PROJECTS

• Code: RD16/0011. CIBERNED groups: G301. Title: RETIC de terapia celular. Other CIBER’s collaboration: No. Principal Investigator: Isabel Fariñas. Type: Intramurales. CIBERNED’s collaboration: Yes. Funding agency: Cytes Biotechnologies. CIBERNED groups: G102, G301, G113, G208, Funding: 450.000€. G105. Duration: 2015-2020 Other CIBER’s collaboration: No. Type: National. • Code: 2014SGR968. Funding agency: Instituto de Salud Carlos III. Title: Fisiopatologia i tractament de les Funding: 284.999 €. malalties neurodegeneratives. Duration: 2016-2020 Principal Investigator: Jordi Alberch. CIBERNED’s collaboration: No. • Code: A12076. CIBERNED groups: G301. Title: Studying Human MSN Differentiation Other CIBER’s collaboration: No. from PSC using Single-Cell RNAseq and Type: CC.AA. Rodent Chimeric Models. Funding agency: Generalitat de Catalunya. Principal Investigator: Josep M. Canals. Funding: 30.000€. CIBERNED’s collaboration: No. Duration: 2014-2017 CIBERNED groups: G301. Other CIBER’s collaboration: No. • Code: TECDTP15-1-0022. Type: International. Title: Centre per a la produccio i validacio Funding agency: CHDI. de teràpies avançades - ub (cat-ub). Funding: 553.417€. Principal Investigator: Josep M. Canals. Duration: 2016-2018 CIBERNED’s collaboration: No. CIBERNED groups: G301. • Code: H2020-MSCA-ITN-2016. Other CIBER’s collaboration: No. Title: European Training Network for Cell- Type: CC.AA. based Regenerative Medicine. Funding agency: Generalitat de Catalunya. Principal Investigator: Jordi Alberch. Funding: 43.767€. CIBERNED’s collaboration: No. Duration: 2016-2017 CIBERNED groups: G301. Other CIBER’s collaboration: No. • Code: COMRDI15-1-0013. Type: European. Title: ADVANCECAT: acceleradora pel Funding agency: Comisión Europea. desenvolupament de teràpies avançades Funding: 247.873 €. a Catalunya (ADVANCECAT: accelerating Duration: 2016-2020 development for advanced therapies in Catalonia). • Code: 713140. Principal Investigator: Josep M. Canals. Title: Custom architecturally defined 3D CIBERNED’s collaboration: No. stem cell derived functional human neural CIBERNED groups: G301. networks for transformative progress in Other CIBER’s collaboration: Yes (CIBERBBN). neuroscience and medicine (MESO_BRAIN). Type: CC.AA. Principal Investigator: Jordi Soriano. Funding agency: Generalitat de Catalunya. CIBERNED’s collaboration: No. Funding: 969.352 €. CIBERNED groups: G301. Duration: 2016-2019 Other CIBER’s collaboration: No. Type: European. • Code: RL000776. Funding agency: Comisión Europea. Title: Mitochondrial outcomes measures in Funding: 3.225.890 €. fibroblasts of HD patients. Duration: 2016-2019 Principal Investigator: Silvia Gines. CIBERNED’s collaboration: No. • Code: FBG 308408. CIBERNED groups: G301. Title: Obtaining, processing an/or preserving Other CIBER’s collaboration: No. non-human tissues, cells and related Type: International. products. Funding agency: HDSA. Principal Investigator: Josep M. Canals. Funding: 75.000 €. CIBERNED’s collaboration: No. Duration: 2017-2018

2017 CIBERNED Annual Report / 115 • Code: FBG 308841. • Code: SAF2015-66505-R. Title: Servicio de Salas Blancas para Title: Desarrollo, diferenciación y maduración produccion de vacuna de HIV en neuronal en la enfermedad de Huntington. condiciones GMP. Principal Investigator: Josep M. Canals. Principal Investigator: Josep M. Canals, CIBERNED’s collaboration: No. Raquel Martín Ibañez. CIBERNED groups: G301. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G301. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Funding: 181.500 €. Funding agency: IDIBAPS. Duration: 2016-2018 Funding: 360.000€. Duration: 2016-2018 • Code: SAF2016-08573-R. Title: Alteraciones de la lámina nuclear y • Code: 20140130/1. de la traducción proteica como nuevos Title: Modulacion del deficit dela plasticidad mecanismos patogénicos en la enfermedad sinaptica como estrategia terapeutica en la de Huntington. enfermedad de Hunttington. Principal Investigator: Esther Pérez Navarro. Principal Investigator: Jordi Alberch. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G301. CIBERNED groups: G301. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: Private. Funding agency: MINECO. Funding agency: Fundació La Marató de Funding: 181.500 €. TV3. Duration: 2016-2019 Funding: 299.375€. Duration: 2015-2018 • Code: SAF2015-67474-R. Title: Cdk5 como nueva diana terapéutica 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: National. Funding agency: MINECO. Funding: 217.800€. Duration: 2016-2018 PhD DISSERTATIONS

• Author: Rafael Alcalá Vida. Title: PKC delta and nuclear lamina alterations: Invovement in Huntington’s disease pathophysiology. Date: 16th June 2017. Supervisor: Esther Pérez Navarro.

2017 CIBERNED Annual Report / 116 Enric I. Canela Campos 201

Principal Investigator Enric I. Canela Campos

Research team

Vicent Casadó Burillo Estefanía Moreno Guillén Iñigo Etayo Labiano Research staff Postdoctoral fellow PhD student

Antonio Cortés Tejedor Gemma Navarro Brugal Patricia C. Homar Ruano Research staff Postdoctoral fellow PhD student

Rafael Franco Fernández David Aguinaga Andrés Mireia Medrano Moya Research staff PhD student PhD student

Carmen Lluís Biset Edgar Angelats Canals Irene Reyes Resina Research staff PhD student PhD student

Josefa Mallol Montero Verónica Casadó Anguera M. Mar Rodríguez Ruiz Research staff PhD student PhD student

Marta Sánchez Soto PhD student

Departamento de Bioquímica y Jasmina Jiménez Cano Biología Molecular Technician Facultad de Biología, Universidad de Barcelona Diagonal 643, planta -2 08028 Barcelona, Catalunya (Spain) Tel.: +34 93 402 15 59 Fax: +34 99 402 15 59 [email protected]

2017 CIBERNED Annual Report / 117 SUMMARY

Our area of interest has been the GPCRs in the We found that both acute and chronic CNS, specially their ability to form homo- and cocaine self-administration blunted mGluR1/5 hetero-oligomers. We focused on discovering effects on cAMP responsive-element binding receptor heteromers as new targets for protein phosphorylation in the striatum, which treatment of CNS disorders. Most relevant correlated with the capacity to induce long- publications in this period were: term depression, an effect that was maintained 60 days after chronic cocaine self-administration We found evidence for selective withdrawal. In the nucleus accumbens, the heteromerization of mu-opioid receptor (MOR) principal brain region mediating the rewarding and the galanin receptor Gal1 (Gal1R). A effects of drugs, chronic cocaine self- synthetic peptide selectively disrupted MOR– administration blunted mGluR1/5 stimulation of Gal1R heteromerization as well as specific extracellular signal-regulated protein kinases 1/2 interactions between MOR and Gal1R ligands: and cAMP responsive-element binding protein. a negative cross talk, by which galanin counteracted MAPK activation induced by We tested cannabidiol (CBD) as a potential the endogenous MOR agonist endomorphin-1, allosteric ligand of CB2R. We confirmed that CBD and a cross-antagonism, by which a MOR does not bind with high affinity to the orthosteric antagonist counteracted MAPK activation site of the human CB2R where the synthetic induced by galanin. These specific interactions cannabinoid, [3H]-WIN 55,212-2, binds. CBD was, could then be identified in situ in slices of however, able to produce minor but consistent rat ventral tegmental area (VTA) with MAPK reduction in the homogeneous binding assays activation and AKT and CREB phosphorylation. in living cells using the fluorophore-conjugated Furthermore, in vivo microdialysis experiments CB2R-selective compound, CM-157. The effect demonstrated a key role of MOR-Gal1R on binding to CB2R-expressing living cells was heteromers localized in the VTA in the direct different to that exerted by the orthosteric control of dopamine cell function and their antagonist, SR144528, which decreased the ability to mediate antagonistic interactions maximum binding without changing the between MOR and Gal1R ligands. The results KD. CBD at nanomolar concentrations was indicate that MOR-Gal1R heteromers should be also able to significantly reduce the effect of targets for the treatment of opioid use disorders. the selective JWH133, on forskolin-induced intracellular cAMP levels and on activation of We also provide a high-resolution expression the MAP kinase pathway. These results may map and a detailed functional characterization help to understand CBD mode of action and of adenosine A2AR-cannabinoid CB1R may serve to revisit its therapeutic possibilities. heteromers in the dorsal striatum. Specifically, our data unveil that the A2AR-CB1R heteromer Biophysical studies in a heterologous system (i) is essentially absent from corticostriatal showed physical interactions and potential projections and striatonigral neurons, and, formation of heterotrimeric complexes of leptin, instead, is largely present in striatopallidal orexin and ghrelin receptors. Functional assays neurons, (ii) displays a striking G protein- showed robust allosteric interactions particularly coupled signaling profile, where co-stimulation different when the three receptors are of both receptors leads to strongly reduced expressed together. Further biochemical and downstream signaling, and (iii) undergoes an pharmacological assays provided evidence unprecedented dysfunction in Huntington’s of heterotrimer functional expression in primary disease, an archetypal disease that affects cultures of hypothalamic neurons. These findings striatal neurons. Altogether, our findings constitute evidence of close relationships in the may open a new conceptual framework to action of the three hormones already starting understand the role of coordinated adenosine- at the receptor level in hypothalamic cells. endocannabinoid signaling in the indirect striatal pathway, which may be relevant We obtained results on the glial-neuronal in motor function and neurodegenerative networks involved in the neuroprotective role diseases. of trans-resveratrol. KEYWORDS

Adenosine receptors, basal ganglia, cannabinoid receptors, dopamine receptors, galanin receptors, G-protein coupled receptors oligomerization, HD, opioid receptor

2017 CIBERNED Annual Report / 118 Program 2 Group: Enric I. Canela Campos PUBLICATIONS

• 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. [ePub2017]. • Martínez-Pinilla E, Varani K, Reyes-Resina I, Angelats E, Vincenzi F, Ferreiro-Vera C et al. Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors.Frontiers in pharmacology. ;8. • Franco R., Martinez-Pinilla E., Navarro G., Zamarbide M. Potential of GPCRs to modulate MAPK and mTOR pathways in Alzheimer’s disease. Progress in Neurobiology. 2017;149-150:21-38. • Grochowska KM, Yuanxiang P, Bär J, Raman R, Brugal G, Sahu G et al. Posttranslational modification impact on the mechanism by which amyloid-β induces synaptic dysfunction. EMBO reports. 2017;18(6). • Cuadrado-Tejedor M., Garcia-Barroso C., Sanchez-Arias J.A., Rabal O., Perez-Gonzalez M., Mederos S. et al. A First-in-Class Small-Molecule that Acts as a Dual Inhibitor of HDAC and PDE5 and that Rescues Hippocampal Synaptic Impairment in Alzheimer’s Disease Mice. Neuropsychopharmacology. 2017;42(2):524-539. • Medrano M, Aguinaga D, Reyes-Resina I, Canela EI, Mallol J, Navarro G et al. Orexin A/ Hypocretin Modulates Leptin Receptor-Mediated Signaling by Allosteric Modulations Mediated by the Ghrelin GHS-R1A Receptor in Hypothalamic Neurons.Molecular neurobiology. 2017. • Elgueta D., Aymerich M.S., Contreras F., Montoya A., Celorrio M., Rojo-Bustamante E. et al. Pharmacologic antagonism of dopamine receptor D3 attenuates neurodegeneration and motor impairment in a mouse model of Parkinson’s disease. Neuropharmacology. 2017;113:110-123. • Franco R., Martinez-Pinilla E. Chemical rules on the assessment of antioxidant potential in food and food additives aimed at reducing oxidative stress and neurodegeneration. Food Chemistry. 2017;235:318-323. • Onatibia-Astibia A., Franco R., Martinez-Pinilla E. Health benefits of methylxanthines in neurodegenerative diseases. Molecular Nutrition and Food Research. 2017;61(6). • Moreno E., Quiroz C., Rea W., Cai N.-S., Mallol J., Cortes A. et al. Functional µ-Opioid-galanin receptor heteromers in the ventral tegmental area. Journal of Neuroscience. 2017;37(5):1176-1186. • Celorrio M, Rojo-Bustamante E, Fernández-Suárez D, Sáez E, Estella-Hermoso de Mendoza A, Müller CE et al. GPR55: A therapeutic target for Parkinson’s disease?. Neuropharmacology. 2017;125. • Hoffmann H.M., Crouzin N., Moreno E., Raivio N., Fuentes S., McCormick P.J. et al. Long-lasting impairment of mGluR5-activated intracellular pathways in the striatum after withdrawal of cocaine self-administration. International Journal of Neuropsychopharmacology. 2017;20(1):72-82. • Navarro G., Franco N., Martinez-Pinilla E., Franco R. The epigenetic cytocrin pathway to the nucleus. Epigenetic factors, epigenetic mediators, and epigenetic traits. A biochemist perspective. Frontiers in Genetics. 2017;8(NOV). • Alkozi H.A., Franco R., Pintor J.J. Epigenetics in the eye: An overview of the most relevant ocular diseases. Frontiers in Genetics. 2017;8(OCT). • Moreno E, Chiarlone A, Medrano M, Puigdellívol M, Bibic L, Howell LA et al. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum.Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology. 2017. • Navarro G., Martinez-Pinilla E., Sanchez-Melgar A., Ortiz R., Noe V., Martin M. et al. A genomics approach identifies selective effects of trans-resveratrol in cerebral cortex neuron and glia gene expression. PLoS ONE. 2017;12(4).

2017 CIBERNED Annual Report / 119 RESEARCH PROJECTS

• Code: PI2016/02. CIBERNED groups: G201, G203. Title: Monitoring the onset and evolution of Other CIBER’s collaboration: No. neuronal dysfunctions in propagative neural Type: Private. disorders using microfluidic devices and Funding agency: Fundació La Marató de translational approaches. TV3. Principal Investigator: José Antonio del Río. Funding: 150.000 €. CIBERNED’s collaboration: Yes. Duration: 2015-2017 CIBERNED groups: G114, G401, G201. Other CIBER’s collaboration: No. • Code: SAF2014-54840-R. Type: Intramurales. Title: Oligomerización de receptores Funding agency: CIBERNED. adrenérgicos y dopaminérgicos en el Funding: 210.000 €. SNC: un nuevo enfoque en el tratamiento Duration: 2016-2018 del transtorno por déficit de atención e hiperactividad. • Code: 20140610. Principal Investigator: Enric I. Canela Campos. Title: Targeting neuronal dopamine D1- CIBERNED’s collaboration: No. histamine H3 receptor heteromers as a new CIBERNED groups: G201. treatment of Huntington’s disease. Other CIBER’s collaboration: No. Principal Investigator: Enric I. Canela Campos. Type: National. CIBERNED’s collaboration: No. Funding agency: MINECO. CIBERNED groups: G201. Funding: 242.000 €. Other CIBER’s collaboration: No. Duration: 2015-2017 Type: Private. Funding agency: Fundació La Marató de • Code: BFU2015-64405-R. TV3. Title: Sigma-1 receptor-containing Funding: 200.000 €. heteromeric complexes in cocaine addiction. Duration: 2015-2017 Principal Investigator: Rafael Franco Fernández. • Code: 20141330. CIBERNED’s collaboration: No. Title: Cannabinoid receptor heteromeric CIBERNED groups: G201. complexes as therapeutic targets in Other CIBER’s collaboration: No. Parkinson’s disease. Type: National. Principal Investigator: Rafael Franco Funding agency: MINECO. Fernández. Funding: 284.350 €. CIBERNED’s collaboration: Yes. Duration: 2016-2018 PhD DISSERTATIONS

• Author: David Aguinaga Andrés. • Author: Mireia Medrano Moya. Title: Interacción molecular y funcional entre Title: Implicación estructural y funcional de receptores involucrados en la ingesta de heterómeros de receptores acoplados a alimentos y el consumo de drogas de abuso. proteínas G de relevancia en adicción, Date: 17th July 2017. enfermedad de Huntington y cáncer. Supervisor: Enric Isidre Canela Campos. Date: 23th June 2017. Supervisor: Josefa Mallol Montero. • Author: Marta Sánchez Soto. Title: Noncanonical neurotransmitter activation of catecholamine receptors. Date: 16th January 2017. Supervisor: Vicente Casadó Burillo.

2017 CIBERNED Annual Report / 120 Valentín Ceña Callejo 106

Principal Investigator Valentín Ceña Callejo

Research team

Inmaculada Posadas Darío Manzanares Ana Belén García Sáez Associate professor Pharmacist Lab technician

Laura Gallego Yerga Pablo Játiva Vanesa Guijarro Research staff Pharmacist Lab technician

Laura Romero Cristina de la Torre Elena Galera Pharmacist Pharmacist Lab technician

Unidad Asociada Neurodeath Universidad de Castilla-La Mancha, Facultad de Medicina Avenida Almansa, 14 02006 Albacete (Spain) Tel.: +34 680 22 23 22 [email protected]

2017 CIBERNED Annual Report / 121 SUMMARY

The research of the Neurodeath group has 3) To study the mechanisms involved in been focused on 3 different areas: nanoparticle blood-brain-barrier crossing and to investigate the different approaches 1) The use of nanoparticles, mainly dendrimers used to facilitate such a crossing. (but also polymers derived from cyclodextrin) to transfect genetic material, mainly siRNA, The use of siRNA as therapeutic agent will into different cell types to knock down specific produce, in a short period of time, new proteins involved in tumoral progression and generations of drugs, based on siRNA, with neurodegenerative diseases. the potential to revolutionize therapeutics in different areas. Our group has focused during 2) To show that certain dendrimers, having this year in the development of dendrimers themselves anti-inflammatory activity, are to transfect neuronal cultures with the aim able to prevent, in mice, the development of targeting the nanoparticles and their of experimental allergic encephalitis, a therapeutic cargo to the brain in a near future. generally accepted animal model for multiple sclerosis. KEYWORDS

Gene therapy, dendrimers, nanoparticles, neurodegeneration, siRNA, transfection, glioblastoma, multiple sclerosis PUBLICATIONS

• Posadas I., Romero-Castillo L., El Brahmi N., Manzanares D., Mignani S., Majoral J.-P. et al. Neutral high-generation phosphorus dendrimers inhibit macrophage-mediated inflammatory response in vitro and in vivo. Proceedings of the National Academy of Sciences of the United States of America. 2017;114(37):E7660-E7669. • Manzanares D., Araya-Duran I., Gallego-Yerga L., Jativa P., Marquez-Miranda V., Canan J. et al. Molecular determinants for cyclo-oligosaccharide-based nanoparticle-mediated effective siRNA transfection. Nanomedicine. 2017;12(13):1607-1621. • Jativa P., Cena V. Use of nanoparticles for glioblastoma treatment: A new approach. Nanomedicine. 2017;12(20):2533-2554. • Gallego-Yerga L, Posadas I, de la Torre C, Ruiz-Almansa J, Sansone F, Ortiz Mellet C et al. Docetaxel-Loaded Nanoparticles Assembled from β-Cyclodextrin/Calixarene Giant Surfactants: Physicochemical Properties and Cytotoxic Effect in Prostate Cancer and Glioblastoma Cells. Frontiers in pharmacology. 8. • Broomfield LM, Alonso-Moreno C, Martin E, Shafir A, Posadas I, Ceña V et al. Aminophosphine ligands as a privileged platform for development of antitumoral ruthenium(ii) arene complexes. Dalton transactions (Cambridge, England : 2003). 2017;46(46). • Posadas I, Alonso-Moreno C, Bravo I, Carrillo-Hermosilla F, Garzón A, Villaseca N et al. Synthesis, characterization, DNA interactions and antiproliferative activity on glioblastoma of iminopyridine platinum(II) chelate complexes.Journal of inorganic biochemistry. 2017;168. • Romero-Castillo L., Posadas I., Cena V. Exploring the in vivo toxicity of nanoparticles. Canadian Journal of Chemistry. 2017;95(9):917-926.

2017 CIBERNED Annual Report / 122 Program 2 Group: Valentín Ceña Callejo RESEARCH PROJECTS

• Code: VIHVACD. • Code: PI2016/05. Title: Desarrollo de una vacuna frente a VIH: Title: Dream inhibitors and Alzheimer´s Disease. estudio de los cambios en la biología de Principal Investigator: José Ramón Naranjo células dendríticas humanas tras interacción Orovio. con distintos sistema de liberación de CIBERNED’s collaboration: Yes. péptidos de VIH. CIBERNED groups: G307, G106, G403. Principal Investigator: Mª Ángeles Muñoz Fdez. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Funding: Intramurales. CIBERNED groups: G106. Funding agency: CIBERNED. Other CIBER’s collaboration: Yes CIBER-BBN. Funding: 210.000 €. Type: National. Duration: 2016-2018 Funding agency: CYTED. Funding: 5.000 €. Duration: 2014-2017 • Code: BFU2014-59009-P. Title: Efecto de dendrímeros antiinflamatorios sobre la patogénesis de la esclerosis múltiple. Principal Investigator: Valentín Ceña. CIBERNED’s collaboration: No. CIBERNED groups: G106. Other CIBER’s collaboration: No. Funding: National. Funding agency: MINECO. Funding: 156.453 €. Duration: 2015-2017

2017 CIBERNED Annual Report / 123 Antonio Cuadrado Pastor 101

Principal Investigator Antonio Cuadrado Pastor

Research team

Isabel Lastres Becker Marta Pajares Cabetas Diego Lastra Martínez PhD lecturer PhD student Grade student

Ana Isabel Rojo Sanchís Natalia Robledinos Antón Sara Castro Sánchez PhD assistant lecturer PhD student Grade student

Ángel Juan García Yagüe Raquel Fernández Ginés Marcos Galán Ganga Postdoctoral fellow PhD student Grade student

Maribel Escoll Guerrero Postdoctoral fellow

Departamento de Bioquímica e Instituto de Investigaciones Biomédicas Facultad de Medicina, Universidad Autónoma de Madrid Arzobispo Morcillo 4, 28029 Madrid (Spain) Tel.: +34 91 585 43 83 Fax: +34 915 85 44 01 [email protected]

2017 CIBERNED Annual Report / 124 SUMMARY

Parkinson’s disease (PD) and Alzheimer’s signaling oriented towards neuroprotection disease (AD) are incurable and their disabling and neurogenesis. In fact, we have described effects can continue for years or even decades. that NRF2 participates in the differentiation of Studies in animal models and in post-mortem neural stem cells from the subgranular zone brain tissues of patients indicate that many of the hippocampus, promoting astrocytosis pathological changes in the brain derive from versus neuronal differentiation. Low-grade various local stress types, such as oxidative stress, chronic inflammation is a key element of closely related to inflammatory and proteotoxic neurodegenerative diseases. We are studying stress. Pathologically modified proteins cause the interaction between NF-kB and NRF2, key local alterations that are amplified by low- elements in the pro and anti-inflammatory grade chronic oxidative and inflammatory microglia phenotypes. We have observed that stress that characterizes these diseases. the enzyme heme oxygenase-1, regulated by NRF2, and PCG1-alpha regulate mitochondrial We are currently studying the role of the biogenesis through the activation of the nicotinic transcription factor NRF2 in the protection receptor Alpha-7. We are also analyzing the against stimuli that induce neurodegeneration. role of the fractalkine quemiokine (CX3CL1) NRF2 is a protein that regulates the expression and the activation of its receptor CX3CR1 in of about 250 genes that participate in adaptive the inflammatory processes associated with responses to oxidative, inflammatory and neurodegeneration. Our observations suggest proteotoxic stress and in the regulation of that the transcriptional signature in peripheral enzymes involved in biotransformation and blood monocytes of NRF2 and NF-kB, closely metabolic reprogramming. Using genetically related to oxidative and inflammatory stress, modified rodents, as well as pharmacological could have value as a prognostic biomarker in approaches, during 2017 we studied the AD and PD. contribution of this transcription factor to the Antonio Cuadrado Pastor protection against oxidative damage and Applicability: neuroinflammation in toxic and genetic models (alpha-synuclein) of PD and in transgenic mice We have developed a new mouse model of AD that possess amyloidopathy (APPV717I) and that produces amyloidopathy and tauopathy tauopathy (TauP301L), which are characteristic in the presence or absence of NRF2. This model of AD. During the year 2017 we have progressed represents an important advance in order to in two directions: understand the homeostatic alterations of this disease and develop more effective drugs. In Generation of knowledge: relation to the regulatory pathway mediated by GSK3 and KEAP1 we have developed a The understanding of the mechanisms new compound with multitarget effects that that regulate NRF2 is fundamental to we are studying in preclinical EA models. On determine its physiological function and its the other hand, in collaboration with EVGEN pathological alterations, as well as to design pharma, we have looked for new regulatory new pharmacological strategies. Through a mechanisms of NRF2 in the brain that can serve transcriptomic analysis we have determined to reinforce their activity against neurotoxic that mice lacking NRF2 present alterations in stimuli in the preclinical models of PD. We gene expression that are very similar to those focused on the ruse of sulforadex (SFX-01). We observed in AD patients or caused by aging. have also delved into the mechanisms involved We have already described the regulation of in the neuroprotection produced by treatment NRF2 by the GSK-3/beta-TrCP pathway. Now with dimethyl fumarate and its use in PD and we are analyzing their participation in cell tauopathies. KEYWORDS

Oxidative stress, Neuroinflammation, Proteinopathy, Neuroprotection, NRF2, WNT, GSK-3, TAU

2017 CIBERNED Annnual Report / 125 PUBLICATIONS

• Escoll M., Gargini R., Cuadrado A., Anton I.M., Wandosell F. Mutant p53 oncogenic functions in cancer stem cells are regulated by WIP through YAP/TAZ. Oncogene. 2017;36(25):3515-3527. • 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 Alzheimer’s Association. 2017. • Ghezzi P, Floridi L, Boraschi D, Cuadrado A, Manda G, Levic S et al. Oxidative Stress and Inflammation Induced by Environmental and Psychological Stressors: A Biomarker Perspective. Antioxidants & redox signaling. 2017. • Egea J, Fabregat I, Frapart YM, Ghezzi P, Görlach A, Kietzmann T et al. Corrigendum to European 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. [ePub2017]. • Nayernia Z., Colaianna M., Robledinos-Anton N., Gutzwiller E., Sloan-Bena F., Stathaki E. et al. Decreased neural precursor cell pool in NADPH oxidase 2-deficiency: From mouse brain to neural differentiation of patient derived iPSC. Redox Biology. 2017;13:82-93. • Egea J, Fabregat I, Frapart YM, Ghezzi P, Görlach A, Kietzmann T et al. European contribution to the study of ROS: A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS).Redox biology. 2017;13. • Pajares M., Cuadrado A., Rojo A.I. Modulation of proteostasis by transcription factor NRF2 and impact in neurodegenerative diseases. Redox Biology. 2017;11:543-553. • Rojo A.I., Pajares M., Rada P., Nunez A., Nevado-Holgado A.J., Killik R. et al. NRF2 deficiency replicates transcriptomic changes in Alzheimer’s patients and worsens APP and TAU pathology. Redox Biology. 2017;13:444-451. • 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. [ePub2017]. • Robledinos-Anton N., Rojo A.I., Ferreiro E., Nunez A., Krause K.-H., Jaquet V. et al. Transcription factor NRF2 controls the fate of neural stem cells in the subgranular zone of the hippocampus. Redox Biology. 2017;13:393-401. • Gameiro I, Michalska P, Tenti G, Cores Á, Buendia I, Rojo AI et al. Discovery of the first dual GSK3β inhibitor/Nrf2 inducer. A new multitarget therapeutic strategy for Alzheimer’s disease. Scientific reports. 2017;7. • Cuadrado A. NRF2 in neurodegenerative diseases. Current Opinion in Toxicology. 2017;2:46-53. RESEARCH PROJECTS

• Code: SAF2015-71304-REDT. • Code: PCIN-2016-071. Title: Red de investigacion en NRF2 como Title: Advanced theranostic approach in nodo del patogenosoma. cancer combining photodynamic therapy Principal Investigator: Antonio Cuadrado. and nanoparticles. CIBERNED’s collaboration: No. Principal Investigator: Luis Felipe Ferreira. CIBERNED groups: G101. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G101. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Nacional. Funding : 35.000 €. Funding agency: MINECO. Duration: 2016-2017 Funding: 49.000 €. Duration: 2016-2019

2017 CIBERNED Annual Report / 126 Program 2 Group: Antonio Cuadrado Pastor

• Code: SAF2016-76520-R. • Code:PI2015/02. Title: Papel de NRF2 en la función y el destino Title: Validación de nuevas dianas del cerebro con alzheimer. terapéuticas y nuevos biomarcadores en Principal Investigator: Antonio Cuadrado enfermedad de Parkinson. Pastor. Principal Investigator: José Luis Labandeira CIBERNED’s collaboration: No. García. CIBERNED groups: G101. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G208, G101, G202, G203. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Funding: 340.000€. Funding agency: CIBERNED. Duration: 2017-2019 Funding: 240.000 €. Duration: 2015-2017 • Code: P_37_732. Title: Knowledge transfer in redox biology for developing advanced molecular tools in neurodegenerative diseases. Principal Investigator: Antonio Cuadrado Pastor. CIBERNED’s collaboration: No. CIBERNED groups: G101. Other CIBER’s collaboration: No. Type: European. Funding agency: Comisión Europea. Funding: : 1.915.000 €. Duration: 2016-2020

2017 CIBERNED Annual Report / 127 José Antonio del Río Fernández 114

Principal Investigator José Antonio del Río Fernández

Research team

Rosalina Gavín Marín Laila Lidón Gil Francina Mesquida Veny Senior Researcher PhD student PhD student

Vanessa Gil Fernández Ana López Mengual Laura Urrea Zazurca Postdoctoral Researcher PhD student PhD studentl

Arnau Hervera Abad Andreu Matamoros Anglès Ágata Mata Rodríguez Postdoctoral Researcher PhD student PhD student

Miriam Segura Feliú Technician

Molecular and Cellular Neurobiotechnology Institute for Bioengineering of Catalonia (IBEC) Baldiri Reixac 10-15 08028 Barcelona, Catalunya (Spain) Tel.: +34 93 403 59 23 / 93 402 02 96 [email protected] [email protected]

2017 CIBERNED Annual Report / 128 SUMMARY

PrPC as new receptor for a-synuclein Modeling comorbid tauopathies in familiareprionopathies using iPSc The cellular prion protein, encoded by the gene Prnp, has been reported to be a Gerstmann-Sträussler-Scheinker (GSS) receptor of β-amyloid. We aimed to explore syndrome is a fatal autosomal dominant whether the cellular prion protein participates neurodegenerative prionopathy clinically in the spreading of α-synuclein. Our results characterized by ataxia, spastic paraparesis, demonstrate that Prnp expression is not extrapyramidal signs and dementia. In some mandatory for α-synuclein spreading. GSS familiar cases carrying point mutations in the PRNP gene, patients also showed comorbid However, although the pathological spreading tauopathy leading to mixed pathologies. of α-synuclein can take place in the absence We developed in collaboration with several of Prnp in vivo, α-synuclein pathology groups of Ciberned and TerCell an induced expanded faster in wild-type and PrPC- pluripotent stem (iPS) cell model derived from overexpressing (Tga20) mice. In addition, in fibroblasts of a GSS patient harboring the Y218N vitro studies reported that α-synuclein binds PRNP mutation, as well as an age-matched strongly on PrPC-expressing cells, suggesting healthy control. This patient presented a role in modulating the effect of α-synuclein neurofibrillary degeneration with relevant fibrils. Indeed, the interaction takes place in the Tau hyperphosphorylation. Y218N iPS-derived charged domain of PrPC. Taken together we cultures showed relevant astrogliosis, increased consider that PrPC play roles as a new receptor phospho-Tau, altered microtubule-associated for α-synuclein. transport and cell death.

José Antonio del Río Fernández KEYWORDS

Cellular prion, α-synuclein, GSS, tauopathy PUBLICATIONS

• Garcia-Esparcia P., Lopez-Gonzalez I., Grau-Rivera O., Garcia-Garrido M.F., Konetti A., Llorens F. et al. Dementia with lewy bodies: Molecular pathology in the frontal cortex in typical and rapidly progressive forms. Frontiers in Neurology. 2017;8(MAR). • Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, García-Aznar JM et al. Erratum to: Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons.Molecular neurobiology. 2017. • Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Aznar JM et al. Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons.Molecular neurobiology. 2017. • Matamoros-Angles A., Gayosso L.M., Richaud-Patin Y., Di Domenico A., Vergara C., Hervera A. et al. iPS Cell Cultures from a Gerstmann-Sträussler-Scheinker Patient with the Y218N PRNP Mutation Recapitulate tau Pathology. Molecular Neurobiology. 2017;:1-16. • Urrea L, Ferrer I, Gavín R, Del Río JA. The cellular prion protein (PrPC) as neuronal receptor for α-synuclein.Prion. 2017;11(4).

2017 CIBERNED Annnual Report / 129 RESEARCH PROJECTS

• Code: 392/U/2014. • Code: AGL2015-71764-REDT. Title: Role of the cellular prion protein as Title: Red nacional de priones. “cross-talk” protein between alpha-syn/ Investigador Principal: José Antonio del Río LRRK2 and p-Tau in sporadic and familiar CIBERNED’s colaboration: Yes. Parkinson´s disease. CIBERNED groups: G114, G509, G503, G609, Principal Investigator: José Antonio del Río. G207. CIBERNED’s colaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G109, G114. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Private. Funding: 39.000 €. Funding agency: Fundació la Marató TV3. Duration: 2016-2017 Funding: 195.646 €. Duration: 2014-2017 • Code: BFU2015-6777-R. Title: Funciones de genes implicados en • Code: PI2016/02. angiogénesis y remodelación vascular Title: Monitoring the onset and evolution of durante el desarrollo cortical y en neuronal dysfunctions in propagative neural neurodegeneración. disorders using microfluidic devices and Principal Investigator: José Antonio del Río. translational approaches. CIBERNED’s colaboration: No. Principal Investigator: José Antonio del Río. CIBERNED groups: G114. CIBERNED’s colaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G114, G401, G201. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Funding: 279.259,2593€. Funding agency: CIBERNED. Duration: 2016-2018 Funding: 210.000 €. Duration: 2016-2018 • Code: TEC2015-72718-EXP. Title: Robots biológicos basados en el • Code: 392/U/2014 (IBEC). control de la unión neuromuscular. Title: Paper de la PrPc com a “cross-talk Principal Investigator: Josep Samitier. protein” entre alpha-sinucleina/LRRK2 i CIBERNED’s colaboration: No. p-Tau en la malaltia de Parkinson de tipus CIBERNED groups: G114. esporàdic i/o familiar. Other CIBER’s collaboration: CIBER-BBN. Principal Investigator: José Antonio del Río. Type: National. CIBERNED’s colaboration: Yes. Funding agency: MINECO. CIBERNED groups: G114, G109. Funding: 55.000€. Other CIBER’s collaboration: No. Duration: 2016-2018 Type: Private. Funding agency: Fundació Marato TV3. Funding: 195.646,25 €. Duration: 2015-2017

2017 CIBERNED Annual Report / 130 Isabel Fariñas Gómez 102

Principal Investigator Isabel Fariñas Gómez

Research team

Francisco Pérez Sánchez Alexandra Bizy Anna Lozano Ureña Senior lecturer PhD fellow PhD student

Martina Kirstein Germán Belenguer Pere Duart Abadía Senior lecturer Sánchez PhD student PhD fellow José Manuel Morante Redolat María José Palop Benlloch PhD assistant lecturer Beatriz Martí Prado Lab technician PhD student Sacramento Rodríguez Ferrón Fabrice Durupt PhD fellow Ana Domingo Muelas Lab technician PhD student Ana Pérez Villalba Elba Barberà Ferragud PhD fellow Pau Carrillo Barberá Lab technician PhD student Salomé Sirerol Piquer Miquel Marí Zaragoza PhD fellow Raquel Montalbán Lab technician PhD student Cristina Andrés Carbonell Lab technician Departamento de Biología Celular Universidad de Valencia Doctor Moliner 50 46100 Burjassot (Spain) Tel.: +34 96 354 37 84, (despacho) 3246 Fax: +34 96 354 34 04 [email protected]

2017 CIBERNED Annual Report / 131 SUMMARY

Our laboratory develops two lines of research in of toxic aggregates of alpha-synuclein protein neurodegeneration and cell therapy: obtained from patients’ Lewy bodies, which result in the death of dopaminergic neurons. 1) The study of cellular and molecular Under these conditions, NSCs stop proliferating alterations underlying the dopaminergic and producing new neurons. On the other neurodegeneration associated with hand, alpha-synuclein is a poorly understood Parkinson’s disease. synaptic protein that appears anomalously aggregated in the neurons of patients with PD 2) The study of neural stem cell (NSC) regulation. and mutations in the gene that codes for this protein are associated with familial cases of the We use experimental mouse models in which disease. to identify and evaluate cellular and molecular mechanisms that may underlie dopaminergic We have analyzed the physiological role of neurodegeneration or that can be used for alpha-synuclein using mice carrying a gene the understanding of adult NSCs and their targeted deletion and we have found that this manipulation for therapeutic purposes. In protein regulates the activation of NSCs through the context of the CIBERNED program, apart its effects on the bioavailability of dopamine. All from the specific studies within each of these these data suggest that alpha-synuclein and lines, we work on aspects that combine both, dopamine are regulators of NSCs and that they i.e. those related with the regulation of adult can be therapeutic targets for the modulation neurogenesis in Parkinsonism and during aging. of adult neurogenesis in neurodegenerative processes or during aging. During 2017, we have characterized the effects of the synaptic regulator involved in Also, in collaboration with the CIBERDEM, PD, alpha-synuclein, in adult neurogenesis in we have characterized the effects of insulin collaboration with the CIBERNED groups led by signaling in adult NSCs, finding that the Miquel Vila (Vall d’Hebrón Hospital) and José activation of the cyclin-dependent kinase Cdk4 López Barneo (Juan José Toledo Aral, IBiS). upon stimulation with insulin has effects on adult We have analyzed the behavior of NSCs in neurogenesis by actions that are independent mice with experimental Parkinsonism, induced of the canonical role of this kinase in the cell either by drugs or by the intracerebral infusion cycle. KEYWORDS

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

• Lissek T., Adams M., Adelman J., Ahissar E., Akaaboune M., Akil H. et al. Building Bridges through Science. Neuron. 2017;96(4):730-735. • 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. [ePub2017]. • Gomez-Pinedo U., Sirerol-Piquer M.S., Duran-Moreno M., Garcia-Verdugo J.M., Matias-Guiu J. Alexander disease mutations produce cells with coexpression of glial fibrillary acidic protein and ng2 in neurosphere cultures and inhibit differentiation into mature oligodendrocytes. Frontiers in Neurology. 2017;8(JUN). • Agusti A., Moya-Perez A., Campillo I., Montserrat-de la Paz S., Cerrudo V., Perez-Villalba A. et al. Bifidobacterium pseudocatenulatum CECT 7765 Ameliorates Neuroendocrine Alterations Associated with an Exaggerated Stress Response and Anhedonia in Obese Mice. Molecular Neurobiology. 2017;:1-16.

2017 CIBERNED Annual Report / 132 Program 2 Group: Isabel Fariñas Gómez

• Moya-Perez A., Perez-Villalba A., Benitez-Paez A., Campillo I., Sanz Y. Bifidobacterium CECT 7765 modulates early stress-induced immune, neuroendocrine and behavioral alterations in mice. Brain, Behavior, and Immunity. 2017;65:43-56. • Chirivella L., Kirstein M., Ferron S.R., Domingo-Muelas A., Durupt F.C., Acosta-Umanzor C. et al. Cyclin-Dependent Kinase 4 Regulates Adult Neural Stem Cell Proliferation and Differentiation in Response to Insulin. Stem Cells. 2017;35(12):2403-2416. • Alarcon-Aris D., Recasens A., Galofre M., Carballo-Carbajal I., Zacchi N., Ruiz-Bronchal E. et al. Selective α-Synuclein Knockdown in Monoamine Neurons by Intranasal Oligonucleotide Delivery: Potential Therapy for Parkinson’s Disease. Molecular Therapy. 2017. • Munoz-Soriano V., Domingo-Muelas A., Li T., Gamero E., Bizy A., Farinas I. et al. Evolutionary conserved role of eukaryotic translation factor eIF5A in the regulation of actin-nucleating formins. Scientific Reports. 2017;7(1). RESEARCH PROJECTS

• Code: Convenio Botin-UV. Principal Investigator: Isabel Fariñas. Title: Estudio de células madre en el ámbito CIBERNED’s collaboration: No. de las investigaciones básicas en terapia CIBERNED groups: G102. celular. Other CIBER’s collaboration: No. Principal Investigator: Isabel Fariñas. Type: National. CIBERNED’s collaboration: No. Funding agency: MINECO. CIBERNED groups: G102. Funding: 544.500 €. Other CIBER’s collaboration: No. Duration: 2015-2017 Type: Private. Funding agency: Fundación Botín-Banco • Code: SAF2016-78845-R. Santander. Title: Papel de la impronta genómica y su Funding: 625.000 €. regulación epigenética en células madre Duration: 2014-2018 neurales: relación con la formación de tumores. • Code: PROMETEO/2017/030. Principal Investigator: Sacramento Rodriguez Title: Efectos directos y remotos de la respuesta Ferron. inflamatoria sobre las células madre. CIBERNED’s collaboration: No. Principal Investigator: Isabel Fariñas. CIBERNED groups: G102. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G102. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO Type: CC.AA. Funding: 181.500€. Funding agency: Generalitat de Catalunya. Duration: 2017-2019 Funding: 384.957€. Duration: 2017-2021 • Code: RD16/0011/0016. Title: Combined protective/restorative cell- • Code: RD12/0019/0008. mediated strategies for neurodegenerative Title: RETIC de terapia celular. diseases. Principal Investigator: Isabel Fariñas. Principal Investigator: José Luis Labandeira CIBERNED’s collaboration: Yes. García. CIBERNED groups: G607 ; G301; G102; G113; CIBERNED’s collaboration Yes. G208; G105; G207. CIBERNED groups: G208 ; G102; G113; G105; Other CIBER’s collaboration: No. G207. Type: National. Other CIBER’s collaboration: No. Funding agency: Instituto de Salud Carlos III. Type: Nacional. Funding: 284.999€. Funding agency: Instituto de Salud Carlos III. Duration: 2016-2020 Funding: 240.245€. Duration: 2017-2021 • Code: SAF2014-54581. Title: Regulación molecular de la quiescencia: células madre neurales adultas.

2017 CIBERNED Annnual Report / 133 • Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz Cánoves. CIBERNED’s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350.000 €. Duration: 2016-2017 PhD DISSERTATIONS

• Author: Beatriz Martí Prado. • Author: Germán Belenguer Sánchez. Title: Estudio de la quiescencia y activación Title: Role of TNF-? receptor 2 (TNFR2) in the de las células madre neurales adultas por regulation of adult neural stem cells. uniones adherentes: papel de la N-cadherina. Date: 25th July 2017. Date: 19th July 2017. Supervisor: Isabel Fariñas Gómez. Supervisor: Isabel Fariñas Gómez.

2017 CIBERNED Annual Report / 134 Rafael Fernández-Chacón 606

Principal Investigator Rafael Fernández-Chacón

Research team

Alejandro Arroyo Saborido José Luis Nieto-González Marina Valenzuela Villatoro Lab technician PhD lecturer PhD student

Casandra Romero Cabrera María del Carmen Rivero Santiago López Begines Lab technician Mena PhD Fellow Lab technician Pablo García-Junco- José Luis Muñoz Bravo Clemente Fátima Rubior Pastor PhD Fellow PhD fellow Researcher María Isabel Reina Basic and Experimental Biomedicine graduated

Instituto de Biomedicina de Sevilla (IBiS) Hospital Universitario Virgen del Rocío. CSIC. Universidad de Sevilla Departamento de Fisiología Médica y Biofísica. Avenida Manuel Siurot s/n, 41013 Sevilla (Spain) Tel.: +34 95 592 30 47 Fax: +34 95 592 31 01 [email protected] www.ibis-sevilla.es

2017 CIBERNED Annual Report / 135 SUMMARY

The understanding of the physiological vivo the phenotypic alterations of neurogenesis mechanisms underlying neuronal maintenance associated with lack of CSP-α. would be a great asset for the design of therapeutic strategies to prevent or alleviate On the other hand, we generated mice that neurodegenerative diseases. Our group is express, in the same neurons lacking CSP-α, a interested in understanding the mechanisms that fluorescent marker and light-activated channels maintain the function and structure of the nerve that allow the study of synaptic transmission terminals that form the synapses, the cellular with optogenetics and electrophysiology specializations where neural communication (“patch-clamp”). Some of these lines of mice takes place. Our studies make use of models of allows to study the role of CSP-α in different life genetically modified mice that lack a molecular stages tracking the evolution of unexpected co-chaperon (Cysteine String Protein-α, CSP-α) synaptic phenotypes. These lines are also essential to maintain synaptic function. CSP-α being used to advance a collaboration with is directly related to autosomal-dominant the laboratory of Prof. Jens Hjerling-Leffler at adult-onset neuronal ceroid lipofuscinosis the Karolinska Institute in Stockholm (Sweden) (NLC-4), a devastating neurodegenerative based on the single-cell sequencing of disease caused by mutations in the human neuronal transcriptomes (). With this approach gene DNAJC5 that encodes CSP-α. we hope to identify genes and signaling pathways that are modified by alterations On the other hand, some recent studies suggest in synaptic function and structure that may that CSP-α participates in the release to the open up new perspectives to understand the extracellular medium of proteins involved in changes that a neuron undergoes during the neurodegeneration such as tau, α-synuclein, neurodegenerative process. In addition to TDP43 and SOD1. In the last year we have our studies at the central nervous system, in advanced in the generation of mice in which the last year we have also advanced in the we can conditionally suppress the expression characterization of the phenotype that mice of CSP-α in specific cell types in a controlled lacking CSP-α present at the peripheral level. manner over time. Using these strategies we As a continuation of a previous study in which have been able to demonstrate an unexpected we described functional changes associated role of CSP-α in the control of neural stem cells with the degeneration of motor neurons and in postnatal neural proliferation, we have (Rozas et al., Neuron, 74: 151-65 (2012) we are identified an important signaling pathway that currently identifing specific transformations in connects CSP-α with neurogenesis and we the maintenance of skeletal muscle fibers in the have been able to control pharmacologically in context of a motorneuron disorder.

KEYWORDS

Synapse, synaptic vesicle cycle, hippocampus, neurogenesis, neuronal ceroid lipofuscinosis, single-cell sequencing, transcriptomic, skeletal muscle PUBLICATIONS

• 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. [ePub2017]. • Garcia-Junco-Clemente P, Ikrar T, Tring E, Xu X, Ringach DL, Trachtenberg JT et al. An inhibitory pull-push circuit in frontal cortex.Nature neuroscience. 2017. • Nieto-Gonzalez J.L., Fernandez-Chacon R. Toward the Inner Nanostructure of a Secretory Vesicle. ACS Nano. 2017;11(4):3429-3432.

2017 CIBERNED Annual Report / 136 Program 2 Group: Rafael Fernández-Chacón RESEARCH PROJECTS

• Code: PI2015-2/06. • Code: UNSE15-CE-3249. Title: Molecular mechanisms of brain and Title: Equipamiento para microscopia muscle stem cell function in aging and multifoton in vivo. neurodegeneration. Principal Investigator: Rafael Fernández- Principal Investigator: Pura Muñoz Cánoves. Chacón. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G604, G102, G606, G111, CIBERNED groups: G606. G306. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: Intramurales. Funding agency: MINECO. Funding agency: CIBERNED. Funding: 543.799,35€. Funding: 350.000 €. Duration: 2017 Duration: 2016-2017 • Code: LRKK2-2016/00000112. • Code: BFU2016-76050-P. Title: LRRK2-mediated endolysosomal Title: Mecanismos moleculares del alterations: mechanistic insights and mantenimiento a largo plazo de las sinapsis validation as pharmacodynamic readout glutamatergicas in vivo. for kinase inhibitor studies. Principal Investigator: Rafael Fernández- Principal Investigator: Sabine Hilfiker. Chacón. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G606. CIBERNED groups: G606. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: National. Funding agency: Michael J. Fox Foundation. Funding agency: MINECO. Funding: 11.832,25€. Funding: 280.000€. Duration: 2015-2017 Duration: 2016-2019

2017 CIBERNED Annnual Report / 137 Javier Fernández Ruiz 303

Principal Investigator Javier Fernández Ruiz

Research team

José Antonio Ramos Atance Mª Concepción García María Ceprián Costoso Emeritus Full Professor García PhD student Assistant Professor Mariluz Hernández Gálvez Laura García Toscano Associate Professor Carmen Rodríguez Cueto PhD student Postdoctoral researcher Eva de Lago Femia Irene Santos-García Sanz Assistant Professor Valentina Satta PhD student Postdoctoral researcher María Sagrario Gómez Ruiz Laura Figuerola Asencio Assistant Professor María Gómez Cañas PhD student Postdoctoral researcher Onintza Sagredo Ezkioga Eva Luna Piñel Assistant Professor Francisco Espejo Porras PhD student PhD student Cristina Alonso Gómez PhD student

Departamento de Bioquímica y Biología Molecular Yolanda García Movellán Facultad de Medicina, Universidad Complutense Administrative Assistant de Madrid. Avenida Complutense s/n Pabellón IV, 4ª planta, laboratorios 9-12 28040 Madrid (Spain) Tel.: +34 91 394 14 50 Fax: +34 91 394 16 91 [email protected] [email protected]

2017 CIBERNED Annual Report / 138 SUMMARY

The work during 2017 focused on the study of disease provokes in the endocannabinoid the neuroprotective potential of cannabinoids system and completed the evaluation of in several chronic neurodegenerative phytocannabinoids and derivatives of the disorders, and, in particular, in the identification companies GW Pharmaceuticals and Vivacell of cellular and molecular mechanisms that Biotechnology, using SOD-1 mutant mice and underlie these effects. During this year, the also TDP-43 transgenic mice, for which we have work has been concentrated in Parkinson’s two different colonies, one of them serving for disease (PD), amyotrophic lateral sclerosis the experimental study of FTD. As in PD, we (ALS), frontotemporal dementia (FTD) and have also focused on CB2, PPAR-Δ and GPR55 spinocerebellar ataxia type-3 (SCA-3). receptors as potential targets. In the case of Vivacell Biotechnology, these studies have In the case of PD, the work has also continued been carried out within a project funded in the the collaboration with the companies GW 2014 call of the Retos-Colaboración program of Pharmaceuticals and VivaCell Biotechnology- the MINECO which finished in 2017. During this Spain, in relation with some cannabinoid year, we have continued our current project derivatives generated in these companies. funded by the MINEICO-Biomedicine National For GW, the work consisted in investigating the Program, concentrated in ALS and FTD, which advantages of the combination of cannabidiol is a joint project with a group of the CSIC and Δ 9-THCV for the symptomatic and Medical Chemistry Institute, and addressed to neuroprotective treatment of this disease, the development of new therapies based on including their antidyskinetic effects which are targeting GPR55, GPR18, CB2 receptors using studied in collaboration with the CIBERNED allosteric modulators, and CB2/PPAR-Δ using group leaded by Rosario Moratalla. In the hybrid compounds, as well as we have initiated case of Vivacell, the work concentrated in a new CIBERNED project with the groups of determining the potential of cannabigerol- Adolfo López de Munaín (coordinator), Isidre Javier Fernández Ruiz quinones (they are PPAR-Δ agonists) and Ferrer and Eduardo Soriano concentrated in cannabidiol-quinones (CB2 and PPAR-Δ the development of new therapies for ALS. agonists) in experimental models of this disease. We were particularly interested in the role of Lastly, in the case of SCA-3 (Machado-Joseph PPAR-Δ receptors and in a novel regulatory disease), the work has been concentrated in binding site in this receptor that may be finishing the biochemical and histopathological activated by certain cannabinoids, and also studies addressed to demonstrate the in a novel endocannabinoid-related receptor, occurrence of an endocannabinoid so-called GPR55, always in relation with the dysregulation in different CNS areas during the control of inflammation in this disease, and by course of the disease, using SCA-3 transgenic comparing different experimental models. mice, and we are carrying out pharmacological studies addressed to correct such dysregulation In the case of ALS, the work has again addressed and to elicit neuroprotective effects, including the identification of the changes that the CB2 and GPR55 as new therapeutic targets. KEYWORDS

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

2017 CIBERNED Annual Report / 139 PUBLICATIONS

• Ceprian M., Jimenez-Sanchez L., Vargas C., Barata L., Hind W., Martinez-Orgado J. Cannabidiol reduces brain damage and improves functional recovery in a neonatal rat model of arterial ischemic stroke. Neuropharmacology. 2017;116:151-159. • Ragusa G., Gomez-Canas M., Morales P., Rodriguez-Cueto C., Pazos M.R., Asproni B. et al. New pyridazinone-4-carboxamides as new cannabinoid receptor type-2 inverse agonists: Synthesis, pharmacological data and molecular docking. European Journal of Medicinal Chemistry. 2017;127:398-412. • Fernández-Trapero M, Espejo-Porras F, Rodríguez-Cueto C, Coates JR, Pérez-Díaz C, de Lago E et al. Up-regulation of CB2 receptors in reactive astrocytes in canine degenerative myelopathy, a disease model of amyotrophic lateral sclerosis.Disease models & mechanisms. 2017. • Gómez-Cañas M, Morales P, García-Toscano L, Navarrete C, Muñoz E, Jagerovic N et al. Corrigendum to Biological characterization of PM226, a chromenoisoxazole, as a selective CB2 receptor agonist with neuroprotective profile Pharmacol. Res. 110 (August 2016) (2016) 205- 215.Pharmacological research. 2017;120. • 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. [ePub2017]. • Rodriguez-Cueto C., Hernandez-Galvez M., Hillard C.J., Maciel P., Valdeolivas S., Ramos J.A. et al. Altered striatal endocannabinoid signaling in a transgenic mouse model of spinocerebellar ataxia type-3. PLoS ONE. 2017;12(4). • Valdeolivas S., Sagredo O., Delgado M., Pozo M.A., Fernandez-Ruiz J. Effects of a sativex-like combination of phytocannabinoids on disease progression in R6/2 mice, an experimental model of huntington’s disease. International Journal of Molecular Sciences. 2017;18(4). • 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.Amyotrophic lateral sclerosis & frontotemporal degeneration. 2018. [ePub2017]. • Mohammed N., Ceprian M., Jimenez L., Pazos M.R., Martinez-Orgado J. Neuroprotective effects of cannabidiol in hypoxic ischemic insult. The therapeutic window in newborn mice. CNS and Neurological Disorders - Drug Targets. 2017;16(1):102-108. • Fernandez-Ruiz J., Gomez-Ruiz M., Garcia C., Hernandez M., Ramos J.A. Modeling Neurodegenerative Disorders for Developing Cannabinoid-Based Neuroprotective Therapies. Methods in Enzymology. 2017;593:175-198. • Morales P, Moreno L, Fernández-Ruiz J, Jagerovic N. Synthesis of a novel CB 2 cannabinoid- porphyrin conjugate based on an antitumor chromenopyrazoledione. Journal of Porphyrins and Phthalocyanines. 2017 Jan;21(01):67-76. • de Lago E., Tolón R., Fernández-Ruiz J. Cannabinoides y enfermedades neurodegenerativas. In: Efectos terapéuticos de los cannabinoides. Ramos JA, editors. 2017. • Sagredo O, Martínez-Orgado J. Papel de los cannabinoides en encefalopatías pediátricas. In: Efectos terapéuticos de los cannabinoides. Ramos JA, editors. 2017. RESEARCH PROJECTS

• Code: PR26/16-20352. CIBERNED groups: G303. Title: Posible desregularización del sistema Other CIBER’s collaboration: No. cannabinoide endógeno en encelopatías Type: Private. epilépticas infantiles: estudio con muestras Funding agency: Banco Santander - UCM. biológicas de pacientes. Funding: 6.000 €. Principal Investigator: Onintza Sagredo. Duration: 2017 CIBERNED’s collaboration: No.

2017 CIBERNED Annual Report / 140 Program 2 Group: Javier Fernández Ruiz

• Code: PR26/16-18B. • Code: NEUROLATAM. Title: Estudio de nuevas dianas en Title: Unraveling the neurobiological sustrate demencias neurodegenerativas basadas of protective cannabinoid actions in the en tratamientos neuroprotectoras y brain. neurogénicos. Principal Investigator: Ismael Galve Roperh, Principal Investigator: Eva De Lago, Maria José A. Ramos Atance. Gómez Ruiz, Felipe Ortega De La O. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G303 ; G305. CIBERNED groups: G303. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: Private. Funding agency: Unión Iberoamericana de Funding agency: Banco Santander - UCM. Universidades. Funding: 30.000 €. Funding: 24.225€. Duration: 2017-2018 Duration: 2017-2018

• Code: GW2015. • Code: 2017/VIVACELL/JFR Title: Preclinical development of Title: Investigation in the anti-inflammatory phytocannabinoid-based therapies for and neuroprotective properties of the the treatment of disease progression in phytocannabinoid derivative VCE003.2 in amyotrophic lateral sclerosis/frontotemporal Parkinson´s disease using LPS-lesioned alpha- dementia using TDP-43 transgenic mice. synuclein transgenic mice. Principal Investigator: Javier Fernández Ruiz, Principal Investigator: Javier Fernández Ruiz, Eva de Lago Femia. María Concepción García. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G303. CIBERNED groups: G303. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: International. Funding agency: GW Research Ltd. (UK). Funding agency: VivaCell Biotechnology Funding: 126.544,5€. España. Duration: 2015-2018 Funding: 80.000€. Duration: 2017-2018 • Code: RTC-2014-1877-1. Title: Desarrollo preclínico de nuevos • Code: PI2016/04. cannabinoides para el tratamiento de Title: The ALS CIBERNED challenge: esclerodermia y esclerosis lateral amiotrófica. Accelerating new drug discovery. Principal Investigator: Javier Fernández Ruiz. Principal Investigator: Adolfo López De CIBERNED’s collaboration: No. Munain Arregui. CIBERNED groups: G303. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G609, G303, G503, G408. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Funding: 330.212 €. Funding agency: CIBERNED Duration: 2015-2017 Funding: 200.000 €. Duration: 2016-2018 PhD DISSERTATIONS

• Author: Sara Valdeolivas Rojas. Title: Towards a neuroprotective therapy with cannabinoids in Huntington´s disease: preclinical and clinical studies. Date: 9th June 2017. Supervisor: Javier Fernández Ruiz.

2017 CIBERNED Annnual Report / 141 José Manuel Fuentes Rodríguez 103

Principal Investigator José Manuel Fuentes Rodríguez

Research team

Rosa Ana González Polo Sokhna Maryama Seydina Guadalupe Martínez Chacón Researcher Yakhine DIOP Lab technician PhD student Mireia Niso Santano María Pura Luceño PhD fellow Mario Rodríguez Arribas Lab technician PhD student Ignacio Casado Naranjo Head of Department Neuro- Elisabet Uribe Carretero logy PhD student

Departamento de Bioquímica y Biología Molecular y Genética Facultad de Enfermería y Terapia Ocupacional Universidad de Extremadura. Avenida de la Universidad s/n 10003 Cáceres (Spain) Tel.: +34 92 725 74 50 (ext 51286) Fax: +34 92 725 74 51 [email protected]

2017 CIBERNED Annual Report / 142 SUMMARY

During 2017, our lab working lines has, as well delay their analysis until late 2018. Nonetheless, as in the previous years, focused on studying first evidences suggest significative differences basic molecular mechanisms related with in the metabolomic profile of our PD human Parkinson´s disease (PD) etiopathogenesis, samples when compared to control group, and namely autophagy regulation and the role we also observe differences in polymorphisms of this mechanism and PARK gene-related appearance between G2019S and R1441G proteins in both processes. LRRK2 groups. On the other hand, we have extended our Additionally, in this frame of collaboration with collaboration and provision of services related CIBERNED groups (Dr. Kulisewski, Gutierrez y to molecular diagnosis of PD to the “Servicio Vicario), we have described, leaded by Dr. Extremeño de Salud”. Besides, We have started Moratalla, an accumulation of autophagosomes connections with “Fundacion Isabel Gemio” and alteration of autophagy flux combined in order to develop scientific projects related with lysosomal dysfunction and cholesterol to elucidate the role of autophagy in certain accumulation in a PD model with N370P neuromuscular diseases. In 2009 we established mutation in GBA1 gene. This mutation is related a collaboration with Dr. Adolfo Lopez de with a diminution in β-glucocerebrosidase in Munain that actually persists. In this year, we protein levels as well as protein activity, and this have deployed our efforts in the molecular mutation is also involved in GBA accumulation characterization of the basal deregulation in Endoplasmic reticulum, thus generating we have already observed in PD fibroblasts unfolded protein response and Golgi apparatus with G2019S and R1441G LRRK2 pathogenic fragmentation. Taken together, this phenotype mutations. modifies mitochondrial exchange capacity as well as oxidative stress and cell death. We have started the expression level analyses of acetyltransferase and deacetylase Finally, we have got a deeper insight in the study activities in both models and acetylation of general mechanisms involved in autophagy status of proteins involved in autophagy, as regulation thanks to our collaboration already this posttranslational modification is arising as established with Dr. Guido Kromer´s lab, who a key regulator in macroautophagy process. work in Apoptosis, Cancer & immunity Laboratory In this sense, we have carried out a complete at INSERM, Cordeliers Research Center in Paris, screening of both enzyme classes and we France. As a result of this collaboration, we have observed significative differences among have published two methodological papers at control, idiopathic and G2019S and R1441G “Methods in Emzymology” journal. Related to mutation carrier groups, which are especially our collaboration with the “Servicio Extremeño relevant in the last one. Theses acetylation de Salud”, we would highlight that we are changes correlates with alterations in selective currently performing our provision of services autophagy, namely mitophagy. We are finishing agreement in relation with the molecular a cooperative project as a collaboration within diagnosis of Parkinson and Alzheimer. CIBERNED (PI2015/03) with Dr. Perez Tur and Dr. Perez Castillo´s groups (in this project Dr. Lopez Besides, we have been a promoter group de Munain is also involved). In this project we in the establishment of the “Instituto de have tackled the characterization of differential Inevestigación Biosanitaria de Extremadura metabolic profiles in PD. (INUBE)”. This organization is currently being evaluated and it already counts with supports In 2017, we have already concluded mouse of public administration such as “Consejería de and human samples preparation and we have Sanidad” and “Políticas Sociales de la Junta de performed metabolomic and transcriptomic Extremadura”, as well as “Servicio Extremeño analyses. Such a high amount of information will de Salud”. 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

2017 CIBERNED Annual Report / 143 PUBLICATIONS

• Aguiar de Sousa D., Canhao P., Crassard I., Coutinho J., Arauz A., Conforto A. et al. Safety of Pregnancy After Cerebral Venous Thrombosis: Results of the ISCVT (International Study on Cerebral Vein and Dural Sinus Thrombosis)-2 PREGNANCY Study. Stroke. 2017;48(11):3130-3133. • Sica V., Pedro J.M.B.-S., Chen G., Marino G., Lachkar S., Izzo V. et al. Inhibitor of growth protein 4 interacts with Beclin 1 and represses autophagy. Oncotarget. 2017;8(52):89527-89538. • Lopez Espuela F., Portilla Cuenca J.C., Leno Diaz C., Parraga Sanchez J.M., Gamez-Leyva G., Casado Naranjo I. Sex differences in long-term quality of life after stroke: Influence of mood and functional status Diferencias de género en la calidad de vida a largo plazo tras un ictus: Influencia del estado funcional y el estado de ánimo. Neurologia. 2017. • Garcia-Sanz P., Orgaz L., Bueno-Gil G., Espadas I., Rodriguez-Traver E., Kulisevsky J. et al. N370S- GBA1 mutation causes lysosomal cholesterol accumulation in Parkinson’s disease. Movement Disorders. 2017;32(10):1409-1422. • Yakhine-Diop S.M.S., Martinez-Chacon G., Gonzalez-Polo R.A., Fuentes J.M., Niso-Santano M. Fluorescent FYVE Chimeras to Quantify PtdIns3P Synthesis During Autophagy. Methods in Enzymology. 2017;587:257-269. • Rodriguez-Arribas M., Yakhine-Diop S.M.S., Gonzalez-Polo R.A., Niso-Santano M., Fuentes J.M. Turnover of Lipidated LC3 and Autophagic Cargoes in Mammalian Cells. Methods in Enzymology. 2017;587:55-70. • López Espuela F, Portilla Cuenca JC, Holguín Mohedas M, Párraga Sánchez JM, Cordovilla- Guardia S, Casado Naranjo I. Nutritional status and the relationship regarding functional status after stroke.Nutricion hospitalaria. 2017;34(5). • Labrador P.J., Gonzalez-Sanchidrian S., Polanco S., Davin E., Gomez-Martino J.R., Fuentes J.M. Detection and classification of chronic kidney disease in Primary Care and importance of albuminuria. Semergen. 2017. • Labrador Gomez P.J., Gonzalez Sanchidrian S., Fuentes Rodriguez J.M., Gomez-Martino Arroyo J.R. Trend of lipid profile in general population from Caceres Health Area. Hipertension y Riesgo Vascular. 2017;34(1):17-23. • Yakhine-Diop SMS, Gómez-Sánchez R, Bravo-San Pedro JM, González Polo RA, Fuentes JM. The Roles of Autophagy in Parkinson’s disease. 2017. 1(3): 62 - 72. RESEARCH PROJECTS

• Code: 2015/03. Type: CC.AA. Title: Perfiles metabólicos diferenciales en Funding agency: Gobierno de enfermedad de Parkinson. Extremadura. Principal Investigator: José Manuel Fuentes Funding: 64.640,36 €. Rodríguez. Duration: 2015-2017 CIBERNED’s collaboration: Yes. CIBERNED groups: G103, G110, G209. • Code: PI16/01840. Other CIBER’s collaboration: No. Title: Influencia del tratamiento Type: Intramurales. anticoagulante sobre la función cognitiva Funding agency: CIBERNED. de los pacientes con fibrilación auricular no Funding: 210.000 €. valvular. Duration: 2015-2017 Principal Investigator: Ignacio Casado Naranjo. • Code: GR15045. CIBERNED’s collaboration: No. Title: Ayudas para el fortalecimiento de los CIBERNED groups: G103. grupos de investigación de Extremadura. Other CIBER’s collaboration: No. Principal Investigator: José Manuel Fuentes Type: National. Rodríguez. Funding agency: Instituto de Salud Carlos III CIBERNED’s collaboration: No. Funding: 55.055€. CIBERNED groups: G103. Duration: 2017-2019 Other CIBER’s collaboration: No.

2017 CIBERNED Annual Report / 144 Program 2 Group: José Manuel Fuentes Rodríguez

• Code: PI14/00170. • Code: PEJ-2014-A-96965. Title: Búsqueda de nuevas dianas Title: Ayudas para la Promoción de Empleo terapéuticas en la enfermedad de Parkinson. Joven e Implantación de la Garantía Juvenil Análisis de los sistemas de reciclaje celular y en I+D+i. estado mitocondrial en células procedentes Principal Investigator: José Manuel Fuentes de pacientes. Rodríguez. Principal Investigator: Rosa Ana González CIBERNED’s collaboration: No. Polo. CIBERNED groups: G103. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G103. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: National. Funding: 39.200 €. Funding agency: Instituto de Salud Carlos III. Duration: 2016-2017 Funding: 110.715 €. Duration: 2015-2017 • Code: BFU2015-71869-REDT. Title: Red de Excelencia para el estudio de • Code: PI15/00034. la Autofagia. Title: Alteraciones del perfil metabolico Principal Investigator: Patricia Boya. inducidas por mutaciones patogénicas CIBERNED’s collaboration: No. de LRRK2 como biomarcadores de la CIBERNED groups: G103. enfermedad de Parkinson. Other CIBER’s collaboration: No. Principal Investigator: José Manuel Fuentes Type: National. Rodríguez. Funding agency: MINECO. CIBERNED’s collaboration: No. Funding: 42.000 €. CIBERNED groups: G103. Duration: 2016-2017 Other CIBER’s collaboration: No. Type: National. Funding agency: Instituto de Salud Carlos III. Funding: 98.615 €. Duration: 2016-2018

PhD DISSERTATIONS

• Author: Guadalupe Martínez Chacón. Title: Evaluación de las propiedades neuroprotectoras de componentes de la jalea real en la enfermedad de Parkinson. Papel de la autofagia. Date: 19th October 2017. Supervisor: José Manuel Fuentes Rodríguez.

2017 CIBERNED Annnual Report / 145 José Manuel García Verdugo 113

Principal Investigator José Manuel García Verdugo

Research team

Vicente Herranz Pérez Susana González Granero Arantxa Cebrián Silla PhD fellow Researcher Lab technician

Laboratorio de Neurobiología Comparada (SS7) Instituto Cavanilles de Biodiversidad y Biología Evolutiva. Universidad de Valencia c/Catedrático José Beltrán, nº2 46980 Paterna, Valencia (Spain) Tel.: +34 96 354 35 87 Fax: +34 96 354 36 70 [email protected]

2017 CIBERNED Annual Report / 146 SUMMARY

The main line of our research group focuses on We have made a finding in this organism about the study of adult neurogenesis in various groups the presence of a characteristic structure at of vertebrates, including the human species, in the level of the nuclear membrane that is addition to the analysis of the behavior of neural specific to quiescent neural stem cells. The stem cells in various pathological conditions of aforementioned structure is a specialization the central nervous system. During the last year, of the nuclear membrane that had previously much of our efforts have been directed towards been identified in neutrophils and was named the study of neurogenesis in the dentate gyrus ELCS (envelope-limited chromatin sheets). of the hippocampus in the human brain. We have observed that ELCS appear in early Previous studies suggested that the dentate stages of brain development, and that they are gyrus incorporates hundreds of new neurons disappear when neural stem cells change from every day, in a process affected to a certain a quiescent to an activated state. This type of extent by stress and physical exercise. structure has also been identified in tumor cells resistant to radio- and chemotherapy, which The main contribution of our work is the is why we think they could be related to the observation of a dramatic decrease in the regulation of the cell cycle. number of proliferative progenitors of this region during the first year of life, being very scarce in On the other hand, in our interest to advance juvenile stages. This raises important questions in the knowledge about pathologies of the about the neurogenic capacity of this region in nervous system, we have collaborated in a the adult human brain. These results have been study of ischemic lesions in mouse neonatal recently published by Nature in 2018. stages. Radial glial cells act as neural stem cells during the embryonic development of the brain. On the other hand, from the point of view of These cells, which have long radial expansions, comparative adult neurogenesis, during the disappear in the first days of life after birth in the José Manuel García Verdugo 2017 annuity we published, among others, mouse and remain, at least, until the last weeks the results of the study of this process in a fish of gestation in humans. In this study, we have species: Austrolebias charrua. Fish are a group found that after an ischemic lesion it is possible of organisms with a high regenerative capacity, to find radial glial cells during longer postnatal even in the nervous system, compared to windows than under normal conditions. The mammals. Therefore, the study of their neural radial fibers of these cells provide a scaffold stem cells can provide relevant information that favors N-cadherin mediated migration of about the signals that regulate the activity of neuroblasts from the ventricular-subventricular these cells. In this sense, we have also described zone to injured areas in the cerebral cortex. the neurogenic activity in the ventricular- We have also found that transplanted gelatin subventricular zone of the brain of dogs, finding or synthetic polymer sponges containing greater similarities with the human brain than N-cadherin can act as a scaffold that favors with that of rodents, so it could represent a good the migration of neuroblasts to areas of animal model to study this process. However, injury, in a similar way as they do along radial the model most used today is still the mouse. glial fibers. KEYWORDS

Neurogenesis, neural stem cells, ventricular-subventricular zone, dentate gyrus PUBLICATIONS

• Baglietto-Vargas D, Sánchez-Mejias E, Navarro V, Jimenez S, Trujillo-Estrada L, Gómez-Arboledas A et al. Dual roles of Aβ in proliferative processes in an amyloidogenic model of Alzheimer’s disease.Scientific reports. 2017;7(1). • Gomez-Pinedo U., Sirerol-Piquer M.S., Duran-Moreno M., Garcia-Verdugo J.M., Matias-Guiu J. Alexander disease mutations produce cells with coexpression of glial fibrillary acidic protein and ng2 in neurosphere cultures and inhibit differentiation into mature oligodendrocytes. Frontiers in Neurology. 2017;8(JUN).

2017 CIBERNED Annual Report / 147 • 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. [ePub2017]. • Nager A.R., Goldstein J.S., Herranz-Perez V., Portran D., Ye F., Garcia-Verdugo J.M. et al. An Actin Network Dispatches Ciliary GPCRs into Extracellular Vesicles to Modulate Signaling. Cell. 2017;168(1-2):252-263.e14. • Jinnou H., Sawada M., Kawase K., Kaneko N., Herranz-Perez V., Miyamoto T. et al. Radial Glial Fibers Promote Neuronal Migration and Functional Recovery after Neonatal Brain Injury. Cell Stem Cell. 2018;22(1):128-137.e9. [ePub2017]. • Mirzadeh Z., Kusne Y., Duran-Moreno M., Cabrales E., Gil-Perotin S., Ortiz C. et al. Bi- and uniciliated ependymal cells define continuous floor-plate-derived tanycytic territories. Nature Communications. 2017;8. • Mendivil-Perez M., Soto-Mercado V., Guerra-Librero A., Fernandez-Gil B.I., Florido J., Shen Y.- Q. et al. Melatonin enhances neural stem cell differentiation and engraftment by increasing mitochondrial function. Journal of Pineal Research. 2017;63(2). • Torres-Vega E., Mancheno N., Cebrian-Silla A., Herranz-Perez V., Chumillas M.J., Moris G. et al. Netrin-1 receptor antibodies in thymoma-associated neuromyotonia with myasthenia gravis. Neurology. 2017;88(13):1235-1242. • Pourabdolhossein F, Gil-Perotín S, Garcia-Belda P, Dauphin A, Mozafari S, Tepavcevic V et al. Inflammatory demyelination induces ependymal modifications concomitant to activation of adult (SVZ) stem cell proliferation.Glia. 2017;65(5). • Cebrian-Silla A., Alfaro-Cervello C., Herranz-Perez V., Kaneko N., Park D.H., Sawamoto K. et al. Unique Organization of the Nuclear Envelope in the Post-natal Quiescent Neural Stem Cells. Stem Cell Reports. 2017;9(1):203-216. • Bruel A.-L., Franco B., Duffourd Y., Thevenon J., Jego L., Lopez E. et al. Fifteen years of research on oral-facial-digital syndromes: From 1 to 16 causal genes. Journal of Medical Genetics. 2017;54(6):371-380. • Marcuzzo S., Bonanno S., Figini M., Scotti A., Zucca I., Minati L. et al. A longitudinal DTI and histological study of the spinal cord reveals early pathological alterations in G93A-SOD1 mouse model of amyotrophic lateral sclerosis. Experimental Neurology. 2017;293:43-52. • Cubas-Nuñez L, Duran-Moreno M, Castillo-Villalba J, Fuentes-Maestre J, Casanova B, García- Verdugo JM et al. In situ RT-PCR Optimized for Electron Microscopy Allows Description of Subcellular Morphology of Target mRNA-Expressing Cells in the Brain.Frontiers in cellular neuroscience. ;11. • Arce-Franco M, Dominguez-Luis M, Pec MK, Martínez-Gimeno C, Miranda P, Alvarez de la Rosa D et al. Functional effects of proinflammatory factors present in Sjögren’s syndrome salivary microenvironment in an in vitro model of human salivary gland.Scientific reports. 2017;7(1). • Fernandez-Gil B., Abdel Moneim A.E., Ortiz F., Shen Y.-Q., Soto-Mercado V., Mendivil-Perez M. et al. Melatonin protects rats from radiotherapyinduced small intestine toxicity. PLoS ONE. 2017;12(4). • Torres-Perez M., Rosillo J.C., Berrosteguieta I., Olivera-Bravo S., Casanova G., Garcia-Verdugo J.M. et al. Stem cells distribution, cellular proliferation and migration in the adult Austrolebias charrua brain. Brain Research. 2017;1673:11-22. • Galan L., Gomez-Pinedo U., Guerrero A., Garcia-Verdugo J.M., Matias-Guiu J. Amyotrophic lateral sclerosis modifies progenitor neural proliferation in adult classic neurogenic brain niches. BMC Neurology. 2017;17(1). • Morais G.B., Viana D.A., Silva F.M.O., Xavier Junior F.A.F., Farias K.M., Pessoa C.D.O. et al. Polarization microscopy as a tool for quantitative evaluation of collagen using picrosirius red in different stages of CKD in cats. Microscopy Research and Technique. 2017;80(5):543-550. • Fujioka T., Kaneko N., Ajioka I., Nakaguchi K., Omata T., Ohba H. et al. β1 integrin signaling promotes neuronal migration along vascular scaffolds in the post-stroke brain. EBioMedicine. 2017;16:195-203.

2017 CIBERNED Annual Report / 148 Program 2 Group: José Manuel García Verdugo RESEARCH PROJECTS

• Code: RD16/0011. • Code: RD16/0011/0016. Title: RETIC de terapia celular. Title: Combined protective/restorative cell- Principal Invetigator: Isabel Fariñas. mediated strategies for neurodegenerative CIBERNED’s collaboration: Yes. diseases. CIBERNED groups: G102, G301, G113, G208, Principal Invetigator: José Luis Labandeira G105, G607. García. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: National. Grupos CIBERNED: G208, G102, G113, G105, Funding agency: Instituto de Salud Carlos III. G207. Funding: 284.999€. Colaboración otros CIBERs/Centros: No. Duration: 2016-2020 Type: National. Funding agency: Instituto de Salud Carlos III. • Code: PROMETEOII/2014/075. Funding: 240.245€. Title: Celulas madre adultas: aplicaciones Duration: 2017-2021. en terapia regenerativa. Principal Invetigator: José Manuel García • Code: PI2015/01. Verdugo. Title: Terapia génica dirigida a neuregulinas CIBERNED’s collaboration: No. para el tratamiento de la degeneración Grupos CIBERNED: G113. de las motoneuronas en la esclerosis lateral Colaboración otros CIBERs/Centros: No. amiotrófica. Type: CC.AA. Principal Invetigator: Xavier Navarro Funding agency: Generalitat Valenciana. Acebes. Funding: 156.265€. CIBERNED’s collaboration: Yes. Duration: 2014-2017 Grupos CIBERNED: G607, G113, G406, G407. Colaboración otros CIBERs/Centros: No. • Code: BFU2015-64207-P. Type: Intramurales. Title: Estudio del cerebro humano: Funding agency: CIBERNED. caracterización de nichos neurogénicos y Funding: 240.000€. nuevas vías de migración. Duration: 2015-2017 Principal Invetigator: José Manuel García Verdugo. CIBERNED’s collaboration: No. Grupos CIBERNED: G113. Colaboración otros CIBERs/Centros: No. Type: National. Funding agency: MINECO. Funding: 166.012 €. Duration: 2016-2018 PhD DISSERTATIONS

• Author: Arantxa Cebrián Silla. • Author: Paula García Belda. Title: Estructura nuclear y quiescencia en Title: Direccionamiento de células madre células madre neurales. mesenquimales adultas para el tratamiento Date: 22th June 2017. del infarto cerebral mediante la aplicación Supervisor: José Manuel García Verdugo. de campos magnéticos. Date: 26th September 2017. Supervisor: José Manuel García Verdugo.

2017 CIBERNED Annnual Report / 149 José González Castaño 104

Principal Investigator José González Castaño

Research team

Raúl Sánchez Lanzas Silvia Carlés González Pre-doctoral researcher Research Techician

Departamento de Bioquímica e Instituto de Investigaciones Biomédicas “Alberto Sols” Univerdad Autónoma de Madid-CSIC. Laboratorio B13. Facultad de Medicina UAM Arzobispo Morcillo s/n. 28029 Madrid (Spain) Tel.: +34 91 497 54 27 Fax: +34 91 585 44 01 [email protected]

2017 CIBERNED Annual Report / 150 SUMMARY

Proteasomes and lysosomes play a critical role alpha-synuclein, is not impaired and there is no in protein degradation and proteostasis in the significant changes in the steady-state protein cell. Three major pathways have been shown to and mRNA levels compared to controls. Our be relevant in protein degradation in Parkinson work and previous work from other authors, (PD) and other neurodegenerative diseases; clearly questioned the role of CMA in lysosomal ubiqutin-proteasome, chaperon mediated protein degradation. Recently it has been autophagy (CMA) and macroautophagy reported that DJ-1, a gene whose mutation is Alpha-synuclein, a protein accumulated and known to be responsible of familiar autosomal aggregated in PD, has been reported to be recessive forms of PD, is also degraded by the degraded by the CMA pathway through its CMA pathway. direct binding to the lysosomal receptor Lamp- 2A. In our ongoing studies of the pathways of degradation of DJ-1 wild type and point mutants We have reported last year that in a we are also analyzing the possible involvement lymphoblastoid cell line derived from a patient of CMA pathway in DJ-1 degradation. Our with Danon disease, lacking the expression of work is contributing to understand and clarify the different Lamp-2 isoforms including Lamp- the role of the different pathaways of protein 2A, the degradation of several selective degradation in neurodegenerative disease, substrates of the CMA pathway including especially in connection with PD pathology. KEYWORDS

Synuclein, DJ-1, proteasome, macroautophagy, chaperon-mediated-autophagy, proteolysis, proteostasis, Parkinson PUBLICATIONS

• Morales-Garcia JA, Gine E, Hernandez-Encinas E, Aguilar-Morante D, Sierra-Magro A, Sanz- SanCristobal M et al. CCAAT/Enhancer binding protein β silencing mitigates glial activation and neurodegeneration in a rat model of Parkinson’s disease.Scientific reports. 2017;7(1). • Sanchez-Lanzas R., Castano J.G. Lysine-less variants of spinal muscular atrophy SMN and SMNΔ7 proteins are degraded by the proteasome pathway. International Journal of Molecular Sciences. 2017;18(12). RESEARCH PROJECTS

• Code: BM1307. • Code: SAF_2012_34556. Title: 2013 COST action proteostasis. Title: Papel de la proteostasis de alfa- Principal Investigator: Manuel S. Rodríguez, sinucleina y DJ-1 en la función celular Rosa Barrios. y su relación con la patogénesis de la CIBERNED’s collaboration: No. enfermedad de Parkinson. CIBERNED groups: G140. Principal Investigator: José González Other CIBER’s collaboration: No. Castaño. Type: European. CIBERNED’s collaboration: No. Funding agency: Comisión Europea. CIBERNED groups: G104. Funding: 13.500 €. Other CIBER’s collaboration: No. Duration: 2014-2017 Type: National. Funding agency: MINECO. Funding: 120.000 €. Duration: 2013-2017

2017 CIBERNED Annual Report / 151 Manuel Guzmán Pastor 305

Principal Investigator Manuel Guzmán Pastor

Research team

Ismael Galve-Roperh Adán de Salas Quiroga José Aguareles Gorines Associate Professor. Principal Postdoctoral fellow PhD student Co-Investigator Andrea Ruiz Calvo Carlos Costas Insua Guillermo Velasco Díez Postdoctoral fellow PhD student Associate professor Daniel García Rincón Eva Resel Mariné Cristina Blázquez Ortiz PhD student Lab manager Associate professor Juan Paraíso Luna Elena García Taboada Justo García de Yébenes Prous PhD student Lab technician Collaborator doctor Irene Berenice Maroto Javier Palazuelos Diego Martínez Postdoctoral fellow PhD student

Departamento de Bioquímica y Biología Molecular Facultad de Ciencias Químicas Universidad Complutense 28040 Madrid (Spain) Tel.: +34 91 394 46 68 Fax: +34 91 394 46 72 [email protected]

2017 CIBERNED Annual Report / 152 SUMMARY

Our research focuses globally on the study of the Huntington’s disease patients and animal molecular and cellular mechanisms underlying models, and, of interest, this loss of CB1 the control of neural cell physiopathology by receptors seems to reflect, at least in part, the endocannabinoid system. Thus, in 2017 we the neuron-damage pattern characteristic have kept on studying how this system tunes of the disease. neural progenitor proliferation, differentiation and survival. 3) This loss of CB1 receptors occurs at early stages of Huntington’s disease and prior to Specifically, in the context of our CIBERNED the appearance of overt clinical symptoms, program, we have evaluated the role of the neurodegeneration and bulk changes in endocannabinoid system in Huntington’s other neurochemical parameters. disease. As a matter of fact, Huntington’s disease constitutes, in our opinion, the best In 2017, our studies on the endocannabinoid currently available model disease to assess the system in Huntington´s disease have aimed at pathophysiological relevance and therapeutic defining: potential of the endocannabinoid system in neurodegenerative diseases. This is due to • Whether stimulation of the endocannabinoid several reasons: system in Huntington’s disease models promotes neuroprotection and therefore delays the onset and/or attenuates the 1) CB1 is the most abundant G protein-coupled progression of the pathology. receptor in the brain; specifically, it is highly expressed in the basal ganglia at synapses • Whether the alterations of CB1 cannabinoid established by neurons containing GABA receptors observed in MSNs and/or [especially medium-sized spiny neurons corticostriatal terminals at early stages of the (MSNs), the cells that primarily degenerate disease are involved in the pathogenesis of in Huntington’s disease] or glutamate Huntington’s disease. (especially corticostriatal projecting neurons, which critically control MSN function) as • Whether the induction of CB2 cannabinoid transmitters, and play a key role in the control receptors in reactive microglial cells of motor behaviour (one of the processes modulates the excitotoxicity processes that is most characteristically affected in associated with Huntington’s disease. Huntington’s disease). We aim at unravelling the molecular 2) A remarkable and dorsolaterally-selective and cellular bases as well as the down-regulation of the CB1 receptor has potential clinical relevance of those been documented in the basal ganglia of processes. KEYWORDS

Endocannabinoid system, Huntington’s disease, neuroprotection, neurogenesis, cell signalling, experimental therapeutics PUBLICATIONS

• Jansen WJ, Ossenkoppele R, Tijms BM, Fagan AM, Hansson O, Klunk WE et al. Association of Cerebral Amyloid-β Aggregation With Cognitive Functioning in Persons Without Dementia. JAMA psychiatry. 2017. • Moreno E, Chiarlone A, Medrano M, Puigdellívol M, Bibic L, Howell LA et al. Singular Location and Signaling Profile of Adenosine A2A-Cannabinoid CB1 Receptor Heteromers in the Dorsal Striatum.Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology. 2017.

2017 CIBERNED Annnual Report / 153 • Mateo Y., Johnson K.A., Covey D.P., Atwood B.K., Wang H.-L., Zhang S. et al. Endocannabinoid Actions on Cortical Terminals Orchestrate Local Modulation of Dopamine Release in the Nucleus Accumbens. Neuron. 2017;96(5):1112-1126.e5. • Díaz-Alonso J, de Salas-Quiroga A, Paraíso-Luna J, García-Rincón D, Garcez PP, Parsons M et al. Loss of Cannabinoid CB1 Receptors Induces Cortical Migration Malformations and Increases Seizure Susceptibility.Cerebral cortex (New York, N.Y. : 1991). 2017;27(11). • Ruiz-Calvo A, Maroto IB, Bajo-Grañeras R, Chiarlone A, Gaudioso Á, Ferrero JJ et al. Pathway- Specific Control of Striatal Neuron Vulnerability by Corticostriatal Cannabinoid CB1 Receptors. Cerebral cortex (New York, N.Y. : 1991). 2017. • Bravo-Ferrer I., Cuartero M.I., Zarruk J.G., Pradillo J.M., Hurtado O., Romera V.G. et al. Cannabinoid type-2 receptor drives neurogenesis and improves functional outcome after stroke. Stroke. 2017;48(1):204-212. • Sanchez A.-M., Quattrone F., Pannese M., Ulisse A., Candiani M., Diaz-Alonso J. et al. The cannabinoid receptor CB1 contributes to the development of ectopic lesions in a mouse model of endometriosis. Human Reproduction. 2017;32(1):175-184. • Alarcon F., Cedeno Y., de Yebenes J.G. Parkinsonism and other movement disorders in 23 cases of neurocysticercosis. Parkinsonism and Related Disorders. 2017. • Campos A.C., Fogaca M.V., Scarante F.F., Joca S.R.L., Sales A.J., Gomes F.V. et al. Plastic and neuroprotective mechanisms involved in the therapeutic effects of cannabidiol in psychiatric disorders. Frontiers in Pharmacology. 2017;8(MAY). RESEARCH PROJECTS

• Code: NEUROLATAM. • Code: PI15/00339. Title: Unraveling the neurobiological sustrate Title: Papel de la autofagia en cáncer: of protective cannabinoid actions in the brain. mecanismos de muerte mediada Principal Investigator: Ismael Galve Roperh, por autofagia en células tumorales y José A. Ramos Atance. participación de genes reguladores de la CIBERNED’s collaboration: Yes. autofagia en el control de la tumorigénesis. CIBERNED groups: G303 ; G305. Principal Investigato: Guillermo Velasco Diez. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: G305. Funding agency: Unión Iberoamericana de Other CIBER’s collaboration: No. Universidades. Type: National. Funding: 24.225€. Funding agency: Instituto de Salud Carlos III. Duration: 2017-2018 Funding: 202.000 €. Duration: 2016-2018 • Code: EUR.TRAIN16. Title: Tribbles Research and Innovation • Code: PI15/00310. Network. Title: Papel del sistema endocannabinoide Principal Investigato: Guillermo Velasco Díez. en malformaciones del desarrollo cortical CIBERNED’s collaboration: No. asociadas a epilepsia refractaria. CIBERNED groups: G305. Principal Investigato: Ismael Galve Roperh. Other CIBER’s collaboration: CIBERONC. CIBERNED’s collaboration: No. Type: European. Grupos CIBERNED: G305. Funding agency: Comisión Europea. Other CIBER’s collaboration: No. Funding: 452.545,92€. Type: National. Duration: 2016-2020 Agencia financiadora: Instituto de Salud Carlos III. Funding: 110.500 €. Duration: 2016-2018

2017 CIBERNED Annual Report / 154 Program 2 Group: Manuel Guzmán Pastor

• Code: RTC-2015-3364-1. • Code: SAF2015-64945-R. Title: Desarrollo preclínico de VCE-003.2 Tíitle: Identificación y caracterización para el tratamiento de la enfermedad de de subpoblaciones del receptor Huntington (CANNADERIV). CB1 cannabinoide con actividad Principal Investigator: Ismael Galve-Roperh. neuroprotectora. CIBERNED’s collaboration: No. Principal Investigator: Manuel Guzmán CIBERNED groups: G305. Pastor. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: G305. Funding agency: Ministerio de Economía, Other CIBER’s collaboration: No. Industria y Competitividad. Type: National. Funding: 80.800 €. Funding agency: MINECO. Duration: 2016-2017 Funding: 330.000 €. Duration: 2016-2018 PhD DISSERTATIONS

• Author: Adán de Salas Quiroga. • Author: Andrea Ruiz Calvo. Title: Alteraciones celulares y funcionales Title: Mecanismos de neurotoxicidad en el derivadas de la manipulación del receptor circuito corticoestriatal: papel protector del cannabinoide CB1 durante el desarrollo receptor CB1 cannabinoide. cerebral. Date: 31st Mat 2017. Date: 26th May 2017. Supervisor: Manuel Guzmán Pastor. Supervisor: Ismael Galve Roperh.

2017 CIBERNED Annual Report / 155 Teresa Iglesias Vacas 111

Principal Investigator Teresa Iglesias Vacas

Research team

Lucía García Guerra Julia Pose Utrilla Guillermo Cano García PhD fellow PhD student Degree student

Ana Mª del Puerto del Pino Alicia Belmonte Alfaro Jacob Deyell PhD fellow Master´s student Visiting student

Álvaro Sebastián Serrano Samantha Benito Hernández PhD fellow Technician student

Instituto de Investigaciones Biomédicas “Alberto Sols” CSIC-UAM y CIBERNED (ISCIII) Departamento de Fisiología Endocrina y del Sistema Nervioso c/Arturo Duperier, nº4, 28029 Madrid (Spain) Tel.: +34 91 585 44 87 Fax: +34 91 585 44 01 [email protected]

2017 CIBERNED Annual Report / 156 SUMMARY

Excitotoxicity is a type of neuronal death transcription factor NF-kB and decreases in that takes place in numerous human manganese superoxide dismutase, the enzyme neuropathologies, such as stroke, traumatic responsible for the elimination of mitochondrial brain injury, epilepsy, amyotrophic lateral ROS. In addition, we have designed a sclerosis, Parkinson´s disease, Huntington strategy for the neurospecific expression of a disease and Alzheimer´s disease. Thus, acting constitutively active mutant of PKD that confers on the molecular mechanisms involved in strong neuroprotection against excitotoxicity excitotoxicity may confer neuroprotection in primary neuronal cultures and animal in a broad range of acute or chronic models. The use this mutant in different models neurodegenerative conditions. of neurodegenerative diseases in our future preclinical research could have important We investigate excitotoxicity and analyse therapeutic implications. Additionally, we how this process affects prosurvival pathways. have explored possible alterations in Kidins220 We are particularly interested in studying in Huntington´s disease (HD), in the context of the role of protein kinase D (PKD) and its cooperative project CIBERNED 2013/07. substrate Kidins220 in excitotoxic death in different neuropathologies. During last year, Previously, we found that this molecule we have shown that PKD is crucial for neuronal accumulates in samples from patients with survival, and that its activity in healthy neurons Alzheimer´s disease (AD), where it shows potentiates a free-radical detoxification a positive correlation with tau and its pathway, decreasing this way neuronal death hyperphosphorylated forms (López-Menéndez under conditions of high oxidative stress (Pose- & Gamir-Morralla et al, Hum Mol Genet 2013; Utrilla & García-Guerra et al, Nat Commun, Gamir-Morralla et al, J Alzheimers Dis, 2017). Using 2017). We believe that this cascade can decay brain necropsies from HD patients, and animal with aging, leaving neurons less protected and and cellular models, we have found significant Teresa Iglesias Vacas exposed to higher oxidative stress damage. changes in Kidins220 from early presyntomatic stages. These results could indicate that the We have also found that excitotoxicity provokes observed modifications could contribute to PKD inactivation, leading to increases in reactive the pathogenesis of this neurodegenerative oxigen species (ROS), neuronal damage disease. We will continue our research to and neurodegeneration. We have identified determine the contribution of the observed the molecular mechanisms participating in modifications in Kidins220 and PKD to the the silencing of this pathway, that involve etiopathology of neurodegenerative diseases, activation of phosphatases, nuclear exit of the and to design novel neuroprotective strategies. KEYWORDS

Kidins220, Protein Kinase D (PKD), Excitotoxicity, Neuroprotection, Huntington, Alzheimer PUBLICATIONS

• Pose-Utrilla J., Garcia-Guerra L., Del Puerto A., Martin A., Jurado-Arjona J., De Leon-Reyes N.S. et al. Excitotoxic inactivation of constitutive oxidative stress detoxification pathway in neurons can be rescued by PKD1. Nature Communications. 2017;8(1). • Gamir-Morralla A, Belbin O, Fortea J, Alcolea D, Ferrer I, Lleó A et al. Kidins220 Correlates with Tau in Alzheimer’s Disease Brain and Cerebrospinal Fluid.Journal of Alzheimer’s disease : JAD. 2017;55(4). • Fernandez A.M., Hernandez-Garzon E., Perez-Domper P., Perez-Alvarez A., Mederos S., Matsui T. et al. Insulin regulates astrocytic glucose handling through cooperation with IGF-I. Diabetes. 2017;66(1):64-74.

2017 CIBERNED Annnual Report / 157 • Tapia M., Dominguez A., Zhang W., Puerto A.D., Ciorraga M., Benitez M.J. et al. Cannabinoid receptors modulate neuronal morphology and ankyring density at the axon initial segment. Frontiers in Cellular Neuroscience. 2017;11. • Alvaro-Blanco J., Urso K., Chiodo Y., Martin-Cortazar C., Kourani O., Gomez-Del Arco P. et al. MAZ induces MYB expression during the exit from quiescence via the E2F site in the MYB promoter. Nucleic Acids Research. 2017;45(17):9960-9975. • Teresa Miras-Portugal P.M., Sebastian-Serrano A., De Diego Garcia L., Diaz-Hernandez M. Neuronal P2X7 receptor: Involvement in neuronal physiology and. Journal of Neuroscience. 2017;37(30):7063-7072. RESEARCH PROJECTS

• Code: SAF2014-52737-P. • Code: PI2015-2/06. Title: Participación y regulación de PKD y Title: Molecular mechanisms of brain and KIDINS220 en los procesos y rutas moleculares muscle stem cell function in aging and de la neurodegeneración. neurodegeneration. Principal Investigator: Teresa Iglesias Vacas. Principal Investigator: Pura Muñoz Cánoves. CIBERNED’s collaboration: No. CIBERNED’s collaboratio: Yes. CIBERNED groups: G111. CIBERNED groups: G604, G102, G606, G111, Other CIBER’s collaboration: No. G306. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Intramurales. Funding: 193.600 €. Funding agency: CIBERNED. Duration: 2015-2017 Funding: 350.000 €. Duration: 2016-2017

2017 CIBERNED Annual Report / 158 Jon Infante Ceberio 601

Principal Investigator Jon Infante Ceberio

Research team

José Berciario Antonio García Miguel A. Lafarga Eloy Rodríguez Full Professor of Clinical Full Professor of Cell Rodríguez Neurology Neurophysiologist Biology Neurologist Carmen Lage María Teresa Berciario Isabel González- Neurologist Pascual Sánchez-Juan Full Professor of Cell Aramburu Neurologist Biology Neurologist José Ignacio Mateo Neurologist Coro Sánchez- María Bravo Andrés González- Quintana Nurse Mandly Oriol Narcís Majos Lab technician Radilogist PhD student Iñigo Casafont Olga Tapia Martínez PhD assistant lecturer Andrea González Ana Lara Pelayo PhD fellow Suarez Neurologist Marta de la Fuente Neurologist María J. Sedano Clerk Ana Pozueta Neurologist Marta Kazimierczak Psychologist Elena Gallardo Nurse María Sierra Radilogist Javier Riancho- Neurologist Zarrabeitia Neurologist

Servicio de Neurología Hospital Universitario Marqués de Valdecilla Universidad de Cantabria 39008 Santander, España [email protected]

2017 CIBERNED Annual Report / 159 SUMMARY

Peripheral Neuropathies which we evaluated the extensive experience of the Cognitive Impairment Unit in the clinical We have established from our clinico- use of amyloid-PET. Within this same line of pathological studies and from a comprehensive amyloid-PET it should be noted that Dr. Sánchez- review of the literature that the predominant Juan was invited in 2017 to write an editorial in lesions in early GBS are located at the spinal Neurology titled: “Dynamic measurements of roots and nerves. Figure 1 illustrates this β-amyloid accumulation: The early effect of question. After a historical review, we have APOE.” We completed the project “MicroRNA stablished that spinal nerve lesions constitute Profile in Patients with Alzheimer’s Disease the historical legacy of Haymaker´s and “funded by CIBERNED, for the study of value Kernohan´s exceptional clinico-pathological diagnosis of the determination of these study (Medicine 1949; 28: 59-141), a question structures in CSF both free and exosomes, and largely unnoticed in the literature. We have whose results were published in JAD. Within established new neurophysiological criteria in the multi-center project PI16/01652 “Study the intermediate form of CMT for considering of rare variants in genes Associated with a neuropathy of the intermediate type, and Alzheimer’s Disease in the Spanish population” neither axonal or demyelinating. The algorithm we gathered a population of 1000 samples in figure 2 summarizes our contribution. We from Early Onset Alzheimer’s disease patients. have proposed the need of modifying the Coordinated by Sánchez-Juan, this is the first OMIM (Online Mendelian Inheritance in Man) large-scale study of dementia genomics in classification. In national and international Spain. collaborative studies, we have contributed to delimit the nosology of CMT associated Amyotrophic lateral sclerosis (ALS) to GDAP mutations, and have stablished potential biomarkers of CMT1A with PMP22 In 2017, we have continued with our previous duplication. research lines: i) RXR agonists as a new therapy, in collaboration with the University of Santiago Parkinson’s disease de Compostela, ii) metabolic dysfunction and ALS (in collaboration with Biodonostia Institute), We have completed a four-year follow-up of a and iii) the role of environmental factors in the cohort of asymptomatic carriers of the G2019S motor neuron degeneration. Also, we have mutation of the LRRK2 gene, through molecular initiated a collaboration with the researcher imaging (DaTSCAN) and olfaction testing. We Ana María Cuervo from the University Albert have defined the utility of the DaTSCAN asa Einstein in New York in order to characterize tool for predicting the risk of early conversion disturbances in the process of autophagy in to PD in asymptomatic carriers (Sierra et al, fibroblasts. Neurology 2017). We have continued our participation in the COPPADIS study, a national- Cell neurobiology based prospective multi-centric study analysing biomarkers in a large cohort of patients with We have demonstrated that SMN deficiency idiopathic PD (Santos García, BMC Neurology in postnatal MNs from the SMA murine model 2016). produces a depletion of functional Cajal bodies (CBs). Disruption of CBs is the earliest nuclear Ataxias sign of MN degeneration. We demonstrate that depletion of CBs, with loss of CB-nucleolus We have continued the prospective evaluation interactions, induces a progressive nucleolar of patients with dominant ataxias included in dysfunction in ribosome biogenesis, which is EUROSCA-NHS and RISCA studies. We are also associated with a disruption of protein synthesis a collaborator centre for recruitment in the machinery. Also, we have demonstrated, in European study ESMI (European spinocerebellar human cell lines and cultured motor neurons, ataxia type 3/Machado Joseph initiative), that SMN is acetylated in the lysine K119 by the financed in the 2015 JPND call. Also, we have acetiltranferase CBP. Furthermore, we have contributed in a multicenter international study observed that SMN acetylation increases its half- to typify the progression of the body mass index life and regulates its subcellular localization and in dominant ataxias. molecular interactions required for the biogenesis of spliceosomal snRNP and CB assembly. Alzheimer’s disease We have completed the project “Utility of amyloid and FDG-PET in clinical practice” in

2017 CIBERNED Annual Report / 160 Program 2 Group: Jon Infante Ceberio KEYWORDS

CMT, Guillain Barré syndrome, LRRK2, ataxia, Spinal muscular atrophy, Alzheimer´s disease, Parkinson´s disease, ALS, Cajal bodies, nucleolus PUBLICATIONS

• Sims R., Van Der Lee S.J., Naj A.C., Bellenguez C., Badarinarayan N., Jakobsdottir J. et al. Rare coding variants in PLCG2, ABI3, and TREM2 implicate microglial-mediated innate immunity in Alzheimer’s disease. Nature Genetics. 2017;49(9):1373-1384. • Jansen WJ, Ossenkoppele R, Tijms BM, Fagan AM, Hansson O, Klunk WE et al. Association of Cerebral Amyloid-β Aggregation With Cognitive Functioning in Persons Without Dementia. JAMA psychiatry. 2017. • Sánchez-Juan P, Seshadri S. Dynamic measurements of β-amyloid accumulation: The early effect of APOE.Neurology. 2017;89(10). • Sierra M., Martinez-Rodriguez I., Sanchez-Juan P., Gonzalez-Aramburu I., Jimenez-Alonso M., Sanchez-Rodriguez A. et al. Prospective clinical and DaT-SPECT imaging in premotor LRRK2 G2019S-Associated Parkinson disease. Neurology. 2017;89(5):439-444. • Fledrich R., Mannil M., Leha A., Ehbrecht C., Solari A., Pelayo-Negro A.L. et al. Biomarkers predict outcome in Charcot-Marie-Tooth disease 1A. Journal of Neurology, Neurosurgery and Psychiatry. 2017;88(11):941-952. • Rossor A.M., Carr A.S., Devine H., Chandrashekar H., Pelayo-Negro A.L., Pareyson D. et al. Peripheral neuropathy in complex inherited diseases: An approach to diagnosis. Journal of Neurology, Neurosurgery and Psychiatry. 2017;88(10):846-863. • 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. 2017;64. • Montal V., Vilaplana E., Alcolea D., Pegueroles J., Pasternak O., Gonzalez-Ortiz S. et al. Cortical microstructural changes along the Alzheimer’s disease continuum. Alzheimer’s and Dementia. 2017. • Lafarga V., Tapia O., Sharma S., Bengoechea R., Stoecklin G., Lafarga M. et al. CBP-mediated SMN acetylation modulates Cajal body biogenesis and the cytoplasmic targeting of SMN. Cellular and Molecular Life Sciences. 2017;:1-20. • Tapia O., Narcis J.O., Riancho J., Tarabal O., Piedrafita L., Caldero J. et al. Cellular bases of the RNA metabolism dysfunction in motor neurons of a murine model of spinal muscular atrophy: Role of Cajal bodies and the nucleolus. Neurobiology of Disease. 2017;108:83-99. • Sivera R., Frasquet M., Lupo V., Garcia-Sobrino T., Blanco-Arias P., Pardo J. et al. Distribution and genotype-phenotype correlation of GDAP1 mutations in Spain. Scientific Reports. 2017;7(1). • 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. [ePub2017]. • Riancho J., Pozueta A., Santos M., Lage C., Carril J.M., Banzo I. et al. Logopenic Aphasia due to a Strategic Stroke: New Evidence from a Single Case. Journal of Alzheimer’s Disease. 2017;57(3):717-721. • Berciano J, García A, Gallardo E, Peeters K, Pelayo-Negro AL, Álvarez-Paradelo S et al. Intermediate Charcot-Marie-Tooth disease: an electrophysiological reappraisal and systematic review.Journal of neurology. 2017;264(8). • Brun S., Abella N., Berciano M.T., Tapia O., Jaumot M., Freire R. et al. SUMO regulates p21Cip1 intracellular distribution and with p21Cip1 facilitates multiprotein complex formation in the nucleolus upon DNA damage. PLoS ONE. 2017;12(6).

2017 CIBERNED Annnual Report / 161 • Santurtun A., Ruiz P.B., Lopez-Delgado L., Sanchez-Lorenzo A., Riancho J., Zarrabeitia M.T. Stroke: Temporal Trends and Association with Atmospheric Variables and Air Pollutants in Northern Spain. Cardiovascular Toxicology. 2017;17(3):360-367. • Vazquez-Bourgon J., Martino J., Sierra Pena M., Infante Ceberio J., Martinez Martinez M.A., Ocon R. et al. Deep brain stimulation and treatment-resistant obsessive-compulsive disorder: A systematic review. Revista de Psiquiatria y Salud Mental. 2017. • Perez-Campo F.M., May T., Zauers J., Sanudo C., Delgado-Calle J., Arozamena J. et al. Generation and characterization of two immortalized human osteoblastic cell lines useful for epigenetic studies. Journal of Bone and Mineral Metabolism. 2017;35(2):150-160. • Berciano J. Spinal nerve involvement in early Guillain-Barré syndrome: The Haymaker and Kernohan’s legacy.Journal of the neurological sciences. 2017;382. • Berciano J. Progressive multifocal leukoencephalopathy in an immunocompetent patient. Neurologia (Barcelona, Spain). 2017. • Perez-Pereda S., Lage-Martinez C., Paredes M.J., Infante J. Recurrent meningoencephalitis due to allopurinol. Neurologia. 2017. • Berciano J., Garcia A. Letter regarding the article: A novel missense variant (Gln220Arg) of GNB4 encoding guanine nucleotide-binding protein, subunit beta-4 in a Japanese family with autosomal dominant motor and sensory neuropathy. European Journal of Medical Genetics. 2017. • Serrano-Cardenas K.M., Sanchez-Rodriguez A., Pozueta A., Pelayo A.L., Riancho J. Mesial encephalitis: an uncommon presentation of neurosyphilis: a case report and review of the literature. Neurological Sciences. 2017;:1-4. • Gazulla J., Almarcegui C., Berciano J. Reversible inflammatory neuropathy superimposed on Charcot-Marie-Tooth type 1A disease. Neurological Sciences. 2017;:1-2. • Fernández-Torre JL, Paramio-Paz A, López-Delgado A, Martín-García M, González-Aramburu I, Hernández-Hernández MA. De novo absence status epilepticus of late onset (DNASLO) precipitated by oral treatment with cefuroxime: description of an ambulatory case.Epileptic disorders : international epilepsy journal with videotape. 2017. • Jimenez-Bonilla J.F., Quirce R., De Arcocha-Torres M., Martinez-Rodriguez I., Martinez-Amador N., Sanchez-Juan P. et al. 11C-PIB retention patterns in white and grey cerebral matter in idiopathic normal pressure hydrocephalus patients. A visual analysis. Revista Espanola de Medicina Nuclear e Imagen Molecular. 2017. • Moreno F., Indakoetxea B., Barandiaran M., Caballero M.C., Gorostidi A., Calafell F. et al. The unexpected co-occurrence of GRN and MAPT p.A152T in Basque families: Clinical and pathological characteristics. PLoS ONE. 2017;12(6). • Diallo A, Jacobi H, Schmitz-Hübsch T, Cook A, Labrum R, Durr A, Brice A, Charles P, Marelli C, Mariotti C, Nanetti L, et al. Body Mass Index Decline Is Related to Spinocerebellar Ataxia Disease Progression. Movement Disorders Clinical Practice. 2017 Sep 1. RESEARCH PROJECTS

• Code: Tau Consortium (UCSF, USA) 2016. Type: International. Title: Tau Consortium. Funding agency: UCSF. Principal Investigator: Suzee Lee. Funding: 98.000€. CIBERNED’s collaboration: No. Duration: 2014-2018 CIBERNED groups: G601. Other CIBER’s collaboration: No.

2017 CIBERNED Annual Report / 162 Program 2 Group: Jon Infante Ceberio

• Code: PI2016/06. Other CIBER’s collaboration: No. Title: Identificación de vías fisiopatológicas y Type: Intramurales. biomarcadores candidatos en la fase pre- Funding agency: IDIVAL. diagnóstica de la enfermedad de Parkinson. Funding: 74.400€. Principal Investigator: Miquel Vila Bover. Duration: 2015-2017 CIBERNED’s collaboration: Yes. CIBERNED groups: G109, G601, G207, G410. • Code: NVAL 16/21. Other CIBER’s collaboration: No. Title: Enfermedad de Alzheimer y sindrome Type: Intramurales. de Down. Una aproximación clínica y Funding agency: CIBERNED. experimental. Funding: 196.000 €. Principal Investigator: Javier Riancho. Duration: 2016 - 2018 CIBERNED’s collaboration: No. CIBERNED groups: G601. • Code: IGOS. Other CIBER’s collaboration: No. Title: International Guillain-Barre syndrome Type: Intramurales. outcome study (IGOS). Funding agency: IDIVAL. Principal Investigator: Bart Jacobs. Funding: 15.000 €. CIBERNED’s collaborationD: No. Duration: 2017 - 2018 CIBERNED groups: G60. Other CIBER’s collaboration: No. • Code: NVAL17/22. Type: International. Title: Efecto del déficit del factor de Funding agency: Erasmus Medical Center. supervivencia de las neuronas motoras Funding: NP. (SMN) sobre la organización estructural Duration: 2012-2019 y molecular de lkos compartimentos nucleares implicados en la biogénesis • Code: NVAL16/21. de RNPs espliceosomales y en el Title: Diseño de nuevas moléculas para procesamiento de RNAs: estudio experim. el tratamiento de la esclerosis lateral Principal Investigator: Olga Tapia Martínez. amiotrofica-Innopharma. CIBERNED’s collaboration: No. Principal Investigator: Javier Riancho. CIBERNED groups: G601. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G601. Type: Intramurales. Other CIBER’s collaboration: No. Funding agency: IDIVAL. Type: Intramurales. Funding: 8.000 €. Funding agency: IDIVAL. Duration: 2017 - 2019 Funding: 15.000 €. Duration: 2017 - 2018 • Code: INNVAL17/20. Title: Diseño de estructuras biopoliméricas • Code: BFU2014-54754P. funcionalizadas con grafeno para el Title: Regulación por acetilación del factor desarrollo de cultivos neuronales en modelos de supervivencia de las neuronas motoras. celulares de patología de la motoneurona. Su importancia en la biogenésis de snRNPs Principal Investigator: Olga Tapia Martínez. y en el ensamblaje de los cuerpos nucleares CIBERNED’s collaboration: No. de Cajal. CIBERNED groups: G601. Principal Investigator: María Teresa Berciano Other CIBER’s collaboration: No. Blanco, Miguel Ángel Lafarga Coscojuela. Type: Intramurales. CIBERNED’s collaboration: No. Funding agency: IDIVAL. CIBERNED groups: G601. Funding: 12.000 €. Other CIBER’s collaboration: No. Duration: 2017 - 2019 Type: National. Funding agency: MINECO. • Code: PT13/0010/0024. Funding: 115.000€. Title: Plataforma de Biobancos, nodo Hospital Duration: 2015-2017 “Universitario Marques de Valdecilla”. Principal Investigator: Pascual Sánchez • Code: WLA (Contrato PostMir Wenceslao Juan. Lopez Albo) 2015. CIBERNED’s collaboration: No. Title: Epigenética y nuevas tecnologías en las CIBERNED groups: G601. demencias. Una aproximación traslacional Other CIBER’s collaboration: No. y de innovación asistencial. Type: National. Principal Investigator: Pascual Sánchez- Funding agency: Instituto de Salud Carlos III. Juan, Javier Riancho. Funding: 98.000 €. CIBERNED’s collaboration: No. Duration: 2017 CIBERNED groups: G601.

2017 CIBERNED Annual Report / 163 • Code: ESMI. • Code: SMA Europe Call 8 grant application. Title: European Spinocerebellar Ataxia Title: Regulation of the survival motor Type 3/Machado Joseph disease initiative. neuron (SMN) protein by acetylation Principal Investigator: Thomas Klockgether. and its importance in snRNP biogenesis CIBERNED’s collaboration: No. and molecular assembly of Cajal bodies. CIBERNED groups: G601. Principal Investigator: Olga Tapia Martinez. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: European. CIBERNED groups: G601. Funding agency: JPND. Other CIBER’s collaboration: No. Funding: 3.000 €. Type: European. Duration: 2016 - 2019 Funding agency: SMA Europe. Funding: 40.000 €. Duration: 2017 - 2018 PhD DISSERTATIONS

• Author: Jorge Mata Garrido. Title: Organización y dinámica de los focos nucleares permanentes de reparación del daño del DNA en neuronas. Date: 12th January 2017. Supervisor: María Teresa Berciano Blanco.

2017 CIBERNED Annual Report / 164 Jaime Kulisevsky Bojarski 202

Principal Investigator Jaime Kulisevsky Bojarsky

Research team

Ignacio Aracil Bolaños Andrea Horta Barba Javier Pagonabarraga Mora PhD student Research collaborator Researcher

Helena Berj Kasem Marco Cristina Izquierdo Barrionuevo Berta Pascual Sedano Researcher Researcher Researcher

Antonia Campolongo Perillo Juan Martín Lahoz Jesús Pérez Pérez Researcher PhD student PhD student

Carmen García Sánchez Saúl Indra Martínez Horta Frederic Sampedro Santaló Researcher PhD student Researcher

Alexandre Gironell Carreró Researcher

Unidad de trastornos de movimiento, Servicio de Neurología Hospital de la Santa Creu i Sant Pau Sant Antoni M. Claret 08025 Barcelona, Catalunya (Spain) Tel.: +34 93 553 76 13 Fax: +34 93 553 78 72 [email protected]

2017 CIBERNED Annual Report / 165 SUMMARY

Parkinson’s Disease (PD): • Utility of remote motor activity measurement system (PKG, Global A) General Kinetics Corporation). • COPPADIS: COhort of Patients with Huntington Disease (HD): PArkinson’s DIsease in Spain. • Coordination of the “Cognitive Phenotype B) Cognition Working Group” of the European Huntington’s Disease Network. • FIS PI14 / 02058: Blood and • Spanish coordination of Enroll-HD: A neurophysiological markers of progression Prospective Registry Study in a Global of cognitive impairment Huntington’s Disease Cohort. (PI in Spain: J. • Validation of alternative version of Kulisevsky) (CHDI) cognitive scale (PD-CRS). • Validation of scale for cognition: Huntington’s • Validation and normative data of PD-CRS Disease Cognitive Rating Scale (HDCRS) by research groups from other countries. • Diabetes in HD: epidemiology, genetics • Difussion, translation and validation of and potential biomarkers as a therapeutic neuropsychological scales designed strategy. for PD in an international web platform • FIS PI17 / 01885 Longitudinal study of clinical (www.movementscales.com). and neuroradiological correlates associated • Functional neuroimaging of cognitive with levels of huntingtin and other biomarkers impairment associated with PD: in CSF and plasma in HD (BCNHD project) evaluation of functional connectivity (PI: Jesús Pérez) and analysis by graph theory in normal • Collaboration in: a) FIS PI14 / 00834: cognition and mild cognitive impairment. Longitudinal changes in functional and • DuoCog Project: a randomized, double- structural connectivity in pre-symptomatic blind, cross-over study comparing patients; b) FIS PI15 / 02227: Phenotype and immediate levodopa vs. intraduodenal in haplotypes associated with intermediate cognition and mood. alleles of the EH gene; c) HDClarity: a multi- • Ciberned intramural project: randomized, site cerebrospinal fluid collection initiative double-blind, placebo-controlled clinical to facilitate therapeutic development for trial of efficacy and safety of candesartan HD (CHDI); d) Overcoming Depression in vs. placebo in mild cognitive impairment. HD: Cdk5 as a potential biomarker and • Diagnostic precision and neuronal pharmacological target (HD’s disease correlates of the Buschke memory test in Society of America); e) The nuclear lamina mild cognitive impairment. in HD: physiopathology and therapeutic applications (Ramón Areces Foundation) C) Neuropsychiatric disorders: • Study of circadian rhythm in HD. Financing: SEN • International collaborative study on the concept of apathy in PD. Tremor: • Apathy in PD: analysis of structural bases. • Continues Marató TV3 project (nº 477, • Development of a hardware-software 2013): Prediction of impulse control system for a portable neurophysiological disorders (ICD) and apathy. study. • FIS (PI15 / 00962): Anatomical-functional • Joint project with SEIDOR company and correlates of ICD and apathy. University of Vic. • Minor hallucinations: analysis of functional • Study of typification of tremoric syndromes in correlates (rEEG) and neuroimaging Parkinson’s. (fMRI) • Minor hallucinations: disintegration Other neurodegenerative diseases: of the default neural network as the pathophysiological basis. • Validation of the diagnostic platform for neurodegenerative diseases through D) Advanced PD: OSCANN oculográfico registry. Multicenter study of patients with Alzheimer’s, PD • Continues the analysis of nutritional status andfrontotemporal dementia vs. controls. in patients with intraduodenal levodopa • Lewy Body dementia: participation in an • Prospective cognitive, behavioral, sleep international multi-center study integrated in and quality of life study in patients with the “Joint Program for Neurodegenerative deep brain stimulation and intraduodenal Diseases” (European Commission). infusion of levodopa. • MSA: Collaborative project within the

2017 CIBERNED Annual Report / 166 Program 2 Group: Jaime Kulisevsky Bojarski

Catalan MSA Registry (CMSAR). Neuroimagen and data engineering: • FXTAS: Assistance and collaborative research with Genetics of Hospital Clínic (Barcelona) • Functional and structural cerebral correlates and Associació Catalana X Fràgil (ACXF). of: apathy in HD, minor hallucinations and mild cognitive impairment in PD. KEYWORDS

Parkinson, Cognition, Mild Cognitive Impairment, Hallucinations, Huntington, FxTAS, AMS, Essential Tremor, Neuroimaging, Data Engineering PUBLICATIONS

• Luquin MR, Kulisevsky J, Martinez-Martin P, Mir P, Tolosa ES. Consensus on the Definition of Advanced Parkinson’s Disease: A Neurologists-Based Delphi Study (CEPA Study).Parkinson’s disease. ;2017. • Cortes-Vicente E., Pradas J., Marin-lahoz J., De Luna N., Clarimon J., Turon-Sans J. et al. Early diagnosis of amyotrophic lateral sclerosis mimic syndromes: pros and cons of current clinical diagnostic criteria. Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration. 2017;18(5- 6):333-340. • Catalan M.J., Antonini A., Calopa M., Bajenaru O., de Fabregues O., Minguez-Castellanos A. et al. Can suitable candidates for levodopa/carbidopa intestinal gel therapy be identified using current evidence?. eNeurologicalSci. 2017;8:44-53. • 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. [ePub2017]. • Ruiz-Idiago J.M., Floriach M., Mareca C., Salvador R., Lopez-Sendon J.L., Mananes V. et al. Spanish validation of the problem behaviors assessment–short (PBA-s) for Huntington’s disease. Journal of Neuropsychiatry and Clinical Neurosciences. 2017;29(1):31-38. • Schapira AH, Fox SH, Hauser RA, Jankovic J, Jost WH, Kenney C et al. Assessment of Safety and Efficacy of Safinamide as a Levodopa Adjunct in Patients With Parkinson Disease and Motor Fluctuations: A Randomized Clinical Trial.JAMA neurology. 2017;74(2). • Spataro N., Roca-Umbert A., Cervera-Carles L., Valles M., Anglada R., Pagonabarraga J. et al. Detection of genomic rearrangements from targeted resequencing data in Parkinson’s disease patients. Movement Disorders. 2016. • Garcia-Gorro C., de Diego-Balaguer R., Martinez-Horta S., Perez-Perez J., Kulisevsky J., Rodriguez-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. [ePub2017]. • Bellosta Diago E., Perez Perez J., Santos Lasaosa S., Viloria Alebesque A., Martinez Horta S., Kulisevsky J. et al. Circadian rhythm and autonomic dysfunction in presymptomatic and early Huntington’s disease. Parkinsonism and Related Disorders. 2017. • Antonini A., Poewe W., Chaudhuri K.R., Jech R., Pickut B., Pirtosek Z. et al. Levodopa-carbidopa intestinal gel in advanced Parkinson’s: Final results of the GLORIA registry. Parkinsonism and Related Disorders. 2017;45:13-20. • Garcia-Sanz P., Orgaz L., Bueno-Gil G., Espadas I., Rodriguez-Traver E., Kulisevsky J. et al. N370S- GBA1 mutation causes lysosomal cholesterol accumulation in Parkinson’s disease. Movement Disorders. 2017;32(10):1409-1422.

2017 CIBERNED Annual Report / 167 • Gelpi E, Botta-Orfila T, Bodi L, Marti S, Kovacs G, Grau-Rivera O et al. Neuronal intranuclear (hyaline) inclusion disease and fragile X-associated tremor/ataxia syndrome: a morphological and molecular dilemma.Brain : a journal of neurology. 2017;140(8). • Ayala A., Trivino-Juarez J.M., Forjaz M.J., Rodriguez-Blazquez C., Rojo-Abuin J.-M., Martinez- Martin P. et al. Parkinson’s disease severity at 3 years can be predicted from non-motor symptoms at baseline. Frontiers in Neurology. 2017;8(OCT). • Fernandez-Bobadilla R., Martinez-Horta S., Marin-Lahoz J., Horta-Barba A., Pagonabarraga J., Kulisevsky J. Development and validation of an alternative version of the Parkinson’s Disease- Cognitive Rating Scale (PD-CRS). Parkinsonism and Related Disorders. 2017. • Colom-Cadena M., Grau-Rivera O., Planellas L., Cerquera C., Morenas E., Helgueta S. et al. Regional overlap of pathologies in lewy body disorders. Journal of Neuropathology and Experimental Neurology. 2017;76(3):216-224. • Pagonabarraga J, Kulisevsky J. Apathy in Parkinson’s Disease.International review of neurobiology. ;133. • Pagonabarraga J., Kulisevsky J. Safinamide from daily clinical practice: First clinical steps Safinamida desde la práctica clínica diaria: Primeros pasos clínicos. Revista de Neurologia. 2017;65(10):433-438. • Rosa-Grilo M, Qamar MA, Taddei RN, Pagonabarraga J, Kulisevsky J, Sauerbier A et al. Rotigotine transdermal patch and sleep in Parkinson’s disease: where are we now?. NPJ Parkinson’s disease. ;3. RESEARCH PROJECTS

• Code: 477/U/2014 MARATO. Other CIBER’s collaboration: No. Title: Prediction of apathy and impulse Type: Private. control disorders in Parkinson Disease based Funding agency: Fundació La Marató de on feedback related negativity. TV3. Principal Investigator: Jaime Kulisevsky. Funding: 199.486,25 €. CIBERNED’s collaboration: No. Duation: 2015-2018 CIBERNED groups: G202. Other CIBER’s collaboration: No. • Code: PI15/00962. Type: Private. Title: Correlatos anatómico-funcionales Funding agency: Fundació La Marató de de los trastornos del control de impulsos y TV3. apatía en la enfermedad de Parkinson. Funding: 199.607,5 €. Principal Investigator: Jaime Kulisevsky. Duation: 2015-2017 CIBERNED’s collaboration: No. CIBERNED groups: G202. • Code: 484/U/2014 MARATO. Other CIBER’s collaboration: No. Title: Blood-based and neurophysiological Type: National. markers of cognitive deterioration and Funding agency: Instituto de Salud Carlos III. dementia in Parkinson’s disease. Funding: 74.415€. Principal Investigator: Javier Pagonabarraga. Duation: 2016-2018 CIBERNED’s collaboration: No. CIBERNED groups: G202.

2017 CIBERNED Annual Report / 168 Program 2 Group: Jaime Kulisevsky Bojarski

• Code: CM15/00071. • Code: PI2015/02. Title: Río Hortega 2015. Title: Validación de nuevas dianas Principal Investigator: Juan Marín Lahoz. terapéuticas y nuevos biomarcadores en CIBERNED’s collaboration: No. enfermedad de Parkinson. CIBERNED groups: G202. Principal Investigator: José Luis Labandeira Other CIBER’s collaboration: No. García. Type: National. CIBERNED’s collaboration: Yes. Funding agency: Instituto de Salud Carlos III. CIBERNED groups: G208, G101, G202, G203. Funding: 53.732 €. Other CIBER’s collaboration: No. Duration: 2016-2017 Type: Intramurales. Funding agency: CIBERNED. Funding: 240.000 €. Duration: 2015-2017 PhD DISSERTATIONS

• Author: Mª Pilar Sanz Cartagena. • Author: Ramón Fernández de Bobadilla. Title: Epidemiology, pathophysiology and Title: Desarrollo y validación de nuevas treatment implications of non-dopaminergic herramientas para la valoración cognitiva y life threatening symptoms in Parkinson funcional del deterioro cognitivo leve en la Disease: Oropharyngeal dysphagia and enfermedad de Parkinson. balance impariment. Date: 22th September 2017. Date: 2nd Dicember 2017. Supervisor: Jaime Kulisevsky Bojarski. Supervisor: Jaime Kulisevsky Bojarski.

2017 CIBERNED Annnual Report / 169 José Luis Labandeira García 208

Principal Investigator José Luis Labandeira García

Research team

María J. Guerra Seijas Carmen Díaz Ruiz Mª Alicia Costa Besada Full professor Associate professor Predoctoral researcher

Ramón Soto Otero Begoña Villar Cheda María García Garrote Full professor Associate professor Predoctoral researcher

Estefanía Méndez Álvarez Pablo Garrido Gil Andrea López López Full professor Postdoctoral researcher Predoctoral researcher

Jannette Rodríguez Pallares Rita Valenzuela Limiñana María Ángeles Pedrosa Associate professor Postdoctoral researcher Predoctoral researcher

Ana M. Muñoz Patiño Juan A. Parga Martín Pilar Aldrey García Associate professor Postdoctoral researcher Specialist Technician

Ana I. Rodríguez Pérez Antonio Domínguez Meijide Iria Novoa Pérez Associate professor Postdoctoral researcher Specialist Technician

Center for Research in Molecular Medicine Mª Cristina Gianzo Quintela and Chronic Diseases (CIMUS) Specialist Technician Universidad de Santiago de Compostela Avenida Barcelona, 22 Santiago de Compostela (Spain) Tel.: +34 88 181 22 23/ 54 71 Fax: +34 88 181 23 78 [email protected]

2017 CIBERNED Annual Report / 170 SUMMARY

In 2017, we have investigated mechanisms and the mechanisms involved in the increase involved in progression of Parkinson’s disease in RAS activity and glial neuroinflammatory (PD) and higher vulnerability of dopaminergic response induced by a decrease in dopamine neurons with aging. We investigated the role levels (Brain Behav Immun 62:277-290). of several factors involved in the progression of dopaminergic degeneration, including IGF- We discovered an intracellular or intracrine RAS 1 (Mol. Neurobiol. 2017 doi:10.1007/s12035- in dopaminergic neurons (Cell Death and Dis 017-0681-5), prostaglandin EP2 receptors (Mol. 8(9):e3044; Mol. Neurobiol. 2017, doi: 10.10007/ Neurobiol. 2017 doi:10.1007/s12035-017-0681- s12035-017-0805). Interestingly, this intracellular 5), or metals such as cooper (J. Neurochem. system buffers the deleterious pro-oxidative 141: 738-749). We have also investigated effects of the extracellular paracrine system, mechanisms involved in L-dopa-induced which must be taken into account for the dyskinesias, particularly the role of BDNF and design of new therapeutical strategies based serotonergic striatal system (Exp. Neurol 297:73- on brain RAS modulation. 81), and CB1/CB2 endocannabinoid receptors (Brain Behav. Immun. 67:139-151). Finally, we developed new laser capture microdissection protocols for gene expression We have particularly investigated the role analysis in the brain (Histochem. Cell Biol. of the brain renin-angiotensin system (RAS) 148:299-311), and we collaborated in the in neuroinflammation and progression of development of new PD models in primates dopaminergic degeneration (Front. Aging (Brain Struct. Func. 223:343-55; J. Neural Transm. Neurosci. 9:129). We have focused on 125:575-589). An important part of the above- identification and functional characterization mentioned studies and experiments undertaken of new RAS components, such as MAS receptors in 2017 have been performed in collaboration or prorrenin, in the basal ganglia of rodents and with different Ciberned research groups: Dr. primates (Brain Struct Funct. 222:2559-71; Mol. Lanciego, Drs R. Franco/Canela/, Dr. Kulisevsky, Neurobiol. 2017, doi: 10.10007/s12035-017-0805), Dr. Cuadrado, Dra. Moratalla. KEYWORDS

Neurodegeneration, neuroprotection, neuroinflammation, Parkinson, aging, angiotensin, cell therapy PUBLICATIONS

• Costa-Besada M.A., Valenzuela R., Garrido-Gil P., Villar-Cheda B., Parga J.A., Lanciego J.L. et al. Paracrine and Intracrine Angiotensin 1-7/Mas Receptor Axis in the Substantia Nigra of Rodents, Monkeys, and Humans. Molecular Neurobiology. 2017;:1-21. • Garrido-Gil P., Rodriguez-Perez A.I., Fernandez-Rodriguez P., Lanciego J.L., Labandeira-Garcia J.L. Expression of angiotensinogen and receptors for angiotensin and prorenin in the rat and monkey striatal neurons and glial cells. Brain Structure and Function. 2017;222(6):2559-2571. • Labandeira-Garcia J.L., Rodriguez-Perez A.I., Garrido-Gil P., Rodriguez-Pallares J., Lanciego J.L., Guerra M.J. Brain renin-angiotensin system and microglial polarization: Implications for aging and neurodegeneration. Frontiers in Aging Neuroscience. 2017;9(MAY). • Pignataro D, Sucunza D, Rico AJ, Dopeso-Reyes IG, Roda E, Rodríguez-Perez AI et al. Gene therapy approaches in the non-human primate model of Parkinson’s disease.Journal of neural transmission (Vienna, Austria : 1996). 2017. Parga J.A., Garcia-Garrote M., Martinez S., Raya A., Labandeira-Garcia J.L., Rodriguez-Pallares J. Prostaglandin EP2 Receptors Mediate Mesenchymal Stromal Cell-Neuroprotective Effects on Dopaminergic Neurons. Molecular Neurobiology. 2017;:1-14.

2017 CIBERNED Annual Report / 171 • Dominguez-Meijide A., Rodriguez-Perez A.I., Diaz-Ruiz C., Guerra M.J., Labandeira-Garcia J.L. Dopamine modulates astroglial and microglial activity via glial renin-angiotensin system in cultures. Brain, Behavior, and Immunity. 2017;62:277-290. • Dopeso-Reyes I.G., Sucunza D., Rico A.J., Pignataro D., Marin-Ramos D., Roda E. et al. Glucocerebrosidase expression patterns in the non-human primate brain. Brain Structure and Function. 2017;:1-13. • Labandeira-Garcia J.L., Costa-Besada M.A., Labandeira C.M., Villar-Cheda B., Rodriguez- Perez A.I. Insulin-like growth factor-1 and neuroinflammation. Frontiers in Aging Neuroscience. 2017;9(NOV). • Garrido-Gil P., Fernandez-Rodriguez P., Rodriguez-Pallares J., Labandeira-Garcia J.L. Laser capture microdissection protocol for gene expression analysis in the brain. Histochemistry and Cell Biology. 2017;148(3):299-311. • Villar-Cheda B, Costa-Besada MA, Valenzuela R, Perez-Costas E, Melendez-Ferro M, Labandeira- Garcia JL. The intracellular angiotensin system buffers deleterious effects of the extracellular paracrine system.Cell death & disease. 2017;8(9). • Tronci E., Napolitano F., Munoz A., Fidalgo C., Rossi F., Bjorklund A. et al. BDNF over-expression induces striatal serotonin fiber sprouting and increases the susceptibility to L-DOPA-induced dyskinesia in 6-OHDA-lesioned rats. Experimental Neurology. 2017;297:73-81. • 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. [ePub2017]. • Cruces-Sande A., Mendez-Alvarez E., Soto-Otero R. Copper increases the ability of 6-hydroxydopamine to generate oxidative stress and the ability of ascorbate and glutathione to potentiate this effect: potential implications in Parkinson’s disease. Journal of Neurochemistry. 2017;141(5):738-749.

RESEARCH PROJECTS

• Code: RD16/0011. • Code: PI 2015/02. Title: Red de Terapia Celular. Title: Validación de nuevas dianas Principal Investigator: Isabel Fariñas. terapéuticas y nuevos biomarcadores en CIBERNED’s collaboration: Yes. enfermedad de Parkinson. CIBERNED groups: G607, G301, G102, G113, Principal Investigator: José Luis Labandeira. G208, G105, G207. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G208, G101, G202, G203. Type: National. Other CIBER’s collaboration: No. Funding agency: Instituto de Salud Carlos III. Type: Intramurales. Funding: 284.999€. Funding agency: CIBERNED. Duration: 2016-2020 Funding: 240.000 €. Duration: 2015-2017 • Code: R2014/050. Title: Red Gallega de Terapia celular. • Code: GRC2014/002. Principal Investigator: José Luis Labandeira Title: Consolidación y estructuración de García. grupos de referencia competitiva. CIBERNED’s collaboration: No. Principal Investigator: José Luis Labandeira CIBERNED groups: G208. García. Other CIBER’s collaboration: CIBEROBN. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: G208. Funding agency: Instituto de Salud Carlos III. Other CIBER’s collaboration: No. Funding: 120.000€. Type: CC.AA. Duration: 2017-2018 Funding agency: Xunta Galicia. Funding: 320.000 €. Duration: 2015-2018

2017 CIBERNED Annual Report / 172 Program 2 Group: José Luis Labandeira García

• Code: RD16/0011/0016. • Code: BFU2015-70523. Title: Combined protective/restorative cell- Title: Sistema renina-angiotensina cerebral mediated strategies for neurodegenerative en neuroinflamación y degeneración diseases. dopaminérgica. Más allá del eje AII/AT1/ Principal Investigator: José luis Labandeira. NADPH-oxidase. CIBERNED’s collaboration: Yes. Principal Investigator: José Luis Labandeira. CIBERNED groups: G208, G102, G113, G105, CIBERNED’s collaboration: No. G207. CIBERNED groups: G208. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: National. Funding agency: Instituto de Salud Carlos III. Funding agency: MINECO. Funding: 240.245€. Funding: 296.500 €. Duration: 2017-2021 Duration: 2016-2018

2017 CIBERNED Annnual Report / 173 José Luis Lanciego Pérez 203

Principal Investigator José Luis Lanciego Pérez

Research team

Gloria González-Aseguinolaza Iria María González-Dopeso Diego Sucunza Guibert Researcher Reyes Researcher PhD fellow Alberto José Rico Martín Elvira Roda Recalde PhD fellow José Diego Pignataro Lab technician PhD student

Fundación para la Investigación Médica Aplicada (FIMA) Centro de Investigación Médica Aplicada (CIMA) Departamento de Neurociencias Laboratorio de Anatomía Funcional de Ganglios Basales Avenida Pío XII, Edificio CIMA 31008 Pamplona, Navarra (Spain) Tel.: +34 94 819 47 00 (2002) Fax: +34 94 819 47 15 [email protected]

2017 CIBERNED Annual Report / 174 SUMMARY

Studies on heteromers made of G protein- these including Parkinson’s disease, demetia coupled receptors (GPCRs). with Lewy bodies (DLB) and multiple system atrophy (MSA). It might be argued that GPCRs have a natural tendency to form heteromeric complexes • NHP models of PD with AAV9-SynA53T. that represent novel molecular entities • NHP models of DLB with AAV2-retro-SynA53T with properties distinct from those of each • NHP models of MSA with AAV-Olig001-SynA53T component receptor, when considered separately. GPCR heteromers represent novel The ultimate goal is to use gene therapy targets for pharmacological developments approaches to further increase the activity in the field of Parkinson’s disease. At present, of glucocerebrosidase (GCase). GCase our main activities focus on GPCR heteromers enhanced activity is expected to induce a safe made of dopaminergic receptors (D1 and D2), and efficient clearance of aggregated alpha- as well as those formed by endocannabinoid synuclein, thus slowing-down (and ideally receptors (CB1, CB2 and GPR55). arrest), the progressive course that typically characterizes these neurodegenerative Gene therapy for Parkinson’s disease. disorders.

We are currently using adeno-associated • Gene therapy with AAV9-GBA1 for PD viral vectors for modeling a number of • Gene therapy with AAV2-retro-GBA1 for DLB synucleinopathies in non-human primates, • Gene therapy with AAV-Olig001-GBA1 for MSA

José Luis Lanciego Pérez KEYWORDS

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

• Dopeso-Reyes I.G., Sucunza D., Rico A.J., Pignataro D., Marin-Ramos D., Roda E. et al. Glucocerebrosidase expression patterns in the non-human primate brain. Brain Structure and Function. 2017;:1-13. • Morabito G., Giannelli S.G., Ordazzo G., Bido S., Castoldi V., Indrigo M. et al. AAV-PHP.B- Mediated Global-Scale Expression in the Mouse Nervous System Enables GBA1 Gene Therapy for Wide Protection from Synucleinopathy. Molecular Therapy. 2017. • Martínez-Pinilla E, Varani K, Reyes-Resina I, Angelats E, Vincenzi F, Ferreiro-Vera C et al. Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors.Frontiers in pharmacology. ;8. • Labandeira-Garcia J.L., Rodriguez-Perez A.I., Garrido-Gil P., Rodriguez-Pallares J., Lanciego J.L., Guerra M.J. Brain renin-angiotensin system and microglial polarization: Implications for aging and neurodegeneration. Frontiers in Aging Neuroscience. 2017;9(MAY). • Pignataro D., Sucunza D., Vanrell L., Lopez-Franco E., Dopeso-Reyes I.G., Vales A. et al. Adeno- associated viral vectors serotype 8 for cell-specific delivery of therapeutic genes in the central nervous system. Frontiers in Neuroanatomy. 2017;11. • Garrido-Gil P., Rodriguez-Perez A.I., Fernandez-Rodriguez P., Lanciego J.L., Labandeira-Garcia J.L. Expression of angiotensinogen and receptors for angiotensin and prorenin in the rat and monkey striatal neurons and glial cells. Brain Structure and Function. 2017;222(6):2559-2571.

2017 CIBERNED Annual Report / 175 • Pignataro D, Sucunza D, Rico AJ, Dopeso-Reyes IG, Roda E, Rodríguez-Perez AI et al. Gene therapy approaches in the non-human primate model of Parkinson’s disease.Journal of neural transmission (Vienna, Austria : 1996). 2017. • Costa-Besada M.A., Valenzuela R., Garrido-Gil P., Villar-Cheda B., Parga J.A., Lanciego J.L. et al. Paracrine and Intracrine Angiotensin 1-7/Mas Receptor Axis in the Substantia Nigra of Rodents, Monkeys, and Humans. Molecular Neurobiology. 2017;:1-21. • 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. [ePub2017]. RESEARCH PROJECTS

• Code: 20141330. • Code: PI 2015/02. Title: Cannabinoid receptor heteromeric Title: Validación de nuevas dianas complexes as therapeutic targets in terapéuticas y nuevos biomarcadores en Parkinson’s disease. enfermedad de Parkinson. Principal Investigator: Rafael Franco Principal Investigator: José Luis Labandeira. Fernández. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G208, G101, G202, G203. CIBERNED groups: G201, G203. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: Private. Funding agency: CIBERNED. Funding agency: Fundació La Marató de Funding: 240.000 €. TV3. Duration: 2015-2017 Funding: 150.000 €. Duration: 2015-2017

• Code: 340527 ERC Advanced. Title: New Experimental therapeutic approaches for Parkinson’s disease by direct DA neuronal reprogramming. Principal Investigator: José Luis Lanciego. CIBERNED’s collaboration: No. CIBERNED groups: G203. Other CIBER’s collaboration: No. Type: European. Funding agency: Comisión Europea. Funding: 2.415.767 €. Duration: 2014-2018

2017 CIBERNED Annual Report / 176 José López Barneo 105

Principal Investigator José López Barneo

Research team

Ana María Muñoz Cabello Gloria Paula García Flores José María Cabeza Fernández PhD fellow Technician Technician

Alejandro Moreno Domínguez Hee-Sool Rho Juan José Toledo Aral PhD fellow PhD fellow Researcher

Candela Caballero Erazo Helia Sarmiento Soto Lin Gao Physician Technician Researcher

Daniel Enterría Morales Hortensia Torres Torrelo Olaia Colinas Miranda PhD student PhD student Technician

Daniel Cabello Rivera Ignacio Arias Mayenco Pablo Mir Rivera PhD student Technician Physician

Francisco Javier Villadiego Ivette López López Patricia Ortega Sáenz Luque Technician Researcher PhD student Pilar Gómez Garre Instituto de Biomedicina de Sevilla Researcher Hospital Universitario Virgen del Rocío CISC, Universidad de Sevilla Victoria Bonilla Henao Avenida Manuel Siurot s/n Technician 41013 Sevilla (Spain) Tel.: +34 95 592 30 00 Fax: +34 95 592 31 01 Xavier d’Anglemont de Tassigny [email protected] PhD fellow

2017 CIBERNED Annual Report / 177 SUMMARY

During 2017 the research activity in our group glomus cells (Sobrino et al., EMBO Reports, has focused on the following main lines. 2018). We have continued the study of the molecular We have continued our study of the GDNF- mechanisms underlying sensitivity to hypoxia of producing parvalbumin (PV) positive striatal carotid body (CB) glomus cells. This is a research gabaergic interneurons. GDNF produced topic in our group for over 30 years that after by these PV positive interneurons (only in the several seminal contributions is now giving rise striatum and not in other locations such as the to a comprehensive model of acute oxygen cerebral cortex) is essential for the maintenance sensing. In recent publications (see Fernández- of the dopaminergic nigrostriatal pathway. We Agüera et al., Cell Metabolism, 2015; Gao et al., have performed a genetic profiling analysis of Journal of Physiology, 2017) we have shown that the GDNF-producing interneurons and selected oxygen-sensitive glomus cells have metabolic a group of genes, which may be responsible for specializations that determine the production the selective production of GDNF. Currently, of NADH and reactive oxygen species (ROS) we are testing pharmacologic compounds during hypoxia; this leads to modulation of that increase endogenous production of plasmalemmal ion channels, calcium influx and GDNF. These compounds could eventually be neurotransmitter release. This model is being of medical interest in the treatment of Parkinson confirmed in new experimental models using disease. genetically modified mice. Some of the data obtained so far is currently under evaluation in Our group has recently carried out a research a leading scientific journal. project on central neurogenesis in the adult brain (d’Anglemont de Tassigny et al., Hypoxia, We have previously shown that the adult CB is 2015), but how hypoxia (which might be a a neurogenic niche that contains multipotent risk factor in neurodegenerative diseases; see neural stem cells. In response to hypoxia, these Serrano-Pozo et al., Plos One, 2017) affects cells proliferate and eventually differentiate neurogenesis and the potential adaptive role into chemoreceptor glomus cells and other cell of the arterial chemoreceptors is poorly known. types (see Platero-Luengo et al., Cell, 2014). During 2017 we have started the study of mice In 2017 we have shown a new form of “fast models lacking a functional CB to investigate neurogenesis” which depends on the existence whether the lack of adaptation to hypoxia of immature cells (neuroblasts) that in few hours alters neurogenesis or stimulates the initiation of can complete their differentiation into mature neurodegenerative pathologies. KEYWORDS

Oxygen sensing, ion channels, hypoxia, carotid body, dopamine, Parkinson disease, nigrostriatal pathway, GDNF, parvalbumin positive interneurons PUBLICATIONS

• Huertas I., Oldehinkel M., van Oort E.S.B., Garcia-Solis D., Mir P., Beckmann C.F. et al. A Bayesian spatial model for neuroimaging data based on biologically informed basis functions. NeuroImage. 2017;161:134-148. • Serrano-Pozo A., Sanchez-Garci M.A., Heras-Garvin A., March-Diaz R., Navarro V., Vizuete M. et al. Acute and Chronic Sustained Hypoxia Do Not Substantially Regulate Amyloid-β Peptide Generation In Vivo. PLoS ONE. 2017;12(1). • Huertas I., Jesus S., Lojo J.A., Garcia-Gomez F.J., Caceres-Redondo M.T., Oropesa-Ruiz J.M. et al. Lower levels of uric acid and striatal dopamine in non-tremor dominant Parkinson’s disease subtype. PLoS ONE. 2017;12(3).

2017 CIBERNED Annual Report / 178 Program 2 Group: José López Barneo

• Schellenberg MJ, Lieberman JA, Herrero-Ruiz A, Butler LR, Williams JG, Muñoz-Cabello AM et al. ZATT (ZNF451)-mediated resolution of topoisomerase 2 DNA-protein cross-links.Science (New York, N.Y.). 2017;357(6358). • Luquin MR, Kulisevsky J, Martinez-Martin P, Mir P, Tolosa ES. Consensus on the Definition of Advanced Parkinson’s Disease: A Neurologists-Based Delphi Study (CEPA Study).Parkinson’s disease. ;2017. • 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 autónomas. Neurologia. 2018. [ePub2017]. • 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). [ePub2017]. • Di Biase L., Brittain J.-S., Shah S.A., Pedrosa D.J., Cagnan H., Mathy A. et al. Tremor stability index: A new tool for differential diagnosis in tremor syndromes. Brain. 2017;140(7):1977-1986. • Huertas I., Jesus S., Garcia-Gomez F.J., Lojo J.A., Bernal-Bernal I., Bonilla-Toribio M. et al. Genetic factors influencing frontostriatal dysfunction and the development of dementia in Parkinson’s disease. PLoS ONE. 2017;12(4). • Romero-Moya D, Santos-Ocaña C, Castaño J, Garrabou G, Rodríguez-Gómez JA, Ruiz-Bonilla V et al. Genetic Rescue of Mitochondrial and Skeletal Muscle Impairment in an Induced Pluripotent Stem Cells Model of Coenzyme Q10 Deficiency.Stem cells (Dayton, Ohio). 2017;35(7). • Villadiego J, Labrador-Garrido A, Franco JM, Leal-Lasarte M, De Genst EJ, Dobson CM et al. Immunization with α-synuclein/Grp94 reshapes peripheral immunity and suppresses microgliosis in a chronic Parkinsonism model.Glia. 2017. • Gao L., Gonzalez-Rodriguez P., Ortega-Saenz P., Lopez-Barneo J. Redox signaling in acute oxygen sensing. Redox Biology. 2017;12:908-915. • Tejera-Parrado C., Jesus S., Huertas-Fernandez I., Bernal-Bernal I., Bonilla-Toribio M., Cordoba- Tevar I. et al. Genetic analysis of CHCHD2 in a southern Spanish population. Neurobiology of Aging. 2017;50:169.e1-169.e2. • López-Barneo J. All for one - O2 -sensitive K+ channels that mediate carotid body activation. The Journal of physiology. 2017. • Gao L, Bonilla-Henao V, García-Flores P, Arias-Mayenco I, Ortega-Sáenz P, López-Barneo J. Gene expression analyses reveal metabolic specifications in acute O2 -sensing chemoreceptor cells.The Journal of physiology. 2017;595(18). • Vulinovic F., Schaake S., Domingo A., Kumar K.R., Defazio G., Mir P. et al. Screening study of TUBB4A in isolated dystonia. Parkinsonism and Related Disorders. 2017;41:118-120. • Garcia-Sanz P., Orgaz L., Bueno-Gil G., Espadas I., Rodriguez-Traver E., Kulisevsky J. et al. N370S- GBA1 mutation causes lysosomal cholesterol accumulation in Parkinson’s disease. Movement Disorders. 2017;32(10):1409-1422. • Lukovic D., Diez Lloret A., Stojkovic P., Rodraiguez-Martainez D., Perez Arago M.A., Rodriguez- Jimenez F.J. et al. Highly efficient neural conversion of human pluripotent stem cells in adherent and animal-free conditions. Stem Cells Translational Medicine. 2017;6(4):1217-1226. • Ayala A., Trivino-Juarez J.M., Forjaz M.J., Rodriguez-Blazquez C., Rojo-Abuin J.-M., Martinez- Martin P. et al. Parkinson’s disease severity at 3 years can be predicted from non-motor symptoms at baseline. Frontiers in Neurology. 2017;8(OCT). • Abdulkadir M., Londono D., Gordon D., Fernandez T.V., Brown L.W., Cheon K.-A. et al. Investigation of previously implicated genetic variants in chronic tic disorders: a transmission disequilibrium test approach. European Archives of Psychiatry and Clinical Neuroscience. 2017;:1-16. • De Tassigny X.D., Jayasena C., Murphy K.G., Dhillo W.S., Colledge W.H. Mechanistic insights into the more potent effect of KP-54 compared to KP-10 in vivo. PLoS ONE. 2017;12(5).

2017 CIBERNED Annual Report / 179 • Ruiz-Idiago J.M., Floriach M., Mareca C., Salvador R., Lopez-Sendon J.L., Mananes V. et al. Spanish validation of the problem behaviors assessment–short (PBA-s) for Huntington’s disease. Journal of Neuropsychiatry and Clinical Neurosciences. 2017;29(1):31-38. • 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. [ePub2017]. • Benitez-Rivero S., Palomar F.J., Martin-Rodriguez J.F., Alvarez de Toledo P., Lama M.J., Huertas- Fernandez I. et al. Abnormal sensorimotor integration correlates with cognitive profile in vascular parkinsonism. Journal of the Neurological Sciences. 2017;377:161-166. • d’Anglemont de Tassigny X. Outlook on the neuroprotective effect of estrogen.Neural regeneration research. 2017;12(11). RESEARCH PROJECTS

• Code: RD16/0011. Type: European. Title: Red de Terapia Celular. Funding agency: Comisión Europea. Principal Investigator: Isabel Fariñas. Funding: 671.167,5 €. CIBERNED’s collaboration: Yes. Duration: 2015-2019 CIBERNED groups: G607, G301, G102, G113, G208, G105, G207. • Code: PIE 2012 Excelencia (P12-CTS-2739 Other CIBER’s collaboration: No. MO). Type: National. Title: Terapia Celular en la Enfermedad de Funding agency: Instituto de Salud Carlos III. Parkinson. Funding: 284.999€. Principal Investigator: Juan José Toledo Duration: 2016-2020 Aral. CIBERNED’s collaborationD: No. • Code: ERC 2014. CIBERNED groups: G105. Title: Molecular mechanisms of acute Other CIBER’s collaborations: No. oxygen sensing (OXYGENSENSING). Type: CC.AA. Principal Investigator: José López Barneo. Funding agency: Junta de Andalucía. CIBERNED’s collaboration: No. Funding: 191.875€. CIBERNED groups: G105. Duration: 2014-2017 Other CIBER’s collaboration: No. Type: European. • Code: PI-0125-2016. Funding agency: Comisión Europea. Title: Complicaciones cardiovasculares Funding: 2.843.750 €. y metabólicas del Sindrome de Apnea Duration: 2016-2020 del Sueño. Patogenia y modulacion farmacológica de la actividad del eje • Code: PROPAG-AGEING 2014. cuerpo carotideo/medula adrenal. Title: The continuum between healthy ageing Principal Investigator: Gracia Patricia and idiopathic Parkinson Disease within a Ortega Saenz. propagation perspective of inflammation CIBERNED’s collaboration: No. and damage: the search for new diagnostic, CIBERNED groups: G105. prognostic and therapeutic targets. Other CIBER’s collaboration: No. Principal Investigator: Pablo Mir Rivera. Type: CC.AA. CIBERNED’s collaboration: No. Funding agency: Junta de Andalucía. CIBERNED groups: G105. Funding: 49.909,91€. Other CIBER’s collaboration: No. Duration: 2017-2019

2017 CIBERNED Annual Report / 180 Program 2 Group: José López Barneo

• Code: FIS 16 (PI16/01575). • Code: SAF2012/39343. Title: Estudio integral de los biomarcadores Title: Sensibilidad al oxígeno y implicados en el desarrollo y evolución de la neurodegeneracion. enfermedad de Parkinson. Principal Investigator: José López Barneo, Lin Principal Investigator: Pablo Mir Rivera. Gao Chen. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G105. CIBERNED groups: G105. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Tipo: National. Funding agency: Instituto de Salud Carlos III. Funding agency: MINECO. Funding: 160.325€. Funding: 468.000 €. Duration: 2017-2019 Duration: 2016 • Code: Retos-Colaboracion 2015. • Code: RD16/0011/0016. Title: Desarrollo de una terapia para el Title: Combined protective/restorative cell- tratamiento de variantes genéticas de alfa- mediated strategies for neurodegenerative sinucleina en la enfermedad de Parkinson. diseases. Principal Investigator: Juan José Toledo Principal Investigator: José Luis Labandeira Aral. García. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: G105. CIBERNED groups: G208, G10, G113, G105, Other CIBER’s collaboration: No. G207. Tipo: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Tipo: National. Funding: 383.095,47 €. Funding agency: Instituto de Salud Carlos III. Duration: 2016-2018 Funding: 240.245€. Duration: 2017-2021 • Code: Retos-Colaboracion 2015. Title: Desarrollo de una terapia para el tratamiento de variantes genéticas de alfa- sinucleina en la enfermedad de Parkinson. Principal Investigator: Pablo Mir Rivera. CIBERNED’s collaboration: No. CIBERNED groups: G105. Other CIBER’s collaboration: No. Tipo: National. Funding agency: MINECO. Funding: 317.528€. Duration: 2016-2018

2017 CIBERNED Annnual Report / 181 Adolfo López de Munain Arregui 609

Principal Investigator Adolfo López de Munain

Research team

Ana Aiastui Pujana Rosalía Dacosta Aguayo Jaione Lasa Elgarresta Anabel Rico Castro PhD Neuropsychologist PhD student PhD student

Garazi Aldanondo Roberto Fernández Torrón Haizpea Lasa Fernández Ana Quiroga Valera Aristizabal PhD student PhD student PhD PhD Belén Gago Calderón José Félix Martí Masso Tatiana Rodríguez Miren Karmele Arnaiz Pérez PhD Neurologist Chinchilla Administrative assistant Federico García Bragado Pablo Martínez-Lage PhD student Myriam Barandiarán Pathologist Álvarez María Cruz Rodríguez Amillano Bachelors degree Oroz Neuropsychologist Francisco Javier Gil Bea PhD Alba Mateos Mateos Neurologist Arantzazu Belloso Ayerdi Javier Ruiz Martínez María Goicoechea PhD Iguerategui Bianchi Neurologist PhD student PhD Leyre Merino Galán PhD student Amets Sáenz Peña Alberto Bergareche Yarza Ana Gorostidi Pagola PhD Neurologist PhD Fermín Moreno Izco Neurologist Andone Sistiaga M. Pilar Camaño González Ian James Holt Berrondo PhD student PhD Juanjo Poza Aldea PhD Leire Casas Fraile Neurologist Ainara Vallejo PhD student Haritz Jiménez Urbieta PhD student Illarramendi Neia Naldaiz Gastezi PhD PhD student Biodonostia-Instituto de Investigación Sanitaria Mónica Zufiría García Área de Neurociencias Irene Navalpotro Gómez PhD student Pº Dr. Beguiristain s/n - Planta 1ª PhD student 20014 San Sebastián (Spain) Miren Zulaica Ijurco Bachelors degree Guipúzcoa-España Nahikari Pastoriza Polo Technician Tel.: +34 94 300 62 95 Fax: +34 943 006 250 [email protected]

2017 CIBERNED Annual Report / 182 SUMMARY

Main research lines: genetic forms (patients and asymptomatic carriers of the LRRK2 R1441G mutation) and • Impulse control disorders (ICD) and dyskinesia: sporadic cases of PD will allow to deepen Physiopathology, risk factors and therapeutic in the relationship between glial cells, approaches in these complications derived neuroinflammation and neuronal death in from chronic dopaminergic treatment in both sporadic and genetic forms of PD. Parkinson’s disease. We focus on clinical and experimental research in animal models with several approaches comprising behavioural, Neuromuscular diseases neuropsychological, molecular, genetic and image studies (PET using different The group is composed by neurologists, radiotracers and multimodal MRI). A neuropsychologists and biologists who develop multicenter clinical trial on ICD coordinated a comprehensive approach to neuromuscular by this group is also under development, as pathology from a clinical and basic aspect well as a unicenter one. (including ELA and hereditary-based muscular diseases), as well as on the role of the muscle • Dementia and mild cognitive impairment in the metabolic control and its influence (MCI) in PD. This line is focused on the on the development of central phenomena identification of biomarkers of MCI that might associated with aging (brain proteinopathies be used in the early diagnosis of dementia in and aging). PD as well as in the implementation of early therapeutic interventions. In this regard, • Myotonic dystrophy: A paper has been cognitive stimulation is also one of the lines published about the incidence of cancer of study at this moment. The approach to this in the disease and the group continues the line of research includes multimodal MRI, PET, study of common metabolic pathways that electrophysiological and molecular studies determine the risk of oncogenicity and brain in CSF and plasma as well as proteomic and disorders. epigenetic studies. • Duchenne dystrophy: Study of alterations • Genetic and premotor biomarkers of PD. This in calcium homeostasis and its rescue with line is focused on the study of the phenotype compounds targeting ryanodine receptor of patients with the LRRK2 R1441G mutation stability. The study has resulted in a patent (frequent in the Baque country) and of for some compounds called generically asymptomatic carriers of this mutation AHULKEN, valuing the creation of a start-up. aiming to know pre-motor biomarkers of PD including clinical, molecular, and image • Facioscapulohumeral dystrophy: The group (DATSCAN and fMRI) studies as well as their continues collaborating with other groups evolution. in the clinical characterization of cases with scapuloperoneal syndrome and methylation • Familiar esencial tremor. This line comprises patterns. clinical, neurophysiológical, genetic (exoming and genetic linkage analysis), and • Limb Girdle Dystrophy type 2 A: study of image (MRI) studies. The objective is to study the influence of calpain 3 in proliferation, genetic, molecular functional abnormalities differentiation and replacement of muscle in families with different clinical phenotypes. satellite cells, as well as the relationship with the extracellular matrix proteins. The group • Inflammation in PD. It comprises studies in have published some collaborative and animal models of parkinsonism and in iPS individual papers. derived from patients with PD. The aim is to study the relationship between inflammation • Amyotrophic Lateral Sclerosis: The group has and dopaminergic death in animal models begun its journey in this field with the creation using in vivo PET and post-mortem studies. On of an intraciber network that has obtained a the other hand, the study of iPS derived from project published a review.

2017 CIBERNED Annual Report / 183 KEYWORDS

Enfermedad de Parkinson, discinesias, trastorno de control de impulsos, demencia, deterioro cognitivo, LRRK2, neuroinflamación, distrofias musculares, demencia frontotemporal, calpaína, progranulina, biomarcadores, neurogenética, ELA PUBLICATIONS

• Voon V., Napier T.C., Frank M.J., Sgambato-Faure V., Grace A.A., Rodriguez-Oroz M. et al. Impulse control disorders and levodopa-induced dyskinesias in Parkinson’s disease: an update. The Lancet Neurology. 2017;16(3):238-250. • Virues-Ortega J., Vega S., Seijo-Martinez M., Saz P., Rodriguez F., Rodriguez-Laso A. et al. A protective personal factor against disability and dependence in the elderly: an ordinal regression analysis with nine geographically-defined samples from Spain. BMC Geriatrics. 2017;17(1):1-10. • Sivera R., Frasquet M., Lupo V., Garcia-Sobrino T., Blanco-Arias P., Pardo J. et al. Distribution and genotype-phenotype correlation of GDAP1 mutations in Spain. Scientific Reports. 2017;7(1). • Moreno F., Indakoetxea B., Barandiaran M., Caballero M.C., Gorostidi A., Calafell F. et al. The unexpected co-occurrence of GRN and MAPT p.A152T in Basque families: Clinical and pathological characteristics. PLoS ONE. 2017;12(6). • Quiroga-Varela A., Aguilar E., Iglesias E., Obeso J.A., Marin C. Short- and long-term effects induced by repeated 6-OHDA intraventricular administration: A new progressive and bilateral rodent model of Parkinson’s disease. Neuroscience. 2017;361:144-156. • Hamel W., Koppen J.A., Alesch F., Antonini A., Barcia J.A., Bergman H. et al. Targeting of the Subthalamic Nucleus for Deep Brain Stimulation: A Survey Among Parkinson Disease Specialists. World Neurosurgery. 2017;99:41-46. • Victor R.G., Sweeney H.L., Finkel R., McDonald C.M., Byrne B., Eagle M. et al. A phase 3 randomized placebo-controlled trial of tadalafil for Duchenne muscular dystrophy. Neurology. 2017;89(17):1811-1820. • Montal V., Vilaplana E., Alcolea D., Pegueroles J., Pasternak O., Gonzalez-Ortiz S. et al. Cortical microstructural changes along the Alzheimer’s disease continuum. Alzheimer’s and Dementia. 2017. • Delgado-Alvarado M, Gago B, Gorostidi A, Jiménez-Urbieta H, Dacosta-Aguayo R, Navalpotro- Gómez I et al. Tau/α-synuclein ratio and inflammatory proteins in Parkinson’s disease: An exploratory study.Movement disorders : official journal of the Movement Disorder Society. 2017;32(7). • Jinnah H.A., Albanese A., Bhatia K.P., Cardoso F., Da Prat G., de Koning T.J. et al. Treatable inherited rare movement disorders. Movement Disorders. 2017. • Ferrero H., Larrayoz I.M., Martisova E., Solas M., Howlett D.R., Francis P.T. et al. Increased Levels of Brain Adrenomedullin in the Neuropathology of Alzheimer’s Disease. Molecular Neurobiology. 2017;:1-7. • Minicozzi P., Innos K., Sanchez M.-J., Trama A., Walsh P.M., Marcos-Gragera R. et al. Quality analysis of population-based information on cancer stage at diagnosis across Europe, with presentation of stage-specific cancer survival estimates: A EUROCARE-5 study. European Journal of Cancer. 2017;84:335-353. • Larrayoz IM, Ferrero H, Martisova E, Gil-Bea FJ, Ramírez MJ, Martínez A. Adrenomedullin Contributes to Age-Related Memory Loss in Mice and Is Elevated in Aging Human Brains. Frontiers in molecular neuroscience. ;10. • Gerenu G., Martisova E., Ferrero H., Carracedo M., Rantamaki T., Ramirez M.J. et al. Modulation of BDNF cleavage by plasminogen-activator inhibitor-1 contributes to Alzheimer’s neuropathology and cognitive deficits. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2017;1863(4):991-1001. • Munoz-Culla M., Irizar H., Gorostidi A., Alberro A., Osorio-Querejeta I., Ruiz-Martinez J. et al. Progressive changes in non-coding RNA profile in leucocytes with age. Aging. 2017;9(4):1202- 1218.

2017 CIBERNED Annual Report / 184 Program 2 Group: Adolfo López de Munain

• Flores-Burgess A, Millón C, Gago B, Narváez M, Borroto-Escuela DO, Mengod G et al. Galanin (1-15) enhancement of the behavioral effects of Fluoxetine in the forced swimming test gives a new therapeutic strategy against depression.Neuropharmacology. 2017;118. • Millon C., Flores-Burgess A., Castilla-Ortega E., Gago B., Garcia-Fernandez M., Serrano A. et al. Central administration of galanin N-terminal fragment 1-15 decreases the voluntary alcohol intake in rats. Addiction Biology. 2017. • Ezquerra-Inchausti M., Barandika O., Anasagasti A., Irigoyen C., Lopez De Munain A., Ruiz- Ederra J. High prevalence of mutations affecting the splicing process in a Spanish cohort with autosomal dominant retinitis pigmentosa. Scientific Reports. 2017;7. • Jaka O., Casas-Fraile L., Azpitarte M., Aiastui A., Lopez de Munain A., Saenz A. FRZB and melusin, overexpressed in LGMD2A, regulate integrin β1D isoform replacement altering myoblast fusion and the integrin-signalling pathway. Expert Reviews in Molecular Medicine. 2017. • Gil-Bea F.J., Aldanondo G., Lasa-Fernandez H., Lopez de Munain A., Vallejo-Illarramendi A. Insights into the mechanisms of copper dyshomeostasis in amyotrophic lateral sclerosis. Expert Reviews in Molecular Medicine. 2017. • Brockmann K, Schulte C, Schneiderhan-Marra N, Apel A, Pont-Sunyer C, Vilas D et al. Inflammatory profile discriminates clinical subtypes in LRRK2-associated Parkinson’s disease. European journal of neurology. 2017;24(2). • Villanueva V., Bermejo P., Montoya J., Toledo M., Gomez-Ibanez A., Garces M. et al. EARLY-ESLI study: Long-term experience with eslicarbazepine acetate after first monotherapy failure. Acta Neurologica Scandinavica. 2017;136(3):254-264. • Matamoros-Angles A., Gayosso L.M., Richaud-Patin Y., Di Domenico A., Vergara C., Hervera A. et al. iPS Cell Cultures from a Gerstmann-Sträussler-Scheinker Patient with the Y218N PRNP Mutation Recapitulate tau Pathology. Molecular Neurobiology. 2017;:1-16. • Arruti M, Piñeiro LD, Salicio Y, Cilla G, Goenaga MA, López de Munain A. Incidence of varicella zoster virus infections of the central nervous system in the elderly: a large tertiary hospital-based series (2007-2014).Journal of neurovirology. 2017;23(3). • Morís G, Wood L, FernáNdez-Torrón R, González Coraspe JA, Turner C, Hilton-Jones D et al. Chronic pain has a strong impact on quality of life in facioscapulohumeral muscular dystrophy. Muscle & nerve. 2017. • Millón C, Flores-Burgess A, Narváez M, Borroto-Escuela DO, Gago B, Santín L et al. The neuropeptides Galanin and Galanin(1-15) in depression-like behaviours.Neuropeptides. 2017;64. • Villanueva V, Garcés M, López-González FJ, Rodriguez-Osorio X, Toledo M, Salas-Puig J et al. Erratum to Safety, efficacy and outcome-related factors of perampanel over 12months in a real-world setting: The FYDATA study Epilepsy Res. 126 (2016) 201-210.Epilepsy research. 2017;129. • Galceran J., Ameijide A., Carulla M., Mateos A., Quiros J.R., Rojas D. et al. Cancer incidence in Spain, 2015. Clinical and Translational Oncology. 2017;19(7):799-825. • Chirlaque M.D., Salmeron D., Galceran J., Ameijide A., Mateos A., Torrella A. et al. Cancer survival in adult patients in Spain. Results from nine population-based cancer registries. Clinical and Translational Oncology. 2017;:1-11. • Gasca-Salas C., Garcia-Lorenzo D., Garcia-Garcia D., Clavero P., Obeso J.A., Lehericy S. et al. Parkinson’s disease with mild cognitive impairment: severe cortical thinning antedates dementia. Brain Imaging and Behavior. 2017;:1-9. • Ayala A., Trivino-Juarez J.M., Forjaz M.J., Rodriguez-Blazquez C., Rojo-Abuin J.-M., Martinez- Martin P. et al. Parkinson’s disease severity at 3 years can be predicted from non-motor symptoms at baseline. Frontiers in Neurology. 2017;8(OCT). • Muiño E, Rubio MA, Navalpotro I, Munteis E. Progressive multifocal leukoencephalopathy in an immunocompetent patient.Neurologia (Barcelona, Spain). 2017;32(5). • Lee A.J., Wang Y., Alcalay R.N., Mejia-Santana H., Saunders-Pullman R., Bressman S. et al. Penetrance estimate of LRRK2 p.G2019S mutation in individuals of non-Ashkenazi Jewish ancestry. Movement Disorders. 2017;32(10):1432-1438.

2017 CIBERNED Annnual Report / 185 • de la Riva P., Zubikarai M., Sarasqueta C., Tainta M., Munoz-Lopetegui A., Andres-Marin N. et al. Nontraditional Lipid Variables Predict Recurrent Brain Ischemia in Embolic Stroke of Undetermined Source. Journal of Stroke and Cerebrovascular Diseases. 2017;26(8):1670-1677. • Dal-Re R., de Munain A.L., Ayuso C. Patients organizations and new drug approval in the us. Eteplirsen and duchenne muscular dystrophy case Asociaciones de pacientes y autorización de nuevos fármacos en estados unidos. El caso del eteplirseno para la distrofia muscular de duchenne. Revista de Neurologia. 2017;65(8):373-380. • Lopez-Alava S., Aliri J., Olascoaga J., Berrondo A.S. Psychosocial factors and cognitive performance in multiple sclerosis: Gender differences Factores psicosociales y rendimiento cognitivo en sclerosis múltiple: Diferencias de sexo. Revista de Neurologia. 2017;65(5):216-222. • Fernández-Torrón R, López de Munain A.Directorio de recursos diagnósticos, centros de referencia, asociaciones, fundaciones y páginas web relacionadas con las enfermedades neuromusculares. Algoritmos diagnósticos en trastornos neuromusculares. In: Manual de enfermedades neuromusculares. Gutiérrez-Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Camaño Gonzalez P, Fernández-Torrón R, Lemmers R, Sacconi S, Sistiaga Berrondo A, López de Munain A. Distrofia facioescapulohumeral y síndromes relacionados. In: Manual de enfermedades neuromusculares. Gutiérrez-Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Rodriguez-Rojas R, Carballo-Barreda M, Alvarez L, Guridi J, Pavon N, Garcia-Maeso I et al. Subthalamotomy for Parkinson’s disease: clinical outcome and topography of lesions.Journal of neurology, neurosurgery, and psychiatry. 2017. • Poza Aldea JJ, Espinal Valencia JB, Sáenz Peña A, Fernández-Torrón R, Jericó Pascual Y, López de Munain A, Urtizberea JA, Cobo Esteban AM, Garcia Bragado F. Distrofias de musculares de cinturas. Introducción. Clasificación. In: Manual de enfermedades neuromusculares. Gutiérrez- Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Saénz Peña A, López de Munain A, Fernández-Torrón R.La Herencia en los trastornos neuromusculares. In: Manual de enfermedades neuromusculares. Gutiérrez-Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Poza Aldea JJ, Fernández-Torrón R, Urtizberea JA, Cobo Esteban AM, Garcia Bragado F, López de Munain A. Miopatías. In: Manual de enfermedades neuromusculares. Gutiérrez-Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Fernández-Torrón R, Maneiro Vicente M, Carrera Martí I, Lafuente Hidalgo M, Cobo Esteban AM, Martorell Sampol L, Sistiaga Berrondo A, Bargiela Schönbrunn, Artero Allepuz R, López de Munain A. Miotonías distróficas. In: Manual de enfermedades neuromusculares. Gutiérrez-Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Fernández-Torrón R, Illa Sendra I, Vílchez Padilla JJ, Berciano Blanco J, López de Munain A. Panorama de la investigación sobre las enfermedades neuromusculares en España. In: Manual de enfermedades neuromusculares. Gutiérrez-Rivas E, editores. Madrid, Ediciones ERGON . 2017. • Ruiz-Martínez J, Azcona LJ, Bergareche A, Martí-Massó JF, Paisán-Ruiz C. Whole-exome sequencing associates novel CSMD1 gene mutations with familial Parkinson disease.Neurology. Genetics. 2017;3(5). RESEARCH PROJECTS

• Code: FPU/04677. Other CIBER’s collaboration: No. Title: Beca Formación de Profesorado Type: National. Universitario (FPU) 2015. Funding agency: MECD. Principal Investigator: Irene Navalpotro. Funding: 16.422€. CIBERNED’s collaboration: No. Duration: 2016-2018 CIBERNED groups: G609.

2017 CIBERNED Annual Report / 186 Program 2 Group: Adolfo López de Munain

• Code: AGL2015-71764-REDT. Funding agency: BIOEF - Eitb Maratoia. Title: Red nacional de priones. Funding: 21.500€. Principal Investigator: José Antonio del Río. Duration: 2017 CIBERNED’s collaboration: Yes. CIBERNED groups: G114, G509, G503, G609, • Code: BIO/ER/022. G207. Title: Análisis del papel de la Calpaina 3 en la Other CIBER’s collaboration: No. regulación de las células satélite musculares. Type: National. Principal Investigator: Adolfo López De Funding agency: MINECO. Munaín. Funding: 39.000 €. CIBERNED’s collaboration: No. Duration: 2016-2017 CIBERNED groups: G609. Other CIBER’s collaboration: No. • Code: DTS15/00141. Type: CC.AA. Title: Evaluación del impacto de la imagen Funding agency: BIOEF - Eitb Maratoia. PET de amiloide en el diagnóstico de Funding: 40.541,34€. los pacientes con deterioro cognitivo Duration: 2017-2019 evaluados por sospecha de Alzheimer. Principal Investigator: Dr. Javier Arbizu. • Code: PI2016/04. CIBERNED’s collaboration: Yes. Title: The ALS CIBERNED Challenge: CIBERNED groups: G504, G502, G609. Accelerating New Drug Discovery. Other CIBER’s collaboration: No. Principal Investigator: Adolfo López De Type: National. Munaín. Funding agency: Instituto de Salud Carlos III. CIBERNED’s collaboration: Yes. Funding: 59.139 €. CIBERNED groups: G609, G303, G503, G408. Duration: 2016 Other CIBER’s collaboration: No. Type: Intramurales. • Code: RTC-2015-3750-1. Funding agency: CIBERNED. Title: Desarrollo de agentes terapéuticos Funding: 200.000€. basados en ácidos nucléicos para el Duration: 2016-2018 tratamiento de enfermedades neuromotoras y neuromusculares. Principal Investigator: Rubén Lépez Vales. • Code: 2017222021. CIBERNED’s collaboration: Yes. Title: Desarrollo pleclínico de una terapia CIBERNED groups: G607, G609. combinada frente a la distrofia miotónica Other CIBER’s collaboration: No. con rycals y compuestos anti envejecimiento. Type: National. Principal Investigator: Adolfo López De Funding agency: MINECO. Munaín. Funding: 204.315 €. CIBERNED’s collaboration: No. Duration: 2015-2018 CIBERNED groups: G609. Other CIBER’s collaboration: No. • Code: ASAP. Type: CC.AA. Title: Aligned and Standardized Funding agency: Gobierno Vasco. Neuroimaging in Atypical Parkinsonism. Funding: 82.125,54€. Principal Investigator: Thilo Van Eimeren. Duration: 2017 CIBERNED’s collaboration: No. CIBERNED groups: G609. • Code: DFG17/003. Other CIBER’s collaboration: No. Title: Terapia combinada con tioredoxinas Type: European. ancestrales y nuevos compuestos rycals Funding agency: JPND. para tratar desequilibrios calcio-redox en Funding: ND. enfermedades neurodegenerativas. Duration: 2017-2019 Principal Investigator: Adolfo López De Munaín. • Code: 2017222027. CIBERNED’s collaboration: No. Title: El sustrato metabólico de la CIBERNED groups: G609. esclerosis lateral amiotrófica: En busca de Other CIBER’s collaboration: No. biomarcadores y mecanismos patogénicos. Type: CC.AA. Principal Investigator: Francisco Javier Gil Funding agency: Gobierno Vasco. Bea. Funding: 74.000€. CIBERNED’s collaboration: No. Duration: 2017-2018 CIBERNED groups: G609. Other CIBER’s collaboration: No. Type: CC.AA.

2017 CIBERNED Annual Report / 187 • Code: Beca formación concedida a Tatiana • Code: PI16/01325. Rodríguez Chinchilla. Title: Regulación de la expresión de FRZB Title: Inicio y progresión en la enfermedad in vitro e in vivo, para el restablecimiento de Parkinson: papel de la activación glial. de la homeostasis en la fibra muscular de Principal Investigator: María Cruz Rodríguez Oroz. pacientes con distrofia de cinturas tipo 2A CIBERNED’s collaboration: No. (LGMD2A). CIBERNED groups: G609. Principal Investigator: Amets Saenz Pena. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: Private. CIBERNED groups: G609. Funding agency: Fundación Jesús Gangoiti Other CIBER’s collaboration: No. Barrera. Type: National. Funding: 10.000€. Funding agency: Instituto de Salud Carlos III. Duration: 2017 Funding: 57.475€. Duration: 2016-2019 • Code: 2015111038. Title: Implicación de la calpaina 3 en la • Code: PI14/00436. senescencia prematura de las células Title: Estudio de la capacidad regenerativa satelite musculares: estudio del ciclo celular de los progenitores musculares derivados del en progenitores musculares derivados de ipss del pacientes con distrofia de cinturas IPSs de pacientes con LGMD2A. tipo 2ª. Estudio in vitro y en un modelo murino Principal Investigator: Adolfo López de Munain. de dano tisular. CIBERNED’s collaboration: No. Principal Investigator: Adolfo López De Munain. CIBERNED groups: G609. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G609. Type: CC.AA. Other CIBER’s collaboration: No. Funding agency: Gobierno Vasco. Type: National. Funding: 53.920 €. Funding agency: Instituto de Salud Carlos III. Duration: 2016-2018 Funding: 131.000 €. Duration: 2015-2017 • Code: 2015111122. Title: La esclerosis lateral amiotrófica como • Code: CM16/00033. enfermedad metabólica: estudio de Title: Contrato Rio Hortega. mecanismos patogénicos y aproximación Principal Investigator: Irene Navalpotro Gómez. terapéutica. CIBERNED’s collaboration: No. Principal Investigator: Francisco Javier Gil Bea. CIBERNED groups: G609. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G609. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: CC.AA. Funding: 53.732€. Funding agency: Gobierno Vasco. Duration: 2017-2018 Funding: 48.000 €. Duration: 2016-2017 PhD DISSERTATIONS

• Author: Alba Judith Mateos Aierdi. • Author: Jesús Alberto Bergareche Yarza. Title: Calpain 3 in skeletal muscle Title: Estudio clínico-epidemiológico y regeneration: Potential role in satellite cells molecular del temblor familiar. and in the myogenic differentiation of iPSC- Date: 25th May 2017. derived muscle progenitors. Supervisor: José Félix Martí Masso. Date: 18th Dicember 2017. Supervisor: Adolfo López de Munain Arregui. • Author: Manuel Delgado Alvarado. Title: Biomarcadores en la detección • Author: Garazi Aldanondo Aristizabal. temprana del deterioro cognitivo de la Title: Effect of novel ryanodine receptor Enfermedad de Parkinson. modulators in mouse and human models of Date: 14th June 2017. Duchenne muscular dystrophy. Supervisor: María Cruz Rodríguez Oroz. Date: 31th May 2017. Supervisor: Adolfo López de Munain Arregui.

2017 CIBERNED Annual Report / 188 José Javier Lucas Lozano 306

Principal investigator José Javier Lucas Lozano

Research team

Ainara Elorza Peregrina Ivó Hernández Hernández Miriam Lucas Santamaría Postdoctoral fellow PhD student Technician

Alberto Parras Rodríguez Sara Picó del Pino María Santos Galindo Postdoctoral fellow PhD student Technician

Centro de Biología Molecular “Severo Ochoa” CSIC/UAM c/Nicolás Cabrera, 1, 28049 Madrid (Spain) Tel.: +34 91 196 45 52 / 91 196 45 82 (Lab) Fax: +34 91 196 44 20 www.cbm.uam.es/lineas/joselucas.htm www.ciberned.es/grupo-lucas-lozano.html

2017 CIBERNED Annual Report / 189 SUMMARY

Huntington’s disease (HD) is a neurodegenerative tau mutations. The appearance of TNRs in this disease caused by an expansion of the model led us to conclude that TNR detection trinucleotide CAG in the huntingtin gene. Being could be used as marker to monitor tau strictly hereditary, monogenic and dominant, it imbalance (Brain Pathol. 2017 May;27(3):314- becomes amenable for modeling and studying 322). Splicing factors such as SRSF6 often act the basis of neurodegeneration in general. on functionally related genes. We have studied In our lab we study the molecular basis of HD in HD the levels and isoforms of MAP2, another through the analysis of cell models and in vivo microtubule-associated protein functionally (by generating and characterizing transgenic and phylogenetically related to tau. We models that could mimic/revert the disease) and found a marked decrease in its levels and an through studies on human tissue from patients. imbalance between high and low molecular weight isoforms in HD. According to our results In the last years we have demonstrated an in cellular models, this mis-splicing could be imbalance in tau isoforms and discovered attributable to SRSF6. Furthermore, MAP2 is a new histopathological hallmark (the Tau restrained to the cell body and doesn’t fill the Nuclear Rods or TNRs) in this disease, consisting dendritic compartment of EH neurons (Brain on the presence of tau in nuclear indentations. Pathol. 2017 Mar;27(2):181-189). The use of transgenic animals with varying levels of tau allowed us to observe how a moderate Another research line in the lab deals with decrease in this protein in HD model mice the alteration of the endoplasmic reticulum resulted in attenuated symptoms. Regarding stress response both in mouse models and the mechanism underlying tau imbalance in postmortem brains of HD patients (Fernandez- HD, we found that the splicing factor SRSF6 Fernandez et al., Neurobiol Dis. 41:23-32, -which was known to modulate tau exon 10 2011), we have previously described that splicing and postulated to bind CAG repeats-, adult neurons do express the splicing factor is altered in HD as it is sequestered into the ATF5 and that it plays a neuroprotective role mutant huntingtin inclusion bodies (Nat Med. for instance in epilepsy (Brain. 2013 Apr;136(Pt 2014, 20(8):881-5). This discovery relates HD 4):1161-76). This year we have reported ATF5 with tauopathies, which include Alzheimer’s alteration in HD, as it is recruited into the mutant disease, thus broadening the possible therapies huntingtin inclusion bodies which correlates applicable to HD to those developed for with diminished levels of soluble ATF% and of its tauopathies. In order to characterize if tau target the antiapoptotic protein MCL1. Besides, alteration is enough for TNR appearance, we in a cellular model of HD, ATF5 overexcpression have tested a mouse model of frontotemporal reduces polyglutamine-induced apoptosis, dementia with Parkinsonism associated to thus indicating a neuroprotective role (Acta chromosome 17 (FTDP-17), a disease caused by Neuropathol. 2017 Dec;134(6):839-850). KEYWORDS

Huntington’s disease, transgenic mice, Tau, MAP2, SRSF6, splicing, ER-stress, ATF5 PUBLICATIONS

• Hernandez I.H., Torres-Peraza J., Santos-Galindo M., Ramos-Moron E., Fernandez-Fernandez M.R., Perez-Alvarez M.J. et al. The neuroprotective transcription factor ATF5 is decreased and sequestered into polyglutamine inclusions in Huntington’s disease. Acta Neuropathologica. 2017;:1-12. • Engel T., Martinez-Villarreal J., Henke C., Jimenez-Mateos E.M., Sanz-Rodriguez A., Alves M. et al. Spatiotemporal progression of ubiquitin-proteasome system inhibition after status epilepticus suggests protective adaptation against hippocampal injury. Molecular Neurodegeneration. 2017;12(1). • Engel T., Lucas J.J., Henshall D.C. Targeting the proteasome in epilepsy. Oncotarget. 2017;8(28):45042-45043.

2017 CIBERNED Annual Report / 190 Program 2 Group: José Javier Lucas Lozano RESEARCH PROJECTS

• Code: PI2015-2/06. Title: Molecular mechanisms of brain and muscle stem cell function in aging and neurodegeneration. Principal Investigator: Pura Muñoz Cánoves. CIBERNED’s collaboration: Yes. CIBERNED groups: G604, G102, G606, G111, G306. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350.000 €. Duration: 2016-2017 PhD DISSERTATIONS

• Author: Alberto Parras Rodríguez. Title: Role of CPEB4 in Huntington’s Disease and in Autism Spectrum Disorder. Date: 7th July 2017. Supervisor: José Javier Lucas Lozano.

2017 CIBERNED Annual Report / 191 Rosario Moratalla Villalba 204

Principal Investigator Rosario Moratalla Villalba

Research team

Noelia Granado Martínez Irene Ruiz De Diego Samuel Alberquilla PhD fellow PhD fellow Researcher

Luz María Suárez Oscar Solís Castrejón Emilia Rubio Rubio PhD fellow PhD fellow Lab technician

Patricia García Sanz Lorena Orgaz Gordillo Beatriz Pro PhD fellow PhD student Lab technician

José Rubén García Montes Guillermo Bueno Gil PhD fellow PhD student

Instituto Cajal (CSIC) Avenida Dr. Arce, 37 28002 Madrid (Spain) Tel.: +34 91 585 47 05 / 47 26 Fax: +34 91 585 47 54 [email protected]

2017 CIBERNED Annual Report / 192 SUMMARY

During this year 2017, we have shown that b-glucocerebrosidase-1, are known to increase D3 dopaminergic receptors (D3Rs) modulate the risk of developing Parkinson’s disease. We dyskinesias induced by L-Dopa treatment have demonstrated using patient specific cell in Parkinson’s disease, through D1 receptor- models of the N370S mutation, that this mutation mediated striatal signaling, suggesting that causes loss of b-glucocerebrosidase-1 function, decreasing this signaling pathway may help accumulation of lysosomal cholesterol, and improve L-Dopa-induced dyskinesias, since disruption of cell homeostasis in Parkinson’s L-Dopa-induced dyskinesias are more common disease (García-Sanz et al., 2017, Mov Disord). in Parkinson’s disease. (Solis et al., 2017, Cereb Thus, we have seen that the N370S mutation Cortex). changes the subcellular location of the glucocerebrosidase, being partially retained In addition, we have demonstrated that the in the endoplasmic reticulum producing stress overexpression of the protein catechol-O- and disorganization of this organelle, as well methyltransferase (COMT), which degrades as the fragmentation of the golgi apparatus. dopamine and its precursor L-DOPA and As a result, the activity of GCase is reduced plays a critical role in the regulation of and cholesterol accumulates in the lysosomes, synaptic dopamine actions, confers increased resulting in multi-sample bodies. This phenotype susceptibility to dyskinesias in mice and alters is associated with a lysosomal dysfunction molecular and neurochemical responses in the that promotes ineffective autophagia with injured striatum of mice in a model of Parkinson’s abnormal removal of damaged mitochondria, disease with 6-hydroxydopamine. (Solis et al., production of ROS and increased cell death. 2017, Neurobiology of Disease). All this could be one of the reasons why dopaminergic neurons are degenerating in In addition, in collaboration with Dr. Obeso’s Parkinson’s patients with this mutation. group, we have detected that striatal activation by optogenetics induces dyskinesias Also this year, we have studied the main in a rat model of Parkinson’s disease with mechanisms responsible for the neurotoxicity 6-hydroxydopamine (F.Hernández et al., 2017, of amphetamine-related drugs in humans and Mov Disord). Heterozygous mutations in the GBA1 experimental animals (Moratalla et al., 2017, gene, which encodes the lysosomal enzyme Prog Neurobiol).

KEYWORDS

Parkinson’s disease, tyrosine, glucocerebrosidase, neurotoxicity PUBLICATIONS

• Garcia-Sanz P., Orgaz L., Bueno-Gil G., Espadas I., Rodriguez-Traver E., Kulisevsky J. et al. N370S- GBA1 mutation causes lysosomal cholesterol accumulation in Parkinson’s disease. Movement Disorders. 2017;32(10):1409-1422. • F. Hernandez L., Castela I., Ruiz-DeDiego I., Obeso J.A., Moratalla R. Striatal activation by optogenetics induces dyskinesias in the 6-hydroxydopamine rat model of Parkinson disease. Movement Disorders. 2017;32(4):530-537. • Solis O., Garcia-Montes J.-R., Garcia-Sanz P., Herranz A.S., Asensio M.-J., Kang G. et al. Human COMT over-expression confers a heightened susceptibility to dyskinesia in mice. Neurobiology of Disease. 2017;102:133-139. • Garcia-Sanz P., Mirasierra M., Moratalla R., Vallejo M. Embryonic defence mechanisms against glucose-dependent oxidative stress require enhanced expression of Alx3 to prevent malformations during diabetic pregnancy. Scientific Reports. 2017;7(1).

2017 CIBERNED Annnual Report / 193 • Moratalla R, Solís O, Suárez LM. Morphological Plasticity in the Striatum Associated With Dopamine Dysfunction. In: Handbook of Behavioral Neuroscience. Heinz Steiner, Kuei Y. Tseng, editors. 2017. RESEARCH PROJECTS

• Code: MHE-200017. Principal Investigator: Rosario Moratalla. Title: Mecanismos involucrados en la CIBERNED’s collaboration: No. generación de disquinesiaspor L-DOPA en CIBERNED groups: G204. un modelo experimental de la enfermedad Other CIBER’s collaboration: No. de Parkinson. Type: National. Principal Investigator: Rosario Moratalla. Funding agency: MINECO. CIBERNED’s collaboration: No. Funding: 350.000€. CIBERNED groups: G204. Duration: 2017-2019 Other CIBER’s collaboration: No. Type: National. • Code: PNSD 2016I033. Funding agency: CSIC. Title: Estudio del consumo de metanfetamina Funding: 25.500€. en la adolescencia como factor de Duration: 2016-2018 riesgo para la adición y la vulnerabilidad dopaminérgica en el adulto: papel de la • Code: Cajal Blue Brain Project. glia y del glutamato. Title: Cajal Blue Brain Project. International Principal Investigator: Rosario Moratalla. Blue Brain Proyect. CIBERNED’s collaboration: No. Principal Investigator: Javier de Felipe. CIBERNED groups: G204. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G204, G403. Type: National. Other CIBER’s collaboration: No. Funding agency: MSSSI. Type: International. Funding: 68.943€. Funding agency: Ecole Polytechnique Duration: 2017-2019 Federale de Lausanne (Suiza). Funding: N.D. • Code: PCIN-2015-098. Duration: 2009-2019 Title: Development of a new in vivo radiotracer for alpha-synuclein. • Code: Fundación Ramón Areces 2017. Principal Investigator: Mireille Dumoilin. Title: Efecto de las mutaciones del gen CIBERNED’s collaboration: No. glucocerebrosidasa-1 en neuronas CIBERNED groups: G204. derivadas de celulas iPS de enfermos Other CIBER’s collaboration: No. de párkinson. Rescate del fenotipo y Type: European. trasplante celular. Área: terapia celular en Funding agency: Comisión Europea. enfermedades neurodegenerativas. Funding: 110.000 €. Principal Investigator: Carlos Vicario Abejón. Duration: 2015-2018 CIBERNED’s collaboration: Yes. CIBERNED groups: G204, G108. • Code: PI2015-2/02. Other CIBER’s collaboration: No. Title: Potencial patológico de los astrocitos: Type: Private. una nueva perspectiva en la enfermedad Funding agency: Fundación Ramón Areces. de Alzheimer. Funding: 120.000€. Principal Investigator: Joan X. Comella Carnice. Duration: 2017-2020 CIBERNED’s collaboration: Yes. CIBERNED groups: G413, G415, G204, G108, G411. • Code: SAF2016-48532-R. Other CIBER’s collaboration: No. Title: Bases moleculares de la plasticidad Type: Intramurales. sináptica estriatal en las disquinesias y el Funding agency: CIBERNED. trastorno de control de impulsos inducidos Funding: 350.000 €. por L-DOPA en la enfermedad de Parkinson. Duration: 2016-2017

2017 CIBERNED Annual Report / 194 Pura Muñoz-Cánoves 604

Principal Investigator Pura Muñoz-Cánoves

Research team

Antonio L. Serrano Sánchez Marina Raya Vanessa López-Polo Senior Researcher Labmanager Master Student

Eusebio Perdiguero Jessica Segales Dalmau Marta Masso Santamaría PhD Fellow Pre-doctoral Fellow Senior Researcher Antonio Martínez David Thomson Mercè Jardí Ripoll Pre-doctoral Fellow Sabbatical researcher Technician Victoria Moiseeva Francesco Paolo Lavarone Vera Lukesova Pre-doctoral Fellow Collaborator researcher Technician Eduardo Álvaro Rodríguez Xiaotong Hong Laura Ortet Technician Pre-doctoral Fellow PhD Fellow Mireia vaca Dempere Ana María Martín de Ana Pre-doctoral Fellow Animal Facility Technician

Cell Biology Group Joan Isern Marín ICREA and Pompeu Fabra University (UPF) PhD Fellow Department of Experimental and Life Sciences (DCEXS) Tel.: +34 93 316 08 91 Fax: +34 93 316 09 01 [email protected]

2017 CIBERNED Annual Report / 195 SUMMARY

Muscle stem cell regenerative decline with Cánoves P. Rejuvenating stem cells to restore aging muscle regeneration in aging. F1000Res. 6:76, 2017. 1. Muscle stem cells are subjected to circadian • García-Prat L, Sousa-Victor P, Muñoz- regulation, and this regulation is altered during Cánoves P. Proteostatic and Metabolic aging. Control of Stemness. Cell Stem Cell 20:593- 608, 2017. We have shown for the first time that muscle • García-Prat L, Muñoz-Cánoves P, Martínez- stem cells, despite being quiescent, and Vicente M. Monitoring Autophagy in Muscle considered practically inactive, are subject to Stem Cells. Methods Mol Biol. 1556:255-280, a strict circadian control throughout the 24 hour 2017. day/night cycle, and this circadian regulation is altered with aging. This finding completely 3. Additional studies on stem cells and striated changes the classical conception of dormancy muscle of adult muscle stem cells and their regenerative functions, thus unveiling the possibility that tissue • Serrano AL, Muñoz-Cánoves P. Fibrosis repair might be influenced by cycles of day/ development in early-onset muscular nigh, light/darkness. The importance of these dystrophies: Mechanisms and translational findings has ample implications in regenerative implications. Semin Cell Dev Biol. 64:181-190, medicine. 2017. • Perdiguero E, Serrano AL, Muñoz-Cánoves • Solanas G, Peixoto FO, Perdiguero E, Jardí P. Cilia Control Fat Deposition during Tissue M, Ruiz-Bonilla V, Datta D, Symeonidi A, Repair. Dev Cell 42:114-116, 2017. Castellanos A, Welz PS, Caballero JM, • Testoni G, Duran J, García-Rocha M, Sassone-Corsi P, Muñoz-Cánoves P*, Benitah Vilaplana F, Serrano AL, Sebastián D, López- SA*. Aged Stem Cells Reprogram Their Daily Soldado I, Sullivan MA, Slebe F, Vilaseca M, Rhythmic Functions to Adapt to Stress. Cell Muñoz-Cánoves P, Guinovart JJ. Lack of 170:678-692, 2017 (* co-corresponding). Glycogenin Causes Glycogen Accumulation and Muscle Function Impairment. Cell 2. Additional studies on stem cells showing that Metab. 26:256-266, 2017. their autophagic activity is lost during aging • López-Luppo M, Catita J, Ramos D, Navarro and their entrance in senescent state. M, Carretero A, Mendes-Jorge L, Muñoz- Cánoves P, Rodriguez-Baeza A, Nacher V, We previously showed (Nature 2016) that Ruberte J. Cellular Senescence Is Associated basal autophagy is indispensable to maintain With Human Retinal Microaneurysm the normal stem-cell quiescent state in mice. Formation During Aging. Invest Ophthalmol Failure of autophagy in physiologically aged Vis Sci. 58:2832-2842, 2017. satellite cells causes senescence entry and • Romero-Moya D, Santos-Ocaña C, Castaño increased mitochondrial dysfunction, resulting J, Garrabou G, Rodríguez-Gómez JA, Ruiz- in numerical and functional satellite cell Bonilla V, Bueno C, González-Rodríguez P, decline. Autophagy reestablishment reverses Giorgetti A, Perdiguero E, Prieto C, Moren- senescence and restores regenerative Nuñez C, Fernández-Ayala DJ, Victoria functions in geriatric satellite cells. These Cascajo M, Velasco I, Canals JM, Montero findings uncover autophagy as a decisive stem R, Yubero D, Jou C, López-Barneo J, cell-fate regulator and have implications for Cardellach F, Muñoz-Cánoves P, Artuch fostering muscle regeneration in sarcopenia. R, Navas P, Menendez P. Genetic Rescue of Mitochondrial and Skeletal Muscle We have further describe this process in three Impairment in an Induced Pluripotent Stem revisions: Cells Model of Coenzyme Q10 Deficiency. Stem Cells 35:1687-1703, 2017 • Bengal E, Perdiguero E, Serrano AL, Muñoz- KEYWORDS

Skeletal muscle, regeneration, muscle stem cells, sarcopenia, aging, inflammation, fibrosis

2017 CIBERNED Annual Report / 196 Program 2 Group: Pura Muñoz-Cánoves PUBLICATIONS

• Romero-Moya D, Santos-Ocaña C, Castaño J, Garrabou G, Rodríguez-Gómez JA, Ruiz-Bonilla V et al. Genetic Rescue of Mitochondrial and Skeletal Muscle Impairment in an Induced Pluripotent Stem Cells Model of Coenzyme Q10 Deficiency.Stem cells (Dayton, Ohio). 2017;35(7). • Garcia-Prat L., Munoz-Canoves P., Martinez-Vicente M. Monitoring autophagy in muscle stem cells. Methods in Molecular Biology. 2017;1556:255-280. • Solanas G., Peixoto F.O., Perdiguero E., Jardi M., Ruiz-Bonilla V., Datta D. et al. Aged Stem Cells Reprogram Their Daily Rhythmic Functions to Adapt to Stress. Cell. 2017;170(4):678-692.e20. • Garcia-Prat L., Sousa-Victor P., Munoz-Canoves P. Proteostatic and Metabolic Control of Stemness. Cell Stem Cell. 2017;20(5):593-608. • Testoni G., Duran J., Garcia-Rocha M., Vilaplana F., Serrano A.L., Sebastian D. et al. Lack of Glycogenin Causes Glycogen Accumulation and Muscle Function Impairment. Cell Metabolism. 2017;26(1):256-266.e4. • Perdiguero E., Serrano A.L., Munoz-Canoves P. Cilia Control Fat Deposition during Tissue Repair. Developmental Cell. 2017;42(2):114-116. • Lopez-Luppo M., Catita J., Ramos D., Navarro M., Carretero A., Mendes-Jorge L. et al. Cellular senescence is associated with human retinal microaneurysm formation during aging. Investigative Ophthalmology and Visual Science. 2017;58(7):2832-2842. • Tedesco F.S., Moyle L.A., Perdiguero E. Muscle interstitial cells: A brief field guide to non-satellite cell populations in skeletal muscle. Methods in Molecular Biology. 2017;1556:129-147. • Cornelison D.D.W., Perdiguero E. Muscle stem cells: A model system for adult stem cell biology. Methods in Molecular Biology. 2017;1556:3-19. • Bengal E., Perdiguero E., Serrano A.L., Munoz-Canoves P. Rejuvenating stem cells to restore muscle regeneration in aging. F1000Research. 2017;6. RESEARCH PROJECTS

• Code: 2014 SGR 102. • Code: ERARE13-fp-149. Title: Understanding the basis of muscle Title: Stimulating Intrinsic Repair for DMD regeneration. (SIRD). Principal Investigator: Pura Muñoz-Cánoves. Principal Investigator: Michael Rudnicki. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G604. CIBERNED groups: G604. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: CC.AA. Type: Intramurales. Funding agency: Generalitat de Catalunya. Funding agency: Comisión Europea. Funding: 40.000 €. Funding: 40.000 €. Duration: 2015-2017 Duration: 2014-2017 • Code: AGAUR SGR 102. • Code: PI2015-2/06. Title: Biología Celular. Title: Molecular mechanisms of brain and Principal Investigator: Pura Muñoz-Cánoves. muscle stem cell function in aging and CIBERNED’s collaboration: No. neurodegeneration. CIBERNED groups: G604. Principal Investigator: Pura Muñoz-Cánoves. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: CC.AA. CIBERNED groups: G604, G102, G606, G111, Funding agency: Generalitat de Catalunya. G306. Funding: 48.800 €. Other CIBER’s collaboration: No. Duration: 2015-2017 Type: Intramurales. Funding agency: CIBERNED. Funding: 350.000 €. Duration: 2016-2017

2017 CIBERNED Annnual Report / 197 • Code: MDA 2016. • Code: : SAF2015-67369-R). Tíitle: Understanding and reversing muscle Tíitle: Understanding skeletal muscle stem cell regenerative decline in DMD. regenerative decline with aging. Principal Investigator: Pura Muñoz Cánoves. Principal Investigator: Pura Muñoz-Cánoves. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G604. CIBERNED groups: G604. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: National. Funding agency: Muscular Dystrophy Funding agency: MINECO. Association (MDA)-USA. Funding: 150.000 €. Funding: 100.000 €. Duration: 2016-2018 Duration: 2016-2019

2017 CIBERNED Annual Report / 198 José Ramón Naranjo Orovio 307

Principal Investigator José Ramón Naranjo Orovio

Research team

Britt Mellström Valentina González M. Paz González Researcher Pre-doctoral Fellow Assistant researcher

Alberto Rábano Daniel Fernández Xosé Manuel Dopazo Researcher Master UAM, student Assistant researcher

Alejandro López-Hurtado Rocio Naranjo Pre-doctoral Fellow TFG UAM, student

CSIC. Centro Nacional de Biotecnología Molecular and Cellular Biology c/Darwin, 3 28049 Madrid (Spain) Tel.: +34 91 585 46 82 [email protected]

2017 CIBERNED Annual Report / 199 SUMMARY

We investigate the nuclear components of with presenilins, has an important role in activity- and Ca2+-dependent transcriptional neurodegenerative diseases (NDD) through responses in neurons to: the control of Ca2+ and protein homeostasis. Importantly, an early reduction in DREAM levels 1) Understand the molecular determinants of is found in mouse models for these pathologies downstream events that are responsible for (AD, DS and HD) and, genetic experiments show early plastic changes in synaptic functionality that this could be part of a neuroprotective during neurodegeneration. mechanism operating in HD. The mechanism involves the protein-protein interaction 2) To design small molecules to intervene between DREAM and ATF6, a key protein in in physiological and pathological the early activation of the unfolded protein output processes including learning response (UPR), as well as the regulation at and memory, motor coordination and the transcriptional level of several cytoskeletal neurodegeneration. proteins that are important for axonal transport and synaptic function. We have developed Early compensatory changes in transcriptional in vitro and in vivo assays to screen for small programs and altered neuronal calcium and molecules able to bind to and modify DREAM protein homeostasis are common features biological activity. The goal is to identify potent of some neurodegenerative pathologies, inhibitors of the DREAM activity in vivo that including Alzheimer’s (AD), Down syndrome could represent new therapeutic venues for (DS) and Huntington’s disease (HD), pathologies the treatment of NDD patients. that are in the focus of our work. Finally, we have comparatively studied the role DREAM (DRE Antagonist Modulator), a Ca2+- of DREAM in cytosolic versus mitochondrial UPR dependent transcriptional repressor, also using amyloidogenic and non-amyloidogenic known as calsenilin because its interaction AD mouse models. KEYWORDS

Calcium pathologies, DREAM, Alzheimer’s disease, Tau, UPR, ATF6, ATF5, DREAM inhibitors PUBLICATIONS

• Bolos M., Llorens-Martin M., Perea J.R., Jurado-Arjona J., Rabano A., Hernandez F. et al. Absence of CX3CR1 impairs the internalization of Tau by microglia. Molecular Neurodegeneration. 2017;12(1). • Dols-Icardo O, García-Redondo A, Rojas-García R, Borrego-Hernández D, Illán-Gala I, Muñoz- Blanco JL et al. Analysis of known amyotrophic lateral sclerosis and frontotemporal dementia genes reveals a substantial genetic burden in patients manifesting both diseases not carrying the C9orf72 expansion mutation.Journal of neurology, neurosurgery, and psychiatry. 2017. • Carro E., Bartolome F., Bermejo-Pareja F., Villarejo-Galende A., Molina J.A., Ortiz P. et al. Early diagnosis of mild cognitive impairment and Alzheimer’s disease based on salivary lactoferrin. Alzheimer’s and Dementia: Diagnosis, Assessment and Disease Monitoring. 2017;8:131-138. • Rodriguez-Rodriguez P, Sandebring-Matton A, Merino-Serrais P, Parrado-Fernandez C, Rabano A, Winblad B et al. Tau hyperphosphorylation induces oligomeric insulin accumulation and insulin resistance in neurons.Brain : a journal of neurology. 2017. • Perez-Canamas A., Benvegnu S., Rueda C.B., Rabano A., Satrustegui J., Ledesma M.D. Sphingomyelin-induced inhibition of the plasma membrane calcium ATPase causes neurodegeneration in type A Niemann-Pick disease. Molecular Psychiatry. 2017;22(5):711-723.

2017 CIBERNED Annual Report / 200 Program 2 Group: José Ramón Naranjo Orovio

• Diack A.B., Boyle A., Ritchie D., Plinston C., Kisielewski D., De Pedro-Cuesta J. et al. Similarities of variant Creutzfeldt-Jakob disease strain in mother and son in Spain to UK reference case. Emerging Infectious Diseases. 2017;23(9):1593-1596. • Lopez-Sanchez N., Fontan-Lozano A., Palle A., Gonzalez-Alvarez V., Rabano A., Trejo J.L. et al. Neuronal tetraploidization in the cerebral cortex correlates with reduced cognition in mice and precedes and recapitulates Alzheimer’s-associated neuropathology. Neurobiology of Aging. 2017;56:50-66. • Pisa D., Alonso R., Fernandez-Fernandez A.M., Rabano A., Carrasco L. Polymicrobial Infections in Brain Tissue from Alzheimer’s Disease Patients. Scientific Reports. 2017;7(1).

RESEARCH PROJECTS

• Code: PI2016/05. Title: Dream inhibitors and Alzheimer´s disease. Principal Investigator: José Ramón Naranjo Orovio. CIBERNED’s collaboration: Yes. CIBERNED groups: G307, G106, G403. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 210.000 €. Duration: 2016-2018

2017 CIBERNED Annnual Report / 201 Xavier Navarro Acebes 607

Principal Investigator Xavier Navarro Acebes

Research team

Esther Udina Bonet Roser Velasco Fargas Natalia De la Oliva María Rodríguez Associate Professor- Assistant Professor Muñoz Cañón Researcher PhD Student PhD Student Jaume del Valle Macià Rubén López Vales Postdoctoral Fellow Nuria Gaja Capdevila David Romeo Guitart Associate Professor- PhD Student PhD Student Researcher Joaquim Hernández Martín Tatiana Leiva Alba Sánchez Caty Casas Louzao Postdoctoral Fellow Rodríguez Fernández Associate Professor- PhD Student PhD Student Researcher Mireia Herrando Grabulosa Víctor Manuel López Jordi Badia Casahuja Assumpció Bosch Postdoctoral Fellow Álvarez Researcher Merino PhD Student Associate Professor- Clara Penas Pérez Miguel Ángel Rubio Researcher Postdoctoral Fellow Clara López Serrano Pérez PhD Student Researcher Guillermo García Alías Jesús Amo Aparicio Ramon y Cajal PhD Student Anna Martínez Muriana Jessica Jaramillo Researcher PhD Student Rodríguez Ariadna Arbat Plana Technician Jordi Cuadras Mas PhD Student Sara Marmolejo Emeritous Professor- Martínez-Artesero Israel Blasco Ruano Researcher Olaia Baylo Marín PhD Student Technician PhD Student Jordi Bruna Escuer María Puigdomenech Neus Hernández Assistant Professor Aina Calls Cobos Poch Solanes PhD Student PhD Student Technician

Dept. Cell Biology, Physiology and Immunology Institute of Neurosciences Universitat Autònoma de Barcelona Avenida Can Domènech, edificio M, campus UAB 08193 Bellaterra, Catalunya (Spain) Tel.: +34 93 581 19 66 Fax: +34 93 581 29 86 [email protected]

2017 CIBERNED Annual Report / 202 SUMMARY

The Group of Neuroplasticity and Regeneration • Etiopathogenic role of lipid mediators of the UAB is a multidisciplinary research group and modulators of the neuroinflammatory working on repair, regeneration and functional response in neurodegeneration induced by recovery after peripheral nerve and spinal cord central nervous system lesions. lesions and in neurodegenerative diseases. • Activity-dependent therapies for enhancing The research activities of the Group of axonal regeneration and functional Neuroplasticity and Regeneration have focused recovery after peripheral nerve lesions and on the study of physiopathological mechanisms for preventing neuropathic pain. of neural lesions, neuropathic pain and neurodegeneration, and on the application • Physiopathologic mechanisms in neuropathic of novel therapeutic strategies for promoting pain induced by nerve and spinal cord neuroprotection and regeneration in traumatic lesions. Study of the relationship between and degenerative lesions of the nervous neuroinflammation and hyperexcitability. system. The active research lines include: • Study of etiopathogenic mechanisms • Cellular and molecular mechanisms and potential neuroprotective treatments implicated in motoneuron degenerative in peripheral neuropathies induced by diseases in experimental models. Search for diabetes and by antitumoral agents. biomarkers. Neuroprotective strategies based on gene and pharmacological therapy. • Neuromodulation of neural plasticity for promoting functional restitution in spinal • Cell therapy by transplantation of cord injuries. mesenchimal stem cells and neural stem cells for the repair of spinal cord injuries and • Design and evaluation of neural interfaces motoneuron degenerative diseases. for the development of neuroprostheses applied to neurorehabilitation. Study of • Repair of spinal root avulsion injuries by surgical new intraneural electrodes for the selective reimplantation and pharmacological stimulation and recording of neural neuroprotection. activity. KEYWORDS

Neurodegeneration, nerve regeneration, spinal cord injury, neuropathic pain, motoneuron diseases, peripheral neuropathies, neuroplasticity, neural interfaces PUBLICATIONS

• Cayuela N, Simó M, Majós C, Rifà-Ros X, Gállego Pérez-Larraya J, Ripollés P et al. Seizure-susceptible brain regions in glioblastoma: identification of patients at risk.European journal of neurology. 2017. • Argyriou A.A., Bruna J., Genazzani A.A., Cavaletti G. Chemotherapy-induced peripheral neurotoxicity: Management informed by pharmacogenetics. Nature Reviews Neurology. 2017;13(8):492-504. • Massó A, Sánchez A, Bosch A, Giménez-Llort L, Chillón M. Secreted αKlotho isoform protects against age-dependent memory deficits.Molecular psychiatry. 2017. • Wurth S., Capogrosso M., Raspopovic S., Gandar J., Federici G., Kinany N. et al. Long- term usability and bio-integration of polyimide-based intra-neural stimulating electrodes. Biomaterials. 2017;122:114-129. • Miralles M., Eixarch H., Tejero M., Costa C., Hirota K., Castano A.R. et al. Clinical and Histopathological Amelioration of Experimental Autoimmune Encephalomyelitis by AAV Vectors Expressing a Soluble Interleukin-23 Receptor. Neurotherapeutics. 2017;14(4):1095-1106.

2017 CIBERNED Annual Report / 203 • Gonzalez-Perez F, Cobianchi S, Heimann C, Phillips JB, Udina E, Navarro X. Stabilization, Rolling, and Addition of Other Extracellular Matrix Proteins to Collagen Hydrogels Improve Regeneration in Chitosan Guides for Long Peripheral Nerve Gaps in Rats.Neurosurgery. 2017;80(3). • 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. [ePub2017]. • Francos-Quijorna I., Santos-Nogueira E., Gronert K., Sullivan A.B., Kopp M.A., Brommer B. et al. Maresin 1 promotes inflammatory resolution, neuroprotection, and functional neurological recovery after spinal cord injury. Journal of Neuroscience. 2017;37(48):11731-11743. • Illa M., Brito V., Pla L., Eixarch E., Arbat-Plana A., Batalle D. et al. Early Environmental Enrichment Enhances Abnormal Brain Connectivity in a Rabbit Model of Intrauterine Growth Restriction. Fetal Diagnosis and Therapy. 2017. • Udina E., Putman C.T., Harris L.R., Tyreman N., Cook V.E., Gordon T. Compensatory axon sprouting for very slow axonal die-back in a transgenic model of spinal muscular atrophy type III. Journal of Physiology. 2017;595(5):1815-1829. • 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. [ePub2017]. • Delgado-Martinez I., Righi M., Santos D., Cutrone A., Bossi S., D’Amico S. et al. Fascicular nerve stimulation and recording using a novel double-aisle regenerative electrode. Journal of Neural Engineering. 2017;14(4). • Mueller M., De La Oliva N., Del Valle J., Delgado-Martinez I., Navarro X., Stieglitz T. Rapid prototyping of flexible intrafascicular electrode arrays by picosecond laser structuring. Journal of Neural Engineering. 2017;14(6). • Velasco R., Navarro X., Gil-Gil M., Herrando-Grabulosa M., Calls A., Bruna J. Neuropathic Pain and Nerve Growth Factor in Chemotherapy-Induced Peripheral Neuropathy: Prospective Clinical-Pathological Study. Journal of Pain and Symptom Management. 2017;54(6):815-825. • Romeo-Guitart D., Fores J., Navarro X., Casas C. Boosted Regeneration and Reduced Denervated Muscle Atrophy by NeuroHeal in a Pre-clinical Model of Lumbar Root Avulsion with Delayed Reimplantation. Scientific Reports. 2017;7(1). • 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. [ePub2017]. • Mòdol-Caballero G, Santos D, Navarro X, Herrando-Grabulosa M. Neuregulin 1 Reduces Motoneuron Cell Death and Promotes Neurite Growth in an in Vitro Model of Motoneuron Degeneration.Frontiers in cellular neuroscience. ;11. [ePub2017]. • Casas C. GRP78 at the centre of the stage in cancer and neuroprotection. Frontiers in Neuroscience. 2017;11(APR). • de la Oliva N., Navarro X., del Valle J. Time course study of long-term biocompatibility and foreign body reaction to intraneural polyimide-based implants. Journal of Biomedical Materials Research - Part A. 2017. • Sánchez-Brualla I, Boulenguez P, Brocard C, Liabeuf S, Viallat-Lieutaud A, Navarro X et al. Activation of 5-HT2A receptors restores KCC2 function and reduces neuropathic pain after spinal cord injury.Neuroscience. 2017. • Cobianchi S., Jaramillo J., Luvisetto S., Pavone F., Navarro X. Botulinum neurotoxin A promotes functional recovery after peripheral nerve injury by increasing regeneration of myelinated fibers. Neuroscience. 2017;359:82-91. • Arbat-Plana A., Cobianchi S., Herrando-Grabulosa M., Navarro X., Udina E. Endogenous modulation of TrkB signaling by treadmill exercise after peripheral nerve injury. Neuroscience. 2017;340:188-200.

2017 CIBERNED Annual Report / 204 Program 2 Group: Xavier Navarro Acebes

• Santos D., Gonzalez-Perez F., Giudetti G., Micera S., Udina E., Del Valle J. et al. Preferential enhancement of sensory and motor axon regeneration by combining extracellular matrix components with neurotrophic factors. International Journal of Molecular Sciences. 2017;18(1). • Arbat-Plana A., Navarro X., Udina E. Effects of forced, passive, and voluntary exercise on spinal motoneurons changes after peripheral nerve injury. European Journal of Neuroscience. 2017. • Mancuso R., Navarro X. Sigma-1 receptor in motoneuron disease. Advances in Experimental Medicine and Biology. 2017;964:235-254. • Bisoni L, Carboni C, Puddu R, Barabino G, Pani D, Raffo L, Mueller M, Stieglitz T, Del Valle J, de la Oliva N, Delgado-Martinez I, Navarro X, Barbaro M. A 64-channels neural interface for biopotentials recording and PNS stimulation. Conference Proc IEEE Eng Med Biol Soc. 2017. • Guiho T., Andreu D., Lopez-Alvarez V.M., Cvancara P., Hiairrassary A., Granata G. et al. Advanced 56 Channels Stimulation System to Drive Intrafascicular Electrodes. Biosystems and Biorobotics. 2017;15:743-747. • Yoshida K., Malec J., Comoglio C., Mosier K., Lontis R., Larsen K. et al. Evaluation of the Effect of Sensory Feedback on Phantom Limb Pain in Multi-center Clinical Trials. Biosystems and Biorobotics. 2017;15:725-730. • Micera S., Raspopovic S., Petrini F., Carpaneto J., Oddo C., Badia J. et al. On the Use of Intraneural Transversal Electrodes to Develop Bidirectional Bionic Limbs. Biosystems and Biorobotics. 2017;15:737-741.

RESEARCH PROJECTS

• Code: RD16/0011. • Code: 2015/01. Title: Red de Terapia Celular. Title: TRANSAUTOPHAGY: European network Principal Investigator: Isabel Fariñas. for multidisciplinar research and translation CIBERNED’s collaboration: Yes. of autophagy knowledge. CIBERNED groups: G607, G301, G102, G113, Principal Investigator: Caty Casas. G208, G105, G207. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G607. Type: National. Other CIBER’s collaboration: No. Funding agency: Instituto de Salud Carlos III. Type: European. Funding: 284.999€. Funding agency: Comisión Europea. Duration: 2016-2020 Funding: 520.000€. Duration: 2016-2020 • Code: CA15138. Title: Terapia génica dirigida a neuregulinas • Code: AC16/00050. para el tratamiento de la degeneración Title: Non-invasive electrical stimulation of de las motoneuronas en la esclerosis lateral the cervical spinal cord to facilitate arm amiotrofica. and hand functional recovery in incomplete Principal Investigator: Xavier Navarro Acebes. traumatic cervical spinal cord injured CIBERNED’s collaboration: Yes. patients (CERMOD). CIBERNED groups: G607, G113, G406, G407. Principal Investigator: Guillermo García-Alias. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: Intramurales. CIBERNED groups: G607. Funding agency: CIBERNED. Other CIBER’s collaboration: No. Funding: 240.000 €. Type: European. Duration: 2015-2017 Funding agency: Comisión Europea. Funding: 750.000€. Duration: 2017-2020

2017 CIBERNED Annnual Report / 205 • Code: FP7-611687. Type: National. Title: Neurocontrolled bidirectional artificial Funding agency: Instituto de Salud Carlos III. upper limb and hand prosthesis (NEBIAS). Funding: 86.515 €. Principal Investigator: Xavier Navarro Duration: 2015-2018 Acebes. CIBERNED’s collaboration: No. • Code: PI15/01271. CIBERNED groups: G607. Title: Estudio molecular y tratamiento de la Other CIBER’s collaboration: No. Mucopolisacaridosis tipo VII. Type: European. Principal Investigator: Assumpcio Bosch. Funding agency: Comisión Europea. CIBERNED’s collaboration: No. Funding: 517.424 €. CIBERNED groups: G607. Duration: 2013-2017 Other CIBER’s collaboration: No. Type: National. • Code: E10668. Funding agency: Instituto de Salud Carlos III. Title: eBIONERVE: personalized nerve grafts Funding: 147.015 €. for treating human nerve injurie. Duration: 2016-2018 Principal Investigator: Xavier Navarro. CIBERNED’s collaboration: No. • Code: SAF2013-48431-R. CIBERNED groups: G607. Title: Participación de los receptores del Other CIBER’s collaboration: No. aácido lisofosfatídico en la fisiopatología de Type: European. la lesión medular. Funding agency: Comisión Europea. Principal Investigator: Rubén Lopez-Vales. Funding: 800.000€. CIBERNED’s collaboration: No. Duration: 2016-2019 CIBERNED groups: G607. Other CIBER’s collaboration: No. • Code: CI17-00019. Type: National. Title: Immunoresolvent lipids for multiple Funding agency: MINECO. . sclerosis and amyotrophic lateral sclerosis. Funding: 157.300 €. Principal Investigator: Rubén López-Vales. Duration: 2014-2017 CIBERNED’s collaboration: No. CIBERNED groups: G607. • Code: RTC-2015-3750-1. Other CIBER’s collaboration: No. Title: Desarrollo de agentes terapéuticos Type: Private. basados en ácidos nucleicos para el Funding agency: Fundación La Caixa. tratamiento de enfermedades neuromotoras Funding: 52.500€. y neuromusculares. Duration: 2017-2019 Principal Investigator: Rubén López Vales. CIBERNED’s collaboration: Yes. • Code: TV32014-2810. CIBERNED groups: G607, G609. Title: Terapia génica dirigida a neuregulinas Other CIBER’s collaboration: No. para el tratamiento de la esclerosis lateral Type: National. amiotrófica. Funding agency: MINECO. Principal Investigator: Xavier Navarro. Funding: 204.315 €. CIBERNED’s collaboration: No. Duration: 2015-2018 CIBERNED groups: G607. Other CIBER’s collaboration: No. • Code: SAF 2014-59701. Type: Private. Title: Reprogramación neuronal para Funding agency: Fundació La Marató-TV3. promover los mecanismos endogenos de Funding: 199.682 €. neuroprotección usando biología sintética Duration: 2015-2017 en un modelo de degeneración retrógrada de motoneuronas. • Code: PI15/01303. Principal Investigator: Caty Casas. Title: Estudio exploratorio sobre la CIBERNED’s collaboration: No. etiopatogenia, biomarcadores de riesgo y CIBERNED groups: G607. mecanismos implicados en la regeneración Other CIBER’s collaboration: No. de la neuropatía inducida por platinos. Type: National. Principal Investigator: Jordi Bruna. Funding agency: MINECO. CIBERNED’s collaboration: No. Funding: 108.900 €. CIBERNED groups: G607. Duration: 2015-2018 Other CIBER’s collaboration: No.

2017 CIBERNED Annual Report / 206 Program 2 Group: Xavier Navarro Acebes

• Code: RTC-2015-3731-1. • Código: SAF2016-79774-R. Title: Optimización y desarrollo de un agente Title: Estudio de los mecanismos de acción terapéutico basado en ácidos nucleicos de la interleucina-37 en el sistema nervioso para el tratamiento de la enfermedad de central lesionado. Huntington. Principal Investigator: Rubén López-Vales. Principal Investigator: Assumpció Bosch, CIBERNED’s collaboration: No. Esther Pérez. CIBERNED groups: G607. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G607, G301. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: National. Funding: 160.000€. Funding agency: MINECO. Duration: 2017-2020 Funding: 1.582.767,92 €. Duration: 2015-2017 • Code: FP7-602547. Title: Natural sensory feedback for phantom • Code: SAF2016-79279-R. limb pain modulation and therapy (EPIONE). Title: Neuromodulación eléctrica del Principal Investigator: Xavier Navarro conectoma cortico-medular para facilitar Acebes. la recuperación de la destreza manual tras CIBERNED’s collaboration: No. una lesión medular. CIBERNED groups: G607. Principal Investigator: Guillermo García- Other CIBER’s collaboration: No. Alias. Type: European. CIBERNED’s collaboration: No. Funding agency: Comisión Europea. CIBERNED groups: G607. Funding: 199.840 €. Other CIBER’s collaboration: No. Duration: 2013-2017 Type: National. Funding agency: MINECO. • Code: WFL-ES-14-17. Funding: 193.600€. Title: Interleukin-37: a novel therapeutic Duration: 2016-2020 approach for the treatment of spinal cord injury. • Code: RYC-2014-16529. Principal Investigator: Rubén Lopez-Vales. Title: Promoting plasticity of reticulospinal CIBERNED’s collaboration: No. axons to recover skilled hand function after CIBERNED groups: G607. spinal cord injury. Other CIBER’s collaboration: No. Principal Investigator: Guillermo García- Type: International. Alias. Funding agency: Wings for Life Foundation. CIBERNED’s collaboration: No. Funding: 193.000€. CIBERNED groups: G607. Duration: 2017-2020 Other CIBER’s collaboration: No. Type: National. Funding agency: MINECO. Funding: 20.000€. Duration: 2016-2021 PhD DISSERTATIONS

• Author: Belén García Lareu. • Author: David Romeo Guitart. Title: Physiopathological and molecular Title: Boosting endogenous mechanisms for characterization of a transgenic mouse neuronal protection and repair after injury in overexpressing TNFa in Schwann cells reveals central nervous system. a model for chronic peripheral neuropathy. Date: 26th October 2017. Date: 14th September 2017. Supervisor: Caty Casas Louzao. Supervisor: Assumpció Bosch Merino. • Author: Víctor M López Álvarez. • Author: Clara López Serrano. Title: Activity-dependent treatments for Title: Role of lysophosphatidic acid receptors neuropathic pain. in the spinal cord injury physiopathology. Date: 30th October 2017. Date: 27th November 2017. Supervisor: Xavier Navarro Acebes. Supervisor: Rubén López Vales.

2017 CIBERNED Annual Report / 207 Jose Ángel Obeso Inchausti 205

Principal Investigator Jose Ángel Obeso Inchausti

Research team

Inés Trigo Damas Álvaro Sánchez Ferro Cristina Calvo González Research staff Research staff Administrative assistant Guillermo Foffani María Ledia Fernández Hdez. José Ángel Pineda Pardo Research staff Research staff Research staff Raúl Martínez Fernández Javier Blesa de los Mozos Gabriela Riederer Research staff Research staff Lab technician Rafael Rodríguez Rojas Laura Arenas Moreno Raquel Márquez López Research staff Lab technician Research assistant Frida Hernández Fernández Iban Castela Muñoz Ignacio Obeso Martín Research staff Research staff Research staff Claudia Ammann Carmen Gasca Salas Natalia López González del Rey PhD fellow Research staff PhD student Marta del Álamo de Pedro Research staff

Centro Integral de Neurociencias AC (CINAC) Alejandro Reinares Sebastián PhD student Hospital Puerta del Sur y Universidad San Pablo CEU David Mata Marín Avenida Carlos V, nº 70 PhD student 28938 Móstoles, Madrid (Spain) Tel.: +34 91 267 32 10 [email protected]

2017 CIBERNED Annual Report / 208 SUMMARY

Dr. Obeso heads HM CINAC, a movement technique which allows performing controlled disorders research center, which is mainly and focused brain lesions without surgery. Thus, focused on the study of Parkinson´s disease mobility, mortality and economic costs are (PD) from a multidisciplinary approach. The greatly reduced compared to current surgical principal objective is the study of the onset treatments. This technique has demonstrated and progression of the disease using basic efficacy and safety in the performance of and clinical experiments aiming to define the thalamotomies aimed to cease essential tremor etiopathogenesis of the disease, optimizing and PD tremor. HM CINAC has been the first the diagnosis and advancing in the treatment center in perform subthalamotomies to treat of neurodegenerative and neuropsiquiatric cardinal manifestations of Parkinson Disease diseases. (rigidity, akinesia, tremor). This study has been published in The Lancet Neurology Journal (IF= Onset of the disease: compensatory 26,28). Martínez-Fernández, R., Rodríguez-Rojas, mechanisms and selective vulnerability R., Del Álamo, M., Hernández-Fernández, F., Pineda-Pardo, J. A., Dileone, M., … Obeso, J. A. So far, treatments for PD have not been (2018). Focused ultrasound subthalamotomy in successful and one of the reasons is because patients with asymmetric Parkinson’s disease: a diagnosis tends to be delayed respect to the pilot study. The Lancet. Neurology. onset of the neurodegenerative process. At CINAC, patients are treated with magnetic Pharmacological treatment of Parkinson´s stimulation (TMS) and neuroimaging directed to disease and motor complications define the onset of the disease. Interestingly, PD is principally characterized by the progressive Dopamine replacement therapy has been degeneration of dopaminergic neurons of the largely used when treating PD. Both dopamine substantia nigra pars compacta (SNc), which agonists and levodopa (LDOPA) are the main are crucial during learning and habit acquisition. treatments. After long term use, this therapy Additionally, since the dopaminergic loss is is also associated not only with involuntary previous to the diagnosis it is assumed that there movements and dyskinesias, but also with with might be a series of compensatory mechanisms motor complications, which make reduce which delay the rise of the symptoms. At CINAC, patients. At CINAC animal models showing in vivo neuronal activity recordings in PD animals progressive dopamine depletion are utilized models are used to define these mechanisms. in order to determine the molecular base Surmeier, D. J., Obeso, J. A., & Halliday, G. M. and mechanisms involved in the onset of (2017). Parkinson’s Disease Is Not Simply a Prion LDOPA dyskinesias (LID) using recording and Disorder. The Journal of Neuroscience : The optogenetic techniques which allow a more Official Journal of the Society for Neuroscience. specific and selective neuronal modulation. F Hernández, L., Castela, I., Ruiz-DeDiego, I., Obeso, J. A., & Moratalla, R. (2017). Striatal Evolution and treatment of the disease activation by optogenetics induces dyskinesias in the 6-hydroxydopamine rat model of HIFU (High Intensity Focused Ultrasound) is a novel Parkinson disease. Movement Disorders. KEYWORDS

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

2017 CIBERNED Annnual Report / 209 PUBLICATIONS

• Cavaliere F, Cerf L, Dehay B, Ramos-Gonzalez P, De Giorgi F, Bourdenx M et al. In vitro α-synuclein neurotoxicity and spreading among neurons and astrocytes using Lewy body extracts from Parkinson disease brains.Neurobiology of disease. 2017;103. • Rodriguez-Rojas R, Carballo-Barreda M, Alvarez L, Guridi J, Pavon N, Garcia-Maeso I et al. Subthalamotomy for Parkinson’s disease: clinical outcome and topography of lesions.Journal of neurology, neurosurgery, and psychiatry. 2017. • Obeso J.A., Stamelou M., Goetz C.G., Poewe W., Lang A.E., Weintraub D. et al. Past, present, and future of Parkinson’s disease: A special essay on the 200th Anniversary of the Shaking Palsy. Movement Disorders. 2017;32(9):1264-1310. • Gasca-Salas C., Garcia-Lorenzo D., Garcia-Garcia D., Clavero P., Obeso J.A., Lehericy S. et al. Parkinson’s disease with mild cognitive impairment: severe cortical thinning antedates dementia. Brain Imaging and Behavior. 2017;:1-9. • Arroyo-Gallego T., Ledesma-Carbayo M.J., Sanchez-Ferro A., Butterworth I., Mendoza C.S., Matarazzo M. et al. Detection of Motor Impairment in Parkinson’s Disease Via Mobile Touchscreen Typing. IEEE Transactions on Biomedical Engineering. 2017;64(9):1994-2002. • Sanchez-Ferro A., Benito-Leon J., Louis E.D., Contador I., Hernandez-Gallego J., Puertas-Martin V. et al. Cognition in non-demented Parkinson’s disease vs essential tremor: A population- based study. Acta Neurologica Scandinavica. 2017. • Surmeier D.J., Obeso J.A., Halliday G.M. Selective neuronal vulnerability in Parkinson disease. Nature Reviews Neuroscience. 2017;18(2):101-113. • 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 Parkinson’s disease: a pilot study. The Lancet Neurology. 2018;17(1):54-63. [ePub2017]. • Manohar A., Foffani G., Ganzer P.D., Bethea J.R., Moxon K.A. Cortex-dependent recovery of unassisted hindlimb locomotion after complete spinal cord injury in adult rats. eLife. 2017;6. • Krack P., Martinez-Fernandez R., del Alamo M., Obeso J.A. Current applications and limitations of surgical treatments for movement disorders. Movement Disorders. 2017;32(1):36-52. • Strafella A.P., Bohnen N.I., Perlmutter J.S., Eidelberg D., Pavese N., Van Eimeren T. et al. Molecular imaging to track Parkinson’s disease and atypical parkinsonisms: New imaging frontiers. Movement Disorders. 2017. • Fernandez-Garcia C., Foffani G., Dileone M., Catalan-Alonso M.J., Gonzalez-Hidalgo M., Barcia J.A. et al. Directional local field potential recordings for symptom-specific optimization of deep brain stimulation. Movement Disorders. 2017;32(4):626-628. • Obeso J.A., Jon Stoessl A., Stamelou M. Editors’ Note: The 200th Anniversary of the Shaking Palsy. Movement Disorders. 2017;32(1):1-. • Voon V., Napier T.C., Frank M.J., Sgambato-Faure V., Grace A.A., Rodriguez-Oroz M. et al. Impulse control disorders and levodopa-induced dyskinesias in Parkinson’s disease: an update. The Lancet Neurology. 2017;16(3):238-250. • Picconi B, Hernández LF, Obeso JA, Calabresi P. Motor complications in Parkinson’s disease: Striatal molecular and electrophysiological mechanisms of dyskinesias.Movement disorders : official journal of the Movement Disorder Society. 2017. • del Rey N.L.-G., Blesa J. Parkinson’s disease: Oh my gut!. Movement Disorders. 2017;32(3):396. • Obeso JA, Martinez-Fernandez R, Del Álamo M, Krack P. To lesion or not to lesion: That was the question (Reply to stereotactic ablative surgery does not just mean ‘adding another lesion’). Movement disorders : official journal of the Movement Disorder Society. 2017;32(7). • Hariz M, Obeso JA. What Would Dr. James Parkinson Think Today? I. The Role of Functional Neurosurgery for Parkinson’s Disease.Movement disorders : official journal of the Movement Disorder Society. 2017;32(1). • Obeso JA, Trenkwalder C. What would Dr. James Parkinson think today? The 21st Annual Congress of the International Parkinson’s disease and movement disorders society.Movement disorders : official journal of the Movement Disorder Society. 2017;32(7).

2017 CIBERNED Annual Report / 210 Program 2 Group: José Ángel Obeso Inchausti

• Foffani G., Dileone M. No modulatory effects by tSMS when delivered during a cognitive task. Brain Stimulation. 2017;10(4):867-. • Obeso I., Wilkinson L., Teo J.T., Talelli P., Rothwell J.C., Jahanshahi M. Theta burst magnetic stimulation over the pre-supplementary motor area improves motor inhibition. Brain Stimulation. 2017;10(5):944-951. • Ayala A., Trivino-Juarez J.M., Forjaz M.J., Rodriguez-Blazquez C., Rojo-Abuin J.-M., Martinez- Martin P. et al. Parkinson’s disease severity at 3 years can be predicted from non-motor symptoms at baseline. Frontiers in Neurology. 2017;8(OCT). • Liu C., Foffani G., Scaglione A., Aguilar J., Moxon K.A. Adaptation of thalamic neurons provides information about the spatiotemporal context of stimulus history. Journal of Neuroscience. 2017;37(41):10012-10021. • James Surmeier D., Obeso J.A., Halliday G.M. Parkinson’s disease is not simply a prion disorder. Journal of Neuroscience. 2017;37(41):9799-9807. • Carrasco-Lopez C., Soto-Leon V., Cespedes V., Profice P., Strange B.A., Foffani G. et al. Static magnetic field stimulation over parietal cortex enhances somatosensory detection in humans. Journal of Neuroscience. 2017;37(14):3840-3847. • Quiroga-Varela A., Aguilar E., Iglesias E., Obeso J.A., Marin C. Short- and long-term effects induced by repeated 6-OHDA intraventricular administration: A new progressive and bilateral rodent model of Parkinson’s disease. Neuroscience. 2017;361:144-156. • Martinez K., Janssen J., Pineda-Pardo J.A., Carmona S., Roman F.J., Aleman-Gomez Y. et al. Individual differences in the dominance of interhemispheric connections predict cognitive ability beyond sex and brain size. NeuroImage. 2017;155:234-244. • F. Hernandez L., Castela I., Ruiz-DeDiego I., Obeso J.A., Moratalla R. Striatal activation by optogenetics induces dyskinesias in the 6-hydroxydopamine rat model of Parkinson disease. Movement Disorders. 2017;32(4):530-537. • Obeso I., Casabona E., Rodriguez-Rojas R., Bringas M.L., Macias R., Pavon N. et al. Unilateral subthalamotomy in Parkinson’s disease: Cognitive, psychiatric and neuroimaging changes. Cortex. 2017;94:39-48. • Pifl C., Reither H., del Rey N.L.-G., Cavada C., Obeso J.A., Blesa J. Early paradoxical increase of dopamine: A neurochemical study of olfactory bulb in asymptomatic and symptomatic MPTP treated monkeys. Frontiers in Neuroanatomy. 2017;11. • Ponsoda V., Martinez K., Pineda-Pardo J.A., Abad F.J., Olea J., Roman F.J. et al. Structural brain connectivity and cognitive ability differences: A multivariate distance matrix regression analysis. Human Brain Mapping. 2017;38(2):803-816. • Dileone M., Carrasco-Lopez M.C., Segundo-Rodriguez J.C., Mordillo-Mateos L., Lopez-Ariztegui N., Alonso-Frech F. et al. Dopamine-dependent changes of cortical excitability induced by transcranial static magnetic field stimulation in Parkinson’s disease. Scientific Reports. 2017;7(1). • Blesa J., Trigo-Damas I., Dileone M., del Rey N.L.G., Hernandez L.F., Obeso J.A. Compensatory mechanisms in Parkinson’s disease: Circuits adaptations and role in disease modification. Experimental Neurology. 2017. • Lozano-Soto E, Soto-León V, Sabbarese S, Ruiz-Alvarez L, Sanchez-Del-Rio M, Aguilar J et al. Transcranial static magnetic field stimulation (tSMS) of the visual cortex decreases experimental photophobia.Cephalalgia : an international journal of headache. 2017. • Pham M.H., Elshehabi M., Haertner L., Heger T., Hobert M.A., Faber G.S. et al. Algorithm for turning detection and analysis validated under home-like conditions in patients with Parkinson’s disease and older adults using a 6 degree-of-freedom inertial measurement unit at the lower back. Frontiers in Neurology. 2017;8(APR). • Pham M.H., Elshehabi M., Haertner L., Din S.D., Srulijes K., Heger T. et al. Validation of a step detection algorithm during straight walking and turning in Patients with Parkinson’s disease and older adults using an inertial measurement unit at the lower back. Frontiers in Neurology. 2017;8(SEP).

2017 CIBERNED Annual Report / 211 • De la Casa-Fages B., Alonso-Frech F., Grandas F. Effect of subthalamic nucleus deep brain stimulation on balance in Parkinson’s disease: A static posturographic analysis. Gait and Posture. 2017;52:374-380. • Blesa J., Trigo-Damas I., Del Rey N.L.-G., Obeso J.A. The use of nonhuman primate models to understand processes in Parkinson’s disease. Journal of Neural Transmission. 2017;:1-11. RESEARCH PROJECTS

• Code: Sin código. • Code: AC16/00044. Title: Selective vulnerability, progression and Title: Harmonisation metabolic FDG brain synuclein toxicity in Parkinson`s pattern characteristics. disease. Principal Investigator: José Ángel Obeso Principal Investigator: José Ángel Obeso Inchausti. Inchausti. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G205. CIBERNED groups: G205. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: Private. Funding agency: Instituto de Salud Carlos III. Funding agency: Fundación BBVA. Funding: 47.069€. Funding: 125.000 €. Duration: 2017-2019 Duration: 2016-2019 • Code: CD15/00092. • Code: H2020-MSCA-IF-2014- ROSNPD-660964. Title: Contrato posdoctoral de Title: Marie Skłodowska-Curie Individual perfeccionamiento Sara Borrell. Grupo Fellowships. habitual. Principal Investigator: Ledia Fernández Principal Investigator: Ignacio Obeso Hernández. Martín. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G205. CIBERNED groups: G205. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: European. Type: National. Funding agency: Comisión Europea. Funding agency: Instituto de Salud Carlos III. Funding: 170.122€. Funding: 80.598 €. Duration: 2016-2017 Duration: 2016-2019

• Code: Sin codigo_Tatiana. • Code: FJCI-2015-25095. Title: Objetivo α-synucleina: entender Title: Programa Juan De La Cierva. la vulnerabilidad celular y detener la Principal Investigator: José Ángel Pineda progresion en la Enfermedad de Pardo. Parkinson. CIBERNED’s collaboration: No. Principal Investigator: Javier Blesa De Los CIBERNED groups: G205. Mozos, José Ángel Obeso Inchausti. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: G205. Funding agency: MINECO. Other CIBER’s collaboration: No. Duration: 2016-2017 Type: Private. Funding agency: Fundación Tatiana Pérez de Guzmán el Bueno. Funding: 84.500€. Duration: 2017-2019

2017 CIBERNED Annual Report / 212 Program 2 Group: José Ángel Obeso Inchausti PhD DISSERTATIONS

• Author: Raúl Martínez Fernández. Title: The subthalamic nucleus and parkinson’s disease: beyond motor control. Date: 12th June 2017. Supervisor: José Ángel Obeso Inchausti.

2017 CIBERNED Annnual Report / 213 Ana Pérez Castillo 110

Principal Investigator Ana Mª Pérez Castillo

Research team

José A. Morales García A. Santos Montes Alejandro Barranco Almohalla PhD fellow Collaborador Bachelor degree student

Rocío Díaz Sánchez Jesús Alarcón Gil Daniel Carnicero Senabre Bachelor degree Bachelor degree Bachelor degree student

Michiel Van Bulck Ana Sierra Magro Xavier Perosanz Hidalgo PhD student Bachelor degree Bachelor degree student

Sandra Alonso Gil Juan Pedro Gómez Martín Danna Jessen-Howard Technician Bachelor degree student Bachelor degree student

Marina Sanz San Cristóbal David Lozano Muñoz Jack Thomas Whittaker Technician Bachelor degree student Bachelor degree student

Instituto de Investigaciones Biomédicas (CSIC-UAM) Dpto. de Modelos Experimentales de Enfermedades Humanas c/Arturo Duperier 4 28029 Madrid (Spain) Tel.: +34 91 585 44 36 Fax: +34 91 585 44 01 [email protected]

2017 CIBERNED Annual Report / 214 SUMMARY

Research is focused on molecular biological vivo neuroprotection of dopaminergic cells approaches to study pathologies of the central concomitant with a significant attenuation of nervous system. the level of the inflammatory response and glial activation. One major area of interest is the identification of new targets for the treatment of several CNS Also, a main focus of the lab concerns research disorders, specifically Parkinson’s disease, as well on neurogenesis and aging. We have recently as in the development of “proof-of concept” published a work describing the neurogenic for novel drugs directed towards these targets. effect in vitro of β-carbolines, present in Ayahuasca. We are currently working in the We have identified several genes involved in role of different new targets which can expand different brain disorders, GSK-3, C/EBP β, and our understanding of the processes that lead to phosphodiesterase 7 (PDE7). In this regard we improved neurogenesis and that can be of use have shown that lentivirus-mediated C/EBPβ for a better understanding and new treatments deprivation conferred marked in vitro and in of aging-related disorders.

KEYWORDS

Neuroprotection, neuroinflammation, neurogenesis, Parkinson, PDE7, C/EBPβ, Ayahuasca PUBLICATIONS

• Palomo V., Perez D.I., Roca C., Anderson C., Rodriguez-Muela N., Perez C. et al. Subtly Modulating Glycogen Synthase Kinase 3 β: Allosteric Inhibitor Development and Their Potential for the Treatment of Chronic Diseases. Journal of Medicinal Chemistry. 2017;60(12):4983-5001. • Morales-Garcia JA, Gine E, Hernandez-Encinas E, Aguilar-Morante D, Sierra-Magro A, Sanz- SanCristobal M et al. CCAAT/Enhancer binding protein β silencing mitigates glial activation and neurodegeneration in a rat model of Parkinson’s disease.Scientific reports. 2017;7(1). • Gine E., Echeverry-Alzate V., Lopez-Moreno J.A., Rodriguez de Fonseca F., Perez-Castillo A., Santos A. The CB1 receptor is required for the establishment of the hyperlocomotor phenotype in developmentally-induced hypothyroidism in mice. Neuropharmacology. 2017;116:132-141. • Morales-Garcia J.A., Echeverry-Alzate V., Alonso-Gil S., Sanz-Sancristobal M., Lopez-Moreno J.A., Gil C. et al. Phosphodiesterase7 Inhibition Activates Adult Neurogenesis in Hippocampus and Subventricular Zone In Vitro and In Vivo. Stem Cells. 2017. • Morales-Garcia J.A., De La Fuente Revenga M., Alonso-Gil S., Rodriguez-Franco M.I., Feilding A., Perez-Castillo A. et al. The alkaloids of Banisteriopsis caapi, the plant source of the Amazonian hallucinogen Ayahuasca, stimulate adult neurogenesis in vitro. Scientific Reports. 2017;7(1). • Garcia A.M., Salado I.G., Perez D.I., Brea J., Morales-Garcia J.A., Gonzalez-Garcia A. et al. Pharmacological tools based on imidazole scaffold proved the utility of PDE10A inhibitors for Parkinson’s disease. Future Medicinal Chemistry. 2017;9(8):731-748. • Morales-Garcia JA, Salado IG, Sanz-San Cristobal M, Gil C, Perez-Castillo A, Martínez A, Perez DI. Biological and Pharmacological Characterization of Benzothiazole-Based CK-1δ Inhibitors in Models of Parkinson’s Disease. ACS Omega. 2017 Aug 1;2(8):5215-20.

2017 CIBERNED Annual Report / 215 RESEARCH PROJECTS

• Code: PI2015/03. • Code: 2015- Proyectos de I+D+I para Title: Perfiles metabólicos diferenciales en jovenes investigadores sin vinculacion o con enfermedad de Parkinson. vinculacion temporal-SOLICITADO. Principal Investigator: José Manuel Fuentes Title: Regulación de la expresión de Rodríguez. fosfodiesterasa 7 en enfermedad de CIBERNED’s collaboration: Yes. Parkinson: posibles implicaciones para CIBERNED groups: G103, G110, G209. futuras terapias. Other CIBER’s collaboration: No. Principal Investigator: José A. Morales- Type: Intramurales. García. Funding agency: CIBERNED. CIBERNED’s collaboration: No. Funding: 210.000 €. CIBERNED groups: G110. Duraction: 2015 Other CIBER’s collaboration: No. Type: National. • Code: 2015- FUNDACIÓN EUGENIO Funding agency: MINECO. RODRÍGUEZ PASCUAL-SOLICITADO. Funding: 170.000€. Title: Regulacion de la expresion de Duraction: 2016-2018 fosfodiesterasa 7 en enfermedad de Parkinson. • Code: Marie Skłodowska-Curie Actions. Principal Investigator: José A. Morales- H2020-MSCA-ITN-2015. García. Title: Neuroregenerative drug discovery. CIBERNED’s collaboration: No. Principal Investigator: María Laura CIBERNED groups: G110. Bolognesi. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: Private. CIBERNED groups: G110. Funding agency: Fundación Eugenio Other CIBER’s collaboration: No. Rodríguez Pascual. Type: European. Funding: 56.712€. Funding agency: Comisión Europea. Duraction: 2016-2017 Funding: N.D. Duraction: 2015-2017 • Code: SAF2014. Title: Papel del factor de transcripción • Code: Marie Skłodowska-Curie Actions. CEBPβ en la enfermedad de Parkinson: H2020-MSCA-ITN-201e. Concedido. Identificación y caracterización de los Title: Blood biomarker-based diagnostic tools genes mediadores de su acción. for early-stage Alzheimer’s disease. Principal Investigator: Ana Pérez Castillo. Principal Investigator: Ana Pérez Castillo. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: G110. CIBERNED groups: G110. Other CIBER’s collaboration: No. Other CIBER’s collaboration: Si (CIBERER). Type: National. Type: European. Funding agency: MINECO. Funding agency: Comisión Europea. Funding: 100.000 €. Funding: 3.465.749,52 €. Duraction: 2015-2017 Duraction: 2016-2020

2017 CIBERNED Annual Report / 216 Jordi Pérez Tur 209

Principal Investigator Jordi Pérez Tur

Research team

Fernando Cardona Serrate Jacek Szymanski Guillermo Rodríguez Casero Researcher Researcher Lab technician

Unitat de Genètica Molecular Institut de Biomedicina de València-CSIC c/Jaume Roig, 11 460101 Valencia (Spain) Tel.: +34 96 339 17 55 Fax: +34 96 339 37 74 [email protected]

2017 CIBERNED Annual Report / 217 SUMMARY

During this year, we have remained focused Parkinson’s disease, the characterization of on the study of genetic factors related to variants or alterations in DNA related to the diseases of the Amyotrophic Lateral Sclerosis- appearance of a familial form of primary Frontotemporal Dementia spectrum and lateral sclerosis as well as in the definition of Parkinson’s disease, taking advantage the mutation that causes a familial form of of the synergies with other groups of Progressive Supranuclear Palsy. CIBERNED through DEGESCO and with other collaborators. Likewise, we have continued the functional characterization of variants that could be As for work in progress, throughout 2017 we have related to the risk of Parkinson’s disease as well concluded our study on the genetic factors as variants that modify the expression of genes involved in the appearance of dementia in related to this disease. KEYWORDS

Alzheimer, Parkinson, Neurodegeneration, Neurogenetics, ALS PUBLICATIONS

• Vazquez-Costa J.F., Tembl J.I., Fornes-Ferrer V., Cardona F., Morales-Caba L., Fortea G. et al. Genetic and constitutional factors are major contributors to substantia nigra hyperechogenicity. Scientific Reports. 2017;7(1). • Vazquez-Costa J.F., Mazon M., Carreres-Polo J., Hervas D., Perez-Tur J., Marti-Bonmati L. et al. Brain signal intensity changes as biomarkers in amyotrophic lateral sclerosis. Acta Neurologica Scandinavica. 2017. • Vazquez-Costa J.F., Arlandis S., Hervas D., Martinez-Cuenca E., Cardona F., Perez-Tur J. et al. Clinical profile of motor neuron disease patients with lower urinary tract symptoms and neurogenic bladder. Journal of the Neurological Sciences. 2017;378:130-136. RESEARCH PROJECTS

• Code: PI2015/03. • Code: 2016-COHORTE BBN-NED. Title: Perfiles metabólicos diferenciales en Title: COHORTE-BBN-NED. Búsqueda enfermedad de Parkinson. de biomarcadores para la detección Principal Investigator: José Manuel Fuentes temprana de la enfermedad de Alzheimer Rodríguez. en la cohorte del “Proyecto Vallecas”. CIBERNED’s collaboration: Yes. Principal Investigator: Miguel Medina. CIBERNED groups: G103, G110, G209. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G209, G401. Type: Intramurales. Other CIBER’s collaboration: CIBER-BBN. Funding agency: CIBERNED. Type: National. Funding: 210.000 €. Funding agency: Instituto de Salud Carlos III. Duration: 2015-2017 Funding: 150.000€. Duration: 2016-2018

2017 CIBERNED Annual Report / 218 Manuel Rodríguez Díaz 206

Principal Investigator Manuel Rodríguez Díaz

Research team

Tomás González Hernández Ingrid Morales Pérez Fernando Isidro Montón Álvarez Research staff PhD fellow Researcher

Magdalena Sabaté Bel Clara Rodríguez Sabaté Jesús Norellis Lorenzo Brito Research staff PhD student Researcher

Alberto Sánchez Fernández Research staff

Departamento de Fisiología Facultad de Medicina Universidad de La Laguna La Laguna, Canarias (Spain) Tel.: +34 92 231 93 61 Fax: +34 92 231 93 97 [email protected]

2017 CIBERNED Annual Report / 219 SUMMARY

Increasing evidence suggests that the debris during the initial stages of Parkinson’s dopaminergic degeneration which disease. characterizes Parkinson’s disease starts in the striatal dopamine terminals and progresses Our group is collaborating in the iPS project retrogradely to the body of dopamine cells with other groups belonging to CIBERNED in the substantia nigra. The role of striatal who facilitate patient samples and the astrocytes in the striatal initiation of the differentiation of astrocytic cells. By using a dopaminergic degeneration is little known. procedure developed to identify and to get We studied the astrocytic response to the representative BOLD-signals of the main basal dopaminergic denervation of the striatum. The ganglia from magnetic resonance images, the injection of 6-hydroxydopamine (25 μg) in the main interactions of these centers were studied lateral ventricle of adult rats induced a fast in the human brain. The partial correlation (4 hours) and selective (unaccompanied by methods showed a circular cortico-subcortical unspecific lesions of striatal tissue or microgliosis) interaction which was globally similar in humans degeneration of the dopaminergic innervation to that previously found in animals with other of the striatum which was followed by a methods. selective astrocytosis unaccompanied by microgliosis. This astrocytosis was severe This procedure was used to analyze the and had a specific profile which included functional connectivity of the intralaminar some (e.g. upregulation of GFAP, GS, S100β, thalamic centers with the main basal ganglia NDRG2, vimentin) but not all (e.g. astrocytic circuits. Data analysis with Multiple Linear proliferation or differentiation from NG2 cells, Regression showed relevant non-linear astrocytic scars, microgliosis) the characteristics components in the basal ganglia interactions. observed after the non-selective lesion of the Thus, a new non-linear method based on the striatum. This astrocytosis is similar to those Multiple Correspondence Analysis was also observed in the parkinsonian striatum. In these applied to basal ganglia interactions. This is a rats, dopaminergic debris were accumulated multivariate method which may identify massive within spheroids (free-spheroids) which retained interacting networks with an “unsupervised” some proteins of dopaminergic neurons data-driven procedure. This method generates (e.g., tyrosine hydroxylase, the dopamine a self-organizing clustering of centers transporter protein, and APP) but not others according to their interactions which is similar to (e.g., α-synuclein). Free-spheroids showed that generated by Independent Component the initial (LC3-autophagosomes) but not the Analysis, but which may be particularly suitable late (Lamp1/Lamp2-lysosomes and activated when non-linear interactions are being studied. Caspase 6) components of autophagia The Multiple Correspondence Analysis isolated (incomplete autophagia), preparing their six networks within the cortico-subcortical motor autophagosomes for an external phagocytosis loop of basal ganglia. The activity of these (accumulation of phosphatidylserine). Free- networks is now being studied in Parkinson’s spheroids were penetrated by astrocyte disease. The membrane transporter involved processes (fenestrated-spheroids) which in the dopamine uptake by dopaminergic made them immunoreactive for GFAP and neurons is a relevant factor for the vulnerability S100β, and which had the elements needed of mesencephalic dopaminergic neurons in to continue the debris degradation (caspase Parkinson’s disease. 6 and Lamp1). Finally, proteins normally found in neurons (TH, DAT and α-synuclein) were We have studied the possible role of D3 dopamine observed within astrocytes 2-5 days after the receptors on the dopamine transporter activity dopaminergic degeneration, suggesting that in mice and cell cultures. D3 agonists decrease the intracellular contents of degenerated cells dopamine uptake, changing the integration of had been transferred to astrocytes. Taken the transporter in the cytoplasmatic membrane together, present data suggest phagocytosis of the dopaminergic synaptic terminals, and as a physiological role of striatal astrocytes, a the phosforilation and ubiquitinization of the role which could be critical for cleaning striatal dopamine transporter.

2017 CIBERNED Annual Report / 220 Program 2 Group: Manuel Rodríguez Díaz KEYWORDS

Astrocytes, IPs, dopaminergic degeneration, dopamine transporter, dopaminergic vulnerability, functional neuroimaging PUBLICATIONS

• Morales I., Sanchez A., Rodriguez-Sabate C., Rodriguez M. Striatal astrocytes engulf dopaminergic debris in Parkinson’s disease: A study in an animal model. PLoS ONE. 2017;12(10). • Rodriguez-Sabate C., Morales I., Sanchez A., Rodriguez M. The multiple correspondence analysis method and brain functional connectivity: Its application to the study of the non-linear relationships of motor cortex and basal ganglia. Frontiers in Neuroscience. 2017;11(JUN).

2017 CIBERNED Annnual Report / 221 Eduard Tolosa Sarró 207

Principal Investigator Eduard Tolosa Sarró

Research team

María José Martí Doménech Carles Gaig Bárbara Segura Consultant Specialist Researcher

Francesc Valldeoriola i Serra Yaroslau Compta Hinrjy Hugo Baggio Consultant Specialist Researcher

Esteban Muñoz Martínez Mario Ezquerra Trabalón Alicia Garrido Pla Consultant Researcher PhD student

Joan Santamaría Cano Rubén Fernández-Santiago Manuel Fernández Sánchez Consultant Researcher Lab technician

Alejandro Iranzo de Riquer Carme Junqué Plaja Laura Maragall Consultant Full professor Administrative assistant

Institut de Neurociencies Unitat de Parkinson i Trastorns Anormals del Moviment c/Villarroel 171, esc 8 - 4 planta 08036 Barcelona (España) Tel.: +34 93 227 57 85 Fax: +34 93 227 57 83 [email protected]

2017 CIBERNED Annual Report / 222 SUMMARY

In 2017 we continued our research on prodromal as crucial molecules in the maintenance of Parkinson’s disease in high-risk cohorts (idiopathic dopaminergic neurons (DAn), and which are, REM-sleep behaviour disorder [iRBD] and in therefore, potentially related to PD (Kim et al., asymptomatic subjects carrying mutations in Mol Neurosci, 2018). We are currently working LRRK2 or ‘LRRK2-PD’) through genetic, clinical, towards deepening our knowledge of existing and biomarker studies. Of note, studies in LRRK2- models of PD epigenetic alterations by using PD showed its low penetration (Lee et al Mov iPSC-DAn models and patient blood samples Disord 2017), a silent prodromal phase (Pont (work in progress). Sunyer et al , Mov Disord 2017), the presence of inflammatory profiles in LRRK2-PD subtypes In 2017 we also continued our work on (Brockmann et al Europ J Neurol 2017), and biomarkers in parkinsonism cerebrospinal fluid an association between age at onset and a (CSF), published a longitudinal study correlating synuclein polymorphism (Fernández-Santiago, tau and amyloid in CSF derived from patients Garrido et al. Mov Disord 2017). with Parkinson’s disease to motor and non-motor dysfunction (Modreanu et al., J Neurol Sci 2017), In studies published this year in journals a cross-sectional study on reduced Coenzyme including Lancet Neurology (Stokoholm et al.), Q10 levels in multiple system atrophy (Compta Annals of Neurology (Iranzo et al.), Neurology et al., Park Rel Dis 2017), and started a study (Iranzo et al.), and Sleep (Iranzo et al.), we to assess changes in a panel of 38 cytokines in showed that RBD occurs during the prodromal patients with multiple system atrophy . These phase of Parkinson disease, and we assessed latter two studies were carried out as part of the its prevalence in our field and the factors that ‘Catalan registry of multiple system atrophy’ determine its early conversion. In addition, we study funded by the TV3 Marató 2017 (PI: Dr. also participated in creating the new diagnostic M.J. Martí). criteria for Lewy body dementia (McKeith et Eduard Tolosa Sarró al. Neurology 2017) and in describing a new In addition, our participation in an international disease characterised by tau deposits which consortium on the genetics of Lewy body we called anti-IgLON5 disease (Gaig et al. dementia contributed to work on genome- Neurology 2017). wide associations in this condition(Guerreiro et al., Lancet Neurol Epub 2017), while our We described a new mutation in the gene collaboration in the Movement Disorders ACMSD, that causes Parkinson’s disease (Vilas Society-endorsed progressive supranuclear et al. J Parkinsons Dis. 2017) and detected palsy study group resulted in a publication on the presence of pathological aggregates of clinical correlations in a cohort of cases with synuclein in gonadal tissue in patients with this confirmed progressive supranuclear palsy disorder (Garrido et al. JAMA Neurol 2017). (Respondek et al., Mov Dis 2017). Funding from Moreover, we participated in a consensus the Fondo de Investigación en Salud (Fund for study on the definition of advanced Parkinson’s Health Research) has also been granted to us disease (Luquin et al. Parkinson Dis. 2017), and for a new study on the aggregation of different contributed to a method for the differential proteins in brain tissues and CSF (PI: Dr. Y. diagnosis of Parkinson’s tremor by using Compta) and the detection of alpha-synuclein smartphones (Barrantes et al. PLoS One. 2017). in the salivary glands of subjects with iRBD (PI: We also described an association between Dr. A. Iranzo). RBD and a MAPT polymorphism (Fernández- Santiago et al. Neurol Genet 2017). In another In studies on the functional and anatomical study we described the presence of numerous substrates iof cognitive deficits, we found transcriptome alterations in dermal fibroblasts that olfactory disorders advance throughout from patients with Parkinson’s disease and PD progression and that this progressive showed that the disease is systemic and affects deficit correlates with a volumetric loss of peripheral non-neuronal tissues (González- basal ganglia (Campabadal et al., Park Rel Casacuberta et al. Neurobiology of Aging Dis 2017). We applied new technologies to 2018). analyse patterns of atrophy by approximating functional connectomics and machine We discovered a novel alpha-synuclein learning and found that both approaches were expression regulatory mechanism (Marchese useful for automatically discriminating patients et al., Nucleic Acids Res. 2017) and used according to the presence or absence of cellular models to identify RAC1 GTPases cognitive deficits (Abos et al., Sci Rep. 2017).

2017 CIBERNED Annual Report / 223 KEYWORDS

Prodromal Parkinson’s disease, RBD, LRRK2, Biomarkers, CSF, neuroimaging, multiple system atrophy, progressive supranuclear palsy PUBLICATIONS

• Tell-Marti G, Puig-Butille JA, Potrony M, Plana E, Badenas C, Antonell A et al. A Common Variant in the MC1R Gene (p.V92M) is associated with Alzheimer’s Disease Risk.Journal of Alzheimer’s disease : JAD. 2017. • 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. [ePub2017]. • Antonini A., Poewe W., Chaudhuri K.R., Jech R., Pickut B., Pirtosek Z. et al. Levodopa-carbidopa intestinal gel in advanced Parkinson’s: Final results of the GLORIA registry. Parkinsonism and Related Disorders. 2017;45:13-20. • Gelpi E, Botta-Orfila T, Bodi L, Marti S, Kovacs G, Grau-Rivera O et al. Neuronal intranuclear (hyaline) inclusion disease and fragile X-associated tremor/ataxia syndrome: a morphological and molecular dilemma.Brain : a journal of neurology. 2017;140(8). • Darling A., Tello C., Marti M.J., Garrido C., Aguilera-Albesa S., Tomas Vila M. et al. Clinical rating scale for pantothenate kinase-associated neurodegeneration: A pilot study. Movement Disorders. 2017;32(11):1620-1630. • Lee A.J., Wang Y., Alcalay R.N., Mejia-Santana H., Saunders-Pullman R., Bressman S. et al. Penetrance estimate of LRRK2 p.G2019S mutation in individuals of non-Ashkenazi Jewish ancestry. Movement Disorders. 2017;32(10):1432-1438. • Giri A., Mok K.Y., Jansen I., Sharma M., Tesson C., Mangone G. et al. Lack of evidence for a role of genetic variation in TMEM230 in the risk for Parkinson’s disease in the Caucasian population. Neurobiology of Aging. 2017;50:167.e11-167.e13. • Brockmann K, Schulte C, Schneiderhan-Marra N, Apel A, Pont-Sunyer C, Vilas D et al. Inflammatory profile discriminates clinical subtypes in LRRK2-associated Parkinson’s disease. European journal of neurology. 2017;24(2). • 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 autónomas. Neurologia. 2018. [ePub2017]. • Fernández-Santiago R, Iranzo A, Gaig C, Serradell M, Fernández M, Pastor P et al. MAPT association with REM sleep behavior disorder.Neurology. Genetics. 2017;3(1). • Stokholm M.G., Iranzo A., Ostergaard K., Serradell M., Otto M., Svendsen K.B. et al. Assessment of neuroinflammation in patients with idiopathic rapid-eye-movement sleep behaviour disorder: a case-control study. The Lancet Neurology. 2017;16(10):789-796. • Compta Y, Martí MJ. Parkinson disease: What goes around comes around: cognitive impairment as prodromal parkinsonism?. Nature reviews. Neurology. 2017;13(12). • Marchese D, Botta-Orfila T, Cirillo D, Rodriguez JA, Livi CM, Fernández-Santiago R etal. Discovering the 3’ UTR-mediated regulation of alpha-synuclein.Nucleic acids research. 2017. • Garrido A., Aldecoa I., Gelpi E., Tolosa E. Aggregation of α-synuclein in the gonadal tissue of 2 patients with Parkinson disease. JAMA Neurology. 2017;74(5):606-607. • Iranzo A., Santamaria J., Valldeoriola F., Serradell M., Salamero M., Gaig C. et al. Dopamine transporter imaging deficit predicts early transition to synucleinopathy in idiopathic rapid eye movement sleep behavior disorder. Annals of Neurology. 2017;82(3):419-428.

2017 CIBERNED Annual Report / 224 Program 2 Group: Eduard Tolosa Sarró

• Iranzo A., Stefani A., Serradell M., Marti M.J., Lomena F., Mahlknecht P. et al. Characterization of patients with longstanding idiopathic REM sleep behavior disorder. Neurology. 2017;89(3):242-248. • Gaig C., Graus F., Compta Y., Hogl B., Bataller L., Bruggemann N. et al. Clinical manifestations of the anti-IgLON5 disease. Neurology. 2017;88(18):1736-1743. • Luquin MR, Kulisevsky J, Martinez-Martin P, Mir P, Tolosa ES. Consensus on the Definition of Advanced Parkinson’s Disease: A Neurologists-Based Delphi Study (CEPA Study).Parkinson’s disease. ;2017. • Mollenhauer B, Caspell-Garcia CJ, Coffey CS, Taylor P, Shaw LM, Trojanowski JQ et al. Longitudinal CSF biomarkers in patients with early Parkinson disease and healthy controls. Neurology. 2017. • Amara A.W., Chahine L.M., Caspell-Garcia C., Long J.D., Coffey C., Hogl B. et al. Longitudinal assessment of excessive daytime sleepiness in early Parkinson’s disease. Journal of Neurology, Neurosurgery and Psychiatry. 2017;88(8):653-662. • 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. [ePub2017]. • Alcolea D., Vilaplana E., Suarez-Calvet M., Illan-Gala I., Blesa R., Clarimon J. et al. CSF sAPPβ, YKL- 40, and neurofilament light in frontotemporal lobar degeneration. Neurology. 2017;89(2):178-188. • Hoglinger G.U., Respondek G., Stamelou M., Kurz C., Josephs K.A., Lang A.E. et al. Clinical diagnosis of progressive supranuclear palsy: The movement disorder society criteria. Movement Disorders. 2017;32(6):853-864. • Iranzo A., Silber M.H. Stop, look, and listen: Sleep in movement disorders. Movement Disorders. 2017;32(5):633-635. • Pont-Sunyer C, Tolosa E, Caspell-Garcia C, Coffey C, Alcalay RN, Chan P et al. The prodromal phase of leucine-rich repeat kinase 2-associated Parkinson disease: Clinical and imaging Studies.Movement disorders : official journal of the Movement Disorder Society. 2017;32(5). • Respondek G, Kurz C, Arzberger T, Compta Y, Englund E, Ferguson LW et al. Which ante mortem clinical features predict progressive supranuclear palsy pathology?. Movement disorders : official journal of the Movement Disorder Society. 2017;32(7). • Perez-Laso C, Cerdan S, Junque C, Gómez Á, Ortega E, Mora M et al. Effects of Adult Female Rat Androgenization on Brain Morphology and Metabolomic Profile.Cerebral cortex (New York, N.Y. : 1991). 2017. • Gaig C., Iranzo A., Pujol M., Perez H., Santamaria J. Periodic limb movements during sleep mimicking REM sleep behavior disorder: A new form of periodic limb movement disorder. Sleep. 2017;40(3). • Mahlknecht P, Iranzo A, Stefani A, Serradell M, Santamaria J, Tolosa E et al. Caveats of Neurodegenerative Risk Stratification in Idiopathic REM Sleep Behavior Disorder by Use of the MDS Research for Prodromal Parkinson’s Disease.Sleep. 2017;40(10). • Fernandez-Arcos A., Iranzo A., Serradell M., Gaig C., Guaita M., Salamero M. et al. Diagnostic Value of Isolated Mentalis Versus Mentalis Plus Upper Limb Electromyography in Idiopathic REM Sleep Behavior Disorder Patients Eventually Developing a Neurodegenerative Syndrome. Sleep. 2017;40(4). • Iranzo A., Serradell M., Santamaria J. Excessive daytime sleepiness does not predict neurodegeneration in idiopathic REM sleep behavior disorder. Sleep. 2017;40(12). • Ballester-Plane J., Schmidt R., Laporta-Hoyos O., Junque C., Vazquez E., Delgado I. et al. Whole-brain structural connectivity in dyskinetic cerebral palsy and its association with motor and cognitive function. Human Brain Mapping. 2017;38(9):4594-4612. • Campabadal A., Uribe C., Segura B., Baggio H.C., Abos A., Garcia-Diaz A.I. et al. Brain correlates of progressive olfactory loss in Parkinson’s disease. Parkinsonism and Related Disorders. 2017;41:44-50.

2017 CIBERNED Annnual Report / 225 • 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. [ePub2017]. • 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. [ePub2017]. • Abos A., Baggio H.C., Segura B., Garcia-Diaz A.I., Compta Y., Marti M.J. et al. Discriminating cognitive status in Parkinson’s disease through functional connectomics and machine learning. Scientific Reports. 2017;7. • Laporta-Hoyos O., Pannek K., Ballester-Plane J., Reid L.B., Vazquez E., Delgado I. et al. White matter integrity in dyskinetic cerebral palsy: Relationship with intelligence quotient and executive function. NeuroImage: Clinical. 2017;15:789-800. • Contarino M.F., Van Den Dool J., Balash Y., Bhatia K., Giladi N., Koelman J.H. et al. Clinical practice: Evidence-based recommendations for the treatment of cervical dystonia with botulinum toxin. Frontiers in Neurology. 2017;8(FEB). • Perez-Carbonell L., Silva C., Matos N., Gaig C., Iranzo A., Santamaria J. Actigraphy: a useful tool to monitor sleep-related hypermotor seizures. Sleep Medicine. 2017;40:1-3. • Matos N., Gaig C., Santamaria J., Iranzo A. Cataplexy causing subdural hematomas. Sleep Medicine. 2017;30:15-16. • Stefani A., Iranzo A., Santamaria J., Hogl B. Description of sleep paralysis in The Brothers Karamazov by Dostoevsky. Sleep Medicine. 2017;32:198-200. • Pujol M., Pujol J., Alonso T., Fuentes A., Pallerola M., Freixenet J. et al. Idiopathic REM sleep behavior disorder in the elderly Spanish community: A primary care center study with a two- stage design using video-polysomnography. Sleep Medicine. 2017. • Perez-Carbonell L., Silva C., Gaig C., Carreno M., Santamaria J., Iranzo A. Periodic neck myoclonus during sleep. Sleep Medicine. 2017;38:71-72. • Ramos-Campoy O., Gaig C., Villas M., Iranzo A., Santamaria J. REM sleep behavior disorder causing subdural hematoma. Sleep Medicine. 2017;30:43-44. • Lees A.J., Ferreira J., Rascol O., Reichmann H., Stocchi F., Tolosa E. et al. Opicapone for the management of end-of-dose motor fluctuations in patients with Parkinson’s disease treated with L-DOPA. Expert Review of Neurotherapeutics. 2017;17(7):649-659. • Ayala A., Trivino-Juarez J.M., Forjaz M.J., Rodriguez-Blazquez C., Rojo-Abuin J.-M., Martinez- Martin P. et al. Parkinson’s disease severity at 3 years can be predicted from non-motor symptoms at baseline. Frontiers in Neurology. 2017;8(OCT). • Obeso J.A., Stamelou M., Goetz C.G., Poewe W., Lang A.E., Weintraub D. et al. Past, present, and future of Parkinson’s disease: A special essay on the 200th Anniversary of the Shaking Palsy. Movement Disorders. 2017;32(9):1264-1310. • Ottino-Gonzalez J., Jurado M.A., Garcia-Garcia I., Segura B., Marques-Iturria I., Sender-Palacios M.J. et al. Allostatic load is linked to cortical thickness changes depending on body-weight status. Frontiers in Human Neuroscience. 2017;11. • Barrantes S., Sanchez Egea A.J., Gonzalez Rojas H.A., Marti M.J., Compta Y., Valldeoriola F. et al. Differential diagnosis between Parkinson’s disease and essential tremor using the smartphone’s accelerometer. PLoS ONE. 2017;12(8). • Tió E, Gaig C, Giner-Soriano M, Romero O, Jurado MJ, Sansa G et al. The prevalence of narcolepsy in Catalunya (Spain).Journal of sleep research. 2017. • Garcia-Diaz A.I., Segura B., Baggio H.C., Marti M.J., Valldeoriola F., Compta Y. et al. Structural Brain Correlations of Visuospatial and Visuoperceptual Tests in Parkinson’s Disease. Journal of the International Neuropsychological Society. 2017;:1-12. • Colom-Cadena M., Grau-Rivera O., Planellas L., Cerquera C., Morenas E., Helgueta S. et al. Regional overlap of pathologies in lewy body disorders. Journal of Neuropathology and Experimental Neurology. 2017;76(3):216-224.

2017 CIBERNED Annual Report / 226 Program 2 Group: Eduard Tolosa Sarró

• Vilas D., Fernandez-Santiago R., Sanchez E., Azcona L.J., Santos-Montes M., Casquero P. et al. A Novel p.Glu298Lys Mutation in the ACMSD Gene in Sporadic Parkinson’s Disease. Journal of Parkinson’s Disease. 2017;7(3):459-463. • Cisternas A, Aguilar F, Montserrat JM, Àvila M, Torres M, Iranzo A et al. Effects of CPAP in patients with obstructive apnoea: is the presence of allergic rhinitis relevant?. Sleep & breathing = Schlaf & Atmung. 2017;21(4). • Modreanu R., Cerquera S.C., Marti M.J., Rios J., Sanchez-Gomez A., Camara A. et al. Cross- sectional and longitudinal associations of motor fluctuations and non-motor predominance with cerebrospinal τ and Aβ as well as dementia-risk in Parkinson’s disease. Journal of the Neurological Sciences. 2017;373:223-229. • Hamel W., Koppen J.A., Alesch F., Antonini A., Barcia J.A., Bergman H. et al. Targeting of the Subthalamic Nucleus for Deep Brain Stimulation: A Survey Among Parkinson Disease Specialists. World Neurosurgery. 2017;99:41-46. • Valldeoriola F., Santacruz P., Rios J., Compta Y., Rumia J., Munoz J.E. et al. l-Dopa/carbidopa intestinal gel and subthalamic nucleus stimulation: Effects on cognition and behavior. Brain and Behavior. 2017;7(11). • Moren C., Gonzalez-Casacuberta I., Navarro-Otano J., Juarez-Flores D., Vilas D., Garrabou G. et al. Colonic Oxidative and Mitochondrial Function in Parkinson’s Disease and Idiopathic REM Sleep Behavior Disorder. Parkinson’s Disease. 2017;2017. • Valldeoriola F, Salvador A, Gómez-Arguelles JM, Marey J, Moya M, Ayuga Á, Ramírez F. The effects of transdermal rotigotine on non-motor symptoms of Parkinson’s disease: a multicentre, observational, retrospective, post-marketing study. International Journal of Neuroscience. 2018. 128, 4: 369-375. [ePub2017]. • Rodriguez-Violante M., Cervantes-Arriaga A., Fahn S., Tolosa E. Two-hundred years later: Is Parkinson’s disease a single defined entity?. Revista de Investigacion Clinica. 2017;69(6):308-313. • Penadés R, González-Rodríguez A, Catalán R, Segura B, Bernardo M, Junqué C. Neuroimaging studies of cognitive remediation in schizophrenia: A systematic and critical review.World journal of psychiatry. 2017;7(1). • Iranzo A. Parasomnias and Sleep-Related Movement Disorders in Older Adults. Sleep Medicine Clinics. 2018;13(1):51-61. [ePub2017]. • Hogl B., Iranzo A. Rapid Eye Movement Sleep Behavior Disorder and Other Rapid Eye Movement Sleep Parasomnias. CONTINUUM Lifelong Learning in Neurology. 2017;23(4, SleepNeurology):1017-1034. RESEARCH PROJECYS

• Code: RD16/0011. • Code: PI2016/06. Title: Red de Terapia Celular. Title: Identificación de vías fisiopatológicasy Principal Investigator: Isabel Fariñas. biomarcadores candidatos en la fase pre- CIBERNED’s collaboration: Yes. diagnóstica de la enfermedad de Parkinson. CIBERNED groups: G607, G301, G102, G113, Principal Investigator: Miquel Vila Bover. G208, G105, G207. Other CIBER’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G109, G601, G207, G410. Type: National. Other CIBER’s collaboration: No. Funding agency: Instituto de Salud Carlos III. Type: Intramurales. Funding: 284.999€. Funding agency: CIBERNED. Duration: 2016-2020 Funding: 196.000 €. Duration: 2016-2018

2017 CIBERNED Annual Report / 227 • Code: 288/C/2014. • Code: ID11849. Title: Catalan Network of Multiple System Title: LRRK2 peripheral blood mononuclear Atrophy: Biomarkers and Pathophysiology. cells and urine biosample collection’. Principal Investigator: M.j. Marti. Principal Investigator: Maria José Marti CIBERNED’s collaboration: Yes. Domenech, Eduardo Tolosa. CIBERNED groups: G109, G207. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: G207. Type: Private. Other CIBER’s collaboration: No. Funding agency: Fundació La Marató de TV3. Type: International. Funding: 299.611 €. Funding agency: Michael J. Fox Foundation. Duration: 2015-2017 Funding: 13.8506€. Duration: 2016-2018 • Code: 2014\FTPGB-LAGUNA. Title: Estudio del perfil transcripcional en • Code: N.D. subgrupos de pacientes prodrómicos con Title: The Parkinson’s Progression Markers enfermedad de Parkinson. Initiative (PPMI). Principal Investigator: Ariadna Laguna Tuset. Principal Investigator: Eduard Tolosa. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G109, G207. CIBERNED groups: G207. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Private. Type: International. Funding agency: Fundación Tatiana Pérez Funding agency: The Michael J. Fox de Guzmán el Bueno. Foundation. Funding: 44.000 €. Funding: 1.830.210 €. Duration: 2014-2017 Duration: 2013-2018 • Code: AGL2015-71764-REDT. • Code: RD16/0011/0016. Title: Red nacional de priones. Title: Combined protective/restorative cell- Principal Investigator: José Antonio del Río. mediated strategies for neurodegenerative CIBERNED’s collaboration: Yes. diseases. CIBERNED groups: G114, G509, G503, G609, Principal Investigator: José Luis Labandeira G207. García. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: National. CIBERNED groups: G208, G102, G113, G105, Funding agency: MINECO. G207. Funding: 39.000 €. Other CIBER’s collaboration: No. Duration: 2016-2017 Type: National. Funding agency: Instituto de Salud Carlos III. • Code: MJFF _DYSK_2014. Funding: 240.245€. Title: Gene-gene interaction analysis of Duration: 2017-2021 the mTOR pathway in L-DOPA induced dyskinesia’. • Code: PI042977. Principal Investigator: María José Martí Title: Mitochondrial DNA copy number in Domènech, Cristina Malagelada. LRRK2 fibroblasts. CIBERNED’s collaboration: No. Principal Investigator: Eduard Soriano. CIBERNED groups: G207. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G410, G207. Type: International. Other CIBER’s collaboration: No. Funding agency: Michael J. Fox Foundation. Type: International. Funding: 125.000€. Funding agency: Michael J. Fox Foundation. Duration: 2015-2018 Funding: 48.000 €. Duration: 2016-2017

2017 CIBERNED Annual Report / 228 Carlos Vicario Abejón 108

Principal Investigator Carlos Vicario Abejón

Research team

Eva Díaz Guerra Elena Moreno Jiménez Pablo Fernández Rodríguez Postdoctoral Fellow Master Student Master Student

Eva Rodríguez Traver Mª José Román Manuel Antonio Oria Muriel PhD Student Laboratory Technician Master Student

CSIC Avenida Doctor Arce 37 28002 Madrid (Spain) Tel.: +34 91 585 47 21 Fax: +34 91 585 47 54 [email protected]

2017 CIBERNED Annual Report / 229 SUMMARY

The study on the effects on N370S and L444P iPSCs into hippocampal neurons and into mutations in GBA1 on dopaminergic neurons astrocyte with the objective of studying APOE- derived from induced pluripotent stem cells (iPS related phenotypes. This is being carried out cells/iPSCs) prepared from Parkinson´s disease in collaboration with Dr. Moratalla. Thanks patients is progressing notably. Thanks to this to this achievement, a Master Thesis carried achievement, a Doctoral Thesis and a Master out in our laboratory has been successfully Thesis carried out in our laboratory have been dissertated. We are participating in ADAPTED, successfully dissertated. Moreover, we have a European project funded by IMI-H2020, presented the results, with the collaboration of performed in collaboration with Dr. Agustín Ruiz Dr. Rosario Moratalla (Instituto Cajal-CSIC and (Fundació ACE), among other investigators. CIBERNED), at the 47th Annual Meeting of the Society for Neuroscience (SfN) and at the XVII We have extended our study on the effects and Meeting of the Spanish Society for Neuroscience/ mechanisms of action of IGF-1 (specifically that SENC (Rodríguez-Traver,…, Moratalla, Vicario, synthetized in the brain) during hippocampal 2017). In collaboration with Dr. Moratalla neurogenesis to the analisis of synaptic we have obtained a grant funded by the plasticity and behaviour in a Nervous System- Fundación Ramón Areces and have published IGF-I conditional knockout mouse. These an article in the journal Movement Disorders studies are performed in collaboration with (García-Sanz,…, Vicario, Moratalla, 2017). Drs. Gertrudis Perea (Instituto Cajal-CSIC) and Ignacio Torres-Alemán (Instituo Cajal-CSIC and We have characterized iPS cells generated in CIBERNED) our laboratory by reprogramming fibroblasts isolated from APOE-associated Alzheimer´s We are completing the characterization disease (AD) or from AD patients carrying a of neural stem and progenitor cells, mutation in the PSEN1 gene. Moreover, we located in the adult mouse olfactory bulb, have optimized protocols to differentiate with the capacity to generate neurons. KEYWORDS

iPS cells, neural stem cells, Alzheimer´s disease, Parkinson´s disease, neurogenesis, adult neurogenesis, IGF-1 PUBLICATIONS

• Martin-Ibanez R., Pardo M., Giralt A., Miguez A., Guardia I., Marion-Poll L. et al. Helios expression coordinates the development of a subset of striatopallidal medium spiny neurons. Development (Cambridge). 2017;144(8):1566-1577. • Garcia-Sanz P., Orgaz L., Bueno-Gil G., Espadas I., Rodriguez-Traver E., Kulisevsky J. et al. N370S- GBA1 mutation causes lysosomal cholesterol accumulation in Parkinson’s disease. Movement Disorders. 2017;32(10):1409-1422.

2017 CIBERNED Annual Report / 230 Program 2 Group: Carlos Vicario Abejón RESEARCH PROJECTS

• Code: 115975-2. • Code: Fundación Ramón Areces 2017. Title: Alzheimer´s disease apolipoprotein Title: Efecto de las mutaciones del gen pathology for treatment elucidation and glucocerebrosidasa-1 en neuronas development. derivadas de células iPS de enfermos Principal Investigator: Carlos Vicario Abejón. de párkinson. Rescate del fenotipo y CIBERNED’s collaboration: No. trasplante celular. Area: Terapia celular en CIBERNED groups: G108. enfermedades neurodegenerativas. Other CIBER’s collaboration: No. Principal Investigator: Carlos Vicario Abejón. Type: European. CIBERNED’s collaboration: Yes. Funding agency: Comisión Europea. CIBERNED groups: G204, G108. Funding: 307.010,12 €. Other CIBER’s collaboration: No. Duration: 2016-2019 Type: Private. Funding agency: Fundación Ramón Areces. • Code: SAF2016-80419-R. Funding: 120.000€. Title: Regulación de la multipotencialidad Duration: 2017-2020 de células madre neurales humanas y la neurogénesis en modelos de alzhéimer • Code: PI2015-2/02. esporádico y alzhéimer familiar. Title: Potencial patológico de los astrocitos: Principal Investigator: Carlos Vicario Abejón. una nueva perspectiva en la enfermedad CIBERNED’s collaboration: No. de Alzheimer. CIBERNED groups: G108. Investigador Principal: Joan X. Comella Other CIBER’s collaboration: No. Carnice. Type: National. CIBERNED’s collaboration: Yes. Funding agency: MINECO. CIBERNED groups: G413, G415, G204, G108, Funding: 157.300 €. G411. Duration: 2016-2019 Colaboración otros CIBERs/Centros: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 350.000 €. Duration: 2016-2017

PhD DISSERTATIONS

• Author: Elena Moreno Jiménez. • Author: Pablo Fernández Rodríguez. Title: Establecimiento de modelos celulares Title:Human iPSCs: Characterization and de alzhéimer, asociado a la ApoE4, protocol optimization for dopaminergic basados en la tecnología de células madre neuronal differentiation as a model for pluripotentes inducidas. Parkinson´s disease. Date: 25th September 2017. Date: 4th September 2017. Supervisor: Carlos Vicario Abejón. Supervisor: Carlos Vicario Abejón.

• Author: Eva Rodríguez Traver. Title: Generation and differentiation of dopaminergic neurons from mouse multipotent and human induced pluripotent stem cells to study Parkinson´s disease. Date: 13th September 2017. Supervisor: Carlos Vicario Abejón.

2017 CIBERNED Annual Report / 231 Miquel Vila Bover 109

Principal Investigator Miquel Vila Bover

Research team

Jordi Bové Badell Thais Cuadros Arasa Núria Peñuelas Peñarroya Miguel Servet Researcher Postdoctoral fellow Predoctoral fellow

Marta Martínez Vicente Ariadna Laguna Tuset Annabelle Parent Miguel Servet Researcher Researcher Technician

Oriol de Fàbregues-Boixar Sandra Franco Iborra Beatriz Rodríguez Galván Nebot Predoctoral fellow Technician Collaborator Albert Torra Talavera Jordi Romero Giménez Iria Carballo Carbajal Predoctoral fellow Technician Postdoctoral fellow Jordi Galiano Landeira Marta González Sepúlveda Predoctoral fellow Postdoctoral fellow

Neurodegenerative Diseases Research Group Vall d’Hebron Research Institute Mediterranean Building, First Floor, Lab. 102 Pg. Vall d’Hebron, 119-129 08035 Barcelona (Spain) Tel.: +34 93 489 45 43 / 38 85 Fax: +34 93 489 40 15 [email protected] www.vilalab.org

2017 CIBERNED Annual Report / 232 SUMMARY

In 2017, as our most important study, we In addition, we have shown that the selective have developed a new therapeutic strategy genetic ablation in dopaminergic neurons of to selectively decrease the brain levels the lysosomal enzyme glucocerebrosidase of α-synuclein in vivo selectively within (GBA), whose mutations are associated with monoaminergic neurons, the type of neurons an increased risk of PD, does not result in most susceptible to neurodegeneration in neurodegeneration, thus indicating that a Parkinson’s disease (PD). This technique is based loss of GBA function is not sufficient by itself to on the intranasal administration of chemically- cause PD. modified oligonucleotides. KEYWORDS

Parkinson’s disease, α-synuclein, glucocerebrosidase PUBLICATIONS

• Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, García-Aznar JM et al. Erratum to: Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons.Molecular neurobiology. 2017. • Urrea L, Segura-Feliu M, Masuda-Suzukake M, Hervera A, Pedraz L, Aznar JM et al. Involvement of Cellular Prion Protein in α-Synuclein Transport in Neurons.Molecular neurobiology. 2017. • 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. [ePub2017]. • Alarcon-Aris D., Recasens A., Galofre M., Carballo-Carbajal I., Zacchi N., Ruiz-Bronchal E. et al. Selective α-Synuclein Knockdown in Monoamine Neurons by Intranasal Oligonucleotide Delivery: Potential Therapy for Parkinson’s Disease. Molecular Therapy. 2017. • Catalan M.J., Antonini A., Calopa M., Bajenaru O., de Fabregues O., Minguez-Castellanos A. et al. Can suitable candidates for levodopa/carbidopa intestinal gel therapy be identified using current evidence?. eNeurologicalSci. 2017;8:44-53. • Cavaliere F, Cerf L, Dehay B, Ramos-Gonzalez P, De Giorgi F, Bourdenx M et al. In vitro α-synuclein neurotoxicity and spreading among neurons and astrocytes using Lewy body extracts from Parkinson disease brains.Neurobiology of disease. 2017;103. • Soria FN, Engeln M, Martinez-Vicente M, Glangetas C, López-González MJ, Dovero S et al. Glucocerebrosidase deficiency in dopaminergic neurons induces microglial activation without neurodegeneration.Human molecular genetics. 2017;26(14). • Antonini A., Poewe W., Chaudhuri K.R., Jech R., Pickut B., Pirtosek Z. et al. Levodopa-carbidopa intestinal gel in advanced Parkinson’s: Final results of the GLORIA registry. Parkinsonism and Related Disorders. 2017;45:13-20. • Garcia-Prat L., Munoz-Canoves P., Martinez-Vicente M. Monitoring autophagy in muscle stem cells. Methods in Molecular Biology. 2017;1556:255-280. • Garcia-Lezana T., Oria M., Romero-Gimenez J., Bove J., Vila M., Genesca J. et al. Cerebellar neurodegeneration in a new rat model of episodic hepatic encephalopathy. Journal of Cerebral Blood Flow and Metabolism. 2017;37(3):927-937. • Martinez-Vicente M. Neuronal mitophagy in neurodegenerative diseases. Frontiers in Molecular Neuroscience. 2017;10.

2017 CIBERNED Annnual Report / 233 RESEARCH PROJECTS

• Code: PI2016/06. Type: Private. Title: Identificación de vías fisiopatológicasy Funding agency: Fundació la Marató de TV3. biomarcadores candidatos en la fase pre- Funding: 191.656 €. diagnóstica de la enfermedad de Parkinson. Duration: 2015-2018 Principal Investigator: Miquel Vila Bover. CIBERNED’s collaboration: Yes. • Code: 2014\FTPGB-LAGUNA. CIBERNED groups: G109, G601, G207, G410. Title: Estudio del perfil transcripcional en Other CIBER’s collaboration: No. subgrupos de pacientes prodrómicos con Type: Intramurales. enfermedad de Parkinson. Funding agency: CIBERNED. Principal Investigator: Ariadna Laguna Tuset. Funding: 196.000 €. CIBERNED’s collaboration: Yes. Duration: 2016-2018 CIBERNED groups: G109, G207. Other CIBER’s collaboration: No. • Code: PI14/01529. Type: Private. Title: Rol patogénico de la disfunción autofágica/ Funding agency: Fundación Tatiana Pérez lisosomal en la enfermedad de Parkinson: de Guzmán el Bueno. diagnóstico y nuevas terapias pre-clínicas. Funding: 44.000 €. Principal Investigator: Marta Martínez Vicente. Duration: 2014-2017 CIBERNED’s collaboration: No. CIBERNED groups: G109. • Code: MJFF_TARGET ADV2016. Other CIBER’s collaboration: Si (CIBER-BBN). Title: Targeting a novel transcriptional pathway Tipo: National. to reduce alpha-synuclein expression. Funding agency: Instituto de Salud Carlos III. Principal Investigator: Miquel Vila Bover/ Funding: 110.715 €. Solange Desagher. Duration: 2014-2017 CIBERNED’s collaboration: No. CIBERNED groups: G109. • Code: 288/C/2014. Other CIBER’s collaboration: No. Title: Catalan Network of Multiple System Type: International. Atrophy: Biomarkers and Pathophysiology. Funding agency: The Michael J. Fox Principal Investigator: M.J. Marti. Foundation. CIBERNED’s collaboration: Yes. Funding: 91.400 €. CIBERNED groups: G109, G207. Duration: 2016-2017 Other CIBER’s collaboration: No. Type: Private. • Code: SAF2016-77541-R. Funding agency: Fundaciò La maratò de TV3. Title: Diagnostic and therapeutic potential Funding: 299.611€. of neuromelanin in Parkinson’s disease using Duration: 2015-2017 novel humanized preclinical in vivo model. Principal Investigator: Miquel Vila Bover. • Code: 392/U/2014. CIBERNED’s collaboration: No. Title: Role of the cellular prion protein as CIBERNED groups: G109. “cross-talk” protein between alpha-syn/ Other CIBER’s collaboration: No. LRRK2 and p-Tau in sporadic and familiar Type: National. Parkinson´s disease. Funding agency: MINECO. Principal Investigator: José Antonio del Río. Funding: 363.000€. CIBERNED’s collaboration: Yes. Duration: 2017-2020 CIBERNED groups: G109, G114. Other CIBER’s collaboration: No. • Code: MJFF TPP2014. Type: Private. Title: A precise approach for nucleoside- Funding agency: Fundació la Marató de TV3. based therapy of neuromuscular disorders Funding: 195.646 €. with defects in mitochondrial dna. Duration: 2014-2017 Principal Investigator: Miguel Ángel Martín Casanueva. • Code: Fundació la Marató de TV3-Malalties CIBERNED’s collaboration: No. del cor (320/U/2015). CIBERNED groups: G109. Title: New molecular functions of apoptotic Other CIBER’s collaboration: Yes (CIBERER). genes in cardiac development and stress. Type: National. Principal Investigator: Daniel Sanchis Morales. Funding agency: MINECO. CIBERNED’s collaboration: No. Funding: 806.223 €. CIBERNED groups: G109. Duration: 2016-2019 Other CIBER’s collaboration: No.

2017 CIBERNED Annual Report / 234 Program 2 Group: Miquel Vila Bover

• Code: SAF2015-73997-JIN. • Code: 392/U/2014 (IBEC). Title: The Gut-Brain axis in Prodromal Parkinson’s Title: Paper de la PrPc com a “cross-talk disease: new possibilities for risk/diagnostic protein” entre alpha-sinucleina/LRRK2 i biomarkers and therapeutic interventions. p-Tau en la malaltia de Parkinson de tipus Principal Investigator: Ariadna Laguna Tuset. esporàdic i/o familiar. CIBERNED’s collaboration: No. Principal Investigator: José Antonio Del Río. CIBERNED groups: G109. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G114, G109. Type: National. Other CIBER’s collaboration: No. Funding agency: MINECO. Type: Private. Funding: 170.000€. Funding agency: Fundaciò La Maratò de TV3. Duration: 2016-2018 Funding: 195.646,25€.. Duration: 2015-2017

2017 CIBERNED Annual Report / 235

Cooperative research

COOPERATIVE RESEARCH

The current 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 benefit of the research activity. The National Evaluation Agency (ANEP), although subject to final approval by the Steering Committee, externally reviews the proposals, so that the allocation of funds is independent and transparent.

The cooperative research program has so far launched eight 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 first of which included 3 collaborative projects with a total allocation of 690,000€. The projects from the fifth call (2015-I) ended by the end of 2017.

The projects from these first five calls are listed below: 2010 CALL

Code Title Coordinator

PI2010/05 Generating a dopaminergic neuronal model from induced Tolosa Sarró, Eduardo pluripotent stem cells from patients with Parkinson’s disease associated to mutations in the LRRK2 gene

PI2010/06 Neuroprotection in Huntington’s disease García de Yébenes Prous, Justo

PI2010/07 Reelin and GSK3 as therapeutic targets in Alzheimer’s Ávila de Grado, Jesús disease

PI2010/08 Glial activation during the neuroinflammatory process: a Vitorica Fernández, potential therapeutic target for Alzheimer’s disease Francisco Javier

PI2010/09 BESAD-P: Biomarkers of early stages of Alzheimer disease - Ferrer Abizanda, Isidro prevention

PI2010/11 Consortium to generate a common database aimed at Illa Sendra, Isabel implementing clinical and basic research on neuromuscular diseases

2017 CIBERNED Annual Report / 239 2011 CALL

Code Title Coordinator PI2011/01 The Nrf2 transcription factor as a new therapeutic target Cuadrado Pastor, for Parkinson’s disease Antonio

PI2011/02 Onset and progression of Parkinson’s disease. Vulnerability Obeso Inchausti, of the nigrostriatal pathway, events at origin and José Ángel destination

PI2011/03 Generation of dopaminergic neurons from somatic cells of Moratalla Villalba, Rosario parkinsonian patients with cognitive failure

PI2011/04 Multicenter study on LCR and neuroimaging biomarkers in Lleó Bisa, Alberto the continuum preclinical- prodromal Alzheimer’s disease (SIGNAL Study)

2013 CALL

Code Title Coordinator

PI2013/01 Emergent properties of the neuron-glia relationship Torres Alemán, Ignacio underlying neurodegeneration and dementia in Alzheimer’s disease

PI2013/05 Identification and molecular characterization Guzmán Pastor, Manuel of cannabinoid receptors subpopulations in poliglutaminopatías

PI2013/07 Role of GSK-3β in the cortical circuits alterations occurring Iglesias Vacas, Teresa in Alzheimer’s disease

PI2013/08 Mitochondrial dynamics and mitophagy as therapeutic Soriano García, Eduardo targets in Parkinson’s and Huntington’s diseases

PI2013/09 Synaptic degeneration and dysregulation of adult Fernández Chacón, neurogenesis in murine models of neurodegeneration Rafael

2014 CALL

Code Title Coordinator

PI2014/02 Epigenetic mechanisms involved in the etiology and Calero Lara, Miguel progression of rapidly progressive neurodegenerative dementias

PI2014/06 Onset and progression of Parkinson’s disease: role of glial Rodríguez Oroz, Mª Cruz activation

2017 CIBERNED Annual Report / 240 2015-1 CALL

Code Title Coordinator

PI2015/01 Neuregulin gene therapy aimed at the treatment of motor Navarro Acebes, Xavier neuron degeneration in ALS

PI2015/02 Validation of new therapeutic targets and biomarkers in Labandeira Garcia, Parkinson’s disease José Luis

PI2015/03 Differential metabolic profiles in Parkinson’s disease Fuentes Rodríguez, José Manuel

Those twenty already finished projects have actively involved 57 CIBERNED research groups overall, in addition to some other external ones, representing 90,4 % of the existing groups.

The 2015-I call for CIBERNED cooperative projects ended in December 2017 and ss stated in the call remit, the groups involved addressed the obtained results in final reports that were provided to CIBERNED’s scientific management.

Below is a brief description of the results obtained by each of the projects from this 2014 call:

2017 CIBERNED Annual Report / 241 • Project PI2015/01: Neuregulin gene therapy aimed at the treatment of motor neuron degeneration in ALS

Principal Investigator Institution

Navarro Acebes, Xavier Autonomous University of Barcelona Rodríguez Álvarez, José Autonomous University of Barcelona García Verdugo, José Manuel Cavanilles Institute, University of Valencia Sáez Valero, Javier Miguel Hernández University, Elche

Recent evidence suggests an association of the neurotrophic factor Neuregulin 1 (Nrg1) and its receptor ErbB4 in Amyotrophic Lateral Sclerosis (ALS) and other neurodegenerative diseases. Thus, loss-of-function mutations in the Nrg1 receptor ErbB4 are associated with late-onset autosomal dominant ALS. Nrg1 expression is reduced both in ALS patients and SOD1G93A transgenic mice. In addition, active Nrg1 requires sequential processing by α-secretase (ADAM proteins) or β- secretase (BACE1) and presenilin (PS)/γ-secretase, proteases also involved in the processing of APP in Alzheimer´s disease. However, the pathogenic role of Nrg1 and its processing in motoneuron survival is not well understood. Our hypothesis is that Nrg1/ErbB signaling could be a relevant pathogenic mechanism underlying neurodegeneration, and targeting Nrg1/ErbB signaling may represent a novel therapeutic approach for ALS and other neurodegenerative diseases.

We first characterized the expression of Nrg1 isoforms and ErbB receptors in spinal cordand skeletal muscles of SOD1G93A mice and ALS patients. We found a significant reduction of ErbB4 levels in muscles of both ALS mice and human patients. In the spinal cord of the ALS mice, Nrg1 type I levels increased with progression of the disease, whereas Nrg1 type III levels were reduced; we also observed progressively reduced levels of ErbB3 and ErbB4 receptors in the spinal cord of ALS mice with age.

We then used an in vitro model to characterize the effect of NRG1 on motoneuron survival under excitotoxicity. Addition of recombinant human NRG1 (rhNRG1) to the medium significantly increased motoneuron survival through activation of ErbB receptors, and reduced microglial reactivity, overcoming the excitotoxicity effects.

To address the role of Nrg1/ErbB4 processing and signaling in neurodegeneration, we found a significant reduction of Nrg1 type I mRNA during aging but not differences in SOD1G93A mice and PS cKO mice, whereas Nrg1 type III mRNA was reduced in PS cKO mice. Biochemical analyses indicate PS/g-secretase-dependent processing of Nrg1 and ErbB4 in the cortex of adult mice, and impaired Nrg1/ErbB4 processing/levels in the two mouse models of neurodegeneration.

In addition, we have characterized a soluble ectodomain fragment of ErbB4 (NTF) in human CSF. Levels of the 55-kDa ecto-ErbB4 were found decreased in CSF and in plasma samples from ALS and ALS plus frontotemporal dementia patients. It also appeared decreased in plasma of SOD1G93A mice, thus suggesting that soluble ecto-ErbB4 fragments could be a tool to evaluate impairment of ErbB4 pathway in ALS. Light and electron microscopy evidenced clear signs of cell damage in the somas of motoneurons of SOD1G93A mice, including reduction in cell volume and of the number and length of active synaptic contacts.

We assayed the overexpression of Nrg1-I in the skeletal muscle of SOD1G93A mice by means of an AAV1 vector, and demonstrated that Nrg1-I promoted collateral reinnervation of denervated endplates, opening a new window for developing novel ALS therapies for functional recovery rather than preservation. Consequently, a more efficient approach has been attempted by i.v. administration of AAV8 coding for Nrg1-I under regulation of desmin promoter that allowed to

2017 CIBERNED Annual Report / 242 infect several muscles in the limbs and diaphragm. The therapeutic assay in SOD1G93A mice, however, did not produce significant improvement in functional and electrophysiological results. Finally, we injected intrathecally AAVrh10-Nrg1-III in SOD1G93A mice, achieving to increase expression in spinal neurons. We found improved electrophysiological results and higher number of surviving motoneurons in the Nrg1-III overexpressing group than in the AAV-mock mice. These data indicate that Nrg1-III promotes preservation of spinal motoneurons, in agreement with the in vitro results.

Finally, we injected intrathecally AAVrh10-Nrg1-III in SOD1G93A mice, achieving to increase expression in spinal neurons. We found improved electrophysiological results and higher number of surviving motoneurons in the Nrg1-III overexpressing group than in the AAV-mock mice. These data indicate that Nrg1-III promotes preservation of spinal motoneurons, in agreement with the in vitro results.

• Project PI2015/02: Validation of new therapeutic targets and biomarkers in Parkinson’s disease

Principal Investigator Institution

Labandeira Garcia, José Luis University of Santiago de Compostela Cuadrado Pastor, Antonio Autonomous University of Madrid Lanciego Pérez, José Luis Center of Applied Medical Research, Univ. Navarra, Pamplona Kulisevsky Bojarski, Jaime Santa Creu i Sant Pau Hospital, Barcelona

The development of new therapeutic strategies against neurodegenerative diseases such as Parkinson’s disease (PD) is a major objective in biomedical research. New strategies going beyond current symptomatic treatments are necessary. Unfortunately, all neuroprotective strategies have failed in clinical trials.

The lack of therapies against cognitive impairment in PD is particularly problematic. The present project have addressed these problems as follows:

1) A new animal model of PD induced by alpha-synuclein introduced with viral vectors has been developed and optimized.

2) We have investigated two new therapeutic targets related to major mechanism of the redox metabolism and neuroinflammation: inhibition of the pro-inflammatory axis angiotensin/NADPH- oxidase/Rho-Kinase (using AT1 receptors antagonists such as candesartan and telmisartan), and activation of the transcription factor Nfr2, a master regulator of the anti-oxidant defense system(using dimethyl fumarate), and we have obtained positive results.

3) Alterations observed in preclinical models have been correlated with observations in postmortem brain tissue from PD patients.

4) Advanced Glycation End products (AGEs) and other potential biomarkers (such as autoantibodies against angiotensin receptors) are being determined in samples from plasma of PD patients and controls to find possible biomarkers of cognitive impairment in PD. Finally, the effects of the suggested therapies (candesartan) on the above mentioned biomarkers and clinical course of PD (particularly cognitive impairment) are being tested in PD patients and the corresponding controls.

2017 CIBERNED Annual Report / 243 • Project PI2015/03: Differential metabolic profiles in Parkinson’s disease

Principal Investigator Institution

Fuentes Rodríguez, José Manuel University of Extremadura, Cáceres Pérez Castillo, Ana María Institute of Biomedical Research CSIC-UAM, Madrid Pérez Tur, Jordi Institute of Biomedicine of Valencia, CSIC

Intermediate metabolism and cell signaling pathways have been considered and studied to date as independent entities. But in reality catabolic and anabolic pathways are which provide cell energy and precursors required for cell survival. On the other hand subcellular organelles with important metabolic functions, such as mitochondria play a key role in the origin of a large number of diseases including Parkinson’s disease.

Therefore the knowledge of metabolic variations to integrate and understand the mechanisms leading to neurodegeneration is essential. The determination of differential metabolic profiles in models of Parkinson’s disease may allow the development of a diagnostic pattern, risk and prognosis of it.

Objectives:

1) Identification of transcriptome and metabolomics differential profiles in:

• Cell models overexpressing pathogenic G2019S and R1441G pathogenic forms of LRRK2.

• In murine models overexpressing G2019S and R1441G pathogenic forms of Lrrk2.

• In human fibroblasts from healthy individuals, idiopathic Parkinson patients and asymptomatic carriers and patients with G2019S and R1441G pathogenic forms of LRRK2.

• In blood plasma from the same donors described in 1.3.

2) Identification of transcriptome profiles and differential models described in 1.1 metabolomics, 1.2 and 1.3 after exposure to the parkinsonian neurotoxin MPP + (cellular models) or MPTP (murine models).

3) Validation of transcriptomic and metabolomic results by determining the variation of the altered proteins involved in metabolic processes (molecular identification of drug targets).

4) Establishment of cellular and animal models of Parkinson’s disease on which test the effect of modulators / metabolic adapters as potential neuroprotective agents for Parkinson’s disease.

This project is therefore the first combined transcriptomic and metabolomics study aimed at identifying specific genetic biomarkers and neurotoxic models and with a simultaneous study in cellular systems, animals and samples from patients with idiopathic Parkinson, patients Parkinson pathogenic mutations for LRRK2 (G2019S and R1441G) and healthy carriers of the same mutation. It is a continuation of work previously done by our research group in the characterization of the deregulation of autophagy in these same cell models from Parkinson’s patients, as a metabolic stress can lead to increased basal levels of autophagy, we hope to observe important differences especially in relation to redox and energy production systems

In addition, during the second part of 2015, the sixth call was launched, which was again evaluated by the ANEP and resolved at the beginning of 2016 with the selection of 2 new projects to be funded:

2017 CIBERNED Annual Report / 244 2015-II CALL

Code Title Coordinator

PI2015-2/02 Pathological potential of astrocytes: a new perspective on Comella Carnicé, Alzheimer’s disease Joan Xavier

PI2015-2/06 Molecular mechanisms of brain and muscle stem cell Muñoz Cánoves, Pura function in aging and neurodegeneration

The funding approved for this sixth call was 350,000€ per year for 2 years, which meant a total allocation of 700,000€ to the ten CIBERNED research groups participating in both projects. Due to an extension that did not imply additional funding, the projects of the 2015-II call will end during 2018. The following is a brief description of the scientific activity carried out by each of the projects during its first year of execution:

• Project PI2015-2/02 Pathological potential of astrocytes: a new perspective on Alzheimer’s disease

Principal Investigator Institution

Comella Carnicé, Joan Xavier Vall d’Hebron University Hospital, Barcelona Vitorica Ferrández, Francisco Javier University of Seville Gutiérrez Pérez, Antonia University of Malaga Vicario Abejón, Carlos Cajal Institute CSIC, Madrid Moratalla Villalba, Rosario 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

2017 CIBERNED Annual Report / 245 learning and memory. We have additionally 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

Muñoz Cánoves, Pura Pompeu Fabra University, ICREA, Barcelona Fariñas Gómez, Isabel University of Valencia Lucas Lozano, José Javier Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid Fernández Chacón, Rafael University of Seville Iglesias Vacas, Teresa Institute of Biomedical Research 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. Nowadays, it is well established that stem cells in specific neural and muscular niches, active throughout life, are essential for the functional maintenance of the brain and skeletal muscles. Stem cell properties, though, are sensitive to aging and to pathogenic Factors such as molecular insults underlying neurodegeneration. Nevertheless, the molecular mechanisms that progressively hinder stem cells regenerative potential are poorly understood.

Our consortium will focus on the analysis of key pathogenic processes Such as: (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-Canoves has recently described key molecular pathways underlying the functional decline and senescence of satellite cells that impair muscle rejuvenation (Sousa-Victor, 2014). Interestingly, a thorough unbiased transcriptomic analysis included in that study has identified the mTOR signaling pathway as a potential culprit in stem cell aging and, remarkably, several other molecular targets that are major subjects of study in the laboratories of the other consortium’s members. Some of these targets include: (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 signalling pathways regulating cellular responses in nervous, vascular and skeletal muscle systems (Iglesias, 2000; Neubrand, 2012) that is present in neural stem cells (unpublished data from collaboration between Dr. Iglesias and Dr. Farinas groups), supporting a potential role of Kidins220 in regulating NSC function; (c) Rbfox1, a neuron and muscle specific splicing factor previously linked to muscle dysfunction in expanded CUG-caused myotonic dystrophy DM1, that is downregulated In Huntington’s disease brain thus leading to a pathogenic alteration of CPEB4 splicing (unpublished work from Dr. Lucas’ group).

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

2017 CIBERNED Annual Report / 246 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.

Likewise, during the last semester of 2016, the seventh call was launched, which was again evaluated by the ANEP and resolved at the end of 2016 with the selection of five new funded projects: 2016 CALL

Code Title Coordinator

PI2016/01 Alterations of gluco-lipid metabolism and development of Torres Alemán, Ignacio Alzheimer’s dementia

PI2016/02 Monitoring the Onset and Evolution of Neuronal Dysfunctions Del Río Fernández, in Propagative Neural Disorders using Microfluidic Devices José Antonio and Translational approaches

PI2016/04 The ALS CIBERNED Challenge: Accelerating New Drug López de Munain Arregui, Discovery Adolfo

PI2016/05 Dream inhibitors and Alzheimer´s Disease Naranjo Orovio, José Ramón

PI2016/06 Identification of pathophysiological pathways and Vila Bover, Miquel candidate biomarkers in the prediagnostic phase of Parkinson’s disease

The funding approved for this seventh call was 508,000€ per year for 2 years, which meant a total allocation of 1,016,000€ to all nineteen participating CIBERNED research groups. Below is a brief description of the scientific activity carried out by each of the projects during its first year of execution:

• Project PI2016/01: Alterations of gluco-lipid metabolism and development of Alzheimer’s dementia

Principal Investigator Institution

Torres Alemán, Ignacio Cajal Institute CSIC, Madrid Wandosell Jurado, Francisco Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid Camins Espuny, Antonio University of Barcelona Carro Díaz, Eva Doce de Octubre University Hospital, Madrid Cantero Lorente, José Luis Pablo de Olavide University, Sevilla The general aim of the Project is to determine whether metabolic disturbances may lead to cognitive alterations as seen in Alzheimer´s disease (AD). We are conducting parallel clinical and experimental studies. During the first year we have been determining the circulating gluco-lipid profile in a clinical cohort that encompasses all the spectrum of the natural history of the disease. In parallel, we are conducting in the same group of patients a brain imaging study to obtain biomarkers that could –ideally, be linked with peripheral metabolism and identify each of the AD stages. We have already gathered all the data to validate them in a wider clinical cohort and will compare them in animal models of the disease. In another AD cohort, we are also assessing possible circulating biomarkers, such as immunomodulators, that can be regulated by metabolic status.

2017 CIBERNED Annual Report / 247 In animal models we are determining whether metabolic impairment elicited by high-fat diets or altered brain glucose metabolism (as seen in AD) impact on progress of the pathology. We are modeling the familial type of the disease using APP/PS1 double mutant mice as a basic platform in where we 1) have deleted all 3 isoforms of JNK kinases, or 2) have deleted insulin or IGF-I receptors in astrocytes. Initially, we have seen that KO mice for JNK2 submitted to high- fat diet develop marked cognitive losses. In control mice under high fat diet we have also obtained the lipid profile. The same study will be done now with APP/PS1 mice, using sex as an independent variable. In this vein, mice lacking megalin in brain endothelium develop cognitive deterioration, as well as obesity, and present a neuroinflammatory phenotype. In mice where the astrocytic insulin receptor is turned down we have seen reduced brain perfusion, while mice lacking IGF-I receptors in astrocytes show elevated markers of amyloidosis and inflammation.

To model the impact of unbalanced diets (most common nowadays) on the disease, we will continue examining the role played by JNK1 (that phosphorylates proteins downstream of the insulin receptor), the ovarian hormone estradiol (as AD has a greater incidence in women), and amyloidosis. For the latter we are using the EMD mouse that lacks endothelial megalin and mimics diverse AD symptoms but without brain amyloidosis. Using these models we are evaluating several AD- (amyloidosis, tauopathy and brain hipoperfusion) and metabolic-related markers (dyslipidemia, insulin resistance). In a fourth experimental group we are beginning to determine the influence of altered brain glucose metabolism in AD pathology (APP/ PS1 mice lacking IR or IGF-IR in astrocytes), and so far, no changes have been documented.

Next year we will continue exploring pathogenic processes underlying the above mentioned central and peripheral disturbances. We will analyze in vivo and in vitro, macroautophagy mechanisms associated with lipid disturbances or insulin resistance by using high-fat diets or mice lacking insulin receptors in astrocytes, respectively. The aim is to determine the role of autophagy in development of altered proteostasis associated to AD.

• Project PI2016/02: Monitoring the Onset and Evolution of Neuronal Dysfunctions in Propagative Neural Disorders using Microfluidic Devices and Translational approaches

Principal Investigator Institution

Del Río Fernández, José Antonio Catalonian Institute de Bioengineering, Barcelona Ávila de Grado, Jesús Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid Canela Campos, Enric Isidre 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 individuals 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

2017 CIBERNED Annual Report / 248 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

López de Munain Arregui, Adolfo Biodonostia Research Institute, San Sebastian Ferrer Abizanda, Isidro Bellvitge Institute of Biomedical Research, Barcelona Fernández Ruiz, Javier Complutense University of Madrid Soriano García, Eduardo University of Barcelona

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive weakness and atrophy of skeletal muscles until paralysis and death. Multi-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 topographical 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 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. Collaborative 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 cortexin post-mortem brains of ALS (in comparison with FTLD-TDP-43).

2017 CIBERNED Annual Report / 249 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.

• Project PI2016/05: DREAM inhibitors and Alzheimer´s disease

Principal Investigator Institution

Naranjo Orovio, José Ramón National Center of Biotechnology, CSIC, Madrid De Felipe Oroquieta, Javier Cajal Institute CSIC; Technical University, Madrid Ceña Callejo, Valentín University of Castilla La Mancha, 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 dephosphorylation of cofilin 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

2017 CIBERNED Annual Report / 250 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.

• Project PI2016/06: Identification of pathophysiological pathways and candidate biomarkers in the prediagnostic phase of Parkinson’s disease

Principal Investigator Institution

Vila Bover, Miquel Vall d’Hebron University Hospital, Barcelona Tolosa Sarró, Eduardo Hospital Clinic of Barcelona Trullas Oliva, Ramón Institute of Biomedical Research IDIBAPS-CSIC , Barcelona Infante Ceberio, Jon Marqués de Valdecilla Research 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 DATSPECT), 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:

2017 CIBERNED Annual Report / 251 2017 CALL

Code Title Coordinator

PI2017/01 Study of the microRNA in the cerebrospinal fluid exosomal Clarimón Echavarria, compartment as a biomarker of frontotemporal dementia Jordi and a tool for understanding the biological basis of the disease

PI2017/02 Glucocerebrosidase and Neurodegenerative Lanciego Pérez, José Luis Proteinopathies

PI2017/04 Glial dysfunction in Alzheimer’s disease: pathogenic Vitoríca Fernández, implications and clinical potential Francisco Javier

The funding approved for this seventh call was 325,000€ per year for 2 years, which meant a total allocation of 650,000€ to the eleven participating CIBERNED research groups. The following is a summary of the objectives set for the projects approved in this seventh call, initiated at the end of 2017:

• 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

Clarimón Echavarria, Jordi Santa Creu i Sant Pau Hospital, Barcelona Rodríguez Álvarez, José Autonomous University of Barcelona Bullido Gómez-Heras, Mª Jesús Autonomous University of 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. Neuropathological 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.

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 subtype.

2017 CIBERNED Annual Report / 252 To achieve our goals, we will perform gene expression analyses using high density microarrays in neuronal cell cultures in which the levels of the candidate miRNAs will be modulated. We will then compare their expression pattern with control cultures. We will also analyze neuronal viability and possible alterations in synaptic function and density. Finally, we will include CSF specimens from patients with amyotrophic lateral sclerosis (ALS) as well as patients with concomitant ALS and FTD (ALS / FTD), characterized by presenting TDP-43 inclusions, to analyze their exosomal miRNA pattern and compare it with different FTD subtypes, Alzheimer’s disease, Lewy body dementia, corticobasal degeneration and progressive supranuclear palsy. These analyses will allow us to assess the sensitivity and specificity of the candidate miRNAs as biomarkers of DFT and, more specifically, their adequacy to distinguish between FTD-TDP43 to other, non-TDP43 forms of disease.

• Project PI2017/2: Glucocerebrosidase and Neurodegenerative Proteinopathies

Principal Investigator Institution

Lanciego Pérez, José Luis Center of Applied Medical Research, Univ. NavarrA, Pamplona Labandeira García, José Luis University of Santiago de Compostela Kulisevsky Bojarski, Jaime Santa Creu i Sant Pau Hospital, 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.

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,

2017 CIBERNED Annual Report / 253 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

Vitorica Ferrández, Francisco Javier University of Seville Gutiérrez Pérez, Antonia University of Malaga Cuadrado Pastor, Antonio Autonomous University, Madrid Sáez Valero, Javier Miguel Hernández University, Elche Comella Carnicé, Joan Xavier Vall d’Hebron University Hospital, Barcelona

The present project proposes a new pathogenic scenario in Alzheimer’s disease. We propose the microglial dysfunction and consequent astroglial hyperactivation as a fundamental mechanism in the neurodegenerative process. This new scenario could give answers to the continuous failure in the translation of the studies from models to the human clinic. Therefore, it highlights the need to reproduce this dysregulation of the innate cerebral immune response into the animal models to increase their predictive value and to assure a greater success of the therapies in humans.

Although microglia may play a cytotoxic role in neurodegenerative pathologies, the role in Alzheimer’s is not well characterized. Recent results by the consortium members demonstrate that the microglial cells undergo a degenerative process in the hippocampus of Braak V-VI patients. This degeneration totally opposites of that observed in the Abeta-producing models, classically used for preclinical studies. Consequently, as first objective, we propose to evaluate the impact on the Alzheimer’s pathology of the microglial dysfunction observed in patients. These data would improve the knowledge of the pathology and, on the other hand, it will help to improve the current murine models.

On the other hand, astroglia is considered a neuroprotective cell, based on its normal (non- pathological) function in trophic support, redox homeostasis, and elimination of toxic molecules, vascularization and control of synaptic activity. However, the astroglial specific role in the development of Alzheimer’s disease is unknown. It is well known that astrocytes are activated around the Abeta plaques. However, their phenotypic changes and gene expression patterns are not known in detail. Also, it is unknown how the factors secreted by active astrocytes may contribute to the development of the pathology. In this project, we propose to approach this problem by evaluating, on the one hand, the repercussion of microglial deficiency on the phenotype of astrocytes and, on the other hand, the role played by the transcription factor Nrf2 as the master regulator of several hemostatic cytoprotective mechanisms.

Finally, and based on the results obtained in the previous aims, we propose the characterization of the possible microglial and / or astroglial “signature” in collections of cerebrospinal fluid (CSF) of subjects with mild amnestic cognitive impairment (prodromic stages), up to subjects with Alzheimer’s disease. On the other hand, the most relevant results obtained in the first two objectives will be confirmed in post mortem samples of patients, available by several members of the consortium.

2017 CIBERNED Annual Report / 254 The cooperative activity program of CIBERNED has so far involved a global budget of approximately 8,874,174€ distributed in the thirty 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 a dozen associated external groups. In addition, 48 groups (76.1%) have participated in more than one cooperative project and projects 2015-II, 2016 and 2017 are still active, with the involvement of 37 CIBERNED research groups (71.1%).

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 for project in years groups projects and year and 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 €

2017 CIBERNED Annual Report / 255

International relations

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 partnerships. 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.

Alzheimer´s Global Summit Lisbon 2017

2017 CIBERNED Annual Report / 259 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 Institute 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 implemented through co-operative activities that realign or link national investments to achieve increased impact, and the provision of new funding. A guiding principle 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 2017, there have been no CIBERNED research groups involved in active projects within the joint European Union program for research on neurodegenerative diseases (JPND).

2017 CIBERNED Annual Report / 260 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 German 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 Neurodegeneration (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 Foundation 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 (MESRS, Slovakia) • CIBERNED-Fundación CIEN (Spain) • Agence Nationale de la Recherche (ANR, France)

2017 CIBERNED Annual Report / 261 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 establish 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 • 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 Centers 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 2017 is described below:

• Protection of neurons in vitro and in vivo from Synuclein toxicity by molecular tweezers

Project partners: Erwan Bezard (Coordinator, France), Richard Wade-Martins United Kingdom), Carlos Matute (CIBERNED).

Parkinson disease (PD) is hallmarked by the deposition of aggregated α-synuclein (α-syn) species into intra-neuronal Lewy bodies (LB) and Lewy neurites (LN) inclusions while in multiple system atrophy (MSA) α-syn aggregates form oligodendroglial inclusions. The development of therapeutic strategies to block cell death in PD and MSA has been limited by a lack of understanding of the mechanisms driving neurodegeneration. However, increasing evidence of multiple pivotal roles of α-syn in the pathogenesis of PD and MSA has led to consider the therapeutic potential of several strategies aimed at reducing α-synuclein toxicity.

Our objective is to build upon the unique experimental cell (Primary rodent neurons and glial cells - Human neurons derived from PD patients) and animal models of PD (SNCAOVX transgenic mouse and human PD-derived α-syn seed-injected mouse) and MSA (PLP-SYN transgenic mouse

2017 CIBERNED Annual Report / 262 and human MSA-derived α-syn seed-injected mouse), developed by this consortium, for testing whether the prototypical molecular tweezer CLR01, shown to inhibit aggregation and toxicity of other amyloidogenic proteins, could protect neurons in vitro and in vivo from PD / MSA-associated synaptotoxicity and reduce PD and MSA’s disease–like α-syn pathology. The project will thus (in) validate the molecular tweezers CLR01 for PD and/or MSA indications.

• 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 groundwork for the development of a novel disease-modifying therapy for PD based on the modulation of NM levels with tFUS.

• Developing preclinical and clinical biomarkers of NRF2 pathway activation for therapeutic application in neurodegenerative diseases

Grupos participantes: 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.

2017 CIBERNED Annual Report / 263 ALZHEIMER’S GLOBAL SUMMIT LISBON 2017

In 2017 CIBERNED held a very special edition of its annual Scientific Forum. This event, which has taken place year after year since 2007, is essential for the proper functioning of CIBERNED, since it allows main researchers, members of their groups, as well as all the rest of attendees, meeting to discuss the findings of their research, present new data and establish collaborations. In 2013, an International Congress of Research and Innovation in Neurodegenerative Diseases (CIIIEN) was first established, with great success that has been consolidated in successive yearly editions (2014-2017).

During the days 20, 21 and 22 September 2017 at the facilities of the Champalimaud Foundation in Lisbon, and coinciding with the celebration of World Alzheimer’s Day (September 21), the Alzheimer’s Global Summit Lisbon 2017 was held, organized jointly by CIBERNED, the CIEN Foundation, the Queen Sofia Foundation and the Champalimaud Foundation.

2017 CIBERNED Annual Report / 264 The scientific program consisted of an opening session, three keynote lectures and ten scientific sessions. Among the speakers who participated in this Summit, highlighted some international researchers who are a world reference in their field of research, such as Nobel Laureates John O’Keefe (University College London, United Kingdom) and Richard Axel (Columbia University, USA), Drs. Hanna and Antonio Damásio (USC Los Angeles, USA), Dr. Kenneth S. Kosik (University of California, Santa Barbara, USA) or Dr. Maria Grazia Spillantini (University of Cambridge, United Kingdom) among others.

Likewise, and responding to the vocation to promote the training of young researchers of CIBERNED, during the Summit, the Young Investigator Awards (basic and clinical) were presented to Elisabeth Sánchez Mejías and Dolores Vilas Rolán respectively, who made a presentation of the studies for which said recognition has been granted. 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.

António Damasio (USC), Alzheimer´s Global Summit Lisbon 2017

2017 CIBERNED Annual Report / 265 OTHER INTERNATIONAL ACTIVITIES

• H2020: Marie Skłodowska-Curie Actions: Innovative Training Networks (ITN)

The Innovative Training Networks (ITN) are actions created by the European Union (within the Horizon 2020 program framework) to support research in the European Research 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 2017, the CIBERNED groups have been part of two of these actions:

1. Blood Biomarker-based Diagnostic Tools for Early-stage Alzheimer’s Disease – BBDiag

Alzheimer’s disease (AD) affects more than 7 million people in Europe and this figure is expected to double every 20 years. Despite intensive efforts, no disease-modifying treatments or preventive strategies are available. The lack of specific, sensitive and minimally invasive diagnostics to identify people with early-stage AD to be included in clinical drug intervention trials is among the main reasons for many notable trial failures. The main challenges in developing the required diagnostics are identification of AD biomarkers and development of their detection techniques. The complex and interdisciplinary nature of the research underlines the need for innovative training of a new generation of researchers in the field. BBDiag responds to such a need and establishes a much-needed ETN for blood based early-AD diagnostics to address these challenges. It brings together leading academic and industrial experts from five major consortia in Europe and uses their synergies to build a triple-i research & training platform with the required multidisciplinary expertise and cutting-edge technologies. BBDiag Fellows will be trained under the Vitae Researcher Development Framework innovatively combined with the BBDiag platform for gaining multidisciplinary scientific and transferable skills as well as personal quality, creative thinking and business mindset.

The ETN has a highly innovative research programme for the discovery of AD biomarkers, development of novel biosensing techniques and point of care tools, and for technological exploitation of the diagnostics. These advances will strongly support improved care provision and development of disease-modifying treatments and preventive strategies for AD patients. More importantly, BBDiag will deliver its first generation of 13 highly-skilled, creative and entrepreneurial Fellows, setting them on a path to successful careers in academia or industry to ensure that the medical and societal challenges imposed by AD are met.

2. Interdisciplinary training network on the purinergic P2X7 receptor to control neuroinflammation and hyperexcitability in brain diseases - PurinesDX

Brain disorders affect ~179 million people and their families in Europe alone, with an annual cost to the taxpayer estimated at €800 billion- a greater economic burden than cardiovascular disease and cancer combined. Despite diverse etiology, overlap in clinical symptoms and comorbidities between brain disorders suggests shared patho-mechanisms. In particular, hyperexcitible states driven by glial activation and neuroinflammation appear near ubiquitous. Targeting these mechanisms offers the potential to ameliorate symptoms and reverse disease progression across a broad span of brain disorders. Functioning as a gatekeeper to neuroinflammation and mechanistic link between neuronal hyperexcitability and glial activation, the ATP-gated, ionotropic purinergic P2X7 receptor (P2X7R) offers the most promising target for pharmacological intervention in the neuroinflammation-hyperexcitability pathway, to date. With breakthroughs in understanding P2X7R function, highly promising effects demonstrated for antagonists in models of brain disease and vast investment in P2X7R-related drug development programmes, now is the perfect time to pool resources.

2017 CIBERNED Annual Report / 266 PurinesDX brings together global leaders in translational research in purinergic signalling, Europe’s leading clinical specialists in a broad range of brain diseases, and industrial partners specializing in drug and biomarker development. Sharing unique genetic tools, newly developed diagnostic devices and novel, selective and brain-stable P2X7R antagonists, the synergism facilitated within PurinesDX will extend to the training of an urgently needed new generation of highly skilled, innovative, creative and entrepreneurial scientists.

Alongside the provision of this interdisciplinary, international and intersectoral environment, an original and high level training in state-of-the-art neuroscience will be provided, nurturing a cohort of highly competitive researchers with potential to drive a new era of neuroscience research.

• The Association for Frontotemporal Degeneration (AFTD)

The Association for Frontotemporal Degeneration (AFTD) is an organization that promotes and funds research on the diagnosis, treatment and cure of the frontotemporal degeneration, the most common form of dementia for people under 60 years of age. During 2017, Dr. María Llorens, member of CIBERNED, received funding from this entity to develop the following project:

2017 CIBERNED Annual Report / 267 1. Tau and adult neurogenesis therapeutic potential for frontotemporal degeneration (AFTD 2016)

Frontotemporal dementia (FTD) refers to a group of disorders caused by progressive neuronal loss in the frontal or temporal lobes of the brain. The most prominent diseases causing frontotemporal degeneration involve Tau protein.

The principal aim of this project is to dissect the cellular mechanism underlying the neurodegeneration triggered by the presence of a form of human Tau protein bearing three mutations known to cause hereditary FTD with parkinsonism linked to chromosome 17 (FTDP-17). To this end, a murine model of FTDP-17, the TauVLW mouse, will be used to perform an exhaustive analysis of one of the most important brain regions involved in learning and memory, namely the hippocampal dentate gyrus (DG). A unique feature of the DG is the occurrence of adult neurogenesis (AHN). We will use novel viral strategies to characterize the morphology, connectivity and innervation of newborn granule neurons in TauVLW mice. Finally, we will examine the therapeutic potential of two distinct strategies to counteract the alterations caused by the presence of mutated forms of Tau. To this end, we will test the effectiveness of different strategies aimed to increase the activation of these neurons. This is a novel therapeutic approach that, to the best of our knowledge, has never been used in a murine model of a neurodegenerative disease to date. Given that alterations in AHN have been described in numerous neurodegenerative diseases such as Alzheimer disease and FTD, understanding the cellular alterations caused by mutated forms of Tau will be crucial to identify therapeutic strategies to treat these diseases.

• 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 2017, 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 2017 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 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. GSK-3B-and adult neurogenesis. Therapeutic potential for Alzheimer disease (2015-NIRG- 340709)

A few years ago, this research group described that the granular neurons of the hippocampal dentate gyrus undergo a morphological transformation in patients suffering from Alzheimer’s disease. On the other hand, overexpression of GSK-3β in the brain of an AD mouse model led to the appearance in hippocampal granular neurons of a morphology highly similar to that found in the cells of the patients.

2017 CIBERNED Annual Report / 268 However, it is well known that in the brain of patients with Alzheimer’s disease as well as in the AD mouse model always takes place the appearance of a series of non-cell autonomous effects concomitantly when overexpressing any toxic protein. These effects include the activation of numerous mechanisms, but perhaps the most important and one of the best known, is the activation of cells of the immune system resident in the brain, the microglia cells. This series of mechanisms associated with greater neuroinflammation have important effects on neuronal development and that is why the present project aimed at the total elimination of these effects in order to study the role of GSK-3ß in the alterations present in neurons from the dentate gyrus of Alzheimer’s disease patients.

To do that, and based on a unique property of the dentate gyrus of the hippocampus (consisting of the continuous generation of new neurons throughout life, a process known as adult neurogenesis), a highly novel method was developed to ensure that only a small population of our cells of interest overexpressed the GSK-3ß protein, relying on the combination of a mouse model (the tto-GSK- 3ß mouse model) and the infection of those cells of interest with a novel retroviral vector. That retroviral vector or retrovirus was used to express the rtTA protein, which allowed over-expression of GSK-3β in the cells that had previously been infected by the retrovirus.

By using this novel approach we have been able to learn that the effects of GSK-3β on the newly generated granular dentate gyrus neurons are direct, and that they do not depend on the activation of indirect mechanisms such as neuroinflammation. This strategy had not been used to date to study adult hippocampal neurogenesis, nor in the field of neurodegenerative diseases. As a result of this research project, six scientific articles have been published in prestigious international journals such as EMBO J and Oncotarget, and another for papers have been sent for publication.

Finally, in November 2017 the Alzheimer’s Association granted a new project to a member of CIBERNED under its latest call for research projects which will begin to be developed in 2018:

• AARG-17-528125: Novel Methods to Interrogate the Subcellular Machinery of AD Models in Vivo

Principal Investigator: María Llorens-Martín

2017 CIBERNED Annual Report / 269

Scientific productivity and other activities

ANALYSIS OF CIBERNED SCIENTIFIC PRODUCTIVITY IN 2017

The table below summarize the bibliometric indicators related to the scientific activity from years of 2016 and 2017. A quick analysis shows a strengthening of the improved scientific production in 2017 (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 and within the first decile or quartiles 1 and 2; percentage of publications led by CIBERNED groups within quartiles 1 and 2; number of publications in high impact journals; or number of national and international collaborations led by CIBERNED groups).

Indicator 2016 2017 Change (%)

(A) Total number of publications1 568,00 571,00 0,53%

(B) Number of publications in quartiles (Q) 1 y 2 467,00 470,00 0,64%

(E) Number of publications in 1st decile 174,00 189,00 8,62%

(F) Number of publications (1st decile) as percentage of total (A) 30,63 33,10 8,05%

(G) Number of publications in Q1 (includes 1st decile) 347,00 372,00 7,20%

(H) Number of publications in Q1 as percentage of total (A) 61,09 65,15 6,64%

(I) Number of publications in Q1 as percentage of total Q1 and Q2 (B) 74,30 79,15 6,52%

(J) Number of publications in Q2 120,00 98,00 -18,33%

(K) Number of publications in Q2 as percentage of total (A) 21,13 17,16 -18,76%

(L) Number of publications (Q1+Q2) led by CIBERNED groups (1st 265,00 267,00 0,75% author or corresponding author)

(M) Number of publications in Q1 and Q2 led by CIBERNED groups 56,75 56,81 0,11% as percentage of total (B)

(N) Number of publications (Q1 and Q2) not led by CIBERNED 202,00 203,00 0,50% groups (1st author or corresponding author)

(O) Number of publications in Q1 and Q2 not led by CIBERNED 43,25 43,19 -0,15% groups as percentage of total (B)

(P) Number of publications in Q1 and Q2 in which “CIBERNED” 79,23 81,70 3,12% appears in the affiliation (as percentage of total (B))

(Q) Number of publications in Q1 and Q2 led by CIBERNED groups 85,28 90,26 5,84% in which “CIBERNED” appears in the affiliation (as percentage of total (L))

(R) Number of publications in Q1 and Q2 not led by CIBERNED 71,29 70,44 -1,18% groups in which “CIBERNED” appears in the affiliation (as percentage of total (N))

(S) Number of publications in Q1 and Q2 with international groups 255,00 260,00 1,96%

(T) Number of publications in Q1 and Q2 with international groups (S) as percentage of total number of publications (B) 54,60 55,32 1,31%

¹ Number of publications indexed in ISI (Web of Knowledge ).

2017 CIBERNED Annual Report / 273 Indicator 2016 2017 Change (%)

(U) Number of publications in Q1 and Q2 with international groups, 111,00 123,00 10,81% and led by CIBERNED groups

(V) Number of publications in Q1 and Q2 with international groups, 43,53 47,31 8,68% and led by CIBERNED groups as percentage of all international publications (S)

(W) Number of publications in Q1 and Q2 with other CIBERS 11,00 18,00 63,64%

(X) Percentage of publications in Q1 and Q2 with other CIBERS led 9,09 27,78 205,56% by CIBERNED groups

(Y) Number Percentage of published in Q1 y Q2 1,00 5,00 400,00%

(Z) Percentage of Percentage of led by CIBERNED groups 40,00 40,00 0,00%

(A2) Addition of impact factors of journals (Q1 y Q2) in which 2636,90 2859,70 8,45% articles have been published

(B2) Average of impact factors of journals (Q1 y Q2) in which 5,11 6,08 19,06% articles have been published

(C2) Number of publications in journals with impact factor >15 36,00 44,00 22,22%

Number of publications in Q1 and Q2 with 2 CIBERNED groups 62,00 57,00 -8,06%

Number of publications in Q1 and Q2 with 3 CIBERNED groups 11,00 20,00 81,82%

Number of publications in Q1 and Q2 with 4 CIBERNED groups 1,00 1,00 0,00%

Number of publications in Q1 and Q2 with 5 CIBERNED groups 0,00 0,00 -

Number of publications in Q1 and Q2 with 6 CIBERNED groups 0,00 2,00 -

Number of publications in Q1 and Q2 with 7 CIBERNED groups 0,00 0,00 -

Number of publications in Q1 and Q2 with 8 CIBERNED groups 0,00 0,00 -

Cooperative projects – Program 1 60,00 68,00 13,33%

Cooperative projects – Program 2 144,00 157,00 9,03%

Projects with other CIBERs/RETICs/CIEN networks 47,00 36,00 -23,40%

Basic Research-type projects 70,00 79,00 12,86%

Translational research-type projects 134,00 146,00 8,96%

Projects with other national research group 108,00 121,00 12,04%

Projects with international research groups 47,00 45,00 -4,26%

Projects with industry 68,00 82,00 20,59%

2017 CIBERNED Annual Report / 274 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 2017 and its evolution since 2011:

• Figure 1: Percentage of indexed CIBERNED 2017 publications distributed by quartiles

8,8% 2% D1: publications in the 1st decile 9,8% 31,9% Q1: publications in the 1st quartile

16,6% Q2: publications in the 2nd quartile Q3: publications in the 3rd quartile Q4: publications in the last quartile No IF: publications for which no impact factor 62,8% is available

2017 CIBERNED Annual Report / 275 • Figure 2: CIBERNED 2011-2017 production by quartiles/1st decile

700 603 588 5925 92 600 553 525 535 500 467 477 457 471 470 4224 422 400

300 189 200 183 165 174 139 152 156 100

0 2011 2012 2013 2014 2015 2016 2017 Number of publications Q1+Q2 D1

Q1+Q2: sum of publications in 1st and 2nd quartiles. D1: publications in the 1st decile.

• Figure 3: Percentage of publications of excellence (1st decile) CIBERNED 2011- 2017

40%

30% 31,1% 31,9% 29,0% 29,2% 29,8% 29,4% 20% 23,1%

10%

0% 2011 2012 2013 2014 2015 2016 2017

2017 CIBERNED Annual Report / 276 • Figure 4: Number of CIBERNED 2011-2017 publications within 1st and 2nd quartiles and 1st decile

400 372 351 361 351 350 329 337 314 300

250

200 183 174 189 165 152 156 150 139 140 116 113 120 108 98 100 96

0 2011 2012 2013 2014 2015 2016 2017 D1 Q1 Q2

• Figure 5. Evolution of the impact factor from 2011 to 2017

IF: impact factor.

3.100,00 5,6 5,5 6,0 5,3 5,4 5,4 5,1 5,1 3.000,00 2.995,05 2.914,55 5,0 2.900,00 2.869,23 2.820,33 2.790,17 4,0 2.800,00

2.700,00 3,0

2.600,00 2.575,57 2.555,72 2,0 2.500,00 1,0 2.400,00

2.300,00 0,0 2011 2012 2013 2014 2015 2016 2017 Cumulative IF Mean IF

2017 CIBERNED Annual Report / 277 • Figure 6: CIBERNED 2011-2017 publications in high impact journals

6 9 10

17 3 38 12

38 6

15 2

5 34 34 8

3 34 17

IF > 30 IF 20-30 44 IF 10-30

2017 CIBERNED Annual Report / 278 • Figure 7: Scientific journals with 5 or more CIBERNED publications in 2017

Movement Disorders 28

Scientific Reports 28 Molecular Neurobiology 15

PLoS ONE 15 Journal of Alzheimer's Disease 13 Neurology 10

Neuropharmacology 9 Parkinsonism and Related Disorders 9

Redox Biology 9

Frontiers in Neurology 8 Journal of Neuroscience 8

Neurobiology of Aging 8 Brain Structure and Function 7 Manual de enfermedades neuromusculares 7 Neurology 7 Cerebral Cortex 6 Journal of the Neurological Sciences 6 Sleep Medicine 6

Brain 5 Experimental Neurology 5 Frontiers in Molecular Neuroscience 5 Journal of Neurology, Neurosurgery and Psychiatry 5 Neurobiology of Disease 5 Oncotarget 5 Sleep 5

2017 CIBERNED Annual Report / 279 • Figure 8: Most frequent SUBJECT categories (WoS-JCR) in CIBERNED 2017 publications

Neurosciences 177 Clinical Neurology 126 Biochemistry Molecular Biology 42 Multidisciplinary Sciences 31 Cell Biology 30 Geriatrics Gerontology 16 Pharmacology Pharmacy 16 Pathology 14 Endocrinology Metabolism 9 Genetics Heredity 9 Biotechnology Applied Microbiology 7 Chemistry Multidisciplinary 7 Engineering Biomedical 7 Medicine Research Experimental 7 Oncology 7 Biophysics 6 Anatomy Morphology 5 Gerontology 5 Immunology 5 Psychiatry 5

CROSS-NETWORK PROGRAMS

• Neurological Tissue Banks

• Neurological Tissue Bank and Biological Samples of the IDIBELL Institute of Neuropathology, Bellvitge Hospital (Head: Dr. Isidro Ferrer).

• CIEN Foundation Tissue Bank for Neurological Research (BT-CIEIN) (Head: Dr. Alberto Rábano).

• 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).

2017 CIBERNED Annual Report / 280 EVENTS AND OTHER ACTIVITIES

• Alzheimer’s Global Summit Lisbon 2017

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.

Alzheimer´s Global Summit Lisbon 2017

2017 CIBERNED Annual Report / 281 • First DEGESCO Symposium: Genetics of dementia

DEGESCO (Spanish Dementia Genetics 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, especially the joint participation in international multicenter consortia. The generation of a stable nation-wide consortium will make it possible to exploit the existing synergies, improve the statistical power and the precision of the studies carried out individually by each group. DEGESCO philosophy is inclusive, that is, it is open to the incorporation of new research groups, whether public or private as long as they can prove their capacity and interest to conduct research in genetics of dementia.

DEGESCO is currently composed of 22 fully-fledged research groups with the institutional coverage of CIBERNED (Center for Biomedical Research in Neurodegenerative Diseases Network). Those member groups are the following:

BARCELONA -Hospital de Sant Pau -Hospital Clinic -Fundació ACE OVIEDO -Fundació Pasqual -Hospital SANTANDER Maragall Central de Asturias -Hospital Marqués de TERRASSA Valdecilla PAMPLONA -Hospital Mutua de -Navarrabiomed Terrassa LLEIDA 1 -IRB-Lleida

MADRID P. MALLORCA -Hospital la Paz -Hospital Son Espases -Hospital 12 de Octubre -Hospital Ramón y Cajal -CIBERNED -Fundación CIEN -CBM

2017 CIBERNED Annual Report / 282 To enhance the visibility of the consortium, the First DEGESCO Symposium: Genetics of Dementia was held at the CIEN Foundation headquarters in Madrid on February 16-17. The Symposium was organized by Drs. Miguel Medina (CIBERNED) and Miguel Calero (Carlos III Institute of Health / CIBERNED / CIEN Foundation). The Opening Session, held on the morning of Thursday 16, was chaired by H.M. Queen Sofia, and was also attended by various representatives of the Queen Sofia Foundation, CIBERNED and the Neurological Diseases Research Center (CIEN) Foundation, partners of the event and who have an active role in the study of these pathologies. The event was highly successful, attracting an audience of approximately 130 attendees and brought together the best national experts on the genetics of neurodegenerative diseases to address a key area of research, reviewing in depth the genetic aspects of the different types of dementia and their implications in the Spanish population.

• Excellence in Neurodegeneration seminar series

During 2017 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: Fred H. Gage, Salk Institute for • Speaker: Richard Mayeux, Columbia Biological Studies, La Jolla, USA. University, Ney York City, USA. Title: Function and regulation of Title: The Alzheimer’s Disease Sequencing Neurogenesis in the Adult Hippocampus. Project. • Speaker: John O’Keefe, University College • Speaker: Li-Hue Tsai, Massachusetts Institute London, UK. of Technology, USA. Title: Spatial Functions of the Hippocampus: Title: Bringing gamma back — using a model system for studying dementia. noninvasive sensory stimulation to modify Alzheimer’s disease. • Speaker: Maria Grazia Spillantini, University of Cambridge, UK. • Speaker: James Surmeier, Northwestern Title: Understanding toxicity in University, Chicago, USA. neurodegenerative diseases. Title: Calcium, bioenergetics and selective neuronal vulnerability in Parkinson’s disease. • Speaker: George Perry, University of Texas, San Antonio, USA. • Speaker: Sam Sisodia, University of Chicago. Title: Oxidative stress as a window to Alzheimer Title: Modulation of Amyloid Deposition and disease. Neuroinflammation by the Microbiome. • Speaker: Sarah Tabrizi, University College • Speaker: Li Gan, Gladstone Institute, San London, UK. Francisco, USA. Title: Meeting the Therapeutic challenge of Title: Converging pathways in AD and FTD. Huntington’s disease. • Speaker: Andrés Lozano, University of Toronto, • Speaker: Ioannis Sotiropoulos, University of Canada. Minho, Braga. Title: The possibility of treating Alzheimer’s by Title: Tau therapeutics in stress-driven electrically stimulating brain memory circuits. brain pathology: exploring the path from depression to Alzheimer’s disease. • Speaker: Alvaro Pascual Leone, Beth Israel Deaconess Hospital, Boston, USA. • Speaker: Kenneth S. Kosik, University of Title: Noninvasive Brain Stimulation in AD. California, Santa Barbara, USA. Title: Liquid-liquid phase states of Tau. • Speaker: Hanna & Antonio Damasio, USC Los Angeles, USA. • Speaker: Richard Axel, Columbia University, Title: Human Memory: The Neurocognitive USA. Perspective. Title: Order from Disorder: Representations of Olfactory Information in the Cortex.

2017 CIBERNED Annual Report / 283 • Speaker: John T. O’Brien, University of • Speaker: Albino Oliveira-Maia, Cambridge, UK. Champalimaud Foundation, Lisboa. Title: Brain imaging of tau and inflammation Title: Non-invasive measurement and in dementia. manipulation of cortical excitability in health and disease. • Speaker: Andrew Stephens, Piramal Imaging, Berlin, Germany. Title: Initial clinical evaluation of PI-2620, a next generation TAU pet tracer.

• 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.

In the case of the Young Investigator of the Year Award, the candidates must be under 35 years- old, members of a CIBERNED research group and main authors of an article published during 2017in an indexed scientific journal. For the Young Clinical Investigator of the Year Award, the candidates must be under 40 years-old, members of a CIBERNED research group and main authors of an article published during the years 2015 and 2016 in an indexed scientific journal.

The call remit was announced by email addressed to all Principal Investigators 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 due time and form.

For both calls, the selection committee was made up of members of the Steering Committee 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.

Once the process for evaluating the applications was completed, the Scientific Directorate of CIBERNED agreed to award the Young Investigator of the Year Award to Elisabeth Sánchez Mejías for her work “Soluble phospho-tau from Alzheimer’s disease hippocampus drives microglial degeneration”, published in the Acta Neuropathologica (2016) 132 (6): 897-916.

In addition, the Young Clinical Investigator of the Year Award was awarded to Dolores Vilas Rolán for her work “Assessment of α-synuclein in submandibular glands of patients with idiopathic rapid- eye-movement sleep behaviour disorder: a case-control study” published in Lancet Neurology (2016) 15(7):708-718.

OTHER EVENTS

• University of Extremadura (Umex) summer course

Between 28th and 30th June, 2017 the course “Advances in neurobiology and neurodegenerative diseases” was held in Cáceres, as part of the XVIII edition of the International Summer Courses of the University of Extremadura, led by Dr. José Manuel Fuentes Rodríguez, Principal Investigator of CIBERNED and member of the Department of Biochemistry and Molecular Biology and Genetics of the Faculty of Nursing and Occupational Therapy of the University of Extremadura.

With a total of 16 teaching hours, the course was addressed at students and professionals of Health Sciences; with the aim of knowing the most current advances at the molecular and cellular level of the Nervous System (with special emphasis on what refers to neurodegenerative diseases),

2017 CIBERNED Annual Report / 284 to integrate this knowledge in the understanding, prevention or treatment of neurodegenerative diseases.

Among other specialists, Drs. Jesús Ávila de Grado, CIBERNED Scientific Director; Miguel Medina, Deputy Scientific Director, and the main researchers at CIBERNED, José Ramón Naranjo Orovio and Miguel Calero Lara delivered lectures during the Course.

• Workshops Current Trends in Biomedicine 2017

As part of the Current Trends in Biomedicine 2017 cycle, during September 25-27, 2017 Dr. Rafael Fernández-Chacón, Principal Investigator of CIBERNED, organized together with the 2013 Nobel Prize in Medicine Prof. Thomas C. Südhof (Department of Molecular and Cellular Physiology and Howard Hughes Medical Institute of the Stanford University) at the International University of Andalusia in Baeza the meeting “Synapse formation, specification and elimination: from molecules to circuits”.

The meeting brought together 16 renowned international speakers with experience in the study of multiple aspects in the molecular mechanisms of synaptic connections and the remodeling of brain circuits in the organization of the nervous system, and consisted of invited papers and moderate poster sessions, including extensive time for discussion both inside and outside the sessions.

• Alpha-synuclein & Parkinson’s Disease conference

On May 11 and 12, the Conference “Alpha-synuclein and Parkinson’s disease Conference: Lessons from the past 20 years” took place in the Aula Magna of the University of Barcelona. This meeting was organized by the groups of Dr. Eduardo Tolosa and Miquel Vila, both Principal Investigators of CIBERNED.

Synucleinopathies are disorders characterized by the accumulation and propagation of the α-synuclein protein in the nervous system that comprise a group of conditions such as multiple system atrophies, Parkinson’s disease or dementia due to Lewy bodies.

In the context of the 20th anniversary of the discovery of the link between the α-synuclein gene and Parkinson’s disease, this conference brought together a group of experts from different fields related to α-synuclein and its pathology, with the aim of providing latest developments on the essential aspects of synucleinopathies, including those of genetic and neuropathological type, and with a strong emphasis on the latest advances in basic science on this protein. In addition, the role of α-synuclein as a current and novel therapeutic biomarker was reviewed.

• Research on Alzheimer’s and other dementias conference

On November 8th, 2017, CIBERNED collaborated in the Scientific Conference on Alzheimer’s and Other Dementias Research held at the Maimonides Institute of Biomedical Research of Córdoba (IMIBIC) where experts from the most prominent research centers in Spain analyzed the most important research challenges in Alzheimer’s and other dementias.

The main objective of this conference was to present the map of Alzheimer’s research in Spain and to learn about the different research aspects of this discipline (basic, clinical, social-health, social and anthropological).

This scientific meeting, aimed at researchers, health professionals, patient associations and citizenship in general, was attended by CIBERNED’s Manager, María Ángeles Pérez, and Scientific Director, Dr. Jesús Ávila de Grado, among other experts.

2017 CIBERNED Annual Report / 285

Finantial report

The Network Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED) 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, Institutions 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.

Royal Legislative Decree 3/2011, of November 14, which approves the revised text of the Public Sector Contracts Act, governs its public procurement system.

CIBERNED is a virtual Research Center consisting of research groups belonging to different Administrations and Institutions in partnership: the member Principal Investigators 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 2017, it consisted of 52 groups, an average of 126.25 hired staff that add up to 137 by December 31, 2017.

• Average staffing for the year 2017:

Category Women Men Total

Administrative assistant 6,00 0,00 6,00 Undergraduate 3,00 0,83 3,83 Doctor (PhD) 27,83 12,75 40,58 Graduate (BSc, MSc) 24,92 18,08 43,00 Technician 25,75 7,08 32,83

Total 87,50 38,75 126,25

• Staff personnel at year end:

Category Women Men Total

Administrative assistant 6 0 6 Undergraduate 3 0 3 Doctor (PhD) 31 14 45 Graduate (BSc, MSc) 28 20 48 Technician 27 8 35

Total 95 42 137

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.

2017 CIBERNED Annual Report / 289 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 2,303,080.75€.

During 2017, the Consortium counts on the funding of the Carlos III Institute of Health as a promoter organization for the execution of the activities and general objectives already described.

On June 22, 2017, 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 Neurodegenerative Diseases of the nominative assignments provided for in the Statement of Expenditure of the 2016 Expanded Budget for 2017 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 30, 2016, modified by agreement of the Council of Ministers of May 12, 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:

- 2.942.430 euros from budget heading 27,107,465A.44605 -122.535 euros con from budget heading 27.107.465.74605

Corresponding to 75% of the total credit existing in the aforementioned budgetary headings, leaving the remaining 25% of the credit subject to the approval of the General State Budget Law for 2017.

Once Law 3/2017 of June 27 was approved, the General State Budget for 2017 adjusted the amounts initially allocated and by resolution of September 4, 2017, the amounts granted were increased in:

- 980.810 euros from budget heading 27.107.465A.44605 - 40.845 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.

2017 CIBERNED Annual Report / 290 BASIS OF PRESENTATION OF THE ACCOUNTS

• True and fair view:

The financial statements have been prepared from the accounting records of the Institution, having applied the existing legal provisions on accounting matters, and especially, to present fairly the assets, financial situation and results of the Institution.

Accounts are displayed in the standard model and following the accounting principles suggested in the General Public Accounting Plan. More specifically, the Adjustment Plan Public Accounting for government agencies whose spending budget is an estimate is applied, as approved by the 28 July, 2011 Order of the General Comptroller of the State Administration.

These financial statements have been prepared by the Managing Director of the Institution and will be submitted to approval by the Governing Council.

del Río Fernández José Antonio - G114

2017 CIBERNED Annual Report / 291 • Balance sheet year 2017

Assets

A) Non-current assets 1.431.817,28€

I. Intangible assets 63.563,33€

1. Investment in research and development 0 2. Industrial and intellectual property 0 3. IT applications 63.563,33€ 4 Investments in assets used under lease or assignment 0 5. Other intangible assets 0

II. Material assets 928.097,00€

1. Lands 0 2. Buildings 0 3. Infrastructures 0 4. Historic heritage assets 0 5. Other material assets 928.097,00€ 6. Fixed assets under construction and advances 0

III. Real state investemnts 0

IV. Long-term financial investments in group companies, joint ventures and associates 0

V. Long-term financial investments 440.156,95€

1. Financial investments in equity 0 2. Credit and debt securities 439.554,25€ 3. Financial derivatives 0 4. Other financial investments 602,7€

VI. Debtors and other long-term accounts receivables 0

B) Current assets 3.940.079,92€

I. Assets assigned for sale 0

II. Inventory 0

III. Debtors and other accounts receivable 963.893,81€

1. Debtors from management operations 650.542,83€ 2. Other accounts receivable 0 3. Public Administrations 313.350,98€

IV.Short-term financial investments in group companies, joint ventures and associates 0

V. Short-term financial investments 6.048,41€

1. Financial investments in equity 0 2. Credits and debt securities 4.975,01€ 3. Financial derivatives 0 4. Other financial investments 1.073,40€

VI. Accrual 9.441,78€

VII. Cash and cash equivalents 2.960.695,92€

1. Treasury 2.960.695,92€

TOTAL ASSETS (A+B) 5.371.897,20€

2017 CIBERNED Annual Report / 292 Equity and liabilities

A) Equity 2.729.814,66€

I. Contributed equity 0

II. Generated equity 2.729.814,66€

1. Results from previous years 3.085.213,53€ 2. Results in the year -355.398,87€ 3. Reserves 0

III. Valuation adjustments 0

IV. Other assets increases pending allocation to results 0

B) Non-current liabilities 626.421,43€

I. Long-term provisions 14.984,41€

II. Long term debts 611.437,02€

1. Debentures and other marketable securities 0 2. Debts with credit institutions 0 3. Financial derivatives 0 4. Other debts 611.437,02€ 5. Debtors for long-term capital leasing agreements 0

III. Debts with group companies, joint ventures and associates 0

IV. Creditors and other long-term payables 0

V. Adjustments for long-term accrual 0

C) Current liabilities 2.015.661,11€

I. Short-term provisions 79.944,54€

II. Short-term debts 1.545.401,49€

1. Debentures and other marketable securities 0 2. Debts with credit institutions 0 3. Financial derivatives 0 4. Other debts 1.545.401,49€ 5. Debtors for short-term capital leasing agreements 0

III. Debts with group companies, joint ventures and associates 0

IV. Accrual 390.315,08€

1. Creditors from management operations 213.299,62€ 2. Other accounts payaable 383,09€ 3. Public Administrations 176.632,37€

V. Adjustments for accrual 0

TOTAL EQUITY AND LIABILITIES (A+B+C) 5.371.897,20€

2017 CIBERNED Annual Report / 293 • Economic assets income statement

2017 economic assets income statement

1. Income tax and social contributions 0

2. Transfers and subsidies received 6.818.694,67€

a) of the year 6.818.694,67€ a.1) subsidies received to finance expenditures for the year 423.993,92€ a.2) transfers 6.394.700,75€

3. Net sales and services 95.616,22€

4. Changes in inventory of finished and in progress of manufacturing goods and impairment loss 0

5. Work by the company for its fixed assets 0

6. Other income from ordinary management 1.643,59€

7. Provisions surpluses 24.861,48€

A) TOTAL REVENUES FROM ORDINARY MANAGEMENT (1+2+3+4+5+6+7) 6.940.815,96€

8. Personnel costs -3.480.928,45€

a) Wages and salaries and similar expenses -2.617.572,13€ b) Social security costs -863.356,32€

9. Transfers and grants awarded -2.389.966,12€

10. Procurements -819.902,86€

a) Consumption of goods and other supplies -819.902,86€

11. Other ordinary management expenses -525.442,70€

a) Supplies and other external services -525.253,99€ b) Taxes -188,71€ c) Others 0

12. Depreciation of fixed assets -104.453,06€

B) TOTAL COSTS OF ORDINARY MANAGEMENT (8+9+10+11+12) -7.320.693,19€

I Results (surplus or deficit) of ordinary management (A+B) -379.877,23€

13. Impairment loss and results on disposals of non-finacial assets and selling state assets 0

14. Other non-ordinary items -46,69€

a) Income 0 b) Expenditure -46,69€

II Results of non-dinancial operations (I +13+14) -379.923,92€

15. Financial income 1.862,38€

16. . Financial expenses -48,24€

17. Financial expenses charged to assets 0

18. Change of fair value of financial assets and liabilities 0

19. Exchange differences 22.710,91€

2017 CIBERNED Annual Report / 294 2017 economic assets income statement

20. Impairment loss, retirements and disposals of financial assets and liabilities 0

21. Grants to fund financial operations 0

III Result of financial operations (15+16+17+18+19+20+21) 24.525,05€

IV Net surplus (or deficit) in the year (II + III) -355.398,87€

(+-) Adjustments to the account of the outcome of the prior year

Adjusted profit for the prior year

2017 CIBERNED Annual Report / 295 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:

Implementation Personnel Amortization Total costs

Administration, management and coordination 160.508,52€ 172.725,36€ 13.588,72€ 346.822,60€

Program 1 “Alzheimer’s disease and other 788.750,42€ 192.103,16€ 40.025,44€ 1.020.879,02€ degenerative dementias”

Program 2 “Parkinson’s disease and other 1.250.287,56€ 216.215,22€ 46.990,73€ 1.513.493,51€ degenerative motor disorders””

Deputy Scientific Director / Cross-network 259.261,40€ 32.177,87€ 267,59€ 291.706,86€ plataforms

Training / Young Investigator Award / Mobility 0,00 17.336,77€ 0,00 17.336,77€

Media service, internationalization, and 0,00 68.076,79€ 0,00 68.076,79€ transfer of knowledge

COOPERATIVE RESEARCH

Cooperative projects, 4th call (PI2014/2013) 0,00 94,90€ 1.219,31€ 1.314,21€

Cooperative projects,5th call (PI2015) 262.594,42€ 147.925,75€ 0,00 410.520,17€

Cooperative projects, 6th call (PI2015-2) 240.569,42€ 80.310,56€ 0,00 320.879,98€

Cooperative projects, 7th call (PI2016) 249.525,40€ 91.126,95€ 194,47€ 340.846,82€

Cooperative projects, 8th call (PI2017) 0,00 209,16€ 0,00 209,16€

COMPETITIVE PROJECTS

PI11/03028 0,00 0,00 1.952,47€ 1.952,47€

COEN Projects - 2013 0,00 0,00 68,16€ 68,16€

COEN Projects - 2015 0,00 7.860,29€ 0,00 7.860,29€

BBVA Proyectos investigación 0,00 24.781,07€ 0,00 24.781,07€

RTC-2014-1877-1 0,00 187,60€ 0,00 187,60€

InDiWIP/ Ramon Areces Foundation 11.666,94€ 0,00 0,00 11.666,94€

FJCI-2014-19673 14.280,02€ 0,00 0,00 14.280,02€

PIE14/00061 24.073,46€ 35.792,51€ 0,00 59.865,97€

ALZ.ASSOC/Mª LLORENS 23.971,48€ 2.829,22€ 0,00 26.800,70€

COHORTE BBN-CIBERNED 0,00 1.935,00€ 0,00 1.935,00€

ALZ.ASSOC/I. FERRER 42.862,65€ 53.193,60€ 0,00 96.056,25€

BBDiag 33.730,59€ 7.311,75€ 0,00 41.042,34€

AC16-00050 9.790,70€ 0,00 104,50€ 9.895,20€

2017 CIBERNED Annual Report / 296 Implementation Personnel Amortization Total costs

AFTD 2016 0,00 47.990,64€ 41,67€ 48.032,31€

SAF2016-786 0,00 4.234,33€ 0,00 4.234,33€

MS16/00041. Miguel Servet program 28.134,80€ 0,00 0,00 28.134,80€

CP16/00041. Associated project 11.703,42€ 18.696,89€ 0,00 30.400,31€

E-RARE - 470,74€ - 470,74€ - - JPND-COURAGE 12.432,00€ 12.432,00€

OTHER PROJECTS

GW PHARMA 25.601,63€ 0,00 0,00 25.601,63€

VIVACELL 20.454,28€ 34.708,55€ 0,00 55.162,83€

HEMP SOLUTION 5.964,78€ - 5.964,78€ - AMO PHARMA 23.161,34€ 8.154,50€ 0,00 31.315,84€

ASSOCIATED INSTITUTIONS AGREEMENTS 0,00 71.402,83€ 0,00 71.402,83€

ANNUAL FORUM 0,00 70.982,14€ 0,00 70.982,14€

CONTRIBUTIONS ASSOCIATED INSTITUTIONS - - - 0,00

ASSOCIATED INSTITUTIONS AGREEMENTS 0,00 2.308.080,75€ 0,00 2.308.080,75€

3.480.928,45€ 3.735.311,68€ 104.453,06€ 7.320.693,19€

The data in the table below show that CIBERNED has allocated 2,534,372.15€ for implementing research programs, which represents 50,56% of the ordinary operating expenses (excluding taxes and without taking into account 2016 2017 the contribution of Associated Institutions). Considering the 2.759.284,88€ 2.534.372,53€ in-kind contribution made by the Associated Institutions comprising CIBERNED, that percentage goes up to 34.62%. -8,15%

The amount allocated in 2017 to cover the costs CIBERNED research groups or through the calls of the internal calls for cooperative projects for cooperative projects, amounts to a total adds to 1,073,770.34€, representing 14.67%- of 3,608,142.87 euros, which represents 71.98% 21.42% of the overall budget (depending of the year’s expenditure (49.29% taking into on whether or not the contribution from the account the contribution of the associated Associated Institutions is considered). institutions).

2016 2017 2016 2017 731.903,76€ 1.073.770,34€ 3.491.188,64€ 3.608.142,87€

-8,15%+46,71% -8,15%+3,35%

2017 CIBERNED Annual Report / 297 The expenditure on personnel dedicated The resources allocated to the Administration to stable research structures (groups and department are maintained one more year cooperative projects) represents 55.69% of total in percentages identical to those of previous expenditure (excluding from the calculation the years, representing 6.92% of the total of valuation of the contribution of the associated ordinary expenses, excluding the valuation of institutions), remaining at levels very similar to the contribution of the associated institutions. the previous year.

2016 2017 2016 2017 2.805.893,10€ 2.791.727,22€ 336.770,40€ 346.822,60€

-8,15%-0,50% -8,15%+2,98% HUMAN RESOURCES

The table below presents the distribution of human resources, grouped into the categories of own and associated staff:

Group Principal Investigator Institution Associated Staff

101 Cuadrado Pastor, Antonio Universidad Autónoma de Madrid 3 2 102 Fariñas Gómez, Isabel Universidad de Valencia 13 3 103 Fuentes Rodríguez, José Manuel Universidad de Extremadura 1 2 104 González Castaño, José Universidad Autónoma de Madrid - 1 105 López Barneo, José Fundación Pública Andaluza para la Gestión 13 3 de la Investigación en Salud en Sevilla 106 Ceña Callejo, Valentín Universidad de Castilla-La Mancha 1 4 108 Vicario Abejón, Carlos CSIC 1 2 109 Vila Bover, Miquel Fundació Instituto de Recerca del Hospital 10 3 Universitari Vall d’Hebrón 110 Pérez Castillo, Ana María CSIC - 5 111 Iglesias Vacas, Teresa CSIC 1 3 113 García Verdugo, José Manuel Universidad de Valencia - 3 114 del Río Fernández, José Antonio Instituto de Bioingeniería de Cataluña (IBEC) 1 1 201 Canela Campos,Enric Isidre Universidad de Barcelona 4 3 202 Kulisevsky Bojarski, Jaime Fundació Institut de Recerca del Hospital 11 2 de la Sant Creu i Sant Pau 203 Lanciego Pérez, José Luis Fundación para la Investigación Médica 4 2 Aplicada (FIMA) 204 Moratalla Villalba, Rosario CSIC 3 3 205 Obeso Inchausti, José Ángel Hospital de Madrid - HM 2 3 206 Rodríguez Díaz, Manuel Universidad de La Laguna 4 2 207 Tolosa Sarró, Eduardo Hospital Clínico y Provincial de Barcelona 7 3

2017 CIBERNED Annual Report / 298 Group Principal Investigator Institution Associated Staff

208 Labandeira García, José Luis Universidad de Santiago de Compostela 8 2 209 Pérez Tur, Jordi CSIC 1 2 301 Alberch Vié, Jordi Universidad de Barcelona 2 3 303 Fernández Ruiz, Javier Universidad Complutense de Madrid 14 4 305 Guzmán Pastor, Manuel Universidad Complutense de Madrid 9 3 306 Lucas Lozano, José Javier CSIC 1 2 307 Naranjo Orovio, José Ramón CSIC - 3 401 Ávila de Grado, Jesús CSIC 3 4 402 Camins Espuny, Antonio Universidad de Barcelona 4 1 403 de Felipe Oroquieta, Javier CSIC 11 3 404 Matute Almau, Carlos Universidad del País Vasco 10 3 406 Rodríguez Álvarez, José Universidad Autónoma de Barcelona 6 2 407 Sáez Valero, Javier Universidad Miguel Hernández de Elche - 4 408 Soriano García, Eduardo Universidad de Barcelona 3 3 409 Torres Alemán, Ignacio CSIC 4 5 410 Trullás Oliva, Ramón CSIC 1 3 411 Vitorica Ferrández, Fco. Javier Universidad de Sevilla 4 1 412 Wandosell Jurado, Francisco CSIC 2 2 413 Comella Carnicé, Joan Xavier Fundació Institut de Recerca del Hospital 4 3 Universitari Vall d’Hebrón 415 Gutíerrez Pérez, Antonia Universidad de Málaga 7 2 502 Carro Díaz, Eva María Fundación para la Investigación Biomédica 1 3 del Hospital Universitario 12 de Octubre 503 Ferrer Abizanda, Isidro Instituto de Investigación Biomédica de 3 8 Bellvitge (IDIBELL) 504 Lleó Bisa, Alberto Fundació Institut de Recerca del Hospital 11 1 de la Sant Creu i Sant Pau 507 Pastor Muñoz, Mª Asunción Fund. para la Investigación Médica Aplicada (FIMA) 9 - 508 Mengod Los Arcos, Guadalupe CSIC 2 2 509 de Pedro Cuesta, Jesús Instituto de Salud Carlos III 8 2 510 Bullido Gómez-Heras, Mª Jesús Universidad Autónoma de Madrid 2 3 511 Cantero Lorente, José Luis Universidad Pablo de Olavide 5 1 601 Infante Ceberio, Jon Fundación Marqués de Valdecilla 14 3 604 Muñoz Cánoves, Pura Universidad Pompeu Fabra 6 7 606 Fernández Chacón, Rafael Universidad de Sevilla 6 3 607 Navarro Acebes, Xavier Universidad Autónoma de Barcelona 27 4 609 López de Munain Arregui, Adolfo Instituto de Investigación Sanitaria Biodonostia 31 3 Oficina técnica 13 TOTAL 298 158

2017 CIBERNED Annual Report / 299 Since its establishment, CIBERNED has considered its human capital as the most important asset of its research center. From the Human Resources department we know that the recruitment and retention of talent is crucial to achieve great goals, so that for us it is just as important to incorporate young researchers with great potential as having researchers with a proven track record who can transfer all their knowledge and know-how. To achieve this, the leadership of our researchers is essential to drive the research groups in the most efficient way. CIBERNED has a high degree of commitment to maintaining investment in research, development and innovation, as a guarantee for the future and always putting its results at the service of society. The evolution of the results and the balance of the research certify a great soundness and solvency of the research carried out by the different groups of CIBERNED, facing the numerous strategic projects with enormous guarantees and providing maximum visibility to the research carried out, being a reference at the national and international level. CIBERNED’s commitment to society, research, scientific progress and quality management leads it to acquire new knowledge and to innovate under the leadership, responsibility, effort and passion of all its members. • Calls for personnel

To incorporate staff, CIBERNED has established a procedure approved by the Steering Committee and the Governing Council. Este procedimiento es acorde al punto 6.2 de la norma ISO 9001:2008. Todas las convocatorias de empleo se publican en las páginas web de los organismos integrantes del consorcio y en cumplimiento de las obligaciones adquiridas con los diversos centros, se informa a los miembros del Consejo Rector de CIBERNED y a los gerentes o responsables de los centros de realización del contenido de las convocatorias.

An integrated management system that provides Principal Investigators with a quick and easy way of managing calls for personnel has been implemented since 2008. This system through CIBERNED’s intranet has allowed streamlining the selection process and resolution of positions available, with a significant reduction of the time between the announcement and resolution of new hirings.

The application of this system facilitates rapid coverage of job positions needs, foreseen or unforeseen, as well as improves and streamlines the organizational running by introducing efficiency and quality criteria in the internal Human Resources management according to CIBERNED Quality Plan guidelines established under ISO 9001:2008, thus ensuring to the Principal Investigator the worker incorporation on the desired date.

As previously mentioned, Human Resources policy at CIBERNED directs its selective processes towards the recruitment and retention of talent, allowing the incorporation of highly qualified personnel whose levels of technical and behavioral skills are suited to the job profiles sought. During the year 2017 a total of eight calls for personnel have been published, with 36 job positions posted, with a total of 607 job applications received.

• List of published calls during 2016 and 2017

Call Numer of Available Numer of Applicants per year calls positions applicants position

2016 8 37 819 22,14 2017 10 36 607 16,86

2017 CIBERNED Annual Report / 300 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 36 positions filled-in during 2017, ten people had a PhD degree, twenty-two were graduates, and four were advanced technicians.

10 Doctor (PhD) 10

22 Graduate (BSc, MSc) 18

4 Advanced technician 9 2017 2016

• CIBERNED welcomes its new workers 2017 2016 A proper incorporation of new personnel to our Center is as important as the selection process itself. In order to providing the new workers a quick integration and knowledge of the Center, they are supplied with the “Welcome Hanbook”. This guide contains all information that will be useful in providing an overview of our organization as well as basic guidelines to ease their working relationship. Its content includes a brief presentation of CIBERNED, its origin and objectives, job classification, organization of work, leaves and vacations, a summary of Occupational Risk Prevention (ORP) and templates to handle vacations and ORP. They are also provided with a “Safety and Health Handbook”, which include information regarding their job position.

In order to ensure all employees health and welfare in all activities undertaken at the workplace, CIBERNED provides a favorable work environment to health 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 Health and Safety at the Workplace protocols, seeing as an opportunity to detect apportunities for improvement in those areas most closely linked to work procedures, professional development, competences, training and communication with researchers.

2017 CIBERNED Annual Report / 301 • Distribution of research groups personnel during 2017

17% During 2017 the distribution of personnel by type of contract was 27 hired permanently and 131 part- time.

83%

Part-time Permanent 6 PhD Graduate 38 49 The distribution of personnel according to their degree was 49 Doctors (PhD), 62 graduates, 4 Undergraduate undergraduates, 38 advanced technicians, and 6 Technician 4 administrative assistants. Administrative 61

CIBERNED workforce during 2017 consisted of 109 women and 49 men.

31% 69%

The geographic distribution of personnel of the research teams by Regions was as follows:

Basque Country 8 Canary Island 2 Galicia 2 Extremadura 2 Valencia 12 Navarra 2 Madrid 66 Catalonia 49 Castilla - La Mancha 4 Cantabria 3 Andalusia 8

2017 CIBERNED Annual Report / 302 TRAINING PROGRAM

With the aim of promoting and developing programs for the scientific training of researchers, CIBERNED has promoted or participated during 2017 in a wide variety of training activities including lectures, presentations, courses and workshops.

• On February 16 and 17, the First DEGESCO Symposium on Dementia Genetics was held at the CIEN Foundation headquarters in Madrid, organized by Drs. Miguel Medina (CIBERNED) and Miguel Calero (Carlos III Health Institute/CIBERNED/CIEN Foundation). The event was a great success of attendance and brought together the best national experts on the genetics of neurodegenerative diseases to address a key area of research, reviewing in depth the genetic aspects of the different types of dementia and their implications in the Spanish population.

• The First Edition of the Eurotau Meeting, was held on April 27 and 28, 2017 at the University of Lille, where European scientists involved in Tau research met to exchange new ideas and hypotheses about the physiological and pathological roles of Tau protein. Prof. Jesús Ávila, Scientific Director of CIBERNED and Dr. Miguel Medina, Deputy Scientific Director of CIBERNED were part of the Scientific Committee of the Meeting, together with Prof. Luc Buée, Head of the Alzheimer & Tauopathies laboratory at the Research Center Jean-Pierre Aubert, University of Lille, and host of the event. Both Dr. Avila and Dr. Medina took also part of the Scientific Program with lectures on “Influence of tau levels and phosphorylation in cognition” and “Tau secretion and toxicity”, respectively.

• On June 16, 2017, CIBERNED participated in the Scientific Workshop on “Autophagy and Neurodegeneration”, which took place at the Faculty of Medicine of the Autonomous University of Madrid. It involved both the Manager, Maria Angeles Perez, and Drs. Ana Pérez-Castillo, José Manuel Fuentes Rodríguez and Francisco Wandosell.

• Between June 28 and 30, 2017, within the International Summer-Autumn Courses of the University of Extremadura, CIBERNED collaborated and organized the Course “Advances in Neurodegenerative Diseases”, directed by Dr. José Manuel Fuentes Rodríguez.

• The Alzheimer’s Association International Conference (AAIC) 2017 is one of the largest international meetings devoted to the advancement of scientific research in dementia. Every year, the AAIC brings together the world’s leading researchers (next-generation researchers, clinicians and the healthcare research community) to share discoveries in basic and translational research that will lead to methods of prevention, treatment and improvements in the diagnosis of Alzheimer’s disease and other dementias. In the 2017 edition, CIBERNED was represented by some of its main principal investigators such as Dr. Alberto Lleó, Dr. Eva Carro, and its scientific director Prof. Jesús Ávila de Grado, who chaired on Tuesday, July 18,the session on “Tau Propagation Mechanisms and Prevention of Toxicity”. In that same session, he delivered the lecture “Tau Secretion and Propagation”.

• The Global Summit on Alzheimer’s Research and Care was held in Lisbon, from September 18 to 22, 2017. This global meeting, key to Alzheimer’s disease, took place at the Champalimaud Foundation headquarters, with the support of the Queen Sofia Foundation. It was based on two main conferences: the Social and Healthcare Research Conference, from September 18 to 19, organized by the Imserso’s State Reference Center (CRE) of Care for people with Alzheimer’s and other dementias; and the Scientific Research Conferece, from September 19 to 22, organized by the Center for Research in Neurological Diseases (CIEN) Foundation and the Network Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED).

2017 CIBERNED Annual Report / 303 • From September 25 to 27, 2017, the “Synapse formation, specification and elimination: from molecules to circuits” meeting took place at the International University of Andalusia in Baeza within the “Current Trends in Biomedicine” series, organized by Dr. Rafael Fernández-Chacón, Principal Investigator of CIBERNED, together with Prof. Thomas C. Südhof, from Stanford University.

• On November 8, 2017, CIBERNED participated in the Scientific Conference “Challenges in Alzheimer’s and Other Dementia Research”, organized by the Maimónides Institute of Biomedical Research of Córdoba (IMIBIC) and the Alzheimer’s Association of Córdoba.

• On the celebration of the Science Week, CIBERNED organized in November 13 the presentation of the book “Alzheimer, prevention from childhood”, authored by Dr. Felix Bermejo Pareja, neurologist, psychologist and researcher. He is currently Director of the Alzheimer Extraordinary Chair of the Department of Medicine of the Complutense University of Madrid, Emeritus Professor of CIBERNED and consultant neurologist of the 12 de Octubre Hospital Research Institute.

Jesús Ávila de Grado, CIBERNED Scientific Director, Alzhéimer’s Global Summit Lisbon 2017

Presentation of the book “Alzheimer, prevention from childhood by Félix Bermejo Pareja, Science week

2017 CIBERNED Annual Report / 304 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 Occupational 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 of Accident Value accident rate rate

Index 1 6,43 0,01 0

Index 2 0,58 0 0

Index 3 0,36 0 0 QUALITY

In this year, the procedures and working tools required by the new version of the ISO 9001: 2015 standard have been developed and implemented.

The Quality policy seeks guaranteeing and optimizing the processes related to: focus on the external and internal customer, leadership, staff participation, process-based approach, continuous improvement and incorporation of the risk-based process, thus providing greater predictive capacity for the evaluation, management, minimization and/or reduction of risks.

The following measures are worth highlighting: the adoption of a process-based approach, the management of internal and external risks and the planning of quality objectives.

During 2017, renewal of the Certification of the Quality System Management under the UNE-EN ISO 9001:2008 standard has been successfully passed.

2017 CIBERNED Annual Report / 305 COLLABORATION AGREEMENTS

In 2017, the following agreements have been kept in force by the institution:

Signing partner institutions Subject

Ramón Areces Foundation-Ciberned XVIII National Call For Research Grants On Life And Materials Sciences

Biocross- Cien Foundation-Ciberned Collaboration Agreement For The Implementation Of Public-Private Cooperation Actions Within The Call Retos Colaboración 2016

Biocross- Cien Foundation -Ciberned Collaboration Agreement For The Implementation Of Public-Private Cooperation Actions Within The Call Retos Colaboración 2017

Ciber ISCIII Regulate The Participation Of The Research Groups Of CIBERNED In The Vallecas Project (Cohortes P. Vallecas)

University of Barcelona Assignment Of Use To The University Of Barcelona

Biomedical Research Clinic Foundation, Clinic Co-Ownership Agreement Related To Patent Hospital of Barcelona, University of Barcelona and “Method For The Subclassification For Patients CSIC Suffering From Parkinson Disease”

Biocross-Isciii Co-Ownership Agreement Related To Patent “A Fast And Cost-Effective Method For Apolipoprotein E Isotyping As An Alternative To Apoe Genotyping”

University of Bordeaux- Oxford University Coen Project. Consortium Agreement. “Protection Of Neurons In Vitro And In Vivo From Synuclein Toxicity By Molecular Tweezers”

Ciber ISCIII Interciber Agreement To Regulate The Participation Of CIBERNED Research Groups In The Research Project “Links Between Diabetes And Neurodegenerative Disorders”

Cic Biomagune Student Secondment Agreement - CIBERNED Cooperative Project 2014/06 “Onset And Progression Of The Parkinson Nurse: Role Of The Glial Activation”

National Evaluation and Prospective Agency ANEP Scientific-Technical Evaluation Of CIBERNED Cooperative Projects

CSIC- Autonomous University of Madrid Co-Ownership Agreement Related To Patent Nº 201431898 “Modulator Compounds Of The Calcium Dream Neural Sensor And Its Therapeutic Uses”

CSIC-UAM Co-Ownership Agreement Related To Patent Nº201431898 “Modulator Compounds Of The Calcium Dream Neural Sensor And Its Therapeutic Uses”

Secretariat of State of Universities Ministery of Management Of Training And Mobility Grants Education, Culture and Sports From The Promotion Of Talent Program And Its Employability Of The State Plan Of Scientific And Technical Research And Of Innovation 2013-2016 And Other Actions Of Grants Calls For The Mobility Of Students And College Teachers

Janssen Cilag-Neuron Bio-CSIC Collaboration Agreement For The Execution Of Public-Private Cooperation Actions Within The 2015 “Retos Colaboracion” Call

University of Extremadura Curricular And Extracurricular Practices Of Students

2017 CIBERNED Annual Report / 306 Signing partner institutions Subject

Complutense University of Madrid Co-Ownership Agreement Related To Patent “Regulation Of Cannabinoid Cb1 Receptors For The Generation Of Cortico-Spinal Neurons From Murine Coronary Stem Cells”

University of Extremadura Curricular And Extracurricular Practices Of Students

CIBERNED Associated Institutions Iicc Collaboration Agreement

Miguel Hernández University of Elche- Foundation for Co-Ownership Agreement Related To Patent the Promotion of Health And Biomedical Research of Co-Ownership Agreement Related To Patent the Region of Valencia P201130290 “Method For Diagnosis And/Or Prognosis Of Alzheimer’s Disease”

Fibio 12 De Octubre Hospital Co-Ownership Agreement Related To Patent Pct/ Es2013/070693 “Animal Model Of Cognitive Deficit, Method For Obtaining And Applications”

CSIC – Cien Foundation Co-Ownership Agreement Related To Patent Ep13382108.2 “Methods For The Prognosis And Diagnosis Of Neurodegenerative Diseases”

Biocross - Cien Foundation Non-Disclosure Agreement Related To Patent Ep13382108.2 “Methods For The Prognosis And Diagnosis Of Neurodegenerative Diseases”

CSIC - Neuron Bio Non-Disclosure Agreement Related To Patent P201231654 “Recombinant Sgg2 Viral Protein And/ Or Sgg2-Fns Binary Compounds For Use In Axonal Growth And/Or Regeneration”

CSIC-Johnson & Johnson Co-Ownership Agreement Related To Patent P201231807 “METHODS FOR THE PROGNOSIS AND DIAGNOSIS OF NEURODEGENERATIVE DISEASES”

CSIC Co-Ownership Agreement Related To Patent P201231654 “Recombinant Sgg2 Viral Protein And/ Or Sgg2-Fns Binary Compounds For Use In Axonal Growth And/Or Regeneration”

ACE Foundation Framework Collaboration Agreement

Teófilo Hernando Institute (UAM) Framework Collaboration Agreement To Promote And Foster Study, Research, And Teaching Activities, As Well As Dissemination And Celebration Of Events Of Scientific, Academic And Cultural Interest

CSIC Co-Ownership Agreement Related To Patent “Mitochondrial Dna As A Marker For The Diagnosis Of Neurodegenerative Diseases”

Fibio Ramón y Cajal Hospital Co-Ownership Agreement Related To Patent “GLIA CONDITIONED MEDIUM AS NEUROPROTECTIVE AGENT”

Miguel Hernández University of Elche Co-Ownership Agreement Related To Patent “METHOD FOR DETERMINING ALZHEIMER’S DISEASE THROUGH THE DETECTION OF GLICOPROTEINS CARRYING GLYCOPEPIDE HNK-1”

Cien Foundation - CSIC – CRG Co-Ownership Agreement Related To Patent P201130033 “Compounds For The Treatment Of Neurodegenerative Diseases”

2017 CIBERNED Annual Report / 307 ACQUISITION OF FIXED ASSETS

The acquisition of assets made during the year are as follows:

Purchase Equipment Invoice Total Responsible IP C. orderd escription date cost

2062141787 Gas central for CO2 bottles 16/01/2017 3.804,64€ M. Rodríguez 206

0012171786 Monitor LG 22 M38A 01/02/2017 119,79€ Servicios generales 1

0012171785 HP pro desk 400 01/02/2017 719,95€ Servicios generales 1

1032141788 Vacusafe Comfort ECO 10/02/2017 1.172,49€ J.M. Fuentes 103

1112171789 Workstation Z240 J9C02EA+Monitor 14/02/2017 1.702,47€ T. Iglesias 111 HP Z23

4012171796 Multi function laserjet pro printer 16/02/2017 796,18€ J. Ávila 401

4012141797 Orbital Shaker 16/02/2017 1.069,64€ J. Ávila 401

1032141793 Powerpack HC, 100-120/220-240 v 24/02/2017 924,08€ J.M. Fuentes 103

1032141791 Mini-P- Tet cell/mini T-BLOT MOD 24/02/2017 1.093,48€ J.M. Fuentes 103

1032141792 Trans blot SD semi-dry 24/02/2017 1.309,89€ J.M. Fuentes 103

2052171799 Dell monitor S2715H 28/02/2017 289,19€ J.A. Obeso 205

2052171800 PC XPS 8910 28/02/2017 1.680,69€ J.A. Obeso 205

6062141790 Switchable ACC with built in intelligence 08/03/2017 1.261,88€ R. Fernández 606

1032141794 Criterion Blotter W/PATE ELEC 10/03/2017 995,31€ J.M. Fuentes 103

0012171795 Monitor LED HP ProDisplay 21,5” 13/03/2017 180,29€ Servicios generales 1

1142171798 BQ Witbox 2 3D printer 15/03/2017 1.690,00€ J.A. del Río 114

2072141809 New upright freezer, 482 litres 27/03/2017 5.772,91€ E. Tolosa 207

3072171801 MacBool Pro Silver 28/03/2017 2.783,00€ J.R. Naranjo 307

6072141819 Innova U535 - 86º C vertical freezer 28/03/2017 8.470,00€ X. Navarro 607

6072171817 W2100 Wireless System. For 28/03/2017 18.080,91€ X. Navarro 607 electrophysiology data acquisition

6012141803 Sigma 1-14k refrigerated 29/03/2017 4.734,73€ J. Infante 601 microcentrifuge

5092141825 Tyssuel Lyser II (bead mil for simple 29/03/2017 5.445,00€ M. Calero 509 disruption)

3072141807 Electroporator 30/03/2017 2.466,46€ J.R. Naranjo 307

2082141811 Liebherr vertical freezer 30/03/2017 2.538,58€ J.L. Labandeira 208

0012171804 HP Prodisplay P222VA LEDBLT monitor 31/03/2017 192,39€ Servicios generales 1

0012171805 HP Prodisplay P222VA LEDBLT monitor 31/03/2017 192,39€ Servicios generales 1

2082141810 Eppendorf Centrifuge 31/03/2017 5.667,64€ J.L. Labandeira 208

4152141806 Leica VT1000S Vibratome 31/03/2017 19.783,79€ A. Gutiérrez 415

2017 CIBERNED Annual Report / 308 Purchase Equipment Invoice Total Responsible IP C. orderd escription date cost

6012171802 Apple Macbook Pro 15” 04/04/2017 2.374,20€ J. Infante 601

4152141808 Digital Stereotaxic instrument KOPF 04/04/2017 15.187,92€ A. Gutiérrez 415 model 940

6012141813 EIA microplate washer 06/04/2017 2.710,40€ J. Infante 601

6012141812 Microplate reader, Mod RT6500 06/04/2017 5.124,35€ J. Infante 601

4112141820 SumpliAmp Thermal Cycler 18/04/2017 3.872,11€ Fco. J. Vitorica 411

3072141816 Histology bath 1.9 l 20/04/2017 4.924,46€ J.R. Naranjo 307

0012061814 Development of app 20/04/2017 5.445,00€ Servicios generales 1

2082141821 Analytical balance 24/04/2017 3.811,50€ J.L. Labandeira 208

1032141818 Cell counter Olympus 24/04/2017 4.437,05€ J.M. Fuentes 103

4152141815 Leica SM2010 R sliding microtome 26/04/2017 11.870,10€ A. Gutiérrez 415

4082141844 miniV/PCR cabin, front closure 28/04/2017 754,68€ E. Soriano 408

4082141843 Digital New Standard Stereotaxi rat 04/05/2017 11.222,33€ E. Soriano 408 and mouse

6072141827 W2100-HS8-ES2-EXT-2.0mA Wireless 09/05/2017 1.863,40€ X. Navarro 607 headstage

6072141828 W2100-HS8-ES2-EXT-2.0mA Wireless 09/05/2017 1.863,40€ X. Navarro 607 headstage

2092171822 Laptop Lenovo Yoga 900-13ISK2 80UE 11/05/2017 1.524,60€ J. Pérez 209

5072171823 Macbook PRO 15” Space Grey 12/05/2017 2.699,00€ Mª A. Pastor 507

3072141826 Luminiscence equipment 17/05/2017 16.403,97€ J.R. Naranjo 307

0121741824 Xerox WorkCentre 6025V_BI printer 24/05/2017 223,37€ Servicios generales 1

4082141832 Laminar flow cabinet 01/06/2017 2.531,57€ E. Soriano 408

4082141833 Thermo Scientific Forma Series 3 01/06/2017 7.186,53€ E. Soriano 408 Water Jacketed

3072171829 Computer PC LENOVO 02/06/2017 349,69€ J.R. Naranjo 307

2082141835 Inverted microscope with camera 07/06/2017 5.445,00€ J.L. Labandeira 208 and accessories

4012141848 Freezer Liebher G5216 09/06/2017 1.391,20€ J. Ávila 401

2052141849 UltraMicroPump III (W/O controller) 09/06/2017 3.350,49€ J.A. Obeso 205

2052061831 DEDACOM V2.0 basic Software basic 13/06/2017 186,34€ J.A. Obeso 205 for data display

4082141834 Analytical Balance ME204T/00 13/06/2017 3.626,56€ E. Soriano 408

2052141830 RotaRod with Touchscreen, 5 mice 13/06/2017 4.812,30€ J.A. Obeso 205

2042061839 Imaris 8.4 for Neuroscientists 29/06/2017 14.000,00€ R. Moratalla 204

3062171836 PC components 12/07/2017 795,87€ J.J. Lucas 306

2017 CIBERNED Annual Report / 309 Purchase Equipment Invoice Total Responsible IP C. orderd escription date cost

3062171837 Laptop HP Envy 17-ae003ns 14/07/2017 1.363,67€ J.J. Lucas 306

2042171878 Micro Intel Core I7-770K 17/07/2017 2.153,80€ R. Moratalla 204

5112141845 Complete semiautomatic defibrillator 21/07/2017 939,00€ J.L. Cantero 511 ZOLL AED PLUS

5072171838 Laptop ASUS VIVOBOOK 26/07/2017 651,00€ Mª A. Pastor 507

4152171840 Pc P646 ORD VSK4000B 27/07/2017 1.161,60€ A. Gutiérrez 415

4152171841 Pc P646 ORD VSK4000B 27/07/2017 1.161,60€ A. Gutiérrez 415

4152171842 Pc P646 ORD VSK4000B 27/07/2017 1.161,60€ A. Gutiérrez 415

1032141846 Concentrator Plus Rotor Included 09/08/2017 7.260,01€ J.M. Fuentes 103

6072171851 GPULPIP5 10/08/2017 3.710,00€ X. Navarro 607

2042141858 Stereotaxic equipment 01/09/2017 9.848,99€ R. Moratalla 204

2062141850 Narishige mouse adapter mod. 05/09/2017 1.247,62€ M. Rodríguez 206 MA6N

2042141857 Dual laser system 473/561 combined 05/09/2017 14.415,94€ R. Moratalla 204 for optogenetics

1032141847 Doigt refrigerant 250 ml + dewar 06/09/2017 568,70€ J.M. Fuentes 103

4112171860 Monitor LED ASUS VS228DE 21,5” VGA 11/09/2017 100,13€ Fco. J. Vitorica 411

4112171859 Computer black side intel core I5 11/09/2017 869,71€ Fco. J. Vitorica 411

4152061862 E11HPPZ440 REN 2, E11Visiopharm 28/09/2017 14.323,98€ A. Gutierrez 415 and E11INSTALL

0012161852 Chair with adjustable armrest, blue 06/10/2017 195,90€ Servicios generales 1 red backrest and basic seat 169 blue

0012161853 Chair with adjustable armrest, blue 06/10/2017 195,90€ Servicios generales 1 red backrest and basic seat 169 blue

0012161854 Chair with adjustable armrest, blue 06/10/2017 195,90€ Servicios generales 1 red backrest and basic seat 169 blue

0012161855 Chair with adjustable armrest, blue 06/10/2017 195,90€ Servicios generales 1 red backrest and basic seat 169 blue

0012161856 Chair with adjustable armrest, blue 06/10/2017 195,90€ Servicios generales 1 red backrest and basic seat 169 blue

1142171863 Workstation 10/10/2017 8.278,43€ J.A. del Río 114

4082171861 MSI GS73VR 7RF 288ES Stealth Pro 4K 17/10/2017 2.724,92€ E. Soriano 408

4092171870 Monitor BENQ 21,5 “ 31/10/2017 123,42€ I. Torres 409

4092171874 Monitor BENQ 21,5” 31/10/2017 123,42€ I. Torres 409

4092171873 Pantalla BENQ 23,8” 31/10/2017 139,15€ I. Torres 409

4092171871 BCB computer 31/10/2017 896,61€ I. Torres 409

4092171872 BCB computer 31/10/2017 984,55€ I. Torres 409

2017 CIBERNED Annual Report / 310 Purchase Equipment Invoice Total Responsible IP C. orderd escription date cost

4092171875 BCB computer 31/10/2017 984,55€ I. Torres 409

2052171882 Apple Macbook AIR 13” core I5 08/11/2017 1.676,99€ J.A. Obeso 205

2052171881 Apple Macbook AIR 13” core I5 08/11/2017 1.677,00€ J.A. Obeso 205

2072141867 5804 R centrifuge 09/11/2017 8.311,25€ E. Tolosa 207

2052171879 UPS 1500VA/1050W 10/11/2017 314,12€ J.A. Obeso 205

2052171880 UPS 1500VA/1050W 10/11/2017 314,12€ J.A. Obeso 205

0012141884 Sorwall 16 RT centrifuge 13/11/2017 7.077,59€ J.L. Cantero 511

4092141866 Refrigerated centrifuge 14/11/2017 5.263,50€ I. Torres 409

2072171869 HP EliteDesk 800 G2 SFF 23/11/2017 1.250,54€ E. Tolosa 207

2072141868 ULTRA-TURRAZ homogenizer 05/12/2017 880,28€ E. Tolosa 207

4012171883 Monitor HP OMEN BY HP 25 DISPLAY 29/12/2017 336,38€ J. Ávila 401

2017 CIBERNED Annual Report / 311

Principal investigator index

PRINCIPAL INVESTIGATOR INDEX

Alberch Vié, Jordi Program 2 Page 112 Ávila de Grado, Jesús Program 1 Page 27 Bullido Gómez-Heras, Mª Jesús Program 1 Page 30 Calero Lara, Miguel Program 1 Page 33 Camins Espuny, Antonio Program 1 Page 38 Canela Campos, Enric Isidre Program 2 Page 117 Cantero Lorente, José Luis Program 1 Page 43 Carro Díaz, Eva Program 1 Page 47 Ceña Callejo, Valentín Program 2 Page 121 Comella Carnicé, Joan Xavier Program 1 Page 52 Cuadrado Pastor, Antonio Program 2 Page 124 de Felipe Oroquieta, Javier Program 1 Page 55 del Río Fernández, José Antonio Program 2 Page 128 Fariñas Gómez, Isabel Program 2 Page 121 Fernández Chacón, Rafael Program 2 Page 135 Fernández Ruiz, Javier Program 2 Page 138 Ferrer Abizanda, Isidro Program 1 Page 59 Fuentes Rodríguez, José Manuel Program 2 Page 142 García Verdugo, José Manuel Program 2 Page 146 González Castaño, José Program 2 Page 150 Gutiérrez Pérez, Antonia Program 1 Page 65 Guzmán Pastor, Manuel Program 2 Page 152 Iglesias Vacas, Teresa Program 2 Page 156 Infante Ceberio, Jon Program 2 Page 159 Kulisevsky Bojarski, Jaime Program 2 Page 165 Labandeira García, José Luis Program 2 Page 170 Lanciego Pérez, José Luis Program 2 Page 174 Lleó Bisa, Alberto Program 1 Page 68 López Barneo, José Program 2 Page 177 López de Munain Arregui, Adolfo Program 2 Page 182 Lucas Lozano, José Javier Program 2 Page 189 Matute Almau, Carlos Program 1 Page 75 Mengod Los Arcos, Guadalupe Program 1 Page 79 Moratalla Villalba, Rosario Program 2 Page 192 Muñoz Cánoves, Pura Program 2 Page 195 Naranjo Orovio, José Ramón Program 2 Page 199 Navarro Acebes, Xavier Program 2 Page 202 Obeso Inchausti, José Ángel Program 2 Page 208 Pastor Muñoz, María Asunción Program 1 Page 81 Pérez Castillo, Ana María Program 2 Page 214 Pérez Tur, Jordi Program 2 Page 217 Rodríguez Álvarez, José Program 1 Page 84 Rodríguez Díaz, Manuel Program 2 Page 219 Sáez Valero, Javier Program 1 Page 87 Soriano García, Eduardo Program 1 Page 90 Tolosa Sarró, Eduardo Program 2 Page 222 Torres Alemán, Ignacio Program 1 Page 93 Trullás Oliva, Ramón Program 1 Page 96 Vicario Abejón, Carlos Program 2 Page 229 Vila Bover, Miquel Program 2 Page 232 Vitorica Ferrández, Fco. Javier Program 1 Page 99 Wandosell Jurado, Francisco Program 1 Page 102

2017 CIBERNED Annual Report / 315

2017 ANNUAL REPORT

MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD C/ Valderrebollo, 528031 Madrid Tel.: + 34 913 852 200 Fax: +34 913 852 118 www.ciberned.es