2015 Annual Report

MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD

C/ Valderrebollo, 528031 Madrid Tel.: + 34 913 852 200 MINISTERIO MINISTERIO Fax: +34 913 852 118 DE ECONOMÍA DE ECONOMÍA Y COMPETITIVIDAD www.ciberned.es Y COMPETITIVIDAD 2015 Annual Report

MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD 2015 Annual Report

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 ...... 1

• Overview...... 2

• Aims ...... 4

• Directory of research groups and associated institutions ...... 5

• Geographic distribution of CIBERNED research groups ...... 7

• Organizational structure ...... 8

• Organization chart ...... 9

• External scientific advisory committee...... 10

• Consortium members ...... 11

• Scientific report ...... 13

• Research programs ...... 15

• Program 1: Alzheimer’s disease and other degenerative dementias...... 17

• Program 2: Parkinson’s disease and other neurodegenerative movement disorders ...... 103

• Cooperative research ...... 235

• International relations ...... 251

• Scientific productivity and other activities ...... 267

• Financial report ...... 283

• Principal investigator index...... 303

Letter from the Scientific Director

I would like to start this letter by thanking the work of our Center research groups, Steering Committee members and the Directorate of the Health Institute Carlos III because, thanks to their work and assistance, CIBERNED continues to grow scientifically in a gradual and orderly manner that maintains another year our institution at the forefront of the spanish centers with highest impact scientific publications.

The work of our 54 groups, grouped into the “Alzheimer’s Disease and other dementias” and the “Parkinson’s disease and other motor neurodegenerative disorders” programs, has led to 553 CIBERNED papers published, with an average impact factor of 5.626. Furthermore, CIBERNED has had 34 active clinical trials during 2015, and the results of some of these trials have been already published.

Some of the results from the activities carried out have been published in journals of the highest impact such as Nature, New England Journal of Medicine, Cell Stem Cell, JAMA, Cell, Science and The Lancet Neurology, being some of these publications the result of collaborative projects between two or more CIBERNED groups through our intramural research program.

Moreover, since our nature as a translational research with a social projection, we have not only delivered scientific publications, clinical trials or patents, but we have also carried out our Social Forum, holding meetings with patient association in Malaga and Barcelona. In addition, CIBERNED has organized several scientific meetings and our training plan is consolidating.

I finish just as I started, by thanking the effort of everyone to keep growing scientifically and to make CIBERNED a competitive and quality international center.

Jesús Ávila de Grado Scientific Director

2015 CIBERNED Annual Report / 1 Overview

The Network Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED) emerges in 2006 as heir to the Center for Research in Neurological Diseases (CIEN), which together with the Spanish National Cancer Research Centre (CNIO), the Spanish National Center for Cardiovascular Research (CNIC) or the Spanish Center for Microbiology, Virology and Health Immunology (CNMVIS) constituted the four centers whose mission is to combat the most prevalent health problems of Spanish society: cancer, cardiovascular diseases, infectious diseases and neurodegeneration. As for the other Centers, a supporting foundation called CIEN Foundation was created in 2003 as the CIEN’s management authority, although unlike the other centers, it was created as the first Network Center in Spain, eventually being renamed as CIBERNED. Both CIEN and CIEN Foundation partnered with the Queen Sofia Foundation in creating the Alzheimer Center, located in Vallecas (Madrid) and where CIEN Foundation is headquartered since 2007. CIBERNED was founded under the auspices of the Carlos III Institute of Health, under a cooperation CIBERNED emerges agreement signed by the Government and other participating institutions with the idea of creating in 2006 with a research center in which basic, clinical and population-based studies are integrated in order to multidisciplinary y develop a single joint research program, focusing on certain pathologies of great importance for the multi-institutional National Health System, either for its prevalence or because of its social impact. It has a multidisciplinary character and multi-institutional character and it is aimed at boosting the impact of cutting research in neurodegenerative disorders through a Network Research Structure, thus contributing to scientifically substantiate the programs and policies of the National Health System in the priority areas of the National R+D+I. CIBERNED is a research organism, endowed with legal status under the Article 6.5 of Law 30/1992 of November 26 on the Legal Regime of Public Administrations and Common Administrative Procedure. It is formed by the association of research groups, with no physical contiguity, belonging to different administrations, institutions and regional governments, from public and private sectors with research lines and goals focused on the specific common area of neurodegenerative diseases and being coordinated to achieve scientific objectives that could hardly be considered in a more restricted execution context. CIBERNED is governed in the internal operating rules, by way of a regulation. Currently, the CIBERNED-CIEN Foundation The CIBERNED’s goals partnership is the only research center in Spain (and one of the few in the world) are focused on the integrated into the international network of Centres of Excellence in Neurodegeneration neurodegenerative (COEN), an initiative arising from the European Union Joint Programme for diseases Research in Neurodegenerative Diseases (JPND). This joint program constitutes an innovative collaborative research initiative created to address the growing challenges posed by this group of diseases. Its goal is to boost the impact of research by aligning existing national research programs and identifying the common objectives whose scope would benefit through joint action. Its own statutes govern CIBERNED and since the end of 2014 its activity is structured around two scientific programs:

• Program 1: Alzheimer’s disease and other degenerative dementias. • Program 2: Parkinson’s disease and other neurodegenerative movement disorders.

2015 CIBERNED Annual Report / 2 During 2015 CIBERNED has consisted of 54 research groups supported by different universities, hospitals and the Research Council (CSIC), each led by a principal investigator or responsible. CIBERNED is a network research center, composed of research groups belonging to different administrations and affiliated institutions: researcher members physically work in their parent institutions they belong to while simultaneously and actively participating on CIBERNED’s own cooperative research agenda. Thus, it is the result of a collaborative partnership between different institutions and the sum of 54 CIBERNED has research groups. Depending on the results of the scientific evaluation of the various research groups, consisted of 54 there is the possibility to agree on the discontinuation of some groups. Depending on budget availability, research groups it may also consider adding some new research group currently outside CIBERNED that satisfy the requirements of scientific quality and translational activity and aligns with CIBERNED priorities. The Scientific Director Office has been set since October 2011 at the Center for Molecular Biology “Severo Ochoa” (CSIC) in Madrid. The headquarters, including the General Manager Office, is located in the Alzheimer Center of the Queen Sofia Foundation, 5 Valderrebollo Street in Madrid.

2015 CIBERNED Annual Report / 3 Aims

CIBERNED’s ultimate goal is to foster scientific and technical research of excellence in the field of human health, with the overall goal of producing results quickly translatable into clinical practice to improve the health and wellbeing of patients with neurodegenerative diseases, as well as their families and caregivers, providing in turn economic and social benefits. Among its objectives, 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 therapeutical 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.

2015 CIBERNED Annual Report / 4 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 502 Bermejo Pareja, Félix Doce de Octubre University Hospital, Madrid 510 Bullido Gómez-Heras, Mª Jesús Autonomous University, 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 106 Ceña Callejo, Valentín University of Castilla La Mancha, Albacete 601 Combarros Pascual, Onofre Marqués de Valdecilla Foundation, Santander 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 509 de Pedro Cuesta, Jesús Institute of Health Carlos III, 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, Cáceres 113 García Verdugo, José Manuel Cavanilles Institute, University of Valencia 104 González Castaño, José Autonomous University of Madrid 415 Gutiérrez Pérez, Antonia University of Málaga 305 Guzmán Pastor, Manuel Complutense University of Madrid 111 Iglesias Vacas, Teresa Institute of Biomedical Research CSIC-UAM, Madrid 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 Rocío University Hospital, University of Sevilla 506 López de Ceballos Lafarga, María Cajal Institute CSIC, Madrid 609 López de Munain Arregui, Adolfo Biodonostia Research Institute, San Sebastián 306 Lucas Lozano, José Javier Center for Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid 404 Matute Almau, Carlos University of the Basque Country, Bilbao 304 Mena Gómez, Mª Ángeles* Ramón y Cajal Hospital, Madrid

*Until Septembre 2015

2015 CIBERNED Annual Report / 5 Group Principal investigator Institution

508 Mengod Los Arcos, Guadalupe Instituto de Investigaciones Biomédicas IDIBAPS-CSIC , Barcelona 204 Moratalla Villalba, Rosario Cajal Institute CSIC, Madrid 604 Muñoz Cánoves, Pura Pompeu Fabra University, ICREA, Barcelona 307 Naranjo Orovio, José Ramón National Center of Biotechnology, CSIC, Madrid 607 Navarro Acebes, Xavier Autonomous 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

22 32

PROGRAM 1 PROGRAM 2 54 RESEARCH GROUPS

2015 CIBERNED Annual Report / 6

22 22 32 32

PROGRAMAPROGRAM 1 PROGRAMA 1 PROGRAM 2 2 54 GRUPOS DE INVESTIGACIÓN54 RESEARCH GROUPS Geographic distribution of CIBERNED research groups

1 2 1 2 16

1 1 4

2015 CIBERNED Annual Report / 7 Organizational structure

CIBERNED governing bodies are the Governing Council, the Permanent Commission and the Scientific Director. • Governing Council: the highest body of CIBERNED, 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 Foundation) 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 Coun- cil, without a right to vote.

• 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.

• 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 Ávila 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 • Dña. Mª 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 Munain Arregui • Dr. Isidro Ferrer Abizanda (until September 2015) • Dr. Eduardo Soriano García (until September 2015) • Dra. Teresa Iglesias Vacas (from September 2015) • Dr. Rafael Fernández Chacón (from September 2015)

2015 CIBERNED Annual Report / 8 Organization chart

In stitute of Health Carlos III

Governing Council

Associated Institutions

Standing Committe

Steering Committee Scientific Director Manager

Deputy Scienfific Director

Research Program Coordinator

Research Groups

2015 CIBERNED Annual Report / 9 External Scientific Advisory Committee

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

Dr. Vincenzo Bonifati Erasmus University, Rotterdam, The Netherlands

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

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

2015 CIBERNED Annual Report / 10 Consortium members

The Central Administration, represented by:

• The Institute of Health Carlos III.

• The Spanish National Research Council (CSIC).

The following Autonomous Regions (Comunidades Autónomas):

• 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 Canarias, 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 and Ramón y Cajal Hospital), Autonomous University of Madrid, Complutense University of Madrid, and Hospitales de Madrid Foundation.

• 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.

• Center for Applied Medical Research (CIMA).

• Hospital Clinic of Barcelona.

2015 CIBERNED Annual Report / 11 Scientific report Scientific report

Research programs

The structure of CIBERNED research groups is organized around two major Scientific Programs, whose goals, composition and structure is maintained as in previous years, and whose budget allocation is based on the scientific results of the internal scientific evaluation.

The distribution of these two thematic programs shows CIBERNED commitment to encouraging translational research and obtaining scientific knowledge resulting in improving the prevention, diagnosis and treatment of neurodegenerative diseases. All this, without undermining the development of transversal research projects withof interest into different areas (see below). The aim of the structure described is to provide a coherent conceptual scheme embodying the real cooperative scientific activity conducting various research groups.

The programs are described below:

54 RESEARCH 703 I GROUPS RESEARCHERS

553 PUBLICATIONS 34 CLINICAL z TRIALS Ñ

Informe Anual CIBERNED 2015 / 15 Program 1 Alzheimer’s disease and other degenerative dementias

Ávila de Grado, Jesús...... 21 Matute Almau, Carlos...... 64 Bermejo Pareja, Félix...... 24 Mengod Los Arcos, Guadalupe...... 68 Bullido Gómez-Heras, Mª Jesús...... 29 Pastor Muñoz, María Asunción...... 71 Camins Espuny, Antonio...... 32 de Pedro Cuesta, Jesús...... 75 Cantero Lorente, José Luis...... 36 Rodríguez Álvarez, José...... 81 Comella Carnicé, Joan Xavier...... 39 Sáez Valero, Javier...... 84 de Felipe Oroquieta, Javier...... 43 Soriano García, Eduardo...... 87 Ferrer Abizanda, Isidro...... 47 Torres Alemán, Ignacio...... 90 Gutiérrez Pérez, Antonia...... 53 Trullás Oliva, Ramón...... 93 Lleó Bisa, Alberto...... 56 Vitorica Ferrández, Francisco Javier...... 96 López de Ceballos Lafarga, María...... 61 Wandosell Jurado, Francisco...... 99 Program 1 Alzheimer’s disease and other degenerative dementias

Program 1: Alzheimer’s disease and other degenerative dementias

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) . The AD is disease, the most common cause of dementia in the elderly and nowadaysagainst which there is no effective treatment against this diseaseto date. Currently, more than 500,000 people suffer from this disease AD 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 +500.000 (it is considered a synaptopathy), mainly between EA patients 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, 22 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.

2015 CIBERNED Annual Report / 19 Principal investigator Institution

Ávila de Grado, Jesús Center of Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid Bermejo Pareja, Félix Doce de Octubre University Hospital, Madrid Bullido Gómez-Heras, Mª Jesús Autonomous University of Madrid Camins Espuny, Antonio University de Barcelona Cantero Lorente, José Luis Pablo de Olavide University, Sevilla Comella Carnice, Joan Xavier Foundation Vall d’Hebron University Hospital, Barcelona De Felipe Oroquieta, Javier Cajal Institute CSIC, Center of Biomedical Technology, Madrid De Pedro Cuesta, Jesús Institute of Health Carlos III, 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 López de Ceballos Lafarga, María Cajal Institute CSIC, Madrid 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, Fco. Javier University of Seville Wandosell Jurado, Francisco Center of Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid

Program 1 is coordinated by Drs. Isidro Ferrer* (IDIBELL-Bellvitge Hospital), Alberto Lleó (Santa Creu i San Pau Hospital) and Eduardo Soriano* (University of Barcelona).

*Until Septembre 2015.

2015 CIBERNED Annual Report / 20 401 GROUP Jesús Ávila de Grado

Jesús Ávila de Grado Vega García-Escudero Noemí Pallas Bazarra Research Professor-Principal Researcher PhD student Investigator Alberto Gómez Ramos Jesús Merchán Robira Félix Hernández Pérez Researcher PhD student UAM Professor – Co-IP Jerónimo Jurado Arjona Raquel Cuadros Catalán Marta Bolos Jurado Researcher Technician Researcher Patricia Martín-Maestro Esther García García María Llorens-Martín PhD student Technician Researcher Nuria de la Torre Alonso Administrative Assistant

Contact details

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

2015 CIBERNED Annual Report / 21 Summary

We have carried out our work doing a further could take place in tauopathies like Alzheimer analysis of the neuronal connections between disease. Also, we have analyzed, in peripheral entorhinal cortex and the CA1 hippocampal tissue from Alzheimer disease patients (where region. In these connections newborn neurons amyloid beta peptide expression takes place), generated by adult neurogenesis, at the if could be mitochondrial defects in those dentate gyrus (DG), may play a role. Indeed, peripheral cells. we have described a novel connection between newborn neurons from DG with the During the last year we have continued with the CA2 hippocampal region. In addition, we collaborations with other groups from CIBERNED, have continued with the characterization of like those of Dr. Soriano continuing the research tau protein function on axonal development. of specific somatic mutations in brain cells from A novel observation, done this year, has been Alzheimer disease patients, and with the groups the role of microglia cells in tau spreading that of Dr. Lucas and De Felipe.

Keywords

Tau, GSK3, Alzheimer disease, neurogenesis

Publications

• Avila J., Gomez-Ramos A., Bolos M. Ad genetic risk factors and tau spreading. Frontiers in Aging Neuroscience. 2015;7(MAY). [Epub ahead of print]. • Calero M., Gomez-Ramos A., Calero O., Soriano E., Avila J., Medina M. Additional mechanisms conferring genetic susceptibility to Alzheimer’s disease. Frontiers in Cellular Neuroscience. 2015;9(APR). [Epub ahead of print]. • Avila J., Perry G., Strange B.A., Hernandez F. Alternative neural circuitry that might be impaired in the development of Alzheimer disease. Frontiers in Neuroscience. 2015;9(APR). [Epub ahead of print]. • Fernández-Nogales M, Hernández F, Miguez A, Alberch J, Ginés S, Pérez-Navarro E et al. Decreased glycogen synthase kinase-3 levels and activity contribute to Huntington’s disease. Human molecular genetics. 2015;24(17): 517-24. • Gomez-Ramos A., Podlesniy P., Soriano E., Avila J. Distinct X-chromosome SNVs from some sporadic AD samples. Scientific Reports. 2015;5. [Epub ahead of print]. • Jurado-Arjona J., Goni-Oliver P., Rodriguez-Prada L., Engel T., Henshall D.C., Avila J. et al. Excitotoxicity induced by kainic acid provokes glycogen synthase kinase-3 truncation in the hippocampus. Brain Research. 2015;1611:84-92. • Llorens-Martín M, Jurado-Arjona J, Bolós M, Pallas-Bazarra N, Ávila J. Forced swimming sabotages the morphological and synaptic maturation of newborn granule neurons and triggers a unique pro-inflammatory milieu in the hippocampus.Brain, behavior, and immunity. 2015. [Epub ahead of print]. • Llorens-Martin M., Jurado-Arjona J., Avila J., Hernandez F. Novel connection between newborn granule neurons and the hippocampal CA2 field. Experimental Neurology. 2015;263:285-292. • Martín-Maestro P, Gargini R, Perry G, Avila J, García-Escudero V, Avila J. PARK2 enhancement is able to compensate mitophagy alterations found in sporadic Alzheimer’s disease.Human molecular genetics. 2015. [Epub ahead of print]. • Sayas C.L., Tortosa E., Bollati F., Ramirez-Rios S., Arnal I., Avila J. Tau regulates the localization and function of End-binding proteins 1 and 3 in developing neuronal cells. Journal of Neurochemistry. 2015;133(5):653-667. • Sebastian-Serrano A., De Diego-Garcia L., Martinez-Frailes C., Avila J., Zimmermann H., Millan J.L. et al. Tissue-nonspecific Alkaline Phosphatase Regulates Purinergic Transmission in the Central Nervous System during Development and Disease. Computational and Structural Biotechnology Journal. 2015;13:95-100.

2015 CIBERNED Annual Report / 22 Program 1 Group: Jesús Ávila de Grado

• Diaz-hernandez M., Hernandez F., Miras-portugal M.T., Avila J. TNAP plays a key role in neural differentiation as well as in neurodegenerative disorders. Sub-Cellular Biochemistry. 2015;76:375- 385. • Bolos M., Llorens-Martin M., Jurado-Arjona J., Hernandez F., Rabano A., Avila J. Direct Evidence of Internalization of Tau by Microglia in Vitro and in Vivo. Journal of Alzheimer’s Disease. 2015;50(1):77-87. • Llorens-Martin M., Rabano A., Avila J. The ever-changing morphology of hippocampal granule neurons in physiology and pathology. Frontiers in Neuroscience. 2016;9(JAN). Epub 2015.

Research projects

• Code: 2013/07A. • Code: S2010_BMD. Title: Papel de GSK-3 β en las alteraciones Title: Redes de senalización y vías efectoras de los circuitos corticales que ocurren en la en modelos celulares y animales de enfermedad de Alzheimer. enfermedades neurodegenerativas. Principal investigator: Teresa Iglesias Vacas. Principal investigator: José González CIBERNED’s collaboration: Yes. Castaño. CIBERNED groups: G504, G401, G403, G111, CIBERNED’s collaboration: Yes. G307. CIBERNED groups: G111, G401, G104, G307, Other CIBER’s collaboration: No. G409, G412. Type: International. Other CIBER’s collaboration: Yes (CIBERER). Funding agency: CIBERNED. Type: CC.AA. Funding: 316.000 €. Funding agency: Comunidad de Madrid. Duration: 2013-2015 Funding: 483.000 €. Duration: 2012-2016

2015 CIBERNED Annual Report / 23 502 GROUP Félix Bermejo Pareja

Félix Bermejo Pareja José Antonio Molina Arjona Alberto Villarejo Galende Principal Investigator Scientific staff Statistics technician

Eva María Carro Díaz Julián Benito León David Andrés Pérez Co-Principal Investigator Scientific staff Martínez Statistics technician Desireé Antequera Tienda Jesús Hernández Gallego Research technician Scientific staff Alejandro Herrero San Martín Fernando Bartolomé Rocío Trincado Soriano Statistics technician Robledo Statistics technician Postdoctoral researcher Sara Llamas Velasco Statistics technician

Marta González Sánchez Statistics technician

Contact details

Instituto de Investigación Hospital Doce de Octubre Avda. de Córdoba s/n 28041 Madrid (Spain) Ph.: +34 913 908 765 Fax: +34 913 908 544 [email protected]

2015 CIBERNED Annual Report / 24 Summary

The scientific activity has focused on the study • Mechanisms of neurodegeneration, of the physiopathological mechanisms of neuroprotection, designing new therapies neurodegenerative diseases, mainly Alzheimer’s and drug testing in experimental models. disease and associated dementias, and to the identification of new therapeutic targets. 2) Epidemiological research in the field of Specific activities are: neurodegenerative diseases, with special attention to risk factors. 1) Studies on new risk factors, biomarkers of disease and new therapeutic strategies in • Publication of results of cohort NEDICES-I. Alzheimer’s disease and other degenerative dementias. • Implementation of the cohort NEDICES-II.

• Research on biomarkers in biological 3) Clinical research in dementia. samples non-invasive. • Determination of the clinical utility of • Cellular and animals models of Alzheimer’s biomarkers of recent introduction. disease and others degenerative dementias. • Development of new molecular biomarkers and imaging. • Molecular pathology in Alzheimer’s disease.

Keywords

Alzheimer’disease, dementia, biomarkers, transgenic mice, choroid plexus, therapeutic drugs, amyloid, mild cognitive impairment

Publications

2015 CIBERNED Annual Report / 25 • Gartziandia O., Herran E., Pedraz J.L., Carro E., Igartua M., Hernandez R.M. Chitosan coated nanostructured lipid carriers for brain delivery of proteins by intranasal administration. Colloids and Surfaces B: Biointerfaces. 2015;134:304-313. • Benito-Leon J., Domingo-Santos A. Comments on “Role of intestinal microbiota in the development of multiple sclerosis”. Neurologia. 2016. Epub 2015. • Contador I., Bermejo-Pareja F., Del Ser T., Benito-Leon J. Effects of education and word reading on cognitive scores in a community-based sample of Spanish elders with diverse socioeconomic status. Journal of Clinical and Experimental Neuropsychology. 2015;37(1):92-101. • Louis E.D., Benito-Leon J., Faust P.L. Essential tremor is a risk factor for Parkinson’s disease. Parkinsonism and Related Disorders. 2015. [Epub ahead of print]. • Leon-Ruiz M., Benito-Leon J., Sierra-Hidalgo F., Garcia-Soldevilla M.A., Izquierdo-Esteban L., Tejeiro-Martinez J. et al. First case described of isolated, complete and fluctuating cranial nerve III palsy heralding multiple myeloma. Revista de Neurologia. 2015;60(3):115-119. • Esteras N, Alquézar C, Bartolomé F, de la Encarnación A, Bermejo-Pareja F, Molina JA et al. G1/S Cell Cycle Checkpoint Dysfunction in Lymphoblasts from Sporadic Parkinson’s Disease Patients.Molecular neurobiology. 2015;52(1): 386-98 . • Petrov D., Pedros I., Artiach G., Sureda F.X., Barroso E., Pallas M. et al. High-fat diet-induced deregulation of hippocampal insulin signaling and mitochondrial homeostasis deficiences contribute to Alzheimer disease pathology in rodents. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2015;1852(9):1687-1699. • Benito-Leon J., Vega-Quiroga S., Villarejo-Galende A., Bermejo-Pareja F. Hypercholesterolemia in elders is associated with slower cognitive decline: A prospective, population-based study (NEDICES). Journal of the Neurological Sciences. 2015;350(1-2):69-74. • Labiano-Fontcuberta A, Mitchell AJ, Moreno-García S, Puertas-Martín V, Benito-León J. Impact of anger on the health-related quality of life of multiple sclerosis patients.Multiple sclerosis (Houndmills, Basingstoke, England). 2015;21(5): 630-41. • Sánchez-Ferro Á, Rábano A, Catalán MJ, Rodríguez-Valcárcel FC, Fernández Díez S, Herreros- Rodríguez J et al. In vivo gastric detection of α-synuclein inclusions in Parkinson’s disease. Movement disorders : official journal of the Movement Disorder Society. 2015;30(4): 517-24. • Anitua E., Pascual C., Perez-Gonzalez R., Orive G., Carro E. Intranasal PRGF-Endoret enhances neuronal survival and attenuates NF-κB-dependent inflammation process in a mouse model of Parkinson’s disease. Journal of Controlled Release. 2015;203:170-180. • Benito-Leon J., Labiano-Fontcuberta A. Linking Essential Tremor to the Cerebellum: Clinical Evidence. Cerebellum. 2015. [Epub ahead of print]. • Llamas-Velasco S., Contador I., Villarejo-Galende A., Lora-Pablos D., Bermejo-Pareja F. Physical Activity as Protective Factor against Dementia: A Prospective Population-Based Study (NEDICES). Journal of the International Neuropsychological Society. 2015;21(10):861-867. • Labiano-Fontcuberta A., Aladro Y., Martinez-Gines M.L., Ayuso L., Mitchell A.J., Puertas Vero. et al. Psychiatric disturbances in radiologically isolated syndrome. Journal of Psychiatric Research. 2015;68:309-315. • Benito-León J, Louis ED, Villarejo-Galende A, Labiano-Fontcuberta A, Bermejo-Pareja F. Self- rated health and risk of incident essential tremor: A prospective, population-based study (NEDICES).Parkinsonism & related disorders. 2015;21(6): 622-8. • Spuch C., Antequera D., Pascual C., Abilleira S., Blanco M., Moreno-Carretero M.J. et al. Soluble megalin is reduced in cerebrospinal fluid samples of alzheimer’s disease patients. Frontiers in Cellular Neuroscience. 2015;9(APR). [Epub ahead of print]. • Benito-Leon J., De La Aleja J.G., Martinez-Salio A., Louis E.D., Lichtman J.H., Bermejo-Pareja F. et al. Symptomatic atherosclerotic disease and decreased risk of cancer-specific mortality: A prospective, population-based study (NEDICES). Medicine (United States). 2015;94(32). [Epub ahead of print].

2015 CIBERNED Annual Report / 26 Program 1 Group: Félix Bermejo Pareja

• Alquezar C., Barrio E., Esteras N., De La Encarnacion A., Bartolome F., Molina J.A. et al. Targeting cyclin D3/CDK6 activity for treatment of Parkinson’s disease. Journal of Neurochemistry. 2015;133(6):886-897. • Agúndez JA, García-Martín E, Martínez C, Benito-León J, Millán-Pascual J, Díaz-Sánchez M et al. The GSTP1 gene variant rs1695 is not associated with an increased risk of multiple sclerosis. Cellular & molecular immunology. 2015;12(6): 777-9. • Olazaran Rodriguez J., Bermejo Pareja F. There is no scientific basis for retiring the MMSE. Neurologia. 2015;30(9):589-591. • Benito-León J, Louis ED, Puertas-Martín V, Romero JP, Matarazzo M, Molina-Arjona JA et al. Cognitive and neuropsychiatric features of orthostatic tremor: A case-control comparison. Journal of the neurological sciences. 2016;361. Epub 2015. • Krzyzanowska A., Garcia-Consuegra I., Pascual C., Antequera D., Ferrer I., Carro E. Expression of regulatory proteins in choroid plexus changes in early stages of alzheimer disease. Journal of Neuropathology and Experimental Neurology. 2015;74(4):359-369. • Ayuso P., Agundez J.A., Alonso-Navarro H., Martinez C., Benito-Leon J., Ortega-Cubero S. et al. Heme Oxygenase 1 and 2 Common Genetic Variants and Risk for Essential Tremor. Medicine. 2015;94(24):e968-. [Epub ahead of print]. • Bermejo-Pareja F., Contador I., Trincado R., Lora D., Sanchez-Ferro A., Mitchell A.J. et al. Prognostic Significance of Mild Cognitive Impairment Subtypes for Dementia and Mortality: Data from the NEDICES Cohort. Journal of Alzheimer’s Disease. 2016;50(3):719-731. Epub 2015. • Benito-León J, Louis ED, Labiano-Fontcuberta A, Bermejo-Pareja F, De Pedro Cuesta Jesús. Serious head trauma preceding essential tremor: A population-based study (NEDICES).Journal of the neurological sciences. ;353(1-2): 116-21.

Research projects

• Code: 2014-0059. • Code: PI12/01602. Title: Analysis of changes in the pattern Title: Cohorte biomédica, cerrada y de origen typed in a sample of patients with Parkinson comunitario integrada con participantes vs. controls. del centro de España. Estudio observacional Principal Investigator: Félix Bermejo Pareja. del temblor esencial y la enfermedad de CIBERNED’s collaboration: No. Parkinson (NEDICES-2). CIBERNED groups: Principal Investigator: Julián Benito León. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: International. CIBERNED groups: Funding agency: Madrid-MIT M+Visión Other CIBER’s collaboration: No. Consortium. Type: National. Funding: No disponible (ND). Funding agency: Instituto de Salud Carlos III. Duration: 2014-2015 Funding: ND. Duration: 2013-2015 • Code: Madrid-MIT M+Visión Fellow. Title: Motor Response to Dopaminergic • Code: RTC-2015-3967-1. Therapy in a Population of De Novo Title: Plataforma para el seguimiento de Parkinson’s Disease Cases Quantified via trastornos del movimiento (NETMD). Typing Analyses-NeuroQWERTY. Principal Investigator: Julián Benito León. Principal Investigator: Félix Bermejo-pareja. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: International. Funding agency: Ministerio de Economía y Funding agency: Madrid-MIT M+Vision Competitividad. Consortium. Funding: ND. Funding: ND. Duration: 2015-2017 Duration: 2014-2015

2015 CIBERNED Annual Report / 27 • Code: MUTUA 2012. Funding: 30.000 €. Title: Estudio de las propiedades Duration: 2012-2015 terapéuticas de fármacos basados en melatonina como tratamiento innovador • Code: FIS PO 11-1508. para enfermedades neurodegenerativas. Title: Estudio basal de la cohorte NEDICES-2. Principal Investigator: Mª Isabel Rodríguez- Cohorte comunitaria en el centro de España Franco. con información clínica y biobanco. CIBERNED’s collaboration: No. Principal Investigator: Félix Bermejo Pareja. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: International. Other CIBER’s collaboration: No. Funding agency: Fundación Investigación Type: National. Biomédica de la MUTUA. Funding agency: FIS. Funding: 20.000 €. Funding: 104.665 €. Duration: 2012-2015 Duration: 2012-2015 • Code: Fundación Ramón Areces 2012. • Code: Cod. Title: Estudio de la capacidad Title: NeuroTREMOR. neuroprotectora de las células epiteliales Principal Investigator: Julián Benito León. de los plexos coroideos como potencial CIBERNED’s collaboration: No. terapia regenerativa de la enfermedad de CIBERNED groups: Alzheimer. Other CIBER’s collaboration: No. Principal Investigator: Eva Carro. Type: International. CIBERNED’s collaboration: Yes. Funding agency: Unión Europea. CIBERNED groups: G502, G509. Funding: 300.000 €. Other CIBER’s collaboration: No. Duration: 2012-2015 yipe: National. Funding agency: PhD dissertations

• Author: Verónica Puertas Martín. • Author: Andrés Labiano Fontcuberta. Title: Estudio del rendimiento cognitivo Title: Cognición y emoción: su impacto en la en pacientes con temblor esencial. esclerosis múltiple. Comparación con controles y enfermedad Date: 4th May 2015. de Parkinson. Supervisor: Jesús Hernández Gallego. Date: 16th Decembre 2015. Supervisor: Alberto Villarejo Galende.

2015 CIBERNED Annual Report / 28 510 GROUP María Jesús Bullido Gómez

María Jesús Bullido María Recuero Vicente Patricia Llorente Ginés Principal Investigator Researcher PhD student

Ana Frank García Isabel Sastre Merlín Julia Terreros Roncal Associate Professor Technician Student

Jesús Aldudo Soto Henrike Kristen Researcher PhD student

Contact details

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]

2015 CIBERNED Annual Report / 29 Summary

To identify genes and mechanisms involved in lysosomal alterations observed in the model. the neurodegeneration proccess characteristic Furthermore, we found that another member of Alzheimer’s disease (AD), we have developed of the alpha herpesvirus family with neurotropic cellular models of oxidative stress (OS) and of properties, HSV-2, is also able to induce the infection by herpes simplex type 1virus (HSV-1). main neurodegeneration markers previously These models present markers characteristic of observed for HSV-1, supporting the potential of AD, as those observed in the traffic, metabolism diverse infectious agents as participants in the and proteolysis of the amyloid precursor protein neurodegeneration associated to Alzheimer’s (APP) favoring the amyloidogenic route or in disease. the phosphorylation of tau protein. As for the studies in patients we continue to The study of the autophagy lysosome pathway, participate in genetic association studies in which we previously described alterations in under the Dementia Genetics Spanish autophagic flux, has shown us during this period Consortium (DEGESCO), as well as with the that infection with HSV-1 and EO profoundly affect EADI and IGAP consortia, still revealing novel the final steps of the route, namely the lysosomal factors and functions potentially relevant in function, causing an increased cellular burden the pathogenesis of AD. In 2015, two works of lysosomes accompanied by a significant by DEGESCO and IGAP both highlighted the reduction in the activity of different cathepsins presence of genetic association markers in or and decreased lysosomal degradation of near the gene encoding tau protein (MAPT), substrates (such as the EGF receptor). The results and that the susceptibility conferred by these of the functional, gene expression and genetic markers seemed to be modulated by APOE association studies reinforce the hypothesis genotype. Among the collaborative projects in that a failure of the lysosomal function could this year, it has been completed an exploration be one of the mechanisms responsible for in whole exome chips coordinated by Dr. J neurodegeneration induced by the virus and Williams (manuscript in preparation) as well as a EO, and point to the lysosomal membrane study of epistasis coordinated by Dr. D Lehman protein LAMP2 as a relevant participant in the whose results are being currently analyzed.

Keywords

Alzheimer, herpesvirus, oxidative stress, neurodegeneration, lysosomal function, functional genomics, genetic association

Publications

• Lovestone S, Boada M, Dubois B, Hüll M, Rinne JO, Huppertz HJ et al. A phase II trial of tideglusib in Alzheimer’s disease.Journal of Alzheimer’s disease : JAD. 2015;45(1):-. • International Genomics of Alzheimer’s Disease Consortium (IGAP), Lleó Bisa Alberto, Combarros Pascual Onofre, Bullido M. Convergent genetic and expression data implicate immunity in Alzheimer’s disease.Alzheimer’s & dementia : the journal of the Alzheimer’s Association. 2015;11(6):-. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1). Epub 2015. • Kristen H, Santana S, Sastre I, Recuero M, Bullido MJ, Aldudo J. Herpes simplex virus type 2 infection induces AD-like neurodegeneration markers in human neuroblastoma cells.Neurobiology of aging. 2015;36(10):-. • Pastor P, Moreno F, Clarimón J, Ruiz A, Combarros O, Calero M et al. MAPT H1 Haplotype is Associated with Late-Onset Alzheimer’s Disease Risk in APOEε4 Noncarriers: Results from the Dementia Genetics Spanish Consortium.Journal of Alzheimer’s disease : JAD. 2015;49(2):-. • Olazaran J., Gil-De-Gomez L., Rodriguez-Martin A., Valenti-Soler M., Frades-Payo B., Marin- Munoz J. et al. A blood-based, 7-metabolite signature for the early diagnosis of Alzheimer’s disease. Journal of Alzheimer’s Disease. 2015;45(4):1157-1173.

2015 CIBERNED Annual Report / 30 Program 1 Group: María Jesús Bullido Gómez

• Jun G, Ibrahim-Verbaas CA, Vronskaya M, Lambert JC, Chung J, Naj AC et al. A novel Alzheimer disease locus located near the gene encoding tau protein.Molecular psychiatry. 2016;21(1) Epub 2015. • Gualix J., Leon-Otegui M., Recuero M., Bullido M.J., Valdivieso F., Miras-Portugal M.T. Functional characterization of P2Y1 and P2X4 receptors in human neuroblastoma SK-N-MC cells. Anales de la Real Academia Nacional de Farmacia. 2015;81(3):247-257.

2015 CIBERNED Annual Report / 31 402 GROUP Antonio Camins Espuny

Antoni Camins Espuny Jaume Folch Miren Ettcheto Full Professor Associate Professor Research Fellow

Merce Pallás Lliberia Carme Auladell Sonia Abad Full Professor Associate Professor Research Fellow

Jordi Vilaplana Anna María Canudas Christian Grañe Farre Associate Professor Teixidó Research Fellow Assistant Professor Francesc Sureda Gemma Manich Associate Professor Andrés Jiménez Research Fellow Assistant Professor Carme Pelegrí Itsaso Cabezón Rodríguez Associate Professor Ester Verdaguer Cardona Research Fellow Lecturer Victor Rimbau Research Fellow

Contact details

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

2015 CIBERNED Annual Report / 32 Summary

The aims of our research is the study in experimental of sirtuin 1. The sirtuins are a family of highly models of Alzheimer’s Disease (APPSwe / conserved proteins in both eukaryotic and PS1dE9) and ageing (SMP8) the molecular prokaryotic organisms that have a very important pathways involved in neurodegeneration and role in the aging process and in the regulation again. We study potential pathways such as of fundamental physiological processes. The mitogen activated protein kinases (JNK1, JNK2 interest of our group is focused on studying , JNK3), cdk5, etc. involved in cell death and the role of sirtuin 1 (SIRT1) and other sirtuins as tau phosphorylation. In addition our interest is SIRT3 in the process of aging and Alzheimer’s the development of therapeutic strategies for disease in APPswe / PS1dE9 and accelerated Alzheimer’s disease treatment. aging mice (SAMP8). In addition we would like to evaluate the role of SIRT1 in cultured neurons Thus, in recent years we have demonstrated to demonstrate the involvement of this enzyme in the APPSwe / PS1dE9 mice model of in the process of neuroprotection preventing Alzheimer’s a relationship between alterations the neuronal process of apoptosis through in the metabolism specifically, type 2 diabetes regulating p53, ATM and other proteins involved mellitus and an advancement and contributes in the process of neuronal death. to development and progression of the Alzheimer’s disease. Thus treatment of APPSwe Another interesting point is the study of the / PS1dE9 mice with a high fat diet favours role of exercise in the prevention of aging and disease worsening and memory loss. Currently, neuronal death in mice SAMP8 and SAMR1. We we have demonstrated changes in transcripts are studding the possible biochemical pathways and proteins involved in the insulin pathway involved in the beneficial effect of exercise in in the hippocampus of the APPSwe / PS1dE9 rodents. Thus, a study has been performed in mice a familial model of Alzheimer’s disease. these mice evaluating the expression of genes Moreover, we have detected disturbances in modulated by the exercise that could have a intracellular signalling pathways downstream potential beneficial effect, as anti-aging and of prolactin and leptin receptors, JAK/STAT also exerts a possibly neuroprotective role. signalling, as well as abnormalities in cholesterol metabolism in the hippocampi of APPSwe / Characterization of a neo-epitope present PS1dE9 mice between the 3 and 6 months of in brain granule structures related to aging in age. Alzheimer’s disease is also associated mice with accelerated senescence SAMP8. with the decrease in insulin levels in the central These structures appear more amount and nervous system and alterations of insulin frequency in the hippocampus of aged receptors and downstream pathway in the animals and especially in animals strain SAMP8. brain. Defects in insulin signaling / IGF-1 affect These granules contain remains of membranes PI3K / Akt causing neurodegeneration process. and cell organelles and signs of degeneration Besides the alteration of glucose transporters at as autophagosomes and large vacuoles. the brain or decrease its expression in the brain The study of the neo-epitope of the granules of Alzheimer’s patients may cause a lower can provide information about the formation glucose metabolism in the brain and cause of these structures during the aging and lower reduced mitochondrial metabolism and neurodegenerative processes. ATP production. Furthermore, we are also working on developing Resveratrol is a natural product that is of great new synthetic molecules for the potential interest as anti-aging and neuroprotective treatment of Alzheimer’s disease. therapy based on is properties as an activator

Keywords

Alzheimer APPswe / PS1dE9, SAMP8, Resveratrol, aging, diabetes

2015 CIBERNED Annual Report / 33 Publications

• Abad S, Camarasa J, Pubill D, Camins A, Escubedo E. Adaptive Plasticity in the Hippocampus of Young Mice Intermittently Exposed to MDMA Could Be the Origin of Memory Deficits.Molecular neurobiology. 2015. [Epub ahead of print]. • Pedros I., Petrov D., Artiach G., Abad S., Ramon-Duaso C., Sureda F. et al. Adipokine pathways are altered in hippocampus of an experimental mouse model of Alzheimer’s disease. Journal of Nutrition, Health and Aging. 2015;19(4):403-412. • Porquet D., Andres-Benito P., Grinan-Ferre C., Camins A., Ferrer I., Canudas A.M. et al. Amyloid and tau pathology of familial Alzheimer’s disease APP/PS1 mouse model in a senescence phenotype background (SAMP8). Age. 2015;37(1). [Epub ahead of print]. • Bayod S., Felice P., Andres P., Rosa P., Camins A., Pallas M. et al. Downregulation of canonical Wnt signaling in hippocampus of SAMP8 mice. Neurobiology of Aging. 2015;36(2):720-729. • Petrov D., Luque M., Pedros I., Ettcheto M., Abad S., Pallas M. et al. Evaluation of the Role of JNK1 in the Hippocampus in an Experimental Model of Familial Alzheimer’s Disease. Molecular Neurobiology. 2015. [Epub ahead of print]. • Ettcheto M, Petrov D, Pedrós I, de Lemos L, Pallàs M, Alegret M et al. Hypercholesterolemia and neurodegeneration. Comparison of hippocampal phenotypes in LDLr knockout and APPswe/ PS1dE9 mice.Experimental gerontology. 2015;65: 69-78. • Garcia-Matas S., Paul R.K., Molina-Martinez P., Palacios H., Gutierrez V.M., Corpas R. et al. In vitro caloric restriction induces protective genes and functional rejuvenation in senescent SAMP8 astrocytes. Aging Cell. 2015;14(3):334-344. • Folch J., Petrov D., Ettcheto M., Pedros I., Abad S., Beas-Zarate C. et al. Masitinib for the treatment of mild to moderate Alzheimer’s disease. Expert Review of Neurotherapeutics. 2015;15(6):587- 596. • Ettcheto M, Junyent F, de Lemos L, Pallas M, Folch J, Beas-Zarate C et al. Mice Lacking Functional Fas Death Receptors Are Protected from Kainic Acid-Induced Apoptosis in the Hippocampus. Molecular neurobiology. 2015;52(1): 120-9. • Manich G., Auge E., Cabezon I., Pallas M., Vilaplana J., Pelegri C. Neo-epitopes emerging in the degenerative hippocampal granules of aged mice can be recognized by natural IgM auto-antibodies. Immunity and Ageing. 2015;12(1). [Epub ahead of print]. • Castro-Torres R.D., Chaparro-Huerta V., Flores-Soto M.E., Jave-Suarez L., Camins A., Armendariz- Borunda J. et al. Pirfenidone Attenuates Microglial Reactivity and Reduces Inducible Nitric Oxide Synthase mRNA Expression After Kainic Acid-Mediated Excitotoxicity in Pubescent Rat Hippocampus. Journal of Molecular Neuroscience. 2015;56(2):245-254. • Gonzalez-Castillo C., Ortuno-Sahagun D., Guzman-Brambila C., Pallas M., Rojas-Mayorquin A.E. Pleiotrophin as a central nervous system neuromodulator, evidences from the hippocampus. Frontiers in Cellular Neuroscience. 2015;8(JAN):1-7. • Soriano-Cantón R, Perez-Villalba A, Morante-Redolat JM, Marqués-Torrejón MÁ, Pallás M, Pérez- Sánchez F et al. Regulation of the p19(Arf)/p53 pathway by histone acetylation underlies neural stem cell behavior in senescence-prone SAMP8 mice.Aging cell. 2015;14(3): 453-62. • Folch J., Ettcheto M., Petrov D., Abad S., Pedros I., Marin M. et al. Review of the advances in treatment for Alzheimer disease: Strategies for combating β-amyloid protein. Neurologia. 2015. [Epub ahead of print]. • Folch J., Patraca I., Martinez N., Pedros I., Petrov D., Ettcheto M. et al. The role of leptin in the sporadic form of Alzheimer’s disease. Interactions with the adipokines amylin, ghrelin and the pituitary hormone prolactin. Life Sciences. 2015;140:19-28. • Cabezon I., Manich G., Martin-Venegas R., Camins A., Pelegri C., Vilaplana J. Trafficking of Gold Nanoparticles Coated with the 8D3 Anti-Transferrin Receptor Antibody at the Mouse Blood-Brain Barrier. Molecular Pharmaceutics. 2015;12(11):4137-4145. • Verdaguer E, Brox S, Petrov D, Olloquequi J, Romero R, de Lemos ML et al. Vulnerability of calbindin, calretinin and parvalbumin in a transgenic/knock-in APPswe/PS1dE9 mouse model of Alzheimer disease together with disruption of hippocampal neurogenesis.Experimental gerontology. 2015;69: 176-88.

2015 CIBERNED Annual Report / 34 Program 1 Group: Antonio Camins Espuny

• Petrov D., Pedros I., Artiach G., Sureda F.X., Barroso E., Pallas M. et al. High-fat diet-induced deregulation of hippocampal insulin signaling and mitochondrial homeostasis deficiences contribute to Alzheimer disease pathology in rodents. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2015;1852(9):1687-1699.

Research projects

• Code: BFU2013-47382-P. Title: Caracterización de un neo-epitopo cerebral relacionado con el envejecimiento y estudio de la presencia de anticuerpos naturales anti-neoepitopo. Principal Investigator: Carme Pelegri Gavalda. CIBERNED’s collaboration: No. CIBERNED groups: Other CIBER’s collaboration: No. Type: National. Funding agency: Ministerio de Economía y Competitividad. Funding: 85.000 €. Duration: 2014-2016

PhD dissertations

• Author: Dnitry Petrov. • Author: José Iván Patraca Fierro. Title: The role of high-fat diet in an APP/PS1 Title: Papel de factores endocrinos como model of familial Alzheimer disease. melatonina y prolactina, y de adipoquinas Date: 13th Novembre 2015. como leptina en la modulación de Supervisor: Antonio Camins Espuny. la inflamación asociada a procesos neurodegenerativos. • Author: Lucía Soliño Alonso. Date: 19th Novembre 2015. Title: Detección de toxinas marinas y Supervisor: Jaume Folch. caracterización de su mecanismo de acción mediante ensayos celulares. Aplicaciones a la identificación de riesgos alimentarios. Date: 3rd July 2015. Supervison: Francesc X. Sureda Batlle.

2015 CIBERNED Annual Report / 35 511 GROUP José Luis Cantero Lorente

José Luis Cantero Lorente Gabriel González Escamilla David García Solís Principal Investigator Postdoctoral fellow Researcher

Mercedes Atienza Ruiz Mayely Sánchez Espinosa Eulogio Gil Néciga Researcher PhD student Researcher

Maité Crespo García Sofía Rodríguez Peñuela Postdoctoral fellow Technician

Contact details

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

2015 CIBERNED Annual Report / 36 Summary

The aims of the work carried out in 2015 atrophies of the prefrontal cortex, while high included evaluating alterations of cortical levels of Aβ42 are associated with a significant functional connectivity in normal aging and thinning of temporal regions, poorer everyday the prodromal stage of Alzheimer’s disease memory, and increased levels of homocysteine. (AD). Results revealed that alterations of These results suggest that circulating levels of cortical functional connectivity not only are Aβ could indicate different trajectories of aging related to plasma amyloid beta (Aβ) levels, in asymptomatic older adults (Lladó-Saz et al., but they also allowed discriminating patients 2015). with mild cognitive impairment (MCI) carriers and noncarriers of the ApoE ε4 genotype Previous studies conducted in our laboratory (González-Escamilla et al., 2015). Our results have identified alterations of sleep structure in further revealed that episodic memory is MCI patients. Although these sleep alterations remarkably impaired in MCI patients with ApoE might affect memory consolidation processes, ε4. This deficit was mainly due to a reduced it is possible that encoding is also negatively capacity to benefit from semantic congruency, affected. To assess this hypothesis, we have which in turn was associated with difficulties to investigated whether acute sleep restriction activate lateral regions of the temporal lobe applied on the pretraining night might have and additional regions to compensate such a an impact on retrieval processes in young deficit (Prieto del Val et al., 2015). Finally, our people. Even though sleep restriction did studies evidenced that semantic knowledge not affect memory performance, patterns not only enhances episodic memory but also of brain activation during memory retrieval improves generalization of learning in semantic were drastically different in the group that dementia (Suárez-González et al., 2015). was partially deprived of sleep (Alberca-Reina et al., 2015). On the basis of these results, the On the other hand, we have investigated the incapacity of MCI ApoE ε4 carriers to recruit relationship between plasma Aβ levels and neural compensatory mechanisms during patterns of cortical thickness in older adults encoding may be linked to sleep alterations cognitively intact. This study revealed that high previously identified. This hypothesis should be concentrations of Aβ40 are related to bilateral assessed in future studies.

Keywords

Cerebral aging, biomarkers of preclinical and prodromal Alzheimer’s disease, anatomical and functional connectivity of human neocortex

Publications

• Prieto del Val L., Cantero J.L., Atienza M. APOE e{open}4 constrains engagement of encoding- related compensatory networks in amnestic mild cognitive impairment. Hippocampus. 2015;25(9):993-1007. • Romero-Garcia R., Atienza M., Cantero J.L. Different Scales of Cortical Organization are Selectively Targeted in the Progression to Alzheimer’s Disease. International Journal of Neural Systems. 2016; 26(2). Epub 2015. • Alberca-Reina E., Cantero J.L., Atienza M. Impact of sleep loss before learning on cortical dynamics during memory retrieval. NeuroImage. 2015;123:51-62. • Gonzalez-Escamilla G., Atienza M., Cantero J.L. Impaired cortical oscillatory coupling in mild cognitive impairment: anatomical substrate and ApoE4 effects. Brain structure & function. 2015;220(3):1721-1737. • Llado-Saz S., Atienza M., Cantero J.L. Increased levels of plasma amyloid-beta are related to cortical thinning and cognitive decline in cognitively normal elderly subjects. Neurobiology of Aging. 2015;36(10):2791-2797. • Suárez-González A, Crutch SJ, Franco-Macías E, Gil-Néciga E. Neuropsychiatric Symptoms in Posterior Cortical Atrophy and Alzheimer Disease.Journal of geriatric psychiatry and neurology. 2016;29(2). Epub 2015.

2015 CIBERNED Annual Report / 37 • Suárez-González A, Green Heredia C, Savage SA, Gil-Néciga E, García-Casares N, Franco- Macías E et al. Restoration of conceptual knowledge in a case of semantic dementia. Neurocase. 2015;21(3): 309-21. • Huertas-Fernandez I., Garcia-Gomez F.J., Garcia-Solis D., Benitez-Rivero S., Marin-Oyaga V.A., Jesus S. et al. Machine learning models for the differential diagnosis of vascular parkinsonism and Parkinson’s disease using [123I]FP-CIT SPECT. European Journal of Nuclear Medicine and Molecular Imaging. 2015;42(1):112-119. • Sueño y procesos de memoria. Atienza M, Cantero JL. En: Tratado de Medicina del Sueño. (pp. 198-205), 2015.A. Bove, F. Cañellas, I. de Andrés, J. Durán-Cantolla, D. García-Borreguero y T. Sagalés (eds.). ISBN: 978-84-9835-203-0 • Anatomía funcional de la memoria. Atienza M, Cantero JL. En: Conectividad funcional y anatómica en el cerebro humano. (161-169), 2015.F. Maestú, E. Pereda, y F. Del Pozo (eds.). ISBN: 978-84-9022-525-7.

Research projects

• Code: PSI2014-55747-R. CIBERNED groups: Title: Estudio de las oscilaciones cerebrales Other CIBER’s collaboration: No. EEG relacionadas con la memoria como Type: International. potencial biomarcador de la enfermedad Funding agency: Consejeria de Economía, de Alzheimer. Innovación y Ciencia, Junta de Andalucía. Principal Investigator: Mercedes Atienza Funding: 225.419 €. Ruiz. Duration: 2014-2017 CIBERNED’s collaboration: No. CIBERNED groups: Code: UNPO13-1E-2299. Other CIBER’s collaboration: No. • Title: Servicio Centralizado de Investigación Type: National. en Neuroimagen. Escaner de Resonancia Funding agency: Ministerio de Economía y Magnética de 3 Teslas para humanos. Competitividad. Principal Investigator: José Luis Cantero Funding: 117.975 €. Lorente. Duration: 2015-2017 CIBERNED’s collaboration: No. CIBERNED groups: • Code: P12-CTS 2327. Other CIBER’s collaboration: No. Title: Caracterización de las fases Type: National. presintomáticas y preclínicas de la Funding agency: Ministerio de Economía y enfermedad de Alzheimer mediante Competitividad. marcadores biológicos y de neuroimagen. Funding: 1.551.000 €. Principal Investigator: José Luis Cantero Duration: 2015 Lorente. CIBERNED’s collaboration: No.

PhD dissertations

• Author: Gabriel González Escamilla. • Author: Esther Alberca Reina. Title: Conectividad funcional cerebral Title: Efectos de la privación de sueño sobre en personas mayores con y sin deterioro las oscilaciones cerebrales asociadas a la cognitivo leve: Correlatos de neuroimagen codificación y consolidación de la memoria y marcadores en sangre. declarativa. Date: 13th February 2015. Date: 29th May 2015. Supervisor: José Luis Cantero Lorente. Supervisor: José Luis Cantero Lorente.

2015 CIBERNED Annual Report / 38 413 GROUP Joan Xavier Comella Carnicé

Joan Xavier Comella Bruna Barneda Zahonero Koen Galenkamp Carnicé PhD PhD Principal Investigator Jorge Urresti Ibáñez María Sánchez Osuna Víctor Yuste Mateos PhD PhD PhD Laura Planells Ferrer Isabel Calleja Yagüe Joaquín López Soriano PhD PhD student PhD Elena Coccia PhD student

Contact details

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]

Informe Anual CIBERNED 2015 / 39 Summary

Our main interest is to characterize the cooperative effects of TNF and FasL on cell mechanisms controlling neuronal death induced death. Both ligands induce a synergic effect on by a group of receptors collectively known as the cell death induction in some of these cell death receptors, and also the relevance of lines, and pre-incubation with TNF is necessary some of their intracellular antagonists. Among to observe this effect, due to the upregulation of those we are most interested in the antagonists Fas receptor (mRNA and membrane exposure expressed in nervous system, such as FAIM-L and of the receptor). This transcriptional effect of TNF Lifeguard. Our main objective is to characterize is mediated by NFkB pathway. In addition, TNF these proteins at the molecular level, their increases cisplatine- and etoposide-induced involvement in neuronal differentiation and death and caspase-8 activation (these two physiology, and their possible role in different drugs are widely used to treat neuroblastoma), pathologies, mainly neurodegenerative and this effect is explained by the induction of diseases. Fas. However, there is heterogeneity among the neuroblastoma cell lines studied, some We are also characterizing molecular partners of them being sensitive and others resistant of these molecules that main explain their to TNF action, which seems to be correlated mechanisms of action. The knowledge of the with endogenous levels of caspase-8 and Fas molecular basis of these receptor antagonists (Galenkamp et al., 2015). and the activation of survival signaling pathways could be of high relevance to understand the About Lifeguard, we characterized its etiology or to open new therapeutic strategies subcellular localization, showing that is most for the treatment of neurodegenerative present in Golgi and reticulum. In addition, diseases. we characterized that its protection on FasL- induced cell death is dependent on calcium We have recently characterized, in collaboration release inhibition at the reticulum, thus showing with different CIBERNED groups, the role of a previously undescribed molecular mechanism FAIM-L in the development of Alzheimer’s of action for this death receptor antagonist disease, both in animal models (APP/PS1) and (Urresti et al., 2015). patients. We have characterized that FAIM-L levels are downregulated in neurons with Our main research lines at present are: the progression of the disease, particularly in hippocampus and entorhinal cortex, and that beta-amyloid reduces FAIM-L levels in neurons 1) To study FAIM-L function in in vitro and in vivo in vitro. Additionally, TNF is able to protect from models of AD, and its relation with TNF (its the beta-amyloid-induced cell death in vitro, duality as a pro-apoptotic molecule and at only if neurons have enough levels of FAIM-L. the same time as a survival promoter) and its Thus, FAIM-L levels can determine the cell fate signaling pathways, particularly NFkB. in a neuroinflammatory process (Carriba et al., 2015). To characterize FAIM-L functional partners, 2) particularly Siva-1 and XIAP, and their We have also studied, in collaboration with implications in neuron physiology (neuronal Fundació ACE (Barcelona), polymorphisms plasticity) and different neurodegenerative of genes related with death receptors, their diseases (neuronal apoptosis, synaptic ligands and antagonists, in Alzheimer’s patients, degeneration). unveiling a likely correlation with the TNF gen (Moreno-Grau et al, 2015), pending further To study Faim promoter and the functional characterization. 3) differences of FAIM isoforms, including the characterization of two new isoforms, one of In neuroblastoma cell lines we studied them neuron-specific.

Keywords

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

2015 CIBERNED Annual Report / 40 Program 1 Group: Joan Xavier Comella Carnicé

Publications

• Garcia-Belinchon M., Sanchez-Osuna M., Martinez-Escardo L., Granados-Colomina C., Pascual- Guiral S., Iglesias-Guimarais V. et al. An early and robust activation of caspases heads cells for a regulated form of necrotic-like cell death. Journal of Biological Chemistry. 2015;290(34):20841- 20855. • Moreno-Grau S., Barneda B., Carriba P., Marin J., Sotolongo-Grau O., Hernandez I. et al. Evaluation of Candidate Genes Related to Neuronal Apoptosis in Late-Onset Alzheimer’s Disease. Journal of Alzheimer’s Disease. 2015;45(2):621-629. • Urresti J., Ruiz-Meana M., Coccia E., Arevalo J.C., Castellano J., Fernandez-Sanz C. et al. Lifeguard inhibits fas ligand-mediated endoplasmic reticulum-calcium release mandatory for apoptosis in type II apoptotic cells. Journal of Biological Chemistry. 2016;291(3):1221-1234. Epub 2015. • Carriba P., Comella J.X. Neurodegeneration and neuroinflammation: Two processes, one target. Neural Regeneration Research. 2015;10(10):1581-1583. • Galenkamp K.M.O., Carriba P., Urresti J., Planells-Ferrer L., Coccia E., Lopez-Soriano J. et al. TNFα sensitizes neuroblastoma cells to FasL-, cisplatin- and etoposide-induced cell death by NF-κB-mediated expression of Fas. Molecular Cancer. 2015;14(1). [Epub ahead of print]. • Carriba P., Jimenez S., Navarro V., Moreno-Gonzalez I., Barneda-Zahonero B., Moubarak R.S. et al. Amyloid-β reduces the expression of neuronal FAIM-L, thereby shifting the inflammatory response mediated by TNFα from neuronal protection to death. Cell Death and Disease. 2015;6(2). [Epub ahead of print]. • Bosch C, Martínez A, Masachs N, Teixeira CM, Fernaud I, Ulloa F et al. FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult- generated neurons.Frontiers in neuroanatomy. 2015;9: 60.

Research projects

• Code: 2014 SGR 1609. • Code: PI2013/01. Title: Apoptosi i Neurodegeneracio (GRC). Title: Propiedades emergentes de la Principal Investigator: Joan Xavier Comella relación neurona-glia que subyacen a Carnicé. neurodegeneración y demencia en la CIBERNED’s collaboration: Yes. enfermedad de Alzheimer. CIBERNED groups: G109, G413. Principal Investigator: Ignacio Torres- Other CIBER’s collaboration: No. Alemán. Type: International. CIBERNED’s collaboration: Yes. Funding agency: AGAUR. CIBERNED groups: G415, G413, G204, G409, Funding: 30.000 €. G108, G411. Duration: 2014-2016 Other CIBER’s collaboration: No. Type: National. • Code: SAF2013-47989-R. Funding agency: CIBERNED. Title: Neuroinflamación, TNF y antagonistas Funding: 350.000 €. de los receptores de muerte (FAIM-L): Duration: 2013-2015 relevancia en neurodegeneración. Principal Investigator: Joan Xavier Comella • Code: 20141431. Carnicé. Title: Deciphering the link between astrocyte CIBERNED’s collaboration: No. reactivity and neuronal damage in CIBERNED groups: Alzheimer´s disease”. Other CIBER’s collaboration: No. Principal Investigator: Elena Galea. Type: National. CIBERNED’s collaboration: Yes. Funding agency: MINECO. CIBERNED groups: G411, G413, G415. Funding: 240.000 €. Other CIBER’s collaboration: No. Duration: 2014-2016 Type: Private. Funding agency: La Maraó de TV3. Funding: 382.250 €. Duration: 2015-2017

2015 CIBERNED Annual Report / 41 • Code: PIE13/00027. Title: Preventing cardiovascular ischemic events and arresting their consequences in type 2 diabetic population: a multidisciplinary clinical and experimental approach. Principal Investigator: David García- Dorado. CIBERNED’s collaboration: No. CIBERNED groups: Other CIBER’s collaboration: No. Type: National. Funding agency: MINECI/ISCIII. Funding: 605.000 €. Duration: 2015

PhD dissertations

• Author: Koen Galenkamp. Title: Priming neuroblastoma for cisplatin and etoposide drug therapy. Date: 4th Decembre 2015. Supervisor: Joan Xavier Comella Carnicé.

2015 CIBERNED Annual Report / 42 403 GROUP Javier de Felipe Oroquieta

Javier de Felipe Ana Isabel García Ramírez Guillermo Aparicio Torres Research Professor-Principal Technician PhD student Investigator Débora Cano Laiseca Marta Turégano Lidia Alonso-Nanclares Technician PhD student Postdoctoral researcher Mari Carmen Álvarez Pérez Concepción Rojo Ruth Benavides-Piccione Technician Visiting scientist Postdoctoral researcher Silvia Tapia González Miguel Miguens Vázquez Rodrigo Rodríguez Sánchez Postdoctoral researcher Visiting scientist CSIC researcher Gonzalo León Espinosa Juncal González Isabel Fernaud Espinosa Postdoctoral researcher Visiting scientist Postdoctoral researcher Mirian Marín Horcajada Pilar Flores Romero Asta Kastanauskaite Lab Technician Research and Project manager Postdoctoral researcher Alejandro Antón Fernández Montserrat Fernández Ángel Merchán-Pérez PhD student Bouzo Postdoctoral researcher Research and Project manager Liulia Diana Furcila Alberto Muñoz PhD student Yago Rodríguez Cela Postdoctoral researcher Office manager Andrea Santuy Muñoz Lorena Valdés Lora PhD student Technician

Contact details

Laboratorio Cajal de Circuitos Corticales, CTB, UPM Campus de Montegancedo Pozuelo de Alcorcón 28223 Madrid (Spain) Tel.: +34 91 336 46 60 [email protected]

2015 CIBERNED Annual Report / 43 Summary

Microorganization of the normal cerebral • Generation of high resolution confocal cortex and alterations of cortical circuits in microscopy stacks of images of pyramidal brain pathologies. cells to analyze the distribution and morphology of dendritic spines in different The CCCL principally focuses on the cortical areas, layers and species. microorganization of the normal cerebral cortex (including hippocampus) in various species 1.3. Intrinsic and extrinsic connectivity of the (particularly humans) and on the alterations cortical column: of cortical circuits in epilepsy and Alzheimer disease. These studies are performed through • Target identification of cortical synapses the use of anatomical tracers, high resolution (FIB/SEM) to determine the proportion of immunocytochemistry and 3D light and electron synapses on dendritic spines and dendritic microscopy. Another major aim is to develop shafts. informatics technologies to examine the brain. In particular, the research lines carried out at • Analysis of synaptic sizes (FIB/SEM) and the CCCL are as follows: development of computer models to explore the possible relationship between synaptic 1.1. Study of the components of the column: morphology and physiology.

• Development and validation of software • Characterization of the afferent and efferent tools for 3D segmentation of cells of the connections of the hindlimb representation complete cortical column in stacks obtained area of the primary somatosensory cortex in by confocal microscopy in samples from the P14 rats in tract-tracing experiments. somatosensory neocortex. 1.4 Quantification of cellular and subcellular • Spatial analysis distribution of the segmented alterations in Alzheimer’s Disease (AD) and cell to determine the distribution patterns of the possible influence of these alterations on different cell types. cognition.

• Analysis of the spatial distribution of synapses • Quantification of specific synaptic in the six cortical layers by means of spatial alterations in the neuropil and in identified 3D statistical tools. Calculation of the volume reconstructed cortical neurons from areas fraction occupied by mitochondria in the six showing early histopathological changes in cortical layers. AD.

1.2. Pyramidal cells: • Application of systematic methods to human tissue from AD patients. • Intracellular injections and 3D-reconstruction of pyramidal cells and to analyse the • Correlation of quantitative results with microanatomy of these cells in different cognitive impairment. cortical areas, layers and species.

Keywords

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

Publications

• Luengo-Sanchez S., Bielza C., Benavides-Piccione R., Fernaud-Espinosa I., Defelipe J., Larranaga P. A univocal definition of the neuronal soma morphology using gaussian mixture models. Frontiers in Neuroanatomy. 2015;9(November):1-11. • Jimenez-Mateos EM, Engel T, Merino-Serrais P, Fernaud-Espinosa I, Rodriguez-Alvarez N, Reynolds J et al. Antagomirs targeting microRNA-134 increase hippocampal pyramidal neuron spine volume in vivo and protect against pilocarpine-induced status epilepticus.Brain structure & function. 2015;220(4): 2387-99.

2015 CIBERNED Annual Report / 44 Program 1 Group: Javier de Felipe Oroquieta

• Mihaljević B, Benavides-Piccione R, Bielza C, DeFelipe J, Larrañaga P. Bayesian network classifiers for categorizing cortical GABAergic interneurons.Neuroinformatics. 2015;13(2): 193-208. • Anton-Fernandez A., Leon-Espinosa G., DeFelipe J., Munoz A. Changes in the Golgi apparatus of neocortical and hippocampal neurons in the hibernating hamster. Frontiers in Neuroanatomy. 2015;9(DEC). [Epub ahead of print]. • Mihaljević B, Benavides-Piccione R, Guerra L, DeFelipe J, Larrañaga P, Bielza C. Classifying GABAergic interneurons with semi-supervised projected model-based clustering.Artificial intelligence in medicine. 2015;65(1): 49-59. • Bosch C, Martínez A, Masachs N, Teixeira CM, Fernaud I, Ulloa F et al. FIB/SEM technology and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult- generated neurons.Frontiers in neuroanatomy. ;9: 60. • Selvas A, Coria SM, Kastanauskaite A, Fernaud-Espinosa I, DeFelipe J, Ambrosio E et al. Rat-strain dependent changes of dendritic and spine morphology in the hippocampus after cocaine self- administration.Addiction biology. 2015. [Epub ahead of print]. • Markram H., Muller E., Ramaswamy S., Reimann M.W., Abdellah M., Sanchez C.A. et al. Reconstruction and Simulation of Neocortical Microcircuitry. Cell. 2015;163(2):456-492. • Íbias J, Soria-Molinillo E, Kastanauskaite A, Orgaz C, DeFelipe J, Pellón R et al. Schedule- induced polydipsia is associated with increased spine density in dorsolateral striatum neurons. Neuroscience. 2015;300: 238-45. • Anton-Fernandez A., Rubio-Garrido P., DeFelipe J., Munoz A. Selective presence of a giant saccular organelle in the axon initial segment of a subpopulation of layer V pyramidal neurons. Brain structure & function. 2015;220(2):869-884. • Blazquez-Llorca L, Woodruff A, Inan M, Anderson SA, Yuste R, DeFelipe J et al. Spatial distribution of neurons innervated by chandelier cells.Brain structure & function. 2015;220(5): 2817-34. • DeFelipe J. The anatomical problem posed by brain complexity and size: a potential solution. Frontiers in neuroanatomy. ;9: 104 . • DeFelipe J. The dendritic spine story: an intriguing process of discovery.Frontiers in neuroanatomy. ;9:14. • Miguens M., Kastanauskaite A., Coria S.M., Selvas A., Ballesteros-Yanez I., DeFelipe J. et al. The effects of cocaine self-administration on dendritic spine density in the rat hippocampus are dependent on genetic background. Cerebral Cortex. 2015;25(1):56-65. • Montes J, Peña JM, DeFelipe J, Herreras O, Merchan-Perez A. The influence of synaptic size on AMPA receptor activation: a Monte Carlo model.PloS one. ;10(6): e0130924. • Ramaswamy S., Courcol J.-D., Abdellah M., Adaszewski S.R., Antille N., Arsever S. et al. The neocortical microcircuit collaboration portal: A resource for rat somatosensory cortex. Frontiers in Neural Circuits. 2015;9(OCT). [Epub ahead of print].

Research projects

• Code: Cajal Blue Brain Project. Funding agency: Ecole Polytechnique Title: Cajal Blue Brain Project. International Federale de Lausanne (Suiza), IBM, Blue Brain Proyect. Universidad Politecnica de Madrid, Consejo Principal investigator: Javier de Felipe. Superior de Investigaciones Cientificas, CIBERNED’s collaboration: Yes. Espana. CIBERNED groups: G204, G403. Funding: ND. Other CIBER’s collaboration: No. Duration: 2009-2019 Type: International.

2015 CIBERNED Annual Report / 45 • Code: ZEN-15-321663. • Code: 2013/07A. Title: The Pyramidal Neuron in Cognition and Title: Papel de GSK-3 β en las alteraciones Alzheimer's Disease. de los circuitos corticales que ocurren en la Principal Investigator: Javier de Felipe. enfermedad de Alzheimer. CIBERNED’s collaboration: No. Principal Investigator: Teresa Iglesias Vacas. CIBERNED groups: CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G504, G401, G403, G111, Type: International. G307. Funding agency: Alzheimer's Association. Other CIBER’s collaboration: No. Funding: ND. Type: International. Duration: 2015-2018 Funding agency: CIBERNED. Funding: 316.000 €. Duration: 2013-2015

2015 CIBERNED Annual Report / 46 503 GROUP Isidro Ferrer Abizanda

Isidro Ferrer Abizanda Belén Ansoleaga Ávila Meritxell Puig Pinos Principal Investigator PhD student Student Ester Asó Mayelín Domínguez Luis Alberto Escobar Research staff González Research visitor PhD student Marta Barrachina Castilla Alejandra Martínez Research staff Paula García-Esparcia Research visitor PhD student Montserrat Olivé Plana Andre Palmeira Research staff Anusha Konetti Research visitor PhD student Karina Hernández-Ortega Katrin Thüne Research staff Irene López-González Research visitor PhD student Franc Llorens Margarita Carmona Murillo Research staff Raissa Camila Alvear Technician Contreras Noemi Vidal Sarró Student Jesús Moreno Research staff Lab Technician Margarida Frau Méndez Pol Andrés Benito Student Ester Bergés Pujol PhD student Office manager María Francisca García- Garrido Student

Contact details

Instituto de Neuropatología, Servicio Anatomía Patológica, Hospital Universitario de Bellvitge Departamento de Patología y Terapeútica Experimental Facultad de Medicina, Universidad de Barcelona C/ Feixa Llarga sn, 08907 Hospitalet de Llobregat Tel.: +34 93 260 74 52 Móvil: 677 82 14 56 [email protected]

2015 CIBERNED Annual Report / 47 Summary

Study of new alterations in metabolic pathways, Forty-four papers were published during 2015: and of clusters and hubs in the aging nervous system and at early stages of neurodegenerative • Seven focused on diagnostic consensus diseases including Alzheimer’s disease (AD), and optimization of methods for the study of Parkinson’s disease (PD), Dementia with Lewy human neurodegenerative diseases. Bodies, tauopathies, other frontotemporal lobar degenerations, prion diseases and amyotrophic • Seven dealt with the identification and analysis lateral sclerosis; combined use of genomics, of altered clusters in neurodegenerative transcriptomics, proteomics, redox proteomics, diseases including inflammation, protein metabolomics, lipidomics and bioinformatic synthesis, purine metabolism, energy processing followed by validation by qRT-PCR, metabolism. western blotting, ELISA, immunohistochemistry and enzymatic studies; all this on human post- • Five were devoted to characterization mortem brain samples from our brain bank of mouse models of senescence, and other centers belonging to the European AD, P301S tauopathy, Huntington, Brain Bank Network Consortium; national and X-adrenoleukodystrophy and Creutzfledt- international collaborations; translation of the Jakob disease (CJD). results to basal transgenic murine models and newly generated double-transgenic animals; • Seven assessed putative treatments in mouse use of these models to test possible treatments; models. and assessment of possible CSF biomarkers mainly based on metabolomes. • Three examined CSF biomarkers in CJD. Our main goal has been the identification of • Two yielded new descriptions of newly new pathways, networks, hubs and clusters identified diseases, one cardiomyopathy which are modified with age and altered at the linked to combined MuRF1 and MuRF3 initial stages of common neurodegenerative mutation, and the other a new familial diseases, in order to: astrocyte-predominant tauopathy linked to a FUS variant. • Gain understanding about pathogenic factors occurring during brain involution the rest of the publications were studies of RNA and at early stages of neurodegenerative expression, proteins and lipids in the central diseases in old age. nervous system in several diseases including AD, PD and prionic diseases. Finally, four chapters • Identify altered interconnected clusters, in Neuropathology of neurodegenerative odes and hubs. diseases: a practical guide, Kovacs GG, editor, Cambridge University Press, Cambridge; and the • Recognize putative targets and treatments chapter Vascular disease, hypoxia and related for therapeutic prevention. As additional conditions, in Greenfield’s Neuropathology, lines, there is study of neuromuscular diseases Love S, Budka H, Ironside JW, Perry A, editors, carried out by Dr. Montse Olivé Plana, and CRC Press, Boca Raton, Florida, were also the development of zebra fish models in published during the year. collaboration with ZeClinics.

Keywords

Neuropathology, epigenetics, transcriptomics, proteomics, metabolomics, lipidomics, Alzheimer’s, Parkinson’s, Creutzfeldt-Jakob disease, frontotemporal lobar degeneration, amyotrophic lateral sclerosis, cromosma X-linked adrenoleukodystrophy, muscle diseases

2015 CIBERNED Annual Report / 48 Program1 Group: Isidro Ferrer Abizanda

Publications

• Fiesel F.C., Ando M., Hudec R., Hill A.R., Castanedes-Casey M., Caulfield T.R. et al. (Patho-) physiological relevance of PINK1-dependent ubiquitin phosphorylation. EMBO Reports. 2015;16(9):1114-1130. • Terni B., Ferrer I. Abnormal expression and distribution of MMP2 at initial stages of alzheimer’s disease-related pathology. Journal of Alzheimer’s Disease. 2015;46(2):461-469. • Hernandez-Ortega K., Garcia-Esparcia P., Gil L., Lucas J.J., Ferrer I. Altered Machinery of Protein Synthesis in Alzheimer’s: From the Nucleolus to the Ribosome. Brain Pathology. 2015. [Epub ahead of print]. • Blanch M., Mosquera J.L., Ansoleaga B., Ferrer I., Barrachina M. Altered mitochondrial DNA methylation pattern in Alzheimer disease-related pathology and in Parkinson disease. American Journal of Pathology. 2016;186(2):385-397. Epub 2015. • Gonzalez De San Roman E., Manuel I., Giralt M.T., Chun J., Estivill-Torrus G., Rodriguez De Fonseca F. et al. Anatomical location of LPA<inf>1</inf> activation and LPA phospholipid precursors in rodent and human brain. Journal of Neurochemistry. 2015;134(3):471-485. • Launay N, Aguado C, Fourcade S, Ruiz M, Grau L, Riera J et al. Autophagy induction halts axonal degeneration in a mouse model of X-adrenoleukodystrophy.Acta neuropathologica. 2015;129(3): 399-415. • Diaz M., Fabelo N., Martin V., Ferrer I., Gomez T., Marin R. Biophysical alterations in lipid rafts from human cerebral cortex associate with increased BACE1/AβPP interaction in early stages of Alzheimer’s disease. Journal of Alzheimer’s Disease. 2015;43(4):1185-1198. • Aso E., Sanchez-Pla A., Vegas-Lozano E., Maldonado R., Ferrer I. Cannabis-based medicine reduces multiple pathological processes in AβPP/PS1 mice. Journal of Alzheimer’s Disease. 2015;43(3):977-991. • Paniagua-Torija B., Arevalo-Martin A., Ferrer I., Molina-Holgado E., Garcia-Ovejero D. CB1 cannabinoid receptor enrichment in the ependymal region of the adult human spinal cord. Scientific Reports. 2015;5. [Epub ahead of print]. • Strobel S., Grunblatt E., Riederer P., Heinsen H., Arzberger T., Al-Sarraj S. et al. Changes in the expression of genes related to neuroinflammation over the course of sporadic Alzheimer’s disease progression: CX3CL1, TREM2, and PPARγ. Journal of Neural Transmission. 2015;122(7):1069- 1076. • Kalia LV, Lang AE, Hazrati LN, Fujioka S, Wszolek ZK, Dickson DW et al. Clinical correlations with Lewy body pathology in LRRK2-related Parkinson disease.JAMA neurology. 2015;72(1):-. • Grau-Rivera O., Gelpi E., Nos C., Gaig C., Ferrer I., Saiz A. et al. Clinicopathological Correlations and Concomitant Pathologies in Rapidly Progressive Dementia: A Brain Bank Series. Neurodegenerative Diseases. 2015;15(6):350-360. • Berjaoui S., Povedano M., Garcia-Esparcia P., Carmona M., Aso E., Ferrer I. Complex inflammation mRNA-related response in ALS is region dependent. Neural Plasticity. 2015;2015. [Epub ahead of print]. • Zafar S., Schmitz M., Younus N., Tahir W., Shafiq M., Llorens F. et al. Creutzfeldt-Jakob Disease Subtype-Specific Regional and Temporal Regulation of ADP Ribosylation Factor-1-Dependent Rho/MLC Pathway at Pre-Clinical Stage. Journal of Molecular Neuroscience. 2015;56(2):329- 348. • Ansoleaga B., Garcia-Esparcia P., Pinacho R., Haro J.M., Ramos B., Ferrer I. Decrease in olfactory and taste receptor expression in the dorsolateral prefrontal cortex in chronic schizophrenia. Journal of Psychiatric Research. 2015;60:109-116. • Ansoleaga B., Jove M., Schluter A., Garcia-Esparcia P., Moreno J., Pujol A. et al. Deregulation of purine metabolism in Alzheimer’s disease. Neurobiology of Aging. 2015;36(1):68-80. • Fuso A., Ferraguti G., Scarpa S., Ferrer I., Lucarelli M. Disclosing bias in bisulfite assay: MethPrimers underestimate high DNA methylation. PLoS ONE. 2015;10(2). [Epub ahead of print].

2015 CIBERNED Annual Report / 49 • Alafuzoff I., Pikkarainen M., Neumann M., Arzberger T., Al-Sarraj S., Bodi I. et al. Erratum to: Neuropathological assessments of the pathology in frontotemporal lobar degeneration with TDP43-positive inclusions: an inter-laboratory study by the BrainNet Europe consortium[Journal of Neural Transmission, (2015), DOI 10.1007/s00702-014-1304-1]. Journal of Neural Transmission. 2015;122(7):973-974. • Krzyzanowska A., Garcia-Consuegra I., Pascual C., Antequera D., Ferrer I., Carro E. Expression of regulatory proteins in choroid plexus changes in early stages of alzheimer disease. Journal of Neuropathology and Experimental Neurology. 2015;74(4):359-369. • Ferrer I., Legati A., Garcia-Monco J.C., Gomez-Beldarrain M., Carmona M., Blanco R. et al. Familial behavioral variant frontotemporal dementia associated with astrocyte-predominant tauopathy. Journal of Neuropathology and Experimental Neurology. 2015;74(4):370-379. • Aso E, Serrano AL, Muñoz-Cánoves P, Ferrer I. Fibrinogen-Derived γ377-395 Peptide Improves Cognitive Performance and Reduces Amyloid-β Deposition, without Altering Inflammation, in AβPP/PS1 Mice.Journal of Alzheimer’s disease : JAD. 2015;47(2): 403-12. • Petrov D., Pedros I., Artiach G., Sureda F.X., Barroso E., Pallas M. et al. High-fat diet-induced deregulation of hippocampal insulin signaling and mitochondrial homeostasis deficiences contribute to Alzheimer disease pathology in rodents. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2015;1852(9):1687-1699. • Naudi A., Cabre R., Jove M., Ayala V., Gonzalo H., Portero-Otin M. et al. Lipidomics of Human Brain Aging and Alzheimer’s Disease Pathology. International Review of Neurobiology. 2015;122:133-189. • Armand-Ugon M., Aso E., Moreno J., Riera-Codina M., Sanchez A., Vegas E. et al. Memory Improvement in the AβPP/PS1 Mouse Model of Familial Alzheimer’s Disease Induced by Carbamylated-Erythropoietin is Accompanied by Modulation of Synaptic Genes. Journal of Alzheimer’s Disease. 2015;45(2):407-421. • O’Callaghan M.M., Emperador S., Pineda M., Lopez-Gallardo E., Montero R., Yubero D. et al. Mutation loads in different tissues from six pathogenic mtDNA point mutations. Mitochondrion. 2015;22:17-22. • Lopez-Gonzalez I., Aso E., Carmona M., Armand-Ugon M., Blanco R., Naudi A. et al. Neuroinflammatory gene regulation, mitochondrial function, oxidative stress, and brain lipid modifications with disease progression in tau P301S transgenic mice as a modelof frontotemporal lobar degeneration-tau. Journal of Neuropathology and Experimental Neurology. 2015;74(10):975-999. • Lopez-Gonzalez I., Schluter A., Aso E., Garcia-Esparcia P., Ansoleaga B., Llorens F. et al. Neuroinflammatory signals in alzheimer disease and APP/PS1 transgenic mice: Correlations with plaques, tangles, and oligomeric species. Journal of Neuropathology and Experimental Neurology. 2015;74(4):319-344. • Alafuzoff I, Pikkarainen M, Neumann M, Arzberger T, Al-Sarraj S, Bodi I et al. Neuropathological assessments of the pathology in frontotemporal lobar degeneration with TDP43-positive inclusions: an inter-laboratory study by the BrainNet Europe consortium.Journal of neural transmission (Vienna, Austria : 1996). 2015;122(7): 973-4. • Olive M., Abdul-Hussein S., Oldfors A., Gonzalez-Costello J., van der Ven P.F.M., Furst D.O. et al. New cardiac and skeletal protein aggregate myopathy associated with combined MuRF1 and MuRF3 mutations. Human Molecular Genetics. 2015;24(13):3638-3650. • Olive M., Abdul-Hussein S., Oldfors A., Gonzalez-Costello J., van der Ven P.F.M., Furst D.O. et al. New cardiac and skeletal protein aggregate myopathy associated with combined MuRF1 and MuRF3 mutations[Human Molecular Genetics, 24, 13, (2015) 3638-3650] DOI: 10.1093/hmg/ ddv108. Human Molecular Genetics. 2015;24(21):6264-. [Epub ahead of print]. • Sola I., Aso E., Frattini D., Lopez-Gonzalez I., Espargaro A., Sabate R. et al. Novel Levetiracetam Derivatives That Are Effective against the Alzheimer-like Phenotype in Mice: Synthesis, in Vitro, ex Vivo, and in Vivo Efficacy Studies. Journal of Medicinal Chemistry. 2015;58(15):6018-6032. • Zelaya M.V., Perez-Valderrama E., De Morentin X.M., Tunon T., Ferrer I., Luquin M.R. et al. Olfactory bulb proteome dynamics during the progression of sporadic Alzheimer’s disease: Identification of common and distinct olfactory targets across Alzheimer-related co-pathologies. Oncotarget. 2015;6(37):39437-39456.

2015 CIBERNED Annual Report / 50 Program 1 Group: Isidro Ferrer Abizanda

• Fourcade S., Ferrer I., Pujol A. Oxidative stress, mitochondrial and proteostasis malfunction in adrenoleukodystrophy: A paradigm for axonal degeneration. Free Radical Biology and Medicine. 2015;88:18-29. • Ho Kim J, Franck J, Kang T, Heinsen H, Ravid R, Ferrer I et al. Proteome-wide characterization of signalling interactions in the hippocampal CA4/DG subfield of patients with Alzheimer’s disease.Scientific reports. 2015;5: 11138. • Garcia-Esparcia P., Hernandez-Ortega K., Ansoleaga B., Carmona M., Ferrer I. Purine metabolism gene deregulation in Parkinson’s disease. Neuropathology and Applied Neurobiology. 2015;41(7):926-940. • Vergara C, Ordóñez-Gutiérrez L, Wandosell F, Ferrer I, del Río JA, Gavín R. Role of PrP(C) Expression in Tau Protein Levels and Phosphorylation in Alzheimer’s Disease Evolution.Molecular neurobiology. 2015;51(3): 1206-20. • Ferrer I. Selection of controls in the study of human neurodegenerative diseases in old age. Journal of neural transmission (Vienna, Austria : 1996). 2015;122(7): 941-7. • Pantano L, Friedländer MR, Escaramís G, Lizano E, Pallarès-Albanell J, Ferrer I et al. Specific small-RNA signatures in the amygdala at premotor and motor stages of Parkinson’s disease revealed by deep sequencing analysis.Bioinformatics (Oxford, England). 2015. [Epub ahead of print]. • Fernandez-Vega I., Perez de Heredia-Goni K., Santos-Juanes J., Goni Imizcoz M., Zaldumbide L., Zarranz J.J. et al. Sporadic adult-onset leucodystrophy with axonal spheroids and pigmented glia with no mutations in the known targeted genes. Histopathology. 2015. [Epub ahead of print]. • Grau-Rivera O., Sanchez-Valle R., Bargallo N., Llado A., Gaig C., Nos C. et al. Sporadic MM2- thalamic+cortical Creutzfeldt-Jakob disease: Utility of diffusion tensor imaging in the detection of cortical involvement in vivo. Neuropathology. 2015. [Epub ahead of print]. • Garcia-Ovejero D., Arevalo-Martin A., Paniagua-Torija B., Florensa-Vila J., Ferrer I., Grassner L. et al. The ependymal region of the adult human spinal cord differs from other species and shows ependymoma-like features. Brain : a journal of neurology. 2015;138:1583-1597. • Porquet D., Andres-Benito P., Grinan-Ferre C., Camins A., Ferrer I., Canudas A.M. et al. Amyloid and tau pathology of familial Alzheimer’s disease APP/PS1 mouse model in a senescence phenotype background (SAMP8). Age. 2015;37(1). [Epub ahead of print]. • Carulla P., Llorens F., Matamoros-Angles A., Aguilar-Calvo P., Espinosa J.C., Gavin R. et al. Involvement of PrP C in kainate-induced excitotoxicity in several mouse strains. Scientific Reports. 2015;5. [Epub ahead of print]. • Llorens F, Zafar S, Ansoleaga B, Shafiq M, Blanco R, Carmona M et al. Subtype and regional regulation of prion biomarkers in sporadic Creutzfeldt-Jakob disease.Neuropathology and applied neurobiology. 2015;41(5): 631-45.

Research projects

• Code: PI14/00268. • Code: FI12/00410. Title: Establecimiento de un algoritmo Title: Ayudas Predoctorales de formación en mitocondrial para predecir el riesgo de investigación (PFIS). progresión a la enfermedad de Alzheimer Principal Investigator: Isidro Ferrer. en pacientes con deterioro cognitivo leve. CIBERNED’s collaboration: No. Principal Investigator: Marta Barrachina. CIBERNED groups : CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: National. Others CIBER’s collaboration: No. Funding agency: FIS - Instituto de Salud Type: National. Carlos III Referencia. Funding agency: Instituto de Salud Carlos III. Funding: 27.000 €. Funding: ND. Duration: 2012-2016 Duration: 2015-2017

2015 CIBERNED Annual Report / 51 • Code: FIS PI/14-757. • Code: FIS PIE/14-0034. Title: Interactomas y marcadores para Title: Epigenetic and environmental discriminar factores de vulnerabilidad factors bracing cognitive impairment and neuronal regional en la neurodegeneración melancholic depression in elderly and asociada al envejecimiento normal y neurodegeneration. patológico en el SNC. Principal Investigator: Isidro Ferrer Abizanda. Principal Investigator: Isidro Ferrer Abizanda. CIBERNED’s collaboration: No. CIBERNED’s investigator: No. CIBERNED groups: CIBERNED groups: Other CIBER’s organization: No. Other CIBER’s collaboration: No. Type: National. Type: National. Funding agency: Insituto de Salud Carlos III. Fundinf agency: Instituto de Salud Carlos III. Funding: 605.000 €. Funding: 150.000 €. Duration: 2014-2016 Duration: 2014-2016 • Code : CIBERNED 2014/02. Title: Mecanismos epigenéticos implicados en la etiología y progresión de las Demencias Neurodegenerativas rápidamente progresivas. Principal Investigator: Miguel Calero. CIBERNED’s collaboration: Yes. CIBERNED groups: G114, G509, G503, G601. Other CIBER’s collaboration: No. Type: Intramurales. Funding agency: CIBERNED. Funding: 252.074 €. Duration: 2015-2017

PhD dissertations

• Author: Anusha Koneti. Title: Altered mitochondria and protein metabolism in Parkinson’s disease with disease progression. Date: 2nd Octobre 2015. Director: Isidro Ferrer Abizanda.

2015 CIBERNED Annual Report / 52 415 GROUP Antonia Gutiérrez Pérez

Antonia Gutiérrez Pérez David Baglietto Vargas Laura Trujillo Estrada Principal Investigator - Full Postdoctoral fellow Postdoctoral fellow Professor Raquel Sánchez Varo Mercedes Aneiros Ferrer José Carlos Dávila Cansino CIBERNED Postdoctoral CIBERNED Staff Technician Full Professor Researcher Cristina Núñez Díaz Zafaruddin Khan Elisabeth Sánchez Mejías PhD student Lecturer Postdoctoral fellow Juan José Fernández Valenzuela PhD student

Contact details

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]

2015 CIBERNED Annual Report / 53 Summary

During 2015 we have continued investigating To better understand the pathogenic some key pathological aspects as synaptic/ mechanisms and develop animal models with axonal dysfunction, neuronal death, and predictive value, this year we continued with inflammatory response (microglial and astroglial the characterization of postmortem samples activation) in various transgenic models of from patients, particularly the region of the Alzheimer’s disease and in postmortem samples hippocampus and parahippocampal gyrus. of patients from Braak II (asymptomatic) to We observed limited microglial activation along Braak V-VI (demented) stages. In relation with a marked degenerative process of this cell to animal models we determined synaptic population in patients in advanced stages of alterations in the hippocampus of APP/ the disease, in contrast to animal models based PS1 mice since early ages at the onset of on the expression of Abeta which display a amyloid pathology. We have also analyzed gradual activation and proliferation of microglia the vulnerability of cholinergic neurons in the during pathology progression. Together, our basal forebrain of APP/PS1, APP, PS1 and Tau data highlight the need to develop therapies models. In addition, we have made progress aimed to restore the microglial function, rather in understanding the role of the inflammatory than suppress the inflammatory response, to response in the progression of the disease by delay the progression of the disease. Finally, genetic manipulation of these models. Deletion this year we have started the phenotypic of IL-4 in the APP model exacerbates amyloid characterization of reactive astrocytes in both accumulation and causes an increase in animal models and human samples. microglial activation and neuronal vulnerability. Using an immunosuppressant treatment, we have found that a deficient immune response contributes to the disease process in the APP/ PS1 model.

Keywords

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

Publications

Research projects

• Code: BFU2013-43458-R. Other CIBER’s collaboration: No. Title: Una estrategia para recuperar y Type: National. prevenir la pérdida de la memoria Funding agency: Ministerio de Economía y Principal Investigator: Zafaruddin Khan. Competitividad. CIBERNED’s collaboration: No. Funding: 205.700 €. CIBERNED groups: Duration: 2014-2015

2015 CIBERNED Annual Report / 54 Program 1 Group: Antonia Gutiérrez Pérez

• Code: PI12/01439. • Code: PI2013/01. Title: Oligomeros toxicos del Abeta como Title: Propiedades emergentes de la agentes causantes de la disfunción del relación neurona-glia que subyacen a citoesqueleto y los procesos proteolíticos en neurodegeneración y demencia en la la enfermedad de Alzheimer. enfermedad de Alzheimer. Principal Investigator: Javier Vitorica. Principal Investigator: Ignacio Torres- CIBERNED’s collaboration: Yes. Alemán. CIBERNED groups: G411, G415. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G415, G413, G204, G409, Type: National. G108, G411. Funding agency: Instituto de Salud Carlos III. Other CIBER’s collaboration: No. Funding: 228.629 €. Type: National. Duration: 2013-2015 Funding agency: CIBERNED. Funding: 350.000 €. Code: CTS 2035. Duration: 2013-2015 • Title: Oligomerización y toxicidad de los peptidos de Abeta: búsqueda de nuevas • Code: 20141431. dianas de interés terapéutico en la Title: Deciphering the link between astrocyte enfermedad de Alzheimer. reactivity and neuronal damage in Principal Investigator: Javier Vitorica. Alzheimer´s disease. CIBERNED’s collaboration: Yes. Principal Investigator: Elena Galea. CIBERNED group: G411, G415. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G411, G413, G415. Type: CC.AA. Other CIBER’s collaboration: No. Funding agency: Junta de Andalucía. Type: Private. Funding: 289.307,5 €. Funding agency: La Marató de TV3. Duration: 2014-2016 Funding: 382.250 €. Duration: 2015-2017

PhD dissertations

• Author: Laura Isabel Trujillo Estrada. Date: 18th Septembre 2015. Title: Caracterización neuropatológica Supervisor: Antonia Gutiérrez Pérez. y evaluación preclínica de potenciales estrategias terapéuticas en modelos animales transgénicos de la enfermedad de • Author: Irene Navarro Lobato. Alzheimer. Title: RGS14 414-mediated prevention of an Date: 11th Septembre 2015. episodic memory loss: a study of molecular Supervisor: Antonia Gutiérrez Pérez. mechanism. Date: 11th Decembre 2015. Supervisor: Zafaruddin Khan. • Author: Elisabeth Sánchez Mejías. Title: Estudio neuropatológico del hipocampo y giro parahipocampal en la enfermedad de Alzheimer: de modelos transgénicos a humanos.

2015 CIBERNED Annual Report / 55 504 GROUP Alberto Lleó Bisa

Alberto Lleó Bisa M. Belén Sánchez-Saudinós Jordi Pegueroles Clinical Head, Neurology, Neuropsychologist PhD student Principal Investigator Roser Ribosa Daniel Acolea Rodríguez Rafael Blesa González Neurologist Neurologist Neurologist. Head of Department Martí Colom María Carmona Juan Fortea PhD student Neurologist Neurologist Oriol Dols Estrella Morenas Jordi Clarimón Echavarría PhD student Neurologist Postdoctoral Fellow Marta Querol Ignacio Illán-Gala Olivia Belbin PhD student Neurologist Postdoctoral Fellow Eduard Vilaplana Laia Muñoz Isabel Sala PhD student Lab technician Neuropsychologist Raúl Núñez Lab technician

Contact details

Unidad de Memoria. Servicio de Neurología Hospital de la Santa Creu i Sant Pau Sant Antoni Mº Claret 167 08025 Barcelona (Spain) Tel.: +34 93 556 59 86 Fax: +34 93 556 56 02 [email protected]

2015 CIBERNED Annual Report / 56 Summary

Our group is composed of a multidisciplinary continuum of AD, dementia with Lewy bodies, team of neurologists, neuropsychologists, FTD and Down syndrome. The group is also biologists, engineers and lab technicians. In involved in different imaging studies, using the last 5 years, the activities of the group have magnetic resonance imaging and amyloid focused on translational and clinical aspects of positron emission tomography (PET) to determine neurodegenerative dementias. the structural correlates of different biomarkers along the AD continnum, Down syndrome and In particular, the group has been working on other neurodegenerative dementias. Finally, the novel genetics factors and novel biomarkers group also has uncovered important aspects of the most common neurodegenerative of the molecular basis of neurodegenerative dementias. We have investigated new genetic dementias by using cell culture models or risk variants in Alzheimer’s disease (AD) and human brain tissue. The group offers a unique frontotemporal dementia (FTD). In parallel, the environment for clinicians and investigators group has conducted different investigations to tackle basic and translational aspects of in cerebrospinal fluid biomarkers along the neurodegeneration.

Keywords

Alzheimer, dementia, amyloid, imaging, biomarkers

Publications

2015 CIBERNED Annual Report / 57 • Hor H., Francescatto L., Bartesaghi L., Ortega-Cubero S., Kousi M., Lorenzo-Betancor O. et al. Missense mutations in TENM4, a regulator of axon guidance and central myelination, cause essential tremor. Human Molecular Genetics. 2015;24(20):5677-5686. • Díaz-Manera J, Alejaldre A, González L, Olivé M, Gómez-Andrés D, Muelas N et al. Muscle imaging in muscle dystrophies produced by mutations in the EMD and LMNA genes.Neuromuscular disorders : NMD. 2015. [Epub ahead of print]. • Duits FH, Martinez-Lage P, Paquet C, Engelborghs S, Lleó A, Hausner L et al. Performance and complications of lumbar puncture in memory clinics: Results of the multicenter lumbar puncture feasibility study.Alzheimer’s & dementia : the journal of the Alzheimer’s Association. 2015. [Epub ahead of print]. • Heilmann S., Drichel D., Clarimon J., Fernandez V., Lacour A., Wagner H. et al. PLD3 in non- familial Alzheimer’s disease. Nature. 2015;520(7545):E3-E5. • Morenas-Rodríguez E, Cervera-Carles L, Vilaplana E, Alcolea D, Carmona-Iragui M, Dols- Icardo O et al. Progranulin Protein Levels in Cerebrospinal Fluid in Primary Neurodegenerative Dementias.Journal of Alzheimer’s disease : JAD. 2015. [Epub ahead of print]. • Cacace R., Van den Bossche T., Engelborghs S., Geerts N., Laureys A., Dillen L. et al. Rare Variants in PLD3 Do Not Affect Risk for Early-Onset Alzheimer Disease in a European Consortium Cohort. Human Mutation. 2015;36(12):1226-1235. • Bufill E., Roura-Poch P., Sala-Matavera I., Anton S., Lleo A., Sanchez-Saudinos B. et al. Reelin signaling pathway genotypes and Alzheimer disease in a Spanish population. Alzheimer Disease and Associated Disorders. 2015;29(2):169-172. • Reijs B.L.R., Teunissen C.E., Goncharenko N., Betsou F., Blennow K., Baldeiras I. et al. The central biobank and virtual biobank of BIOMARKAPD: A resource for studies on neurodegenerative diseases. Frontiers in Neurology. 2015;6(OCT). [Epub ahead of print]. • Olde Rikkert M.G.M., Verhey F.R., Blesa R., Von Arnim C.A.F., Bongers A., Harrison J. et al. Tolerability and safety of souvenaid in patients with mild Alzheimer’s disease: Results of multicenter, 24-week, open-label extension study. Journal of Alzheimer’s Disease. 2015;44(2):471- 480. • Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agúndez JA, Jiménez-Jiménez FJ, Ross OA et al. TREM2 R47H variant and risk of essential tremor: a cross-sectional international multicenter study.Parkinsonism & related disorders. 2015;21(3): 306-9. • Alcolea D., Martinez-Lage P., Sanchez-Juan P., Olazaran J., Antunez C., Izagirre A. et al. Amyloid precursor protein metabolism and inflammation markers in preclinical Alzheimer disease. Neurology. 2015;85(7):626-633. • Dols-Icardo O., Iborra O., Valdivia J., Pastor P., Ruiz A., de Munain A.L. et al. Assessing the role of TUBA4A gene in frontotemporal degeneration. Neurobiology of Aging. 2015. [Epub ahead of print]. • International Genomics of Alzheimer’s Disease Consortium (IGAP), Lleó Bisa Alberto, Combarros Pascual Onofre, Bullido M. Convergent genetic and expression data implicate immunity in Alzheimer’s disease.Alzheimer’s & dementia : the journal of the Alzheimer’s Association. 2015;11(6): 658-71. • Cuyvers E, van der Zee J, Bettens K, Engelborghs S, Vandenbulcke M, Robberecht C et al. Genetic variability in SQSTM1 and risk of early-onset Alzheimer dementia: a European early- onset dementia consortium study.Neurobiology of aging. 2015;36(5): 2005.e15-22. • Ossenkoppele R, Jansen WJ, Rabinovici GD, Knol DL, van der Flier WM, van Berckel BN et al. Prevalence of amyloid PET positivity in dementia syndromes: a meta-analysis.JAMA. 2015;313(19):1924-38. • Jansen WJ, Ossenkoppele R, Knol DL, Tijms BM, Scheltens P, Verhey FR et al. Prevalence of cerebral amyloid pathology in persons without dementia: a meta-analysis.JAMA. 2015;313(19): 1939-49. • Alcolea D., Vilaplana E., Pegueroles J., Montal V., Sanchez-Juan P., Gonzalez-Suarez A. et al. Relationship between cortical thickness and cerebrospinal fluid YKL-40 in predementia stages of Alzheimer’s disease. Neurobiology of Aging. 2015;36(6):2018-2023.

2015 CIBERNED Annual Report / 58 Program1 Group: Alberto Lleó Bisa

• Bocchetta M, Galluzzi S, Kehoe PG, Aguera E, Bernabei R, Bullock R et al. The use of biomarkers for the etiologic diagnosis of MCI in Europe: an EADC survey.Alzheimer’s & dementia : the journal of the Alzheimer’s Association. 2015;11(2): 195-206. • Kruse N., Persson S., Alcolea D., Bahl J.M.C., Baldeiras I., Capello E. et al. Validation of a quantitative cerebrospinal fluid alpha-synuclein assay in a European-wide interlaboratory study. Neurobiology of Aging. 2015;36(9):2587-2596.

Research projects

• Code: MRTVE/P29. CIBERNED groups: Title: A comprehensive genomic analysis Other CIBER’s collaboration: No. of patients with motor neuron disease and Type: International. frontotemporal dementia to disentangle the Funding agency: Fondo de Investigación missing genetic architecture of amyotrophic Sanitario, ISCIII. lateral sclerosis. Funding: 122.815 €. Principal Investigator: Jordi Clarimón. Duration: 2015 CIBERNED’s collaboration: No. CIBERNED groups: • Code: PI13/01532. Other CIBER’s collaboration: No. Title: Enfermedad de Alzheimer y síndrome Type: National. de Down. Estudios multimodales de líquido Funding agency: Maratón Radio Televisión cefalorraquideo, resonancia magnética y Española. PET de amiloide. Funding: 99.822 €. Principal Investigator: Rafael Blesa. Duration: 2015-2017 CIBERNED’s collaboration: No. CIBERNED groups: • Code: PI12/01311. Other CIBER’s collaboration: No. Title: Variantes genéticas raras y su implicación Type: International. en la enfermedad de Alzheimer: uso de Funding agency: Fondo de Investigación nuevas tecnologías de ultrasecuenciación Sanitario. ISCIII. para el estudio de genes implicados en la Funding: 130.075 €. enfermedad. Duration: 2015 Principal Investigator: Jordi Clarimón. CIBERNED’s collaboration: No. • Code: PI14/01561. CIBERNED groups: Title: Marcadores sinápticos en la Other CIBER’s collaboration: No. enfermedad de Alzheimer. Type: International. Principal Investigator: Alberto Lleó. Fundinf agency: ISCIII. CIBERNED’s collaboration: No. Funding: 110.110 €. CIBERNED groups: Duration: 2013-2016 Other CIBER’s collaboration: No. Type: National. • Code: PI11/030206. Funding agency: Fondo de Investigación Title: Biomarkers for Alzheimer’s disease and Sanitaria. ISCIII. Parkinson’s disease (BIOMARKAPD). Funding: 134.915 €. Principal Investigator: Javier Sáez Valero. Duration: 2015-2017 CIBERNED’s collaboration: Yes. CIBERNED groups: G407, G504. • Code: 531/U/2014. Other CIBER’s collaboration: No. Title: Alzheimer's disease in Down´s syndrome. Type: European. CSF, MRI, EEG and PET multimodal studies. Funding agency: Instituto de Salud Carlos III. Principal Investigator: Juan Fortea. Funding: 117.370 €. CIBERNED’s collaboration: No. Duration: 2012-2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: PI14/01126. Type: International. Title: La estructura cerebral y el metabolismo Funding agency: Fundació La Marato TV3. en la enfermedad de Alzheimer preclínica. Funding: 199.000 €. Interacciones entre "nuevos y viejos" Duration: 2015 biomarcadores. Principal Investigator: Juan Fortea. CIBERNED’s collaboration: No.

2015 CIBERNED Annual Report / 59 • Code: 20142610. • Code: 2013/07A. Title: Synaptic markers in preclinical Title: Papel de GSK-3 β en las alteraciones Alzheimer's disease. de los circuitos corticales que ocurren en la Principal Investigator: Alberto Lleó. enfermedad de Alzheimer. CIBERNED’s collaboration: No. Principal Investigator: Teresa Iglesias Vacas. CIBERNED groups: CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G504, G401, G403, G111, Type: International. G307. Funding agency: Fundacio Marato TV3. Other CIBER’s collaboration: No. Funding: 199.878,75 €. Type: International. Duration: 2015 Funding agency: CIBERNED. Funding: 316.000 €. Duration: 2013-2015 PhD dissertations

• Author: Daniel Alcolea Rodríguez. Title: Cerebrospinal fluid biomarkers for the study of the pathophysiological pathways in Alzheimer’s disease. Date: 30th Novembre 2015. Supervisor: Alberto Lleó Bisa.

2015 CIBERNED Annual Report / 60 506 GROUP María López de Ceballos Lafarga

María López de Ceballos Alba Garcimartín Álvarez Sahba Seddhigi Lafarga Student Student Tenure Scientist, CSIC Nora Kadiri Moreno Clara González Martínez Student Researcher

Contact details

Instituto Cajal, CSIC Dr. Arce, 37 28002 Madrid (Spain) Tel.: +34 91 585 47 16 Fax: +34 91 585 47 54 [email protected]

2015 CIBERNED Annual Report / 61 Summary

Alzheimer Disease (AD) is characterized have been analyzed. In addition, the possible by aberrant protein accumulation like relationship between neuroinflammation in AD β-Amyloid and neurofibrillary tangles, by and viral infection was investigated. neuronal death and by neuroinflammation. This neuroinflammation is caused by glial The levels of different proteins were assessed by cell (astrocytes and microglia) activation Western blotting in frontal cortex from control that will release inflammatory substances. subjects, devoid of pathology, and AD patients, However, “alternative” microglial activation as defined in Braak stages (Neurologic Tissue may be a possible mechanism to counteract Brain Bank, Bellevitge Hospital, Barcelona). neuroinflammation, by increasing the release HSV-1 titre was measured by PCR. Indeed, of anti-inflammatory substances. On the other there was a statistically significant increase in hand, it is known that an increase in inflamm- cyclooxygenase-2, the astrocyte marker GFAP, ation caused by recurrent infections enhances the anti-inflammatory cytokine IL4 and HSV-1 the risk of suffering AD, and some systemic titre. No changes were observed in arginase-1, infections worsen cognitive impairment of AD characteristic of M2 alternative activation, or patients. Herpes Simplex Virus Type 1 (HSV-1) is the microglial cell marker Iba1. A positive and one of those infections known to increase the significant correlation was observed between risk for suffering the neurologic condition. An GFAP and IL4 (p=0.0003), that suggests an enhancement in neuroinflammation due to astrocytic origin rather than microglial. On the infections is a possible mechanism that may other hand, a positive correlation between age explain such increased risk. and GFAP levels (p=0.036), and between age and HSV-1 (p=0.0041) was found. Our results Therefore, in this project the evolution of levels show a large increase in neuroinflammation in in glial cell markers, pro- and anti-inflammatory the latest Braak stages which could be related parameters in frontal cortex of control subjects with the HSV-1 infection. and patients categorized by their Braak stages

Keywords

Alzheimer’s disease, Braak stages, infection, HSV-1, neuroinflammation

Publications

• Nieto E, Delgado M, Sobrado M, de Ceballos ML, Alajarín R, García-García L et al. Preliminary research on 1-(4-bromo-2-nitroimidazol-1-yl)-3-[(18)F]fluoropropan-2-ol as a novel brain hypoxia PET tracer in a rodent model of stroke.European journal of medicinal chemistry. 2015;101: 604- 15

Research projects

• Code: BFU2011-30217-C03-01. • Code: S2010/BMD-2349. Title: Senalización de NOTCH/NGN3 en las Title: Imágen Molecular Multimodal de la acciones neuroprotectoras de compuestos. Inflamación. Principal Investigator: Luis Miguel García Principal Investigator: S Cerdan. Segura. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: CC.AA. Type: National. Funding agency: CAM. Funding agency: Ministerio de Economía y Funding: 829.610 €. Competitividad. Duration: 2012-2015 Funding: 395.000 €. Duration: 2012-2015

2015 CIBERNED Annual Report / 62 Program 1 Group: María López de Ceballos Lafarga

PhD dissertations

• Author: Silvia de Vidania Ballesteros. Title: Efecto de los moduladores estrogénicos sobre funciones gliales: relevancia para la enfermedad de Alzheimer. Date: 18th Decembre 2015. Supervisor: María López de Ceballos Lafarga.

2015 CIBERNED Annual Report / 63 404 GROUP Carlos Matute Almau

Carlos Matute Almau Susana Mato Santos Mónica Benito Full Professor-Principal Investigator Ramón y Cajal Researcher PhD student

Alberto Pérez Samartín Fabio Cavaliere Tania Quintela Associate Professor Research Associate PhD student

Fernando Pérez Cerdá Asier Ruiz Núñez Andrea Manterola Juaristi Associate Professor Postdoctoral Fellow PhD student

Elena Alberdi Alfonso Abraham Cisneros Hazel Gómez Manrique Assistant Professor Mejorado Technician CIBERNED Postdoctoral Fellow María Domercq García Saioa Marcos Ezquerro Assistant Professor Ane Wissenbach Technician CIBERNED PhD student María Victoria Sánchez Juan Carlos Chara Technician CIBERNED Gómez Manuel Canedo Assistant Professor PhD student

Contact details

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]

2015 CIBERNED Annual Report / 64 Summary

The main goal of our laboratory is to unveil found that Aβ1-42 facilitates myelin repair after the pathological mechanisms of glial cells in demyelination via activation of Fyn kinase, a Alzheimer disease (AD) and their contribution member of Src kinase family, and that there to disease progression. Recently, we have is substantial hypermyelination in the APP/ described a novel mechanism by which the PS1/tau triple transgenic mice as well as in AD toxic soluble amyloid beta (Aβ1-42) peptide postmortem brain. In addition, the myelin sheath modulates integrin β1 activity and triggers is thicker while compound action potentials astrogliosis by reactive oxygen species (ROS) have lower amplitude, indicating defective generation through NOX-2 activation. In the conduction along axonal tracts. Together, last year, we have observed that the expression these findings suggest a role for Aβ1-42 and Src levels of key intermediaries in that pathway kinases in the pathophysiology of myelin in AD. (integrin β1, NOX2, GFAP and others) increase in astrocytes located close and distant to amyloid Lastly, we are using microfluidics to examine plaques during AD progression. Preliminary data propagation of pathogenic human α-synuclein at the electron microscope level in APP/PS1/ in Lewy bodies from Parkinson´s disease brains. tau transgenic mice shows that these markers Initial results indicate that this protein is taken are located in astrocytes which are severely up by neurons and astroctyes, and transported damaged, display perisynaptic changes and intracellularly, being astrocytes resistant to its hypertrophia. These results suggest that Aβ1- toxicity. In addition, we have observed neuronal 42 may cause primary astrogliosis in AD and apoptosis folliwing astrocyte exposure to Lewy alter synaptic communication and astrocyte bodies. These findings point to a relevant role of volume. astrocytes in disease propagation. Therefore, we are generating human astrocytes differentiated On the other hand, we have observed that form iPSCs derived from fibroblasts of patients Aβ1-42 favors oligodendrocyte maturation as with sporadic Parkinson disease and with LRRK2 it increases the expression of myelin proteins mutations. (MBP, CNPase and PLP). In the last year, we also

Keywords

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

Publications

• Gu B.J., Field J., Dutertre S., Ou A., Kilpatrick T.J., Lechner-Scott J. et al. A rare P2X7 variant Arg307Gln with absent pore formation function protects against neuroinflammation in multiple sclerosis. Human Molecular Genetics. 2015;24(19):5644-5654. • Rodriguez-Arellano J.J., Parpura V., Zorec R., Verkhratsky A. Astrocytes in physiological aging and Alzheimer’s disease. Neuroscience. 2015. [Epub ahead of print]. • Pekny M., Pekna M., Messing A., Steinhauser C., Lee J.-M., Parpura V. et al. Astrocytes: a central element in neurological diseases. Acta Neuropathologica. 2015;:1-23. • Zorec R., Verkhratsky A., Rodriguez J.J., Parpura V. Astrocytic vesicles and gliotransmitters: Slowness of vesicular release and synaptobrevin2-laden vesicle nanoarchitecture. Neuroscience. 2015. [Epub ahead of print]. • Verkhratsky A., Zorec R., Rodriguez J.J., Parpura V. Astroglia dynamics in ageing and Alzheimer’s disease. Current Opinion in Pharmacology. 2016;26:74-79. Epub 2015. • Verkhratsky A., Parpura V. Astrogliopathology in neurological, neurodevelopmental and psychiatric disorders. Neurobiology of Disease. 2016;85:254-261. Epub 2015. • Cisneros-Mejorado A., Perez-Samartin A., Gottlieb M., Matute C. ATP signaling in brain: release, excitotoxicity and potential therapeutic targets. Cellular and molecular neurobiology. 2015;35(1):1-6. • Arellano RO, Sánchez-Gómez MV, Alberdi E, Canedo-Antelo M, Chara JC, Palomino A et al. Axon-to-Glia Interaction Regulates GABAA Receptor Expression in Oligodendrocytes.Molecular pharmacology. 2016;89(1). Epub 2015.

2015 CIBERNED Annual Report / 65 • Martinez-Cengotitabengoa M., Macdowell K.S., Alberich S., Diaz F.J., Garcia-Bueno B., Rodriguez-Jimenez R. et al. BDNF and NGF Signalling in Early Phases of Psychosis: Relationship with Inflammation and Response to Antipsychotics after 1 Year. Schizophrenia Bulletin. 2016;42(1):142-151. Epub 2015. • Bernal-Chico A., Canedo M., Manterola A., Victoria Sanchez-Gomez M., Perez-Samartin A., Rodriguez-Puertas R. et al. Blockade of monoacylglycerol lipase inhibits oligodendrocyte excitotoxicity and prevents demyelination in vivo. GLIA. 2015;63(1):163-176. • Cisneros-Mejorado A, Gottlieb M, Cavaliere F, Magnus T, Koch-Nolte F, Scemes E et al. Blockade of P2X7 receptors or pannexin-1 channels similarly attenuates postischemic damage.Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism. 2015;35(5): 843-50. • Stenovec M., Trkov S., Lasic E., Terzieva S., Kreft M., Rodriguez Arellano J.J. et al. Expression of familial Alzheimer disease presenilin 1 gene attenuates vesicle traffic and reduces peptide secretion in cultured astrocytes devoid of pathologic tissue environment. GLIA. 2016;64(2):317- 329. Epub 2015. • Cipriani R., Chara J.C., Rodriguez-Antiguedad A., Matute C. FTY720 attenuates excitotoxicity and neuroinflammation. Journal of Neuroinflammation. 2015;12(1). [Epub ahead of print]. • Moriya T., Shibasaki R., Kayano T., Takebuchi N., Ichimura M., Kitamura N. et al. Full-length transient receptor potential vanilloid 1 channels mediate calcium signals and possibly contribute to osmoreception in vasopressin neurones in the rat supraoptic nucleus. Cell Calcium. 2015;57(1):25-37. • Verkhratsky A., Marutle A., Rodriguez-Arellano J.J., Nordberg A. Glial Asthenia and Functional Paralysis: A New Perspective on Neurodegeneration and Alzheimers Disease. Neuroscientist. 2015;21(5):552-568. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1). Epub 2015. • Martin A., Vazquez-Villoldo N., Gomez-Vallejo V., Padro D., Soria F.N., Szczupak B. et al. In vivo imaging of system xc- as a novel approach to monitor multiple sclerosis. European Journal of Nuclear Medicine and Molecular Imaging. 2015;:1-15. • Martin A., Szczupak B., Gomez-Vallejo V., Domercq M., Cano A., Padro D. et al. In vivo PET imaging of the α4β2 nicotinic acetylcholine receptor as a marker for brain inflammation after cerebral ischemia. Journal of Neuroscience. 2015;35(15):5998-6009. • Heneka M.T., Carson M.J., Khoury J.E., Landreth G.E., Brosseron F., Feinstein D.L. et al. Neuroinflammation in Alzheimer’s disease. The Lancet Neurology. 2015;14(4):388-405. • Perez-Cerda F., Sanchez-Gomez M.V., Matute C. Pío del Río hortega and the discovery of the oligodendrocytes. Frontiers in Neuroanatomy. 2015;9(July). [Epub ahead of print]. • Cavaliere F., Donno C., D’Ambrosi N. Purinergic signaling: A common pathway for neural and mesenchymal stem cell maintenance and differentiation. Frontiers in Cellular Neuroscience. 2015;9(JUNE). [Epub ahead of print]. • Gonzalez-Ortega I., Alberich S., Echeburua E., Aizpuru F., Millan E., Vieta E. et al. Subclinical depressive symptoms and continued cannabis use: Predictors of negative outcomes in first episode psychosis. PLoS ONE. 2015;10(4). [Epub ahead of print]. • Peng L., Verkhratsky A., Gu L., Li B. Targeting astrocytes in major depression. Expert Review of Neurotherapeutics. 2015;15(11):1299-1306.

2015 CIBERNED Annual Report / 66 Program 1 Group: Carlos Matute Almau

Research projects

• Code: 2014.0550. Other CIBER’s collaboration: No. Title: Neurogenic potential of dimethyl- Type: International. fumarate. Funding agency: N.D. Principal Investigator: Carlos Matute Almau. Funding: 280.000 €. CIBERNED’s collaboration: No. Duration: 2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: Coen Pathfinder 2015. Type: International. Title: Protection of neurons in vitro and in Funding agency: Biogen Pharma. vivo from Synuclein toxicity by molecular Funding: 50.000 €. tweezers. Duration: 2014-2016 Principal Investigator: Erwan Bezard. CIBERNED’s collaboration: No. • Code: ARSEP2015. CIBERNED groups: Title: Evaluating the potential of targeting OtherCIBERs/Centros: No. 2-AG hydrolytic enzymes MAGL and ABHD6 Type: International. as novel therapeutic strategy in multiple Funding agency: CoEN Pathfinder. sclerosis. Funding: 280.000 €. Principal Investigator: Carlos Matute Almau. Duration: 2015-2017 CIBERNED’s collaboration: No. CIBERNED groups: • Code: SAF2013-45084-R. Other CIBER’s collaboration: No. Title: Therapeutic potential of neurotransmitter Type: International. receptors in glial cells. Funding agency: ARSEP Foundation. Principal Investigator: Carlos Matute Almau. Funding: 50.000 €. CIBERNED’s collaboration: No. Duration: 2015-2016 CIBERNED groups: Other CIBER’s collaboration: No. • Code: Merck KGaA. Type: International. Title: Could reeducating microglia by P2X4R Funding agency: Ministerio de Economía y manipulation constitute an alternative Competitividad. therapy for MS?. Funding: 325.000 €. Principal Investigator: María Domercq Duration: 2014-2016 Garcia. CIBERNED’s collaboration: No. Code: IT702-13. CIBERNED groups: • Title: Bases moleculares y celulares de la Other CIBER’s collaboration: No. neurodegeneración. Type: International. Principal Investigator: Carlos Matute Almau. Funding agency: Merck KGaA. CIBERNED’s collaboration: No. Funding: 250.000 €. CIBERNED groups: Duration: 2014-2017 Other CIBER’s collaboration: No. Type: CC.AA. • Code: 2014/06. Funding agency: Gobierno Vasco. Title: Inicio y Progresión de la Enfermedad de Funding: 444.000 €. Parkinson: Papel de la Activación Glial. Duration: 2013-2018 Principal Investigator: María Cruz Rodríguez Oroz. CIBERNED’s collaboration: Si. CIBERNED groups: G206, G404, G205.

PhD dissertations

• Author: Ane Wysenbach. Title: Mecanismos moleculares implicados en la astrogliosis en la enfermedad de Alzheimer. Date: 2nd June 2015. Supervisor: Elena Alberdi Alfonso.

2015 CIBERNED Annual Report / 67 508 GROUP Guadalupe Mengod Los Arcos

Guadalupe Mengod Silvia Serrano Alba Robles Almenta Full Professor, CSIC Researcher Masters Researcher

Roser Cortés Marina Di Bari Laura Ramiro Pascual Tenure Scientist, CSIC Postdoctoral Fellow Student

Mª Teresa Vilaró Giovanni Di Pinto Melanis Muriel Tenure Scientist, CSIC Erasmus Researcher Lab Technician

Rocío Martín-Álvarez Yasmine Bejarano Condezo Lab Technician Masters Researcher

Contact details

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

2015 CIBERNED Annual Report / 68 Summary

Our group is composed of two research sublines, On the other hand, in the other subline of which have the main and common research work, neuroinflammation, we are analyzing theme of neurodegeneration. the molecular regulation of cAMP signaling in the murine experimental autoimmune In one of the sublines, we investigated the encephalomyelitis (EAE) model of multiple activation of 5-HT4 serotonin receptors as an sclerosis and looking for innovative drugs approach to interfere with Aß amyloid peptide for multiple sclerosis treatment through deposition in the 3xTg-AD mouse model of the selective inhibition of cAMP-specific Alzheimer’s disease. We observed that continued phosphodiesterases. stimulation for 28 days of these receptors resulted Within this line we have established a in very marked improvements in learning and collaboration with a research group from the memory, in parallel with a decrease of both University of Rome within the framework of a intraneuronal Aß and extracellular amyloid project funded by the Fondazione Italiana load. The treatment attenuated neophobia Esclerosi Multipla. We have analyzed the and anxiety and increased cell proliferation cholinergic system in the spinal cord of mice in the two neurogenic areas of the adult EAE by in situ hybridization histochemical and brain. We are currently analyzing whether this enzymatic. increase in cell proliferation does result in true neurogenesis, i.e. whether the proliferating cells differentiate into neurons.

Keywords

Neurodegeneration, neuroinflammation, molecular neuropharmacology

Publications

• Mengod G., Palacios J.M., Cortes R. Cartography of 5-HT1A and 5-HT2A Receptor Subtypes in Prefrontal Cortex and Its Projections.. ACS Chemical Neuroscience. 2015;6(7):1089-1098. • Alvarez-Salas E., Mengod G., Garcia-Luna C., Soberanes-Chavez P., Matamoros-Trejo G., de Gortari P. Mct8 and trh co-expression throughout the hypothalamic paraventricular nucleus is modified by dehydration-induced anorexia in rats. Neuropeptides. 2015. [Epub aheadof print]. • Penas-Cazorla R., Vilaro M.T. Serotonin 5-HT4 receptors and forebrain cholinergic system: receptor expression in identified cell populations. Brain Structure and Function. 2015;220(6):3413- 3434. • Marin C, Bonastre M, Mengod G, Cortés R, Rodríguez-Oroz MC. From unilateral to bilateral parkinsonism: Effects of lateralization on dyskinesias and associated molecular mechanisms. Neuropharmacology. 2015;97: 365-75.

Research projects

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

2015 CIBERNED Annual Report / 69 • Code: 2013/R/25. Title: Relationship between cholinergic dysfuntion and inflammation: study in multiple sclerosis; studies in EAE mice and RRMS patients. Principal Investigator: Ada María Tata. CIBERNED’s collaboration: No. CIBERNED groups: Other CIBER’s collaboration: No. Type: International. Funding agency: Fondazione Italiana Sclerosis Multipla (FISM). Funding: 133.000 €. Duration: 2014-2015

PhD dissertations

• Author: Nuria Paúl Fernández. Title: El papel de las fosfodiesterasas del AMPc en la evolución y tratamiento de la esclerosis múltiple en el modelo de EAE en ratón. Date: 2nd February 2015. Supervisor: Guadalupe Mengod Los Arcos.

2015 CIBERNED Annual Report / 70 507 GROUP María A. Pastor Muñoz

María Asunción Pastor Javier Bernacer María Gonzalo Arrondo Muñoz PhD PhD student Principal investigator María Asunción Fernández Luis Eudave Ramos Pau Pastor Muñoz Seara PhD student Co-principal investigator PhD Silvia Zarraluqui López Miquel Aguilar Barberá Elisa Mengual Poza PhD student PhD PhD Beatriz Echeveste Maite Aznarez Sanado Silvia Romero Contreras PhD student PhD Nurse Sara Ortega Cubero Elkin Luis García Juan Pablo Tartari PhD student PhD Specialist

Contact details

Center for Applied Medical Research Universidad de Navarra 31008 Pamplona (Spain) Tel.: +34 94 825 54 00 Fax: +94 829 65 00 [email protected]

2015 CIBERNED Annual Report / 71 Summary

The research of Neuroimaging/Neurogenetics an increase risk when carrying the MAPT H1/H1 group at the CIMA is focused in the study of polymorphism. genetic markers of neurodegenerative diseases such as dementia or parkinsonisms either We also identified frontotemporal regions familial or sporadic. We are currently analyzing specially impaired among SQSTM1 carriers, as last generation sequencing data of the human well as the accurate characterization of loss of exome to identify novel genes involved in such substantia nigra neural cells among subjects with disorders. We want to highlight the finding of sporadic and monogenic Parkinson disease, the association of variants of TENM4 gene with that can be used as a diagnostic marker. familial essential tremor. With regards to most recent neuroimaging As a result of this work, and from national and sequences, we have updated with success international collaborations. In the genomics perfusion MRI Arterial Spin Labeling (ASL) and core, we have published 16 articles with many BOLD sequences. The result of this work resulted groups of CIBERNED, most of them in peer- on four articles detailing the optimization of reviewed International journals located at the an ASL MRI protocol in the mark of EU COST first 25% of the IF ranking. Most publications program. report new knowledge on the genetic risk Many publications are fruit of a close linked to different kinds of dementia and collaboration with international groups and parkinsonism. In three of the article we reported CIBERNED Spanish groups in the context of the association of certain genetic variants with the DEGESCO Consortium. The DEGESCO specific neuroimaging degenerative patterns. Consortium has already published five articles. In fact, as an example of our activity, we The lines of our group will continue focusing performed a multicentre study analyzing the role on neuroimaging and genetic markers with of the TREM2 R47H variant in the risk of essential diagnostic practical utility that will allow to tremor. In addition, we led in the context of design more specific clinical trials and hopefully DEGESCO consortium, a study showing that new effective therapies. subjects with late-onset Alzheimer disease have

Keywords

Neuroimaging, functional neuroimaging, genetics, SNPs, gene

Publications

• Jun G, Ibrahim-Verbaas CA, Vronskaya M, Lambert JC, Chung J, Naj AC et al. A novel Alzheimer disease locus located near the gene encoding tau protein.Molecular psychiatry. 2016;21(1): 108-117. Epub 2015. • Zamurs LK, Idoate MA, Hanssen E, Gomez-Ibañez A, Pastor P, Lamandé SR. Aberrant mitochondria in a Bethlem myopathy patient with a homozygous amino acid substitution that destabilizes the collagen VI α2(VI) chain.The Journal of biological chemistry. 2015;290(7): 4272-81. • Dols-Icardo O, Nebot I, Gorostidi A, Ortega-Cubero S, Hernández I, Rojas-García R et al. Analysis of the CHCHD10 gene in patients with frontotemporal dementia and amyotrophic lateral sclerosis from Spain.Brain : a journal of neurology. 2015;138(Pt 12): e400. • Dols-Icardo O., Iborra O., Valdivia J., Pastor P., Ruiz A., de Munain A.L. et al. Assessing the role of TUBA4A gene in frontotemporal degeneration. Neurobiology of Aging. 2015. [Epub ahead of print]. • Jiménez-Jiménez FJ, García-Martín E, Alonso-Navarro H, Martínez C, Zurdo M, Turpín-Fenoll L et al. Association Between Vitamin D Receptor rs731236 (Taq1) Polymorphism and Risk for Restless Legs Syndrome in the Spanish Caucasian Population.Medicine. 2015;94(47): e2125. • Castellanos G., Fernandez-Seara M.A., Lorenzo-Betancor O., Ortega-Cubero S., Puigvert M., Uranga J. et al. Automated Neuromelanin Imaging as a Diagnostic Biomarker for Parkinson’s Disease. Movement Disorders. 2015;30(7):945-952.

2015 CIBERNED Annual Report / 72 Program 1 Group: María Asunción Pastor Muñoz

• Ortega-Cubero S, Pagola I, Luquin MR, Viteri C, Pastor P, Gállego Pérez-Larraya J et al. Clinical and neuroimaging characteristics of 14 patients with prionopathy: a descriptive study. Neurologia (Barcelona, Spain). 2015;30(3): 144-52. • Cervera-Carles L., Pagonabarraga J., Pascual-Sedano B., Pastor P., Campolongo A., Fortea J. et al. Copy number variation analysis of the 17q21.31 region and its role in neurodegenerative diseases. American Journal of Medical Genetics, Part B: Neuropsychiatric Genetics. 2015. [Epub ahead of print]. • Arrondo G, Aznárez-Sanado M, Fernández-Seara MA, Goñi J, Loayza FR, Salamon-Klobut E et al. Dopaminergic modulation of the trade-off between probability and time in economic decision-making.European neuropsychopharmacology : the journal of the European College of Neuropsychopharmacology. 2015;25(6): 817-27. • Buendia J., Loayza F.R., Luis E.O., Celorrio M., Pastor M.A., Hontanilla B. Functional and anatomical basis for brain plasticity in facial palsy rehabilitation using the masseteric nerve. Journal of Plastic, Reconstructive and Aesthetic Surgery. 2016;69(3):417-426. Epub 2015. • Cuyvers E, van der Zee J, Bettens K, Engelborghs S, Vandenbulcke M, Robberecht C et al. Genetic variability in SQSTM1 and risk of early-onset Alzheimer dementia: a European early- onset dementia consortium study.Neurobiology of aging. 2015;36(5): 2005.e15-22. • Ayuso P., Agundez J.A., Alonso-Navarro H., Martinez C., Benito-Leon J., Ortega-Cubero S. et al. Heme Oxygenase 1 and 2 Common Genetic Variants and Risk for Essential Tremor. Medicine. 2015;94(24):e968-. [Epub ahead of print]. • Garcia-Martin E., Jimenez-Jimenez F.J., Alonso-Navarro H., Martinez C., Zurdo M., Turpin-Fenoll L. et al. Heme Oxygenase-1 and 2 Common Genetic Variants and Risk for Restless Legs Syndrome. Medicine (United States). 2015;94(34):e1448-. [Epub ahead of print]. • Pastor P, Moreno F, Clarimón J, Ruiz A, Combarros O, Calero M et al. MAPT H1 Haplotype is Associated with Late-Onset Alzheimer’s Disease Risk in APOEε4 Noncarriers: Results from the Dementia Genetics Spanish Consortium.Journal of Alzheimer’s disease : JAD. 2015;49(2): 343- 52. • Hor H., Francescatto L., Bartesaghi L., Ortega-Cubero S., Kousi M., Lorenzo-Betancor O. et al. Missense mutations in TENM4, a regulator of axon guidance and central myelination, cause essential tremor. Human Molecular Genetics. 2015;24(20):5677-5686. • Fachal L., Mosquera-Miguel A., Pastor P., Ortega-Cubero S., Lorenzo E., Oterino-Duran A. et al. No evidence of association between common European mitochondrial DNA variants in Alzheimer, Parkinson, and migraine in the Spanish population. American Journal of Medical Genetics, Part B: Neuropsychiatric Genetics. 2015;168(1):54-65. • Schulte EC, Fukumori A, Mollenhauer B, Hor H, Arzberger T, Perneczky R et al. Rare variants in β-Amyloid precursor protein (APP) and Parkinson’s disease.European journal of human genetics : EJHG. 2015;23(10): 1328-33. • Fernández-Seara MA, Mengual E, Vidorreta M, Castellanos G, Irigoyen J, Erro E et al. Resting state functional connectivity of the subthalamic nucleus in Parkinson’s disease assessed using arterial spin-labeled perfusion fMRI.Human brain mapping. 2015;36(5):-. • Luis EO, Arrondo G, Vidorreta M, Martínez M, Loayza F, Fernández-Seara MA et al. Successful Working Memory Processes and Cerebellum in an Elderly Sample: A Neuropsychological and fMRI Study.PloS one. ;10(7): 1937-50 . • Guerreiro R., Escott-Price V., Darwent L., Parkkinen L., Ansorge O., Hernandez D.G. et al. Genome-wide analysis of genetic correlation in dementia with Lewy bodies, Parkinson’s and Alzheimer’s diseases. Neurobiology of Aging. 2015. [Epub ahead of print]. • Cacace R., Van den Bossche T., Engelborghs S., Geerts N., Laureys A., Dillen L. et al. Rare Variants in PLD3 Do Not Affect Risk for Early-Onset Alzheimer Disease in a European Consortium Cohort. Human Mutation. 2015;36(12):1226-1235. • Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agúndez JA, Jiménez-Jiménez FJ, Ross OA et al. TREM2 R47H variant and risk of essential tremor: a cross-sectional international multicenter study.Parkinsonism & related disorders. 2015;21(3): 306-9.

2015 CIBERNED Annual Report / 73 Research projects

• Code: PI 13/02211. Other CIBER’s collaboration: No. Title: Marcadores de neuroimagen de la Type: National. enfermedad de Parkinson geneticamente Funding agency: Ministerio de Economia y determinada. Neuroimage markers in Competitividad. Parkinson's disease genetically determined. Funding: 169.400 €. Principal Investigator: María A. Pastor. Duration: 2015-2016 CIBERNED’s collaboration: No. CIBERNED groups: • Code: RYC-2010-07161. Other CIBER’s collaboration: No. Title: Medida de perfusión cerebral en Type: National. patología usando la técnica de imagen por Funding agency: Fondo de Investigaciones resonancia magnética. de la Seguridad Social FIS. Principal Investigator: María A. Fernández Funding: 38.600 €. Seara. Duration: 2015 CIBERNED’s collaboration: No. CIBERNED groups: • Code: SAF2013-47939-R. Other CIBER’s collaboration: No. Title: Identificación de genes implicados Type: National. en enfermedad de Parkinson y temblor Funding agency: Ministerio de Economía y esencial mediante secuenciacion de nueva Competitividad - Ramón y Cajal. generacion y analisis de haplotipos. Funding: 243.626,09 €. Principal Investigator: Pau Pastor. Duration: 2010-2015 CIBERNED’s collaboration: No. CIBERNED groups:

PhD dissertations

• Author: Gabriel Castellanos Castañeda. Date: 2nd July 2015. Title: Visualización de la degeneración del Supervisor: María Asunción Pastor Muñoz. Locus Coeruleus y la Sustancia Negra en la enfermedad de Parkinson, a través de la • Author: Elkin O. Luis García. resonancia magnética. Title: Función mnésica operativa en Date: 21th July 2015. la enfermedad de Parkinson: estudio Supervisor: María Asunción Fernández neuropsicológico y de neuroimagen Seara. con múltiples condiciones y cargas de estimulación. • Author: Martín Martínez Villar. Date: 10th Decembre 2015. Title: Movimientos de locomoción en Supervisor: María Asunción Pastor Muñoz. la Enfermedad de Parkinson: análisis cinemático y mediante resonancia magnética funcional.

2015 CIBERNED Annual Report / 74 509 GROUP Jesús de Pedro Cuesta

Jesús de Pedro Cuesta Miguel Calero Lara Pablo Martínez Martín Head of Unit / Principal Researcher Lab Head Staff Researcher Investigator Olga Calero Rueda María José Medrano Albero Enrique Alcalde Cabero Postdoctoral fellow CIBERNED Staff Researcher PhD student Javier Damián Moreno María del Carmen Javier Almazán Isla Postdoctoral fellow Rodríguez Blázquez Technician Research Assistant Belén Frades Payo Fuencisla Avellanal Technician CIBERNED María Ruiz Tovar Calzadilla Postdoctoral fellow Technician

Contact details

Centro Nacional de Epidemiología Instituto de Salud Carlos III Monforte de Lemos, 5 28029 Madrid (Spain) Tel.: +34 91 822 26 50 Fax: +34 91 387 78 15 [email protected]

2015 CIBERNED Annual Report / 75 Summary

Subgroup Public Health/Aging In a study to estimate costs and determining A critical review of studies on the incidence variables of it over a period of four years in of neurodegenerative diseases supports the patients with Parkinson’s disease, a 92.5% idea that this is a group in which there is a increase in mean values of the latter was continuity of lesions when late-onset as well as a observed, reaching the mean and standard relationship between age of onset, magnitude deviations in total costs to €2,082.17 (€2,889.86) of incidence and survival. This pattern appears in the first year up to €4,008.6 (€7,757.35) in to correspond to the type of protein deposit. the fourth. Both the progression and severity of the disease as well as the presence of A study on the impact of detection or non- motor dysfunction and cognitive contribute detection status of depression in institutionalized significantly to increased costs. elderly residents from a network of residences in the Madrid Region shows a significant differences Subgroup Molecular Biology in the group of those with identified depression. Compared with a group without depression, In collaboration with various CIBERNED detected depression was associated with groups and the DEGESCO consortium, an increase in life expectancy in those with several genetic risk factors shared by different depression identified of 1.8 years (95% CI: -3.1- neurodegenerative diseases such as Alzheimer’s 6.7 years), whereas in those with undetected disease and Creutzfeldt-Jakob disease have depression decreased up to 6.3 years (95% CI: been studied. Similarly, in collaboration with 2.6-10.1). In a study with institutionalized elderly various international groups, we have continued on health status evaluation by the primary with the study of rapidly progressive Alzheimer’s physician compared with objective measures, disease and other neurodegenerative assessment of health status of institutionalized disorders, mainly focused on the analysis of residents by their doctor appeared to be too biomarkers in rapidly progressive dementia, high. In another study, a low level of coping was started with funding from the European Union associated with moderate disability according Joint Research Programme (JPND). In addition, to Barthel. In CHRODIS studies pathologies with the coordinated project with CIEN Foundation higher impact on disability-accompanying QoL and the October 12 Hospital on age-associated were mental and musculoskeletal. vascular dysfunction in Alzheimer’s disease is being finished. In the context of the CIBERNED Subgroup Results collaborative project launched in 2015 on “Epigenetic mechanisms involved in the Being essential the ability to detect etiology and progression of rapidly progressive neuropsychiatric symptoms in Parkinson’s neurodegenerative dementias”, our group disease, a review of the literature for the period is focused on the study of CpG dinucleotide 2005-2014 found 11 valid scales or questionnaires hypermutability as a common pathogenetic for depression, two for anxiety and one for mechanism for various neurodegenerative apathy. There were other for impulse control diseases, focusing as an object of study in disorders and hypersexuality. However, since Alzheimer’s, Parkinson’s and Creutzfeldt-Jakob, there were no valid scales or questionnaires for by using in silico approaches, analysis of routes psychosis, the need for its development was or susceptible genetic variations indicative of proposed. points of instability.

Keywords

Alzheimer disease, prion diseases, Creutzfeldt-Jakob, neurodegenerative disorders, conformational disorders, biomarkers, genetic susceptibility markers, risk genes, vascular factors, aging

2015 CIBERNED Annual Report / 76 Program 1 Group: Jesús de Pedro Cuesta

Publications

• Olazaran J., Gil-De-Gomez L., Rodriguez-Martin A., Valenti-Soler M., Frades-Payo B., Marin- Munoz J. et al. A blood-based, 7-metabolite signature for the early diagnosis of Alzheimer’s disease. Journal of Alzheimer’s Disease. 2015;45(4):1157-1173. • Sanchez-Juan P., Bishop M.T., Kovacs G.G., Calero M., Aulchenko Y.S., Ladogana A. et al. A genome wide association study links glutamate receptor pathway to sporadic Creutzfeldt- Jakob disease risk. PLoS ONE. 2015;10(4). [Epub ahead of print]. • Aguera-Ortiz L., Gil-Ruiz N., Cruz-Orduna I., Ramos-Garcia I., Osorio R.S., Valenti-Soler M. et al. A novel rating scale for the measurement of apathy in institutionalized persons with dementia: The APADEM-NH. American Journal of Geriatric Psychiatry. 2015;23(2):149-159. • Lovestone S, Boada M, Dubois B, Hüll M, Rinne JO, Huppertz HJ et al. A phase II trial of tideglusib in Alzheimer’s disease.Journal of Alzheimer’s disease : JAD. 2015;45(1): 75-88. • Fernandez-Mayoralas G., Rojo-Perez F., Martinez-Martin P., Prieto-Flores M.-E., Rodriguez- Blazquez C., Martin-Garcia S. et al. Active ageing and quality of life: Factors associated with participation in leisure activities among institutionalized older adults, with and without dementia. Aging and Mental Health. 2015;19(11):1031-1041. • Alcolea D., Martinez-Lage P., Sanchez-Juan P., Olazaran J., Antunez C., Izagirre A. et al. Amyloid precursor protein metabolism and inflammation markers in preclinical Alzheimer disease. Neurology. 2015;85(7):626-633. • Dafsari H.S., Reddy P., Herchenbach C., Wawro S., Petry-Schmelzer J.N., Visser-Vandewalle V. et al. Beneficial Effects of Bilateral Subthalamic Stimulation on Non-Motor Symptoms in Parkinson’s Disease. Brain Stimulation. 2015. [Epub ahead of print]. • Zea-Sevilla M.A., Bermejo-Velasco P., Serrano-Heranz R., Calero M. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) associated with a Novel C82R Mutation in the NOTCH3 Gene. Journal of Alzheimer’s Disease. 2015;43(2):363-367. • Forjaz M.J., Rodriguez-Blazquez C., Ayala A., Rodriguez-Rodriguez V., De Pedro-Cuesta J., Garcia-Gutierrez S. et al. Chronic conditions, disability, and quality of life in older adults with multimorbidity in Spain. European Journal of Internal Medicine. 2016;26(3):176-181. • Damian J., Pastor-Barriuso R., Valderrama-Gama E., De Pedro-Cuesta J. Discordance between physician-rated health and an objective health measure among institutionalized older people. BMC Geriatrics. 2015;15(1). [Epub ahead of print]. • Biundo R., Weis L., Fiorenzato E., Gentile G., Giglio M., Schifano R. et al. Double-blind Randomized Trial oft-DCSVersus Sham in Parkinson Patients With Mild Cognitive Impairment Receiving CognitiveTraining. Brain Stimulation. 2015. [Epub ahead of print]. • Lopez-Alvarez J., Zea Sevilla M.A., Aguera Ortiz L., Fernandez Blazquez M.A., Valenti Soler M., Martinez-Martin P. Effect of anticholinergic drugs on cognitive impairment in the elderly. Revista de Psiquiatria y Salud Mental. 2015;8(1):35-43. • Martinez-Martin P, Reddy P, Katzenschlager R, Antonini A, Todorova A, Odin P et al. EuroInf: a multicenter comparative observational study of apomorphine and levodopa infusion in Parkinson’s disease.Movement disorders : official journal of the Movement Disorder Society. 2015;30(4): 510-6. • Trenkwalder C., Chaudhuri K.R., Garcia Ruiz P.J., LeWitt P., Katzenschlager R., Sixel-Doring F. et al. Expert Consensus Group report on the use of apomorphine in the treatment of Parkinson’s disease - Clinical practice recommendations. Parkinsonism and Related Disorders. 2015;21(9):1023-1030. • Broen M.P.G., Moonen A.J.H., Kuijf M.L., Dujardin K., Marsh L., Richard I.H. et al. Factor analysis of the Hamilton Depression Rating Scale in Parkinson’s disease. Parkinsonism and Related Disorders. 2015;21(2):142-146. • Martinez-Martin P., Rodriguez-Blazquez C., Forjaz M.J., Kurtis M.M. Impact of Pharmacotherapy on Quality of Life in Patients with Parkinson’s Disease. CNS Drugs. 2015;29(5):397-413. • Timpka J., Fox T., Fox K., Honig H., Odin P., Martinez-Martin P. et al. Improvement of dyskinesias with l-dopa infusion in advanced Parkinson’s disease. Acta Neurologica Scandinavica. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 77 • Rieu I., Martinez-Martin P., Pereira B., De Chazeron I., Verhagen Metman L., Jahanshahi M. et al. International validation of a behavioral scale in Parkinson’s disease without dementia. Movement Disorders. 2015;30(5):705-713. • Forjaz M.J., Ayala A., Martinez-Martin P., Dujardin K., Pontone G.M., Starkstein S.E. et al. Is the Parkinson anxiety scale comparable across raters?. Movement Disorders. 2015;30(4):545-551. • Chaudhuri K.R., Rizos A., Trenkwalder C., Rascol O., Pal S., Martino D. et al. King’s Parkinson’s disease pain scale, the first scale for pain in PD: An international validation. Movement Disorders. 2015;30(12):1623-1631. • Broen M.P., Kohler S., Moonen A.J.H., Kuijf M.L., Dujardin K., Marsh L. et al. Modeling anxiety in Parkinson’s disease. Movement Disorders. 2015. [Epub ahead of print]. • Martinez-Martin P., Rodriguez-Blazquez C., Forjaz M.J., Frades-Payo B., Aguera-Ortiz L., Weintraub D. et al. Neuropsychiatric symptoms and caregiver’s burden in Parkinson’s disease. Parkinsonism and Related Disorders. 2015;21(6):629-634. • Erro R., Picillo M., Amboni M., Moccia M., Vitale C., Longo K. et al. Nonmotor predictors for levodopa requirement in de novo patients with Parkinson’s disease. Movement Disorders. 2015;30(3):373-378. • Zis P., Martinez-Martin P., Sauerbier A., Rizos A., Sharma J.C., Worth P.F. et al. Non-motor symptoms burden in treated and untreated early Parkinson’s disease patients: Argument for non-motor subtypes. European Journal of Neurology. 2015;22(8):1145-1150. • Martinez-Martin P., Rodriguez-Blazquez C., Paz S., Forjaz M.J., Frades-Payo B., Cubo E. et al. Parkinson symptoms and health related quality of life as predictors of costs: A longitudinal observational study with linear mixed model analysis. PLoS ONE. 2015;10(12). [Epub ahead of print]. • Martinez-Martin P., Rodriguez-Blazquez C., Alvarez Mario, Arakaki T., Arillo V.C., Chana P. et al. Parkinson’s disease severity levels and MDS-Unified Parkinson’s Disease Rating Scale. Parkinsonism and Related Disorders. 2015;21(1):50-54. • Leon-Salas B., Ayala A., Blaya-Novakova V., Avila-Villanueva M., Rodriguez-Blazquez C., Rojo- Perez F. et al. Quality of life across three groups of older adults differing in cognitive status and place of residence. Geriatrics and Gerontology International. 2015;15(5):627-635. • Rodriguez-Blazquez C., Martin-Garcia S., Frades-Payo B., Paris M.S., Martinez-Lopez I., Forjaz M.J. Quality of life and health status in institutionalized elderly with dementian. Revista Espanola de Salud Publica. 2015;89(1):51-60. • Gonzalez-Velez A.E., Forjaz M.J., Giraldez-Garcia C., Martin-Garcia S., Martinez-Martin P., Rodriguez-Blazquez C. et al. Quality of life by proxy and mortality in institutionalized older adults with Dementia. Geriatrics and Gerontology International. 2015;15(1):38-44. • Marventano S., Prieto-Flores M.-E., Sanz-Barbero B., Martin-Garcia S., Fernandez-Mayoralas G., Rojo-Perez F. et al. Quality of life in older people with dementia: A multilevel study of individual attributes and residential care center characteristics. Geriatrics and Gerontology International. 2015;15(1):104-110. • Castro-Monteiro E., Alhayek-Ai M., Diaz-Redondo A., Ayala A., Rodriguez-Blazquez C., Rojo- Perez F. et al. Quality of life of institutionalized older adults by dementia severity. International Psychogeriatrics. 2016;28(1):83-92. Epub 2015. • Storch A., Schneider C.B., Klingelhofer L., Odin P., Fuchs G., Jost W.H. et al. Quantitative assessment of non-motor fluctuations in Parkinson’s disease using the Non-Motor Symptoms Scale (NMSS). Journal of Neural Transmission. 2015;122(12):1673-1684. • Cruz-Orduna I., Aguera-Ortiz L.F., Montorio-Cerrato I., Leon-Salas B., Cristina Valle De Juan M., Martinez-Martin P. Reliability and validity of the severe impairment battery, short form (SIB-s), in patients with dementia in Spain. Revista de Neurologia. 2015;60(1):1-9. • Ambrosio L., Portillo M.C., Rodriguez-Blazquez C., Martinez-Castrillo J.C., Rodriguez-Violante M., Serrano-Duenas M. et al. Satisfaction with Life Scale (SLS-6): First validation study in Parkinson’s disease population. Parkinsonism and Related Disorders. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 78 Program 1 Group: Jesús de Pedro Cuesta

• Giglio RE, Rodriguez-Blazquez C, de Pedro-Cuesta J, Forjaz MJ. Sense of coherence and health of community-dwelling older adults in Spain.International psychogeriatrics / IPA. 2015;27(4): 621-8. • Benito-León J, Louis ED, Labiano-Fontcuberta A, Bermejo-Pareja F, De Pedro Cuesta Jesús. Serious head trauma preceding essential tremor: A population-based study (NEDICES).Journal of the neurological sciences. ;353(1-2): 116-21. • Valenti Soler M., Aguera-Ortiz L., Olazaran Rodriguez J., Mendoza Rebolledo C., Perez Munoz A., Rodriguez Perez I. et al. Social robots in advanced dementia. Frontiers in Aging Neuroscience. 2015;7(JUN). [Epub ahead of print]. • Chaudhuri K.R., Sauerbier A., Rojo J.M., Sethi K., Schapira A.H.V., Brown R.G. et al. The burden of non-motor symptoms in Parkinson’s disease using a self-completed non-motor questionnaire: A simple grading system. Parkinsonism and Related Disorders. 2015;21(3):287-291. • Olazaran J., Valenti M., Belen Frades, Zea-Sevilla M.A., Avila-Villanueva M., Fernandez-Blazquez M.A. et al. The Vallecas Project: A cohort to identify early markers and mechanisms of Alzheimer’s disease. Frontiers in Aging Neuroscience. 2015;7(SEP). [Epub ahead of print]. • Schmitz M., Ebert E., Stoeck K., Karch A., Collins S., Calero M. et al. Validation of 14-3-3 Protein as a Marker in Sporadic Creutzfeldt-Jakob Disease Diagnostic. Molecular Neurobiology. 2015. [Epub ahead of print]. • Heller S., Rebolledo C.M., Blazquez C.R., Chillon L.C., Munoz A.P., Perez I.R. et al. Validation of the multimodal assessment of capacities in severe dementia: a novel cognitive and functional scale for use in severe dementia. Journal of Neurology. 2015;262(5):1198-1208. • Hopfner F., Nebel A., Lyons K.E., Troster A.I., Kuhlenbaumer G., Deuschl G. et al. Validation of the QUEST for German-speaking countries. International Journal of Neuroscience. 2016;126(2):127- 134. Epub 2015. • Calero M., Gomez-Ramos A., Calero O., Soriano E., Avila J., Medina M. Additional mechanisms conferring genetic susceptibility to Alzheimer’s disease. Frontiers in Cellular Neuroscience. 2015;9(APR). [Epub ahead of print]. • Martinez-Martin P., Chaudhuri K.R., Rojo-Abuin J.M., Rodriguez-Blazquez C., Alvarez-Sanchez M., Arakaki T. et al. Assessing the non-motor symptoms of Parkinson’s disease: MDS-UPDRS and NMS Scale. European Journal of Neurology. 2015;22(1):37-43. • Odin P., Ray Chaudhuri K., Slevin J.T., Volkmann J., Dietrichs E., Martinez-Martin P. et al. Collective physician perspectives on non-oral medication approaches for the management of clinically relevant unresolved issues in Parkinson’s disease: Consensus from an international survey and discussion program. Parkinsonism and Related Disorders. 2015;21(10):1133-1144. • Zea-Sevilla M.A., Fernandez-Blazquez M.A., Calero M., Bermejo-Velasco P., Rabano A. Combined Alzheimer’s disease and cerebrovascular staging explains advanced dementia cognition. Alzheimer’s and Dementia. 2015;11(11):1358-1366. • De Pedro-Cuesta J., Rabano A., Martinez-Martin P., Ruiz-Tovar M., Alcalde-Cabero E., Almazan- Isla J. et al. Comparative incidence of conformational, neurodegenerative disorders. PLoS ONE. 2015;10(9. [Epub ahead of print]. • Trenkwalder C., Chaudhuri K.R., Martinez-Martin P., Rascol O., Ehret R., Valis M. et al. Prolonged- release oxycodone-naloxone for treatment of severe pain in patients with Parkinson’s disease (PANDA): A double-blind, randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(12):1161-1170.

2015 CIBERNED Annual Report / 79 Research projects

• Code: Fundación Ramón Areces 2012. • Code: CIBERNED 2014/02. Title: Estudio de la capacidad Title: Mecanismos epigenéticos implicados neuroprotectora de las células epiteliales en la etiología y progresión de las Demencias de los plexos coroideos como potencial Neurodegenerativas rapidamente terapia regenerativa de la enfermedad de progresivas. Alzheimer. Principal Investigator: Miguel Calero. Principal Investigator: Eva Carro. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G114, G509, G503, G601. CIBERNED groups: G502, G509. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: Intramurales. Type: National. Funding agency: CIBERNED. Funding agency: N.D. Funding: 252.074 €. Funding: 30.000 €. Duration: 2015-2017 Duration: 2012-2015

2015 CIBERNED Annual Report / 80 406 GROUP José Rodríguez Álvarez

José Rodríguez Álvarez Sergi Marco Martín Dolores Siedlecki Associate Professor. Principal Postdoctoral fellow PhD student Investigator Alfredo Jesús Miñano Meng Chen José Aguilera Ávila Postdoctoral fellow PhD student Associate Professor Laura Ortega Hernández Wenwen Cheng Ana María Candalija Iserte Postdoctoral fellow PhD student PhD student Laura Rubio Ferrarons Rut Fadó Andrés Judit Catala Solsona Technician Postdoctoral fellow PhD student Carlos Saura Antolín Guillem Sánchez Opazo Lilian Enríquez Barreto Associate Professor PhD student Postdoctoral fellow

Contact details

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

2015 CIBERNED Annual Report / 81 Summary

Multiple evidences have supported a role activation underlies associative memory deficits for beta amyloid (Aβ) in the etiology and and dendritic degeneration. In support of this, pathogenesis of AD. It is believed that the hippocampal CRTC1 gene therapy reverses progressive accumulation of self-aggregates of dendritic degeneration, transcriptome changes Aβ as oligomers (oAβ) would mediate synaptic and hippocampal-dependent memory loss dysfunction, leading to the initial cognitive in a faithful neurodegeneration mouse model deficits observed in MCI and earlier AD stages. lacking the PS genes5 (PS cDKO mice) despite Changes in gene expression programs and ongoing cortical degeneration. synaptic receptors and/or scaffolding proteins function could be underlying the early synaptic Moreover we have been exploring the dysfunction associated to cognitive impairment underlying molecular mechanisms, related to in MCI and early AD. glutamate AMPA receptors (AMPAR) regulation, involved in early learning and memory In the last years we have reported that synaptic dysfunction associated to AD and to identify activity and memory training activate a novel therapeutic targets and biomarkers for transcriptional program dependent on the promoting prevention and diagnosis at MCI cAMP-responsive element binding protein and early stages of the disease. Building upon (CREB)-regulated transcription coactivator-1 preliminary data obtained from our laboratory (CRTC1). However, the role of CRTC1 and we believe that oAβ-mediated synaptic its regulatory mechanisms in hippocampal- dysfunction associated to early learning and dependent learning and memory encoding and memory deficits in transgenic AD mice is due loss, and specifically during neurodegeneration, to an alteration in the presence of functional are unknown. GluA1-AMPARs in the synaptic membrane as a consequence of a combination of different During the last year, our studies have revealed for processes including miRNAs regulation, the first time that CRTC1 is specifically activated alteration of AMPARs regulatory proteins or in the hippocampus during associative memory alterations in the neurovascular unit affecting encoding, and that disruption of CRTC1 the reléase of angioneurins.

Keywords

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

Publications

• Sole M., Minano-Molina A.J., Unzeta M. A cross-talk between Aβ and endothelial SSAO/VAP-1 accelerates vascular damage and Aβ aggregation related to CAA-AD. Neurobiology of Aging. 2015;36(2):762-775. • Xifro X., Rodriguez-Alvarez J. Delineating the Factors and Cellular Mechanisms Involved in the Survival of Cerebellar Granule Neurons. Cerebellum. 2015;14(3):354-359. • Saura C.A., Parra-Damas A., Enriquez-Barreto L. Gene expression parallels synaptic excitability and plasticity changes in Alzheimer’s disease. Frontiers in Cellular Neuroscience. 2015;9(AUGUST). [Epub ahead of print]. • Fado R., Soto D., Minano-Molina A.J., Pozo M., Carrasco P., Yefimenko N. et al. Novel regulation of the synthesis of α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid (ampa) receptor subunit glua1 by carnitine palmitoyltransferase 1C (CPT1C) in the Hippocampus. Journal of Biological Chemistry. 2015;290(42):25548-25560. • Patricio-Martinez A., Mendieta L., Martinez I., Aguilera J., Limon I.D. The recombinant C-terminal fragment of tetanus toxin protects against cholinotoxicity by intraseptal injection of β-amyloid peptide (25-35) in rats. Neuroscience. 2016;315:18-30. Epub 2015.

2015 CIBERNED Annual Report / 82 Program 1 Group: José Rodríguez Álvarez

Research projects

• Code: 2014 SGR 0984. Principal Investigator: José Rodríguez Title: Recerca Biomedica en Álvarez. Neurodegeneracio (REBINE). CIBERNED’s collaboration: No. Principal Investigator: José Rodríguez CIBERNED groups: Alvarez. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: Funding agency: Fundación La Maraton Other CIBER’s collaboration: No. TV3. Type: CC.AA. Funding: 199.875 €. Funding agency: AGAUR-Generalitat Duration: 2015-2017 Catalunya. Funding: 30.000 €. • Code: SAF2013-43900. Duration: 2014-2016 Title: Transcriptional mechanisms underlying memory loss in Alzheimer´s disease transgenic • Code: SAF2011-30281. mouse models (TRANSMECAD). Title: Regulación de la señalización mediada Principal Investigator: Carlos A. Saura. por los receptores con dominios de muerte CIBERNED’s collaboration: No. por antagonistas y miRNAs como estrategia CIBERNED groups: protectora en la isquemia cerebral. Other CIBER’s collaboration: No. Principal Investigator: José Rodríguez Type: National. Álvarez. Funding agency: Ministerio de Economía y CIBERNED’s collaboration: No. Competitividad. CIBERNED groups: Funding: 230.000 €. Other CIBER’s collaboration: No. Duration: 2014-2016 Type: International. Funding agency: Ministerio de Ciencia e • Code: 2015/01-1. Innovación. Title: Terapia génica dirigida a neuregulinas Funding: 169.400 €. para el tratamiento de enfermedades Duration: 2012-2015 degenerativas de las motoneuronas. Principal Investigator: Xavier Navarro. • Code: SAF2014-59697-R. CIBERNED’s collaboration: Si. Title: Mecanismos moleculares implicados en CIBERNED groups: G607, G113, G406, G407. la disfunción de la sinapsis glutamatergicas Other CIBER’s collaboration: No. en fases tempranas de la enfermedad de Type: Intramurales. Alzheimer. Funding agency: Instituto de Salid Carlos III, Principal Investigator: José Rodríguez CIBERNED. Álvarez. Funding: 240.000 €. CIBERNED’s collaboration: No. Duration: 2015-2017 CIBERNED groups: Other CIBER’s collaboration: No. • Code: A2014417S. Type: National. Title: Transcriptional mechanisms of memory Funding agency: Ministerio de Economía loss in Alzheimer´s disease. y Competitividad. Programa Nacional de Principal Investigator: Carlos A. Saura. Biomedicina. CIBERNED’s collaboration: No. Funding: 181.500 €. CIBERNED groups: Duration: 2015-2017 Other CIBER’s collaboration: No. Type: International. • Code: Marato TV3 2013-343. Funding agency: BrightFocus Foundation. Title: Searching new biomarkers and Funding: 249.000 €. therapeutic targets related to cognitive Duration: 2014-2016 deficits in early stages of Alzheimer's Disease: Role of AKAP79/150, CPT1C and SSAO/VAP-1 in Ab-mediated AMPAR dysfunction.

PhD dissertations

• Author: Ana Candalija Iserte. Title: Interacción de las neurotoxinas clostridiales con los receptores de neurotrofina. Identificación de TrkB como receptor de la toxina tetánica. Date: 6th Novembre 2015. Supervisor: José Aguilera Ávila.

2015 CIBERNED Annual Report / 83 407 GROUP Javier Sáez Valero

Javier Sáez Valero Trinidad Mata Balaguer Aitana Sogorb Esteve Associate Professor. Principal Postdoctoral Fellow PhD student Investigator María Salud García Ayllón Claudia P. Boix Inmaculada Cuchillo Postdoctoral Fellow PhD student Ibáñez Postdoctoral Fellow Jordi Alom Poveda Esther Llorens Álvarez Head of Neurology department Lab technician Inmaculada López Font Postdoctoral Fellow

Contact details

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]

2015 CIBERNED Annual Report / 84 Summary

The aim of our research group stills focus into We also reviewed recent findings that suggest Alzheimer’s disease (AD) from a basic point of that most of these CSF soluble transmembrane view, but also regarding translational benefits proteins, including APP, but also secretase including therapy and diagnostic applications. proteins such as presenilins and BACE1, could In the basic research line we are focusing in form heteromeric complexes (López-Font et al., understand whether Reelin signaling pathway 2015 Front Neurol). is impaired in the brain of Alzheimer’s patients, in spite of the increasing brain protein levels. The oligomerization state of these potential Two research articles and one review have new biomarkers should be contemplated for been submitted. Nonetheless, in 2015 our assessing their real potential as CSF biomarkers publications have focused on cerebrospinal for AD by adequate molecular tools. It should fluid (CSF) biomarkers. We have been be mentioned that, recently, we demonstrated pioneers in demonstrating that the complete the presence of PS1 as heteromeric complexes form (unprocessed) of the amyloid protein in human CSF. In the context of our participation precursor, or APP, is present in CSF together with in the JPND UE project, BIOMARKAPD, regarding long fragments, sAPPα and sAPPβ; potential the validation of biomarkers for Alzheimer’s and diagnostic biomarkers for AD. All these forms Parkinson’s diseases, we have collaborated of soluble APP species form heteromeric in the inter-laboratory validation of alpha- complexes compromising their determination synuclein study as a biomarker for Parkinson’s by current ELISA assays (see Cuchillo-Ibanez disease (Kruse et al., Neurobiol Aging 2015). et al., Mol Neurodegener 2015). The presence Finally, studies on new glycoproteins altered of sAPP heteromers should be taken into in Alzheimer brain, as well the continuation of consideration when exploring the levels of the research lines in presenilin and APP as AD sAPPα and sAPPβ as potential CSF biomarkers. biomarkers, are ongoing.

Keywords

Alzheimer, β-amyloid, APP, presenilin, secretase, P-tau, Reelin, biomarker

Publications

• Lovestone S, Boada M, Dubois B, Hüll M, Rinne JO, Huppertz HJ et al. A phase II trial of tideglusib in Alzheimer’s disease.Journal of Alzheimer’s disease : JAD. 2015;45(1): 75-88. • Garre-Olmo J., Vilalta-Franch J., Calvo-Perxas L., Monserrat-Vila S., Lopez-Pousa S., J. Alom. Dependence scale for Alzheimer’s disease: Relationship with other clinical indicators and psychometric properties. Journal of Geriatric Psychiatry and Neurology. 2015;28(2):117-125. • Cuchillo-Ibañez I, Lopez-Font I, Boix-Amorós A, Brinkmalm G, Blennow K, Molinuevo JL et al. Heteromers of amyloid precursor protein in cerebrospinal fluid.Molecular neurodegeneration. 2015;10:2 . • Grasso S., Martinez-Lacaci I., Barbera V.M., Castillejo A., Soto J.L., Gallego-Plazas J. et al. HGUE- C-1 is an atypical and novel colon carcinoma cell line. BMC Cancer. 2015;15(1). [Epub ahead of print]. • Hallaq R., Volpicelli F., Cuchillo-Ibanez I., Hooper C., Mizuno K., Uwanogho D. et al. The Notch intracellular domain represses CRE-dependent transcription. Cellular Signalling. 2015;27(3):621- 629. • Lopez-Font I., Cuchillo-Ibanez I., Sogorb-Esteve A., Garcia-Ayllon M.-S., Saez-Valero J. Transmembrane amyloid-related proteins in CSF as potential biomarkers for Alzheimer’s disease. Frontiers in Neurology. 2015;6(JUN). [Epub ahead of print]. • Kruse N., Persson S., Alcolea D., Bahl J.M.C., Baldeiras I., Capello E. et al. Validation of a quantitative cerebrospinal fluid alpha-synuclein assay in a European-wide interlaboratory study. Neurobiology of Aging. 2015;36(9):2587-2596.

2015 CIBERNED Annual Report / 85 Research projects

• Code: SEP-210140568. CIBERNED’s collaboration: No. Title: A natural compensation ovine model CIBERNED groups: of a RELN-knockout, opening new insights Other CIBER’s collaboration: No. into compensation mechanisms to gain Type: International. knowledge into mental disorders. Funding agency: Resarch Grant Council, Principal Investigator: Beatriz Gutiérrez-Gil, Hong Kong. Universidad de León. Funding: 92.005 €. CIBERNED’s collaboration: No. Duration: 2012-2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: Fundación Ramón Areces. Type: International. Title: Regulación epigenética de Reelina en Funding agency: Horizon 2020, Excellent la enfermedad de Alzheimer. Science. Principal Investigator: Javier Sáez Valero. Funding: N.D. CIBERNED’s collaboration: No. Duration: 2015 CIBERNED groups: Other CIBERs collaboration: No. • Code: SAF2015-63204-CIN. Type: International. Title: 1st IN PhD Student & Postdoc Meeting Funding agency: Fundación Ramón Areces “Building Neuroscience: The future of a (XVI Concurso Nacional para la Adjudicación multidisciplinary field”. de Ayudas a la Investigación Científica y Principal Investigator: Javier Sáez Valero. Técnica). CIBERNED’s collaboration: No. Funding: 95.804 €. CIBERNED groups: Duration: 2012-2015 Other CIBER’s collaboration: No. Type: National. • Code: PI11/030206. Funding agency: MEC. Title: Biomarkers for Alzheimer’s disease and Funding: 4.000 €. Parkinson’s disease (BIOMARKAPD). Duration: 2015-2016 Principal Investigator: Javier Sáez Valero. CIBERNED’s collaboration: Yes. • Code: UGP-14-159. CIBERNED groups: G407, G504. Title: Análisis de las variantes de AChE en Other CIBER’s collaboration: No. cerebro control y con enfermedad de Type: European. Alzheimer. Funding agency: Instituto de Salud Carlos III. Principal Investigator: María Salud García Funding: 117.370 €. Ayllon. Duration: 2012-2015 CIBERNED’s collaboration: No. CIBERNED groups: • Code: PI12/00593. Other CIBER’s collaboration: No. Title: Alteración en la vía de senalización Type: International. de Reelina en la enfermedad de Alzheimer: Funding agency: FISABIO, Comunidad implicaciones patologicas. Valenciana. Principal Investigator: Sáez Valero, J. Funding: 7.500 €. CIBERNED’s collaboration: No. Duration: 2015-2016 CIBERNED groups: Other CIBER’s collaboration: No. • Code: PI14/00566. Type: International. Title: Procesamiento por secretasas de la Funding agency: Instituto de Saud Carlos III. forma colinérgica de acetilcolinesterasa, su Funding: 222.035 €. actividad transcripcional e implicación en Duration: 2013-2015 la enfermedad de Alzheimer. Principal Investigator: María Salud García • Code: 2015/01-1. Ayllon. Title: Terapia génica dirigida a neuregulinas CIBERNED’s collaboration: No. para el tratamiento de enfermedades CIBERNED groups: degenerativas de las motoneuronas. Other CIBER’s collaboration: No. Principal Investigator: Xavier Navarro. Type: National. CIBERNED’s collaboration: Yes. Funding agency: Instituto de Salud Carlos III. CIBERNED groups: G607, G113, G406, G407. Funding: 90.144,99 €. Other CIBER’s collaboration: No. Duration: 2015-2017 Type: Intramurales. Funding agency: Instituto de Salud Carlos III, • Code: RGC-663012. CIBERNED. Title: The trafficking of PRiMA-linked tetrameric Funding: 240.000 €. AChE: Probing roles of glycosylation in Duration: 2015-2017 determining the degradation of AChE in the brain. Principal Investigator: Karl Tsim.

2015 CIBERNED Annual Report / 86 408 GROUP Eduardo Soriano García

Eduardo Soriano García Ashraf Muhaisen Cristina Roselló Busquets Full professor. Principal Postdoctoral fellow PhD student investigatorl Serena Mirra Marc Hernaiz Llorens Marta Pascual Sánchez Postdoctoral fellow PhD student Associate Professor. Researcher Daniela Rossi Irene Lobón García Jesús Mariano Ureña Bares Postdoctoral fellow PhD student Associate Professor. Researcher Yasmina Manso Sanz Alba del Valle Vilchez Ferran Burgaya Márquez Postdoctoral fellow Acosta Associate Professor. Researcher PhD student Mónica Pardo Muñoz Fausto Alexánder Ulloa Postdoctoral fellow Marco Solís Benites Darquea Office manager Associate Professor. Researcher Jonatan Dorca Arévalo Postdoctoral fellow Núria Masachs Janoher Tiziana Cotrufo PhD student Postdoctoral fellow Antoni Parcerisas Mosqueda Pablo Barrecheguren Lluís Pujadas Postdoctoral fellow Manero Puigdomènech PhD student Postdoctoral fellow

Contact details

Universidad de Barcelona Facultad de Biología. Departamento de Biología Celular Avenida Diagonal 643, Edificio Prevosti 1r piso Tel.: +34 93 403 71 17 Fax: +34 93 403 71 16 [email protected]

2015 CIBERNED Annual Report / 87 Summary

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

• If the Reelin improves AD in mice, once • Integrating these data in defined the disease has developed, and in mice molecular pathways through systems that exhibit AD pathology related to Tau biology; and (Tau-GSK3 WLW -OE and mice); • We have begun to understand how • Whether a downregulation of Reelin specific somatic genetic events SAD specifically in the adult (using conditional may contribute to the pathogenesis KO mice, FlReelin / AD mice) accelerates of the disease. We believe these data AD pathology; reveal important new molecular and genetic information to understand the • Started to understand the physiological mechanisms that trigger the SAD and to and molecular mechanisms by which open new therapeutic avenues.

Keywords

Reelin, Synapses, Alzheimer disease, somatic mutations

Publications

• Calero M., Gomez-Ramos A., Calero O., Soriano E., Avila J., Medina M. Additional mechanisms conferring genetic susceptibility to Alzheimer’s disease. Frontiers in Cellular Neuroscience. 2015;9(APR). [Epub ahead of print].

• Ulloa F., Gonzalez-Junca A., Meffre D., Barrecheguren P.J., Martinez-Marmol R., Pazos I. et al. Blockade of the SNARE protein syntaxin 1 inhibits glioblastoma tumor growth. PLoS ONE. 2015; 10(3). [Epub ahead of print].

2015 CIBERNED Annual Report / 88 Program 1 Group: Eduardo Soriano García

• Gomez-Ramos A., Podlesniy P., Soriano E., Avila J. Distinct X-chromosome SNVs from some sporadic AD samples. Scientific Reports. 2015; 5. [Epub ahead of print]. Bosch C, Martínez A, Masachs N, Teixeira CM, Fernaud I, Ulloa F et al. FIB/SEM technology • and high-throughput 3D reconstruction of dendritic spines and synapses in GFP-labeled adult- generated neurons.Frontiers in neuroanatomy. 2015;9: 60. • Ros O., Cotrufo T., Martinez-Marmol R., Soriano E. Regulation of patterned dynamics of local exocytosis in growth cones by Netrin-1. Journal of Neuroscience. 2015;35(13):5156-5170. Martínez-Mármol R, Comes N, Styrczewska K, Pérez-Verdaguer M, Vicente R, Pujadas L et al. • Unconventional EGF-induced ERK1/2-mediated Kv1.3 endocytosis.Cellular and molecular life sciences: CMLS. 2015. [Epub ahead of print].

Research projects

• Code: 2014 SGR 1300. • Code: PI2013_08-1. Title: Neurobiologia del Desenvolupament i Title: La dinámica mitocondrial y mitofagia Regeneració Neuronal (SGR 2014-2016). como dianas terapeuticas en las Principal Investigator: Eduardo Soriano Enfermedades de Parkinson y Huntington. García. Principal Investigator: Eduardo Soriano. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G408, G301, G207, G410, Other CIBER’s collaboration: No. G109. Type: Private. Other CIBER’s collaboration: No. Funding agency: Agència d’Ajuts Universitari Type: National. i de Recerca (AGAUR)- Proyectos de Funding agency: Instituto de Salud Carlos III. investigación para potenciar los grupos de Funding: 300.000 €. investigación consolidados. Duration: 2013-2015 Funding: 30.000 €. Duration: 2014-2016 • Code: 613/C/2013. Title: Lipotoxicity, hepatic steatosis and • Code: PI12/02108. hepatocarcinoma: Role of the ARMC10/ Title: Participación de las Semaforinas ARMCX family of mitochondrial proteins. transmembranales y la cinasa Fak en Principal Investigator: Eduardo Soriano. patologías relacionadas con la sinapsis y la CIBERNED’s collaboration: No. actividad neuronal. CIBERNED groups: Principal Investigator: Ferrán Burgaya Other CIBER’s collaboration: Si (CIBER Marquez. Fisiopatologia de la Obesidad y Nutricion). CIBERNED’s collaboration: No. Type: National. CIBERNED groups: Funding agency: Fundació la Marato de Other CIBER’s collaboration: No. TV3. Type: International. Funding: 318.950 €. Funding agency: Instituto de Salud Carlos III. Duration: 2014-2016 Funding: 56.500 €. Duration: 2013-2015 • Code: SAF2013-42445-R. Title: Nuevas aproximaciones a la patogénesis • Code: 20143330-31. de la enfermedad de Alzheimer. Title: The role of reelin at the crossroads of Principal Investigator: Eduardo Soriano alzheimer's disease mechanisms: Tauopathy, Garcia. amyloid toxicity and transmissibility. CIBERNED’s collaboration: No. Principal Investigator: Lluis Pujadas CIBERNED groups: Puigdomenech. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: International. CIBERNED groups: Funding agency: Ministerio de Economía y Other CIBER’s collaboration: No. Competitividad. Type: Private. Funding: 496.100 €. Funding agency: Fundació la Marato de Duration: 2015 TV3. Funding: 187.635,5 €. Duration: 2015-2018

2015 CIBERNED Annual Report / 89 409 GROUP Ignacio Torres Alemán

Ignacio Torres Alemán Laura Genis Ángel Trueba Principal investigator Researcher Researcher

Manuel Domínguez Dolores Guinea Angélica Stein Researcher Researcher Researcher

Ana M. Fernández Víctor Munive James Knight Researcher Researcher Researcher

Miguel García Andrea Santi Researcher Researcher

Contact details

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

2015 CIBERNED Annual Report / 90 Summary

Our group is interested in the role played by this vein, we keep analyzing the role of IGF-I insulin peptides in the adult brain. We must in mood control and we aim to establish a link analyze a wide variety of processes as the with pathologies such as post-traumatic stress actions of these peptides are multifaceted. disorder in collaboration with a clinical study During 2015 we completed analysis of the with a very good sample size. essential mechanisms underlying control of brain glucose metabolism by IGF-I and insulin. Further, we continue studying mechanisms of In brief, we have found that the IGF-I receptor serum IGF-I entrance into the brain in pathology plays a dual role whereby glucose transporter and in the normal brain. Our observations 1, the main facilitative transporter at the blood- support the use of IGF-I for treatment of acute brain-barrier, is regulated through a concerted conditions (trauma or ischemia) presenting with action of IGF-I and insulin. low IGF-I levels. In relation to neuro-inflammatory processes associated to Alzheimer´s disease we We have also characterized gender differences have developed a proof-of-concept study that in mood homeostasis where an interaction of shows a new treatment addressing astrocytes IGF-I with gonadal hormones is involved. Our that ameliorates amyloid beta neurotoxicity. data indicate that the high incidence of mood We are also determining the relationship of co-morbidities in Alzheimer patients may be this disease with diabetes in collaboration with related to these mechanisms, which appear other labs. disturbed early at pre-menopausic stages. In

Keywords

Role of Insulin like growth factors in neuronal plasticity and degenerative diseases, therapeutic targets in neurodegeneration, the aging brain and cognition, blood brain barriers

Publications

• Trueba A, Torres-Aleman I. Insulin-like peptides signaling in Alzheimer’s disease: on the road to alternative therapeutics . Current Opinion in Behavioral Sciences. 2016. 9:15-25. Epub 2015.

Research projects

• Code: PI2013/01. • Code: SAF2013-40710-R. Title: Propiedades emergentes de la Title: Interacción del receptor IGF-I con co- relación neurona-glia que subyacen a receptores y su impacto en la demencia de neurodegeneración y demencia en la alzhéimer. enfermedad de Alzheimer. Principal Investigator: Ignacio Torres Principal Investigator: Ignacio Torres- Alemán. Alemán. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G415, G413, G204, G409, Other CIBER’s collaboration: No. G108, G411. Type: National. Other CIBER’s collaboration: No. Funding agency: Ministerio de Economía y Type: National. Competitividad. Funding agency: CIBERNED. Funding 387.000 €. Funding: 350.000 €. Duration: 2015 Duration: 2013-2015

2015 CIBERNED Annual Report / 91 • Code: S2010_BMD. • Code: Inter-Cibers. Title: Redes de senalización y vías efectoras Title: Molecular links between diabetes and en modelos celulares y animales de neurodegenerative disorders. enfermedades neurodegenerativas. Principal Investigator: Ángel Raya. Principal Investigator: José González CIBERNED’s collaboration: Yes. Castaño. CIBERNED groups: G409, G604. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: Yes CIBERNED groups: G111, G401, G104, G307, (Metabolismo). G409, G412. Type: National. Other CIBERs’s collaboration: Yes (CIBERER). Funding agency: Instituto de Salud Carlos III. Type: CC.AA. Funding: 660.000 €. Funding Agency: Comunidad de Madrid. Duration: 2015 Funding: 483.000 €. Duration: 2012-2016

PhD dissertations

• Author: Ángel Trueba Saiz. Title: Neurotrophic uncoupling of IGF-1 in Alzheimer´s disease: translation into early diagnosis and involvement of lifestyle risk factors. Date: 4th February 2015. Supervisor: Ignacio Torres Alemán.

2015 CIBERNED Annual Report / 92 410 GROUP Ramón Trullás Oliva

Ramón Trullás Oliva Manuel Rodríguez Nuria Serra CSIC Research Professor-Principal Assistant Professor Technician Investigator Petar Podlesniy Laura Mañas Nicole Mahy Research Scientist Technician Professor of Biochemistry Margalida Puigròs Predoctoral student

Contact details

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]

2015 CIBERNED Annual Report / 93 Summary

During 2015 we have continued to investigate for Neurodegenerative Diseases (DZNE) of the mechanisms that regulate mitochondrial Gottingen in Germany and the Department of dynamics in neurons. The results we have Molecular Pathology and Neuropathology of obtained within this period show that the Medical University of Lodz in Poland, we have presence of Neuronal Pentraxin 1 (NP1) investigated the CSF content of mtDNA in two is absolutely required for the processes of populations of patients diagnosed with sporadic mitochondrial fusion and fission. We have Creutzfeldt-Jakob disease, with dementia of also observed that the oligomers of beta the Alzheimer type or with dementias of other amyloid peptide (Ab1-42) cause mitochondrial types. The results confirm again that patients fragmentation in neurons and that this toxic with Alzheimer-type dementia have a decrease effect of Ab1-42 disappears in the absence of of mtDNA content in the CSF. By contrast, in NP1. patients with other types of dementia or with Creutzfeldt-Jakob disease the concentration We have also continued to investigate the of mtDNA in CSF is unchanged. These results mechanisms that regulate the concentration indicate that mtDNA concentration in CSF is of circulating extracellular mitochondrial DNA not a consequence of neuronal damage and (mtDNA) in cerebrospinal fluid (CSF) in several provide further evidence that confirms the neurodegenerative diseases. hypothesis that the decrease of mtDNA in CSF is a fundamental pathophysiological mechanism In collaboration with the German Center in Alzheimer’s disease.

Keywords

Synaptic neurodegeneration, Alzheimer’s disease biomarkers, apoptosis, mitochondrial dynamics, mitochondrial transport in neurodegeneration, neuronal death, neuronal pentraxins, molecular mechanisms of Alzheimer’s disease

Publications

• Popik P, Holuj M, Nikiforuk A, Kos T, Trullas R, Skolnick P. 1-aminocyclopropanecarboxylic acid (ACPC) produces procognitive but not antipsychotic-like effects in rats.Psychopharmacology. 2015;232(6): 1025-38. • Neus Bosch M., Pugliese M., Andrade C., Gimeno-Bayon J., Mahy N., Rodriguez M.J. Amyloid-β immunotherapy reduces amyloid plaques and astroglial reaction in aged domestic dogs. Neurodegenerative Diseases. 2015;15(1):24-37. • Batlle M., Ferri L., Andrade C., Ortega F.-J., Vidal-Taboada J.M., Pugliese M. et al. Astroglia- microglia cross talk during neurodegeneration in the rat hippocampus. BioMed Research International. 2015;2015. [Epub ahead of print]. • Figueiro-Silva J., Gruart A., Clayton K.B., Podlesniy P., Abad M.A., Gasull X. et al. Neuronal pentraxin 1 negatively regulates excitatory synapse density and synaptic plasticity. Journal of Neuroscience. 2015;35(14):5504-5521. • Xifró X, Vidal-Sancho L, Boadas-Vaello P, Turrado C, Alberch J, Puig T et al. Novel epigallocatechin- 3-gallate (EGCG) derivative as a new therapeutic strategy for reducing neuropathic pain after chronic constriction nerve injury in mice.PloS one. ;10(4): e0123122. • Espinosa-Parrilla J.F., Martinez-Moreno M., Gasull X., Mahy N., Rodriguez M.J. The L-type voltage- gated calcium channel modulates microglial pro-inflammatory activity. Molecular and Cellular Neuroscience. 2015;64:104-115. • Vidal-Taboada J.M., Lopez-Lopez A., Salvado M., Lorenzo L., Garcia C., Mahy N. et al. UNC13A confers risk for sporadic ALS and influences survival in a Spanish cohort. Journal of Neurology. 2015;262(10):2285-2292.

2015 CIBERNED Annual Report / 94 Program 1 Group: Ramón Trullás Oliva

• Gomez-Ramos A., Podlesniy P., Soriano E., Avila J. Distinct X-chromosome SNVs from some sporadic AD samples. Scientific Reports. 2015;5. [Epub ahead of print].

Research projects

• Code: LRRK2 Resource RFA 2013. • Code: IPT-2012-0614-010000. Title: Alpha-synuclein and other biomarkers Title: Nuevas aproximaciones terapéuticas in biological samples of LRKK2 PD. basadas en derivados de ácidos grasos Principal Investigator: Eduardo Tolosa. esenciales dirigidos a enfermedades CIBERNED’s collaboration: Yes. oncológicas y neurológicas no cubiertas. CIBERNED groups: G410, G207. Principal Investigator: Manuel José Other CIBER’s collaboration: No. Rodríguez-Allue. Type: International. CIBERNED’s collaboration: No. Funding agency: Michael J. Fox Foundation. CIBERNED groups: Funding: 60.000 €. Other CIBER’s collaboration: No. Duration: 2014-2016 Type: National. Funding agency: Ministerio de Economía y • Code: SAF2014-56644-R. Competividad, Programa INNPACTO. Title: Mecanismos de regulacion del número Funding: 189.523 €. de copias de ADN mitocondrial para el Duration: 2012-2015 tratamiento del Alzheimer. Principal Investigator: Ramón Trullás. • Code: PI2013_08-1. CIBERNED’s collaboration: No. Title: La dinámica mitocondrial y mitofagia CIBERNED groups: como dianas terapéuticas en las Other CIBER’s collaboration: No. enfermedades de Parkinson y Huntington. Type: National. Principal Investigator: Eduardo Soriano. Funding agency: Ministerio de Economía y CIBERNED’s collaboration: Yes. Competitividad, Secretaría de Estado de CIBERNED groups: G408, G301, G207, G410, Investigación. G109. Funding: 160.000 €. Other CIBER’s collaboration: No. Duration: 2015-2017 Type: National. Funding agency: Instituto de Salud Carlos III. Funding: 300.000 €. Duration: 2013-2015

2015 CIBERNED Annual Report / 95 411 GROUP Javier Vitorica Ferrández

Javier Vitorica Ferrández Victoria Navarro Garrido Sebastián Jiménez Muñoz Full Professor and Principal FPI fellow Technician Investigator MªVirtudes Sánchez Mica Mª Luisa García-Calvo Marisa Vizuete Chacón FPU fellow Technician Associate professor

Contact details

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]

2015 CIBERNED Annual Report / 96 Summary

During 2015, we have continued with the pharmacologically (immunosuppression) or characterization of the inflammatory response genetically (IL-4 KO, Galectin3 KO; DREAM in postmortem human samples classified using KO). We are now analyzing the impact of the Braak classification, from BraakII to Braak V-VI microglial manipulation on the pathology (in collaboration with Dr. A. Gutierrez, University (Abeta accumulation, tau phosphorylation, of Malaga ). Braak V-VI group were patients neurodegeneration) of the transgenic models. diagnosed as Alzheimer’s type dementia. On the other hand, we have continues with This activity is part of the objectives of the the characterized synaptic toxicity of Abeta collaborative project CIBERNED we have been plaques in animal models. Initially, we studied developing during 2013-2015. Within this context, in vitro the effect of various soluble oligomeric we have isolated and characterized the soluble forms, using primary neuronal cultures. fractions (extracellular and/or cytosolic) of various human samples. The presence of soluble During this year we have also continue the Abeta and/or tau forms was analyzed by collaborative project (coordinated by Dr. E western blots, immunoprecipitations and ELISAs. Galea) funded by the TV3 on the properties of We have also analyzed, in vitro, the toxicity and astrocytes and their role in the development of /or immunogenicity of these fractions. Alzheimer’s pathology. On the other hand, we are currently Finally, for the characterization of the PS1/APP determined the role of the microglial activation transgenic models, we have established a in the development of Alzheimer’s pathology collaboration with Dr. JL Gómez-Ariza (University using APP or PS1xAPP models. Specifically, of Huelva) for a metabolomic study in transgenic the microglial response in the models is mice PS1xAPP. being manipulated (inhibited or enhanced)

Keywords

Alzheimer’s disease, neurodegeneration, neuroprotection, inflammation, Abeta, oligomers, transgenic models, neuropathology

Publications

• Carriba P., Jimenez S., Navarro V., Moreno-Gonzalez I., Barneda-Zahonero B., Moubarak R.S. et al. Amyloid-β reduces the expression of neuronal FAIM-L, thereby shifting the inflammatory response mediated by TNFα from neuronal protection to death. Cell Death and Disease. 2015;6(2) [Epub ahead of print]. • González-Domínguez R, García-Barrera T, Vitorica J, Gómez-Ariza JL. Application of metabolomics based on direct mass spectrometry analysis for the elucidation of altered metabolic pathways in serum from the APP/PS1 transgenic model of Alzheimer’s disease.Journal of pharmaceutical and biomedical analysis. 2015;107: 378-85. • Gonzalez-Dominguez R., Garcia-Barrera T., Vitorica J., Gomez-Ariza J.L. Deciphering metabolic abnormalities associated with Alzheimer’s disease in the APP/PS1 mouse model using integrated metabolomic approaches. Biochimie. 2015;110:119-128. • Cardona C., Sanchez-Mejias E., Davila J.C., Martin-Rufian M., Campos-Sandoval J.A., Vitorica J. et al. Expression of Gls and Gls2 glutaminase isoforms in astrocytes. GLIA. 2015;63(3):365-382. • Gonzalez-Dominguez R., Garcia-Barrera T., Vitorica J., Gomez-Ariza J.L. High throughput multiorgan metabolomics in the APP/PS1 mouse model of Alzheimer’s disease. Electrophoresis. 2015;36(18):2237-2249. • Gonzalez-Dominguez R., Garcia-Barrera T., Vitorica J., Gomez-Ariza J.L. Metabolomic investigation of systemic manifestations associated with Alzheimer’s disease in the APP/PS1 transgenic mouse model. Molecular BioSystems. 2015;11(9):2429-2440.

2015 CIBERNED Annual Report / 97 • Gonzalez-Dominguez R., Garcia-Barrera T., Vitorica J., Gomez-Ariza J.L. Metabolomic screening of regional brain alterations in the APP/PS1 transgenic model of Alzheimer’s disease by direct infusion mass spectrometry. Journal of Pharmaceutical and Biomedical Analysis. 2015;102:425- 435. • Gonzalez-Dominguez R., Garcia-Barrera T., Vitorica J., Gomez-Ariza J.L. Metabolomics reveals significant impairments in the immune system of the APP/PS1 transgenic mice of Alzheimer’s disease. Electrophoresis. 2015;36(4):577-587. • Heneka M.T., Carson M.J., Khoury J.E., Landreth G.E., Brosseron F., Feinstein D.L. et al. Neuroinflammation in Alzheimer’s disease. The Lancet Neurology. 2015;14(4):388-405.

Research projects

• Code: PI12/01439. • Code: PI2013/01. Title: Oligómeros tóxicos del Abeta como Title: Propiedades emergentes de la agentes causantes de la disfunción del relación neurona-glia que subyacen a citoesqueleto y los procesos proteolíticos en neurodegeneración y demencia en la la enfermedad de Alzeimer. enfermedad de Alzheimer. Principal Investigator: Javier Vitorica. Principal Investigator: Ignacio Torres- CIBERNED’s collaboration: Yes. Aleman. CIBERNED groups: G411, G415. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G415, G413, G204, G409, Type: National. G108, G411. Funding agency: Instituto de Salud Carlos III. Other CIBER’s collaboration: No. Funding: 228.629 €. Type: National. Duration: 2013-2015 Funding agency: CIBERNED. Funding: 350.000 €. • Code: CTS 2035. Duration: 2013-2015 Title: Oligomerización y toxicidad de los peptidos de Abeta: búsqueda de nuevas • Code: 20141431. dianas de interes terapéutico en la Title: Deciphering the link between astrocyte enfermedad de Alzheimer. reactivity and neuronal damage in Principal Investigator: Javier Vitorica. Alzheimer´s disease”. CIBERNED’s collaboration: Si. Principal Investigator: Elena Galea. CIBERNED groups: G411, G415. CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G411, G413, G415. Type: CC.AA. Other CIBER’s collaboration: No. Funding agency: Junta de Andalucía. Type: Private. Funding: 289.307,5 €. Funding agency: La Marató de TV3. Duration: 2014-2016 Funding: 382.250 €. Duration: 2015-2017

PhD dissertations

• Author: Victoria Navarro Garrido. Title: Caracterización de la respuesta inflamatoria en tejido cerebral post-mortem de pacientes de alzhéimer. Date: 11th February 2015. Supervisor: Francisco Javier Vitorica Ferrández.

2015 CIBERNED Annual Report / 98 412 GROUP Francisco Wandosell Jurado

Francisco Wandosell María José Pérez Álvarez María José Benítez Jurado Prof. UAM Associate Professor.UAM CSIC Staff Scientist- Principal Investigator Juan José Garrido Jurado Lara Ordóñez CSIC Staff Scientist-Researcher Postdoctoral Fellow Inés M. Antón CSIC Staff Scientist-Researcher Irene Benito PhD student

Contact details

Centro de Biología Molecular “Severo Ochoa” CSIC-UAM & CIBERNED 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] http://www.ciberned.es/grupofranciscowandosell.aspx

2015 CIBERNED Annual Report / 99 Summary

Our group, “Molecular mechanisms of the activation of PI3K-Akt pathway, at least Neurodegeneration “, is generated from an in part (CoIPs: Dr. J.J.Garrido & F Wandosell). established line in the CBM over several years. At Second, we are analysing the role of the Akt present, this CIBERNED line is formally composed downstream element, mTORC1. The kinase by three sub-groups (IP´s-F. Wandosell (CBM), I. activity of this protein complex control, at Anton (CNB) and J.J. Garrido, Cajal Inst.). least in part, protein synthesis and autophagy; whereas its dysfunction has been considering Our group mostly interested in a key factor of beta amyloid generation and/ neurodegenerative disorders is now focusing or degradation in Alzheimer disease (IPs: Dr. F in a group of brain disorders associated with Wandosell). Third, we are analysing some Akt aging, such as Alzheimer, disease, Stroke and upstream and downstream elements (such some type of brain tumours. as FoxO and Bim), and our data indicated that they are responsible of the cell division of Our work for several years had been focused on cancer-stem cells and the maintenance of its the signalling pathway of IGF1/Insulin-PI3K-Akt, phenotype. Our work is now focusing on the in neuronal differentiation, degeneration and conversion from astrocyte-astrocytoma-glioma more recently in cell growth. It is well known that recently defined the role new Akt-downtream in neuronal development IGF-1/Insulin through elements and how these elements regulated their tyrosine kinase receptors controls the glial cell growth (CoIPs: Drs. F. Wandosell & I. activity of Class1 phosphatidylinositol 3-kinase Anton). (PI3K) and several serine-threonine kinases such as Akt. Our initial studies were focused on GSK3 In summary, we are focusing on track PI3K- and in Akt, we defined the role of GSK3 and Akt Akt signalling and elements that control some isoforms in the neuronal morphogenesis and physiological process, from cell division to how they are modified after Ischemia. Besides, differentiation, and how some of these proteins we described how the neuroprotective role are modified in pathology. of neurosteroids, such as estradiol, is due to

Keywords

Neurodegeneration, cell signaling, neuroprotection, estrogen, Alzheimer’s disease, neuritogenesis, axonal polarity glia growth, astrocyte conversion / glioma, cytoskeleton, actin

Publications

• Del Puerto A., Fronzaroli-Molinieres L., Perez-Alvarez M.J., Giraud P., Carlier E., Wandosell F. et al. ATP-P2X7 Receptor Modulates Axon Initial Segment Composition and Function in Physiological Conditions and Brain Injury. Cerebral Cortex. 2015;25(8):2282-2294. • Gargini R, Cerliani JP, Escoll M, Antón IM, Wandosell F. Cancer stem cell-like phenotype and survival are coordinately regulated by Akt/FoxO/Bim pathway.Stem cells (Dayton, Ohio). 2015;33(3): 46-60. • Perez-Alvarez MJ, Mateos L, Alonso A, Wandosell F. Estradiol and Progesterone Administration After pMCAO Stimulates the Neurological Recovery and Reduces the Detrimental Effect of Ischemia Mainly in Hippocampus.Molecular neurobiology. 2015;52(3): 1690-703. • Franco-Villanueva A., Wandosell F., Anton I.M. Neuritic complexity of hippocampal neurons depends on WIP-mediated mTORC1 and Abl family kinases activities. Brain and Behavior. 2015;5(11). [Epub ahead of print]. • Ordonez-Gutierrez L., Anton M., Wandosell F. Peripheral amyloid levels present gender differences associated with aging in AβPP/PS1 mice. Journal of Alzheimer's Disease. 2015;44(4):1063-1068. • Ordóñez-Gutiérrez L, Re F, Bereczki E, Ioja E, Gregori M, Andersen AJ et al. Repeated intraperitoneal injections of liposomes containing phosphatidic acid and cardiolipin reduce amyloid-β levels in APP/PS1 transgenic mice.Nanomedicine : nanotechnology, biology, and medicine. 2015;11(2): 421-30. • Cabrera JR, Viejo-Borbolla A, Martinez-Martín N, Blanco S, Wandosell F, Alcamí A. Secreted herpes simplex virus-2 glycoprotein G modifies NGF-TrkA signaling to attract free nerve endings to the site of infection.PLoS pathogens. 2015;11(1): e1004571.

2015 CIBERNED Annual Report / 100 Program 1 Group: Francisco Wandosell Jurado

• Mancini S, Minniti S, Gregori M, Sancini G, Cagnotto A, Couraud PO et al. The hunt for brain Aβ oligomers by peripherally circulating multi-functional nanoparticles: Potential therapeutic approach for Alzheimer disease.Nanomedicine : nanotechnology, biology, and medicine. 2016;12(1). Epub 2015. • Reginensi D., Carulla P., Nocentini S., Seira O., Serra-Picamal X., Torres-Espin A. et al. Increased migration of olfactory ensheathing cells secreting the Nogo receptor ectodomain over inhibitory substrates and lesioned spinal cord. Cellular and Molecular Life Sciences. 2015;72(14):2719- 2737. • Vergara C, Ordóñez-Gutiérrez L, Wandosell F, Ferrer I, del Río JA, Gavín R. Role of PrP(C) Expression in Tau Protein Levels and Phosphorylation in Alzheimer's Disease Evolution.Molecular neurobiology. 2015;51(3): 1206-20.

Research projects

• Code: S2010_BMD. Title: Redes de senalización y vías efectoras en modelos celulares y animales de enfermedades neurodegenerativas. Principal Investigator: José González Castaño. CIBERNED’s collaboration: Yes. CIBERNED groups: G111, G401, G104, G307, G409, G412. Other CIBER’s collaboration: Yes (CIBERER). Type: CC.AA. Funding agency: Comunidad de Madrid. Funding: 483.000 €. Duration: 2012-2016

2015 CIBERNED Annual Report / 101 Program 2 Parkinson’s disease and other neurodegenerative movement disorders

Alberch Vie, Jordi...... 109 López Barneo, José...... 174 Canela Campos, Enric Isidre...... 113 López de Munain Arregui, Adolfo...... 179 Ceña Callejo, Valentín...... 118 Lucas Lozano, José Javier...... 186 Combarros Pascual, Onofre...... 121 Mena Gómez, Mª Angeles...... 189 Cuadrado Pastor, Antonio...... 129 Moratalla Villalba, Rosario...... 191 Fariñas Gómez, Isabel...... 133 Muñoz Cánoves, Pura...... 195 Fernández Chacón, Rafael...... 136 Naranjo Orovio, José Ramón...... 198 Fernández Ruiz, Javier...... 139 Navarro Acebes, Xavier...... 201 Fuentes Rodríguez, José Manuel...... 144 Obeso Inchausti, José Ángel...... 207 García Verdugo, José Manuel...... 148 Pérez Castillo, Ana María...... 211 González Castaño, José...... 152 Pérez Tur, Jordi...... 215 Guzmán Pastor, Manuel...... 154 del Río Fernández, José Antonio...... 217 Iglesias Vacas, Teresa...... 158 Rodríguez Díaz, Manuel...... 220 Kulisevsky Bojarski, Jaime...... 161 Tolosa Sarró, Eduardo...... 223 Labandeira García, José Luis...... 167 Vicario Abejón, Carlos...... 229 Lanciego Pérez, José Luis...... 171 Vila Bover, Miquel...... 232 Program 2 Parkinson’s disease and other neurodegenerative movement disorders

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

CIBERNED’s Program 2 brings together 27 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 70,000 people in Spain, a figure that is expected to be increasing due to the +70.000 progressive aging of the population. EP patients

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

a) Key aspects related to the ethiopathogenesis in PD. b) Physiopatological mechanisms related to disease onset and progression. c) Development of symptomatic treatments, especially neuroprotective and curative. Thus, a truly translational research is sought, having the disease and the patients as the main targets.

De esta manera, se busca conseguir una verdadera investigación de carácter traslacional, cuyos principales objetivos sean la enfermedad y el paciente.

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.

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

2015 CIBERNED Annual Report / 105 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 (QMA) 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.

2015 CIBERNED Annual Report / 106 Principal investigator Institution

Alberch Vié, Jordi University of Barcelona Canela Campos, Enric Isidre Marqués de Valdecilla Research Institute, Santander Ceña Callejo, Valentín University of Barcelona Combarros Pascual, Onofre 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 Biomedic al Research Institute CSIC-UAM, Madrid 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 Mena Gómez, Mª Ángeles Ramón y Cajal University Hospital, 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. José J. Lucas (Center for Molecular Biology, CSIC, Madrid), Eduardo Tolosa (Clinic Hospital, Barcelona), Adolfo López de Munain Arregui (Biodonostia Research Institute, San Sebastián), Rafael Fernández Chacón* (University of Seville) y Teresa Iglesias Vacas* (Biomedical Research Institute CSIC-UAM, Madrid).

*Since Septembre 2015.

2015 CIBERNED Annual Report / 107 301 GROUP Jordi Alberch Vié

Jordi Alberch Vié Xavier Xifró Collsamata Núria Suelves Caballol Laura Clua Full Professor Postdoctoral researcher PhD student Degree´s student Josep M Canals Coll Rafael Alcalá Vida Laura Vidal Sancho Marc Espina Assistant Professor PhD student PhD student Degree´s student Silvia Ginés Padrós Elena Álvarez Periel Pedro Belio Pere Llull Assistant Professor PhD student Master’s student Degree´s student

Esther Pérez Navarro Marta Cherubini Beatriz Blasco Elena Meléndez Assistant Professor PhD student Master’s student Degree´s student Glòria Blázquez Andrea Comella Bolla Sílvia Cases Georgina Bombau Postdoctoral researcher PhD student Master’s student Martínez Personal técnico Verónica Brito Jordi Creus Muncunill Marta García Postdoctoral researcher PhD student Master’s student Jordi Carrere Molina Technician Raquel Martín Ibáñez Sara Fernández García David López Postdoctoral researcher PhD student Master’s student Ester González Guerrero Technician Mercè Massana Alfonso Gerardo García Luis Marte Postdoctoral researcher Díaz Barriga Master’s student Cristina Herranz Sotoca PhD student Technician Andrés Míguez González Sara Martínez Postdoctoral researcher Inés Guardia Pena Master’s student Ana López Alonso PhD student Technician Ana Cristina Saavedra Blanca Peguera Postdoctoral researcher Andreas Mezger Master’s student Mª Teresa Muñoz PhD student Technician Phil Sanders Raquel Badillos Postdoctoral researcher Mónica Pardo Muñoz Degree´s student Unai Perpiñá Martín PhD student Technician Marco Straccia Carla Castellar Postdoctoral researcher Degree´s student

Contact details

Departament de Biologia Cellular, Inmunologia i Neurociències 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]

2015 CIBERNED Annual Report / 108 Program 2 Group: Jordi Alberch Vié

Summary

Our goal is to identify new molecular mechanisms the cerebral cortex but not in the striatum that affected by mutant huntingtin (mhtt) to develop might explain the deficiencies in the cortico- treatments that stop or slow the progression of striatal transmission characteristic of this disease. Huntington’s disease (HD). During 2015 we have (Puigdellivol et al., 2015). identified three pharmacological treatments In addition to studies related to HD, we were that improve cognitive function in the R6/1 also interested in the analysis of mechanisms mouse model of HD: (1) the simultaneous that could modulate STEP (Striatal-enriched antagonism of D1 and A2A receptors (Tyebji protein tyrosine phosphatase) protein levels. We et al, 2015), (2) the administration of fingolimod have described a reciprocal regulation of STEP (Míguez et al., 2015) and (3) the administration and BDNF: BDNF induces the degradation STEP of misoprostol, an activator of prostaglandin through proteasome (Saavedra et al., 2015) EP2 receptors (Anglada- Huguet et al., 2015). and, in turn, STEP regulates BDNF expression We have also defined two new targets (Xu et al., 2015). To model HD by using human participating in the major vulnerability of striatal cells, in collaboration with Cardiff University, we neurons to mhtt-induced toxicity. We have stablished a differentiation protocol to obtain determined the role that RTP801 protein plays striatal neurons that show electrophysiological in striatal cell death induced by mhtt (Martin- functionality from human pluripotent stem cells Flores et al., 2015) and a new role of Cdk5 in (Telezhkin et al., 2015). We have also developed dopamine-mediated toxicity by modulating a system for gene characterization through the mitochondrial fragmentation (Cherubini et al, OpenArray technique that allows us to monitor 2015 ). Finally, we have determined a critical both the human striatum development and role of cortical pathology in the cognitive the processes of differentiation of human cells impairment occurring in HD. We have shown (Straccia et al., 2015). very early alterations of the Kalirin protein in

Keywords

BDNF, cognition, motor, kinases, phosphatases, neurogenesis, regenerative medicine, iPSCs Control

Publications

• Puigdellívol M, Cherubini M, Brito V, Giralt A, Suelves N, Ballesteros J et al. A role for Kalirin-7 in corticostriatal synaptic dysfunction in Huntington’s disease.Human molecular genetics. 2015;24(25): 7265-85. • Fernandez-Santiago R., Carballo-Carbajal I., Castellano G., Torrent R., Richaud Y., Sanchez- Danes A. et al. Aberrant epigenome in iPSC-derived dopaminergic neurons from Parkinson’s disease patients. EMBO Molecular Medicine. 2015. [Epub ahead of print]. • Saavedra A., Puigdellivol M., Tyebji S., Kurup P., Xu J., Gines S. et al. BDNF Induces Striatal- Enriched Protein Tyrosine Phosphatase 61 Degradation Through the Proteasome. Molecular Neurobiology. 2015. [Epub ahead of print]. • Cherubini M., Puigdellivol M., Alberch J., Gines S. Cdk5-mediated mitochondrial fission: A key player in dopaminergic toxicity in Huntington’s disease. Biochimica et Biophysica Acta - Molecular Basis of Disease. 2015;1852:2145-2160. • Puigdellivol-Sanchez A., Giralt A., Casanovas A., Alberch J., Prats-Galino A. Cryostat Slice Irregularities May Introduce Bias in Tissue Thickness Estimation: Relevance for Cell Counting Methods. Microscopy and Microanalysis. 2015;21(4):893-901. • Xu J, Kurup P, Azkona G, Baguley TD, Saavedra A, Nairn AC et al. Down-regulation of BDNF in cell and animal models increases striatal-enriched protein tyrosine phosphatase 61 (STEP 61) levels.Journal of neurochemistry. 2015. [Epub ahead of print]. • Miguez A, García-Díaz Barriga G, Brito V, Straccia M, Giralt A, Ginés S et al. Fingolimod (FTY720) enhances hippocampal synaptic plasticity and memory in Huntington’s disease by preventing p75NTR up-regulation and astrocyte-mediated inflammation.Human molecular genetics. 2015;24(17): 4958-70.

2015 CIBERNED Annual Report / 109 • Tyebji S., Saavedra A., Canas P.M., Pliassova A., Delgado-Garcia J.M., Alberch J. et al. Hyperactivation of D1 and A2A receptors contributes to cognitive dysfunction in Huntington’s disease. Neurobiology of Disease. 2015;74:41-57. • Pont L, Benavente F, Jaumot J, Tauler R, Alberch J, Ginés S et al. Metabolic profiling for the identification of huntington biomarkers by on-line solid-phase extraction capillary electrophoresis mass spectrometry combined with advanced data analysis tools.Electrophoresis. 2015. [Epub ahead of print]. • Anglada-Huguet M, Vidal L, Giralt A, García-Díaz Barriga G, Xifró X, Alberch J. Prostaglandin E2 EP2 activation reduces memory decline in R6/1 mouse model of Huntington’s disease by the induction of BDNF-dependent synaptic plasticity.Neurobiology of disease. 2015. [Epub ahead of print]. • Straccia M, Garcia-Diaz Barriga G, Sanders P, Bombau G, Carrere J, Mairal PB et al. Quantitative high-throughput gene expression profiling of human striatal development to screen stem cell-derived medium spiny neurons.Molecular therapy. Methods & clinical development. ;2: 15030. • Martin-Flores N., Romani-Aumedes J., Rue L., Canal M., Sanders P., Straccia M. et al. RTP801 Is Involved in Mutant Huntingtin-Induced Cell Death. Molecular Neurobiology. 2015. [Epub ahead of print]. • Herranz C., Fernandez F., Martin-Ibanez R., Blasco E., Crespo E., De la Fuente C. et al. Spontaneously Arising Canine Glioma as a Potential Model for Human Glioma. Journal of Comparative Pathology. 2015. [Epub ahead of print]. • Azkona G., Saavedra A., Aira Z., Aluja D., Xifro X., Baguley T. et al. Striatal-enriched protein tyrosine phosphatase modulates nociception: Evidence from genetic deletion and pharmacological inhibition. Pain. 2016;157(2):377-386. Epub 2015. • van der Kooij M.A., Masana M., Rust M.B., Muller M.B. The stressed cytoskeleton: How actin dynamics can shape stress-related consequences on synaptic plasticity and complex behavior. Neuroscience and Biobehavioral Reviews. 2016;62:69-75. Epub 2015. • Fernández-Nogales M, Hernández F, Miguez A, Alberch J, Ginés S, Pérez-Navarro E et al. Decreased glycogen synthase kinase-3 levels and activity contribute to Huntington’s disease. Human molecular genetics. 2015;24(17): 5040-52. • Xifro X., Rodriguez-Alvarez J. Delineating the Factors and Cellular Mechanisms Involved in the Survival of Cerebellar Granule Neurons. Cerebellum. 2015;14(3):354-359. • Xifró X, Vidal-Sancho L, Boadas-Vaello P, Turrado C, Alberch J, Puig T et al. Novel epigallocatechin- 3-gallate (EGCG) derivative as a new therapeutic strategy for reducing neuropathic pain after chronic constriction nerve injury in mice.PloS one. ;10(4): e0123122. • European Commitee (Partial Agreement) on Organ Transplantation (CD-P-TO); Josep M. Canals as member. Guide to the quality and safety of tissues and cells for human application (2nd edition). 2015.European directorate for the quality of medicines and healthcare, Council of Europe.ISBN: 978-92-871-8116-8.

Research projects

• Code: 1619.14. Other CIBER’s collaboration: No. Title: Dual therapeutic benefits of isotype- Type: International. selective HDAC inhibition in Huntington’s Funding agency: EHDN. disease. Funding: 25.000 €. Principal Investigator: Silvia Ginés. Duration: 2015-2016 CIBERNED’s collaboration: No. CIBERNED groups:

2015 CIBERNED Annual Report / 110 Program 2 Group: Jordi Alberch Vié

• Code: La lámina nuclear en la enfermedad Type: National. de Huntington: papel en la fisiopatología y Funding agency: Instituto de Salud Carlos III. aplicaciones terapeúticas. Funding: 300.000 €. Title: La lámina nuclear en la enfermedad Duration: 2013-2015 de Huntington: papel en la fisiopatología y aplicaciones terapeúticas. • Code: RTC-2015-3731-1. Principal Investigator: Esther Pérez Navarro. Title: Optimización y desarrollo de un agente CIBERNED’s collaboration: Yes. terapéutico basado en ácidos nucleicos CIBERNED groups: G202, G301. para el tratamiento de la enfermedad de Other CIBER’s collaborarion: No. Huntington. Type: National. Principal Investigator: Assumpcio Bosch Funding agency: Fundación Ramón Areces. (G607)/Esther Pérez (G301). Funding: 121.440 €. CIBERNED’s collaboration: Yes. Duration: 2015 CIBERNED groups: G607, G301. Other CIBER’s collaboration: No. • Code: RD12/0019. Type: International. Title: RETIC de Terapia Celular. Funding agency: Ministerio de Economía y Principal INnvestigator: Coordinador de red: Competitividad. José María Moraleda. Funding: 1.582.767,92 €. CIBERNED’s collaboration: Yes. Duration: 2015-2017 CIBERNED groups: G102, G301, G113, G208, G105, G607. • Code: A4530. Other CIBER’s collaboration: No. Title: Stem Cell Differentiation JSC – Joint Type: National. collaborative effort between the IDIBAPS Funding agency: Fondo de Investigaciones (Barcelona), the University of Milano and the Sanitarias. University of Cardiff. Funding: 296.700 €. Principal Investigator: Josep M. Canals. Duration: 2013-2016 CIBERNED’s collaboration: No. CIBERNED groups: • Code: PI2013_08-1. Other CIBER’s collaboration: No. Title: La dinámica mitocondrial y mitofagia Type: International. como dianas terapéuticas en las Funding agency: CHDI Foundation Inc. enfermedades de Parkinson y Huntington. Funding: 1.845.000 €. Principal Investigator: Eduardo Soriano. Duration: 2012-2016 CIBERNED’s collaboration: Yes. CIBERNED groups: G408, G301, G207, G410, G109. Other CIBER’s collaboration: No.

PhD dissertations

• Author: Andreas Mezger. • Author: Mónica Pardo Muñoz. Title: Development of new methods for the Title: Characterization of morphogens differentiation of hPSCs for Huntington’s and new transcription factors involved in disease. the development of striatal Medium Spiny Date: 18th Novembre 2015. Neurons. Supervisor: Josep Mª. Canals Coll. Date: 12th Novembre 2015. Supervisor: Josep Mª. Canals Coll.

2015 CIBERNED Annual Report / 111 201 GROUP Enric I. Canela Campos

Enric I. Canela Campos Josefa Mallol Montero Verónica Casadó Anguera Full professor and Principal Research staff Phd Student investigator Estefanía Moreno Guillén Iñigo Etayo Labiano Vicent Casadó Burillo Postdoctoral fellow Phd Student Research staff Gemma Navarro Brugal Mireia Medrano Moya Antonio Cortés Tejedor Postdoctoral fellow Phd Student Research staff David Aguinaga Andrés Irene Reyes Resina Rafael Franco Fernández Phd Student Phd Student Research staff Edgar Angelats Canals Mar Rodríguez Ruiz Carmen Lluís Biset Phd Student Phd Student Research staff Jasmina Jiménez Cano Technician

Contact Details

Departamento de Bioquímica y Biología Molecular 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]

2015 CIBERNED Annual Report / 112 Summary

Our area of interest has been the GPCRs in the drug of a receptor to alter the action of the CNS, specially their ability to form homo- and other receptor and underlines that the success hetero-oligomers. We focused on discovering of this type of treatments relies on administering receptor heteromers as new targets for an appropriate dose of the drug. treatment of CNS disorders. Most relevant results in this period were: We revealed that the analgesic and amnesic effects of THC are independent of each other: We reviewed the ability of the enzyme while amnesia induced by THC disappears in the adenosine deaminase (ADA) to act as an mutant mice, THC can still promote analgesia in allosteric modulator of adenosine receptors. these animals. In subsequent molecular studies, Thus, ADA, being a single chain protein, performs we showed that in specific brain regions more than one function, consistent with the involved in memory formation, the receptors definition of a moonlighting protein. We have for THC and the 5-HT2A receptors work together proposed a mechanism by which some of their by physically interacting with each other. moonlighting functions can be coordinated. Experimentally interfering with this interaction We have suggested that drugs modulating prevented the memory deficits induced by ADA properties may act as modulators of the THC, but not its analgesic properties. Our results moonlighting functions of ADA, giving them highlight a novel mechanism by which the additional potential medical interest. We also beneficial analgesic properties of THC can be reviewed the role of activated microglia and dissociated from its cognitive side effects. macrophages in the central nervous system (CNS) and its potential in CNS diseases. In this line, we described an unprecedented role of the cannabinoid CB2 receptor as a pivotal We provided evidence for pharmacologically regulator of HER2 pro-oncogenic signaling in significant interactions between CRF and breast cancer, and they suggest that CB2 may orexin-A that depend on oligomerization be a biomarker with prognostic value in these of CRF1 receptor (CRF1R) and orexin OX1 tumors. receptors (OX1R). CRF1R–OX1R heteromers are the conduits of a negative crosstalk We applied computational tools and high- between orexin-A and CRF as demonstrated throughput screening assays to identify a new in transfected cells and rat VTA, in which they synthesized ligand for dopamine receptors significantly modulate dendritic dopamine with nanomolar affinity and agonist activity at release. The cocaine target sigma1 receptor dopamine D2 receptor. Our results validate (s1R) also associates with the CRF1R–OX1R the application of indoloquinolizidine-peptide heteromer. Cocaine binding to the s1R–CRF1R– combinatorial libraries to fine-tune the OX1R complex promotes a long-term disruption pharmacological profiles of multiple ligands at of the orexin-A–CRF negative crosstalk. Through D1 and D2 dopamine receptors. this mechanism, cocaine sensitizes VTA cells to the excitatory effects of both CRF and orexin-A, Based on PET studies in humans with [11C] thus providing a mechanism by which stress raclopride (dopamine D2/D3 receptor induces cocaine seeking. radioligand sensitive to endogenous dopamine), we showed that caffeine-induced We also investigated the allosteric modulations increases in D2/D3 receptor availability in the of A2AR antagonists that mimic those of A2AR ventral striatum were associated with caffeine- agonists, challenging the traditional view of induced increases in alertness. Our findings antagonists as inactive ligands. These allosteric indicate that in the human brain, caffeine, modulations disappear when agonist and at doses typically consumed, increases the antagonist are coadministered, however. availability of dopamine D2/D3 receptors, A model was proposed that considers which indicates that caffeine does not increase A2AR-D2R heteromers as heterotetramers, dopamine in the striatum for this would have constituted by A2AR and D2R homodimers, decreased D2/D3 receptor availability. Instead, based on radioligand dissociation and we interpret our findings to reflect an increase double complementation of BRET donor and in D2/D3 receptor levels in striatum with caffeine acceptor units. The model predicted that high (or changes in affinity). concentrations of A2AR antagonists would behave as A2AR agonists and decrease We demonstrated that angiotensin II type (AT1) D2R function in the brain. Thus, the binding receptors are expressed in striatum and form of receptors of hormones, neurotransmitters heteromers with dopamine D2 receptors that and neuromodulators forming tetramers may enable drugs selective for the AT1 receptor to explain the contradictory results obtained in alter the functional response of D2 receptors. therapies based on the use of an antagonist

2015 CIBERNED Annual Report / 113 Keywords

Adenosine receptors, allosteric interactions, cannabinoids, cocaine, dopamine receptors, G-protein coupled receptors, receptor oligomerization, two-state dimer receptor model

Publications

• Molero A, Vendrell M, Bonaventura J, Zachmann J, López L, Pardo L et al. A solid-phase combinatorial approach for indoloquinolizidine-peptides with high affinity at D(1) and D(2) dopamine receptors.European journal of medicinal chemistry. 2015;97: 173-80. • Naval-Macabuhay I., Casanova V., Navarro G., Garcia F., Leon A., Miralles L. et al. Adenosine deaminase regulates Treg expression in autologous T cell-dendritic cell cocultures from patients infected with HIV-1. Journal of Leukocyte Biology. 2016;99(2):349-359. Epub 2015. • Bonaventura J, Navarro G, Casadó-Anguera V, Azdad K, Rea W, Moreno E et al. Allosteric interactions between agonists and antagonists within the adenosine A2A receptor-dopamine D2 receptor heterotetramer.Proceedings of the National Academy of Sciences of the United States of America. 2015; 112(27): E3609-18. • Ferré S, Bonaventura J, Tomasi D, Navarro G, Moreno E, Cortés A et al. Allosteric mechanisms within the adenosine A2A dopamine Dreceptor heterotetramer.Neuropharmacology. 2015. [Epub ahead of print]. • Franco R., Fernandez-Suarez D. Alternatively activated microglia and macrophages in the central nervous system. Progress in Neurobiology. 2015;131:65-86. • Volkow ND, Wang GJ, Logan J, Alexoff D, Fowler JS, Thanos PK et al. Caffeine increases striatal dopamine D2/D3 receptor availability in the human brain.Translational psychiatry. 2015;5: e549. • Vinals X., Moreno E., Lanfumey L., Cordomi A., Pastor A., De La Torre R. et al. Cognitive impairment induced by delta9- tetrahydrocannabinol occurs through heteromers between cannabinoid CB<inf>1</inf> and serotonin 5-HT<inf>2A</inf> receptors. PLoS Biology. 2015;13(7). [Epub ahead of print]. • Martinez-Pinilla E., Rodriguez-Perez A.I., Navarro G., Aguinaga D., Moreno E., Lanciego J.L. et al. Dopamine D2 and angiotensin II type 1 receptors form functional heteromers in rat striatum. Biochemical Pharmacology. 2015;96(2):131-142. • Marada S., Navarro G., Truong A., Stewart D.P., Arensdorf A.M., Nachtergaele S. et al. Functional Divergence in the Role of N-Linked Glycosylation in Smoothened Signaling. PLoS Genetics. 2015;11(8). [Epub ahead of print]. • Cortes A., Gracia E., Moreno E., Mallol J., Lluis C., Canela E.I. et al. Moonlighting Adenosine Deaminase: A Target Protein for Drug Development. Medicinal Research Reviews. 2015;35(1):85- 125. • Aymerich MS, Rojo-Bustamante E, Molina C, Celorrio M, Sánchez-Arias JA, Franco R. Neuroprotective Effect of JZL184 in MPP⁺-Treated SH-SY5Y Cells Through CB2 Receptors.Molecular neurobiology. 2015. [Epub ahead of print]. • Navarro G., Quiroz C., Moreno-Delgado D., Sierakowiak A., McDowell K., Moreno E. et al. Orexin–corticotropin-releasing factor receptor heteromers in the ventral tegmental area as targets for cocaine. Journal of Neuroscience. 2015;35(17):6639-6653. Gomez-Vallejo V., Ugarte A., Garcia-Barroso C., Cuadrado-Tejedor M., Szczupak B., Dopeso- • Reyes I.G. et al. Pharmacokinetic investigation of sildenafil using positron emission tomography and determination of its effect on cerebrospinal fluid cGMP levels. Journal of Neurochemistry. 2016;136(2):403-415. Epub 2015. • Navarro G., Borroto-Escuela D.O., Fuxe K., Franco R. Purinergic signaling in Parkinson’s disease. Relevance for treatment. Neuropharmacology. 2015. [Epub ahead of print]. • Perez-Gomez E., Andradas C., Blasco-Benito S., Caffarel M.M., Garcia-Taboada E., Villa-Morales M. et al. Role of cannabinoid receptor CB2 in HER2 pro-oncogenic signaling in breast cancer.. Journal of the National Cancer Institute. 2015;107(6). [Epub ahead of print].

2015 CIBERNED Annual Report / 114 Program 2 Group: Enric I. Canela Campos

• Cordomi A., Navarro G., Aymerich M.S., Franco R. Structures for G-Protein-Coupled Receptor Tetramers in Complex with G Proteins. Trends in Biochemical Sciences. 2015;40(10):548-551. • Franco R., Petrovic M. Suggesting a way to understand the actual potential of anti-Alzheimer’s disease drugs that show promise in transgenic mouse models. Frontiers in Neurology. 2015;6(OCT). [Epub ahead of print]. • Onatibia-Astibia A., Martinez-Pinilla E., Franco R. The potential of methylxanthine-based therapies in pediatric respiratory tract diseases. Respiratory Medicine. 2015. [Epub ahead of print]. • Martinez-Pinilla E., Onatibia-Astibia A., Franco R. The relevance of theobromine for the beneficial effects of cocoa consumption. Frontiers in Pharmacology. 2015;6(FEB). [Epub ahead of print]. • Franco R, Casadó-Anguera V, Muñoz A, Petrovic M, Navarro G, Moreno E et al. Hints on the Lateralization of Dopamine Binding to D1 Receptors in Rat Striatum..Molecular neurobiology. 2015. [Epub ahead of print]. • Allosteric mechanisms in the adenosine A2A-dopamine D2 receptor heteromer.Ferré S, Navarro G, Bonaventura J, Moreno E, Volkow ND, Lluís C, Casadó V. En: The Adenosinergic System: A non-dopaminergic target in Parkinson’s disease. (pp. 27-38), 2015.Springer International Publishing, Switzerland.ISBN: 978-331920273-0.

Research projects

• Code: 2014-SGR-1236. • Code: SAF2012-39875-C02-01. Title: Neurobiología Molecular. Title: Heteromerización de receptores GPR55, Principal Investigator: Enric I. Canela CB1 y CB2: senalización celular, farmacología Campos. y análisis de su potencial como dianas de CIBERNED’s collaboration: No. enfermedades neurodegenerativas". CIBERNED groups: Principal Investigator: Rafael Franco Other CIBER’s collaboration: No. Fernández. Type: CC.AA. CIBERNED’s collaboration: No. Funding agency: Generalitat de Catalunya: CIBERNED groups: Programas: Ajuts a Grups de recerca Other CIBER’s collaboration: No. consolidats. Type: International. Funding: 24.000 €. Funding agency: convocatoria 2012 de Duration: 2014-2016 Proyectos de Investigación Fundamental no Orientada. • Code: 20141330. Funding: 178.856,13 €. Title: Cannabinoid receptor heteromeric Duration: 2014-2015 complexes as therapeutic targets in Parkinson's disease. • Code: 20140610. Principal Investigator: Rafael Franco Title: Targeting neuronal dopamine D1- Fernández. histamine H3 receptor heteromers as a new CIBERNED’s collaboration: Yes. treatment of Huntington’s disease. CIBERNED groups: G201, G203. Principal Investigator: Enric I. Canela Other CIBER’s collaboration: No. Campos. Type: Private. CIBERNED’s collaboration: No. Funding agency: Fundació La Marató de CIBERNED groups: TV3. Other CIBER’s collaboration: No. Funding: 150.000 €. Type: International. Duration: 2015-2017 Funding agency: Fundació La Marató de TV3. Funding: 200.000 €. Duration: 2015-2017

2015 CIBERNED Annual Report / 115 • Code: CIBERNED 2013/05. • Code: SAF2014-54840-R. Title: Identificación y caracterización Title: Oligomerización de receptores molecular de subpoblaciones de receptores adrenérgicos y dopaminérgicos en el cannabinoides en poliglutaminopatias. SNC: un nuevo enfoque en el tratamiento Principal Investigator: Manuel Guzmán del transtorno por déficit de atención e Pastor. hiperactividad. CIBERNED’s collaboration: Yes. Principal Investigator: Enric I. Canela CIBERNED groups: G305, G201, G303, G304. Campos. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: International. CIBERNED groups: Funding agency: CIBERNED. Other CIBER’s collaboration: No. Funding: 200.000 €. Type: National. Duration: 2013-2015 Funding agency: Ministerio de Economía y Competitividad. Funding: 242.000 €. Duration: 2015-2017

2015 CIBERNED Annual Report / 116 106 GROUP Valentín Ceña Callejo

Valentín Ceña Ana Belén García Sáez Elena Galera Principal investigator Lab technician Lab technician

Inmaculada Posadas Vanesa Guijarro Laura Romero Associate professor Lab technician Bachelor degree

Contact details

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]

2015 CIBERNED Annual Report / 117 Summary

The research of the Neurodeath group has drugs. One limitation to drug effectivity is the been focused on two different aspects: lack of selectivity on similar proteins (receptor types and subtypes, ionic channels, isoenzymes, 1) The use of nanoparticles, mainly dendrimers etc.) producing an important number of side (but also polymers derived from cyclodextrin) effects that limit treatment efficiency. to transfect genetic material, mainly siRNA, into different cell types to knock down specific An innovative therapeutic approach, that proteins involved in tumoral progression and solves this limitation, is the use of siRNA to neurodegenerative diseases. remove selectively certain proteins whose function is modified in several illness which are 2) The role of the endoplasmic reticulum and difficult to treat because the lack of effective the transcription factor HIF in the molecular therapies, drug toxicity and/or the presence mechanisms involved in neuronal death of resistance to treatment (cancer, TB, Etc.). during hypoxia and neurodegenerative This approach will produce, in a short period diseases. of time, new generations of drugs, based on siRNA, with the potential to revolutionize the Related to the use of nanoparticles to remove, therapeutics. Our group has focused during in a selective way, signaling proteins, it is this year in the development of dendrimers important to note that most drug targets are to transfect neuronal cultures with the idea of proteins that are inhibited or modulated by transfecting in vivo in a near future.

Keywords

Gene therapy, dendrimers, neurodegeneration, siRNA, transfection, endoplasmic reticulum, PERK

Publications

• Lopez-Hernandez B., Cena V., Posadas I. The endoplasmic reticulum stress and the HIF-1 signalling pathways are involved in the neuronal damage caused by chemical hypoxia. British Journal of Pharmacology. 2015;172(11):2838-2851. • Dumitru A.C., Herruzo E.T., Rausell E., Cena V., Garcia R. Unbinding forces and energies between a siRNA molecule and a dendrimer measured by force spectroscopy. Nanoscale. 2015;7(47):20267-20276. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1):-. Epub 2015.

2015 CIBERNED Annual Report / 118 Program 2 Group: Valentín Ceña Callejo

Research projects

• Code: VIHVACD. • Code: BFU2014-59009-P. Title: Desarrollo de una vacuna frente a VIH: Title: Efecto de dendrímeros antiinflamatorios estudio de los cambios en la biología de sobre la patogénesis de la esclerosis células dendríticas humanas tras interacción múltiple. con distintos sistema de liberación de Principal Investigator: Valentín Cena. péptidos de VIH. CIBERNED’s collaboration: No. Principal Investigator: María Ángeles Muñoz CIBERNED groups: Fernández. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: Funding agency: Ministerio de Economía y Other CIBER’s collaboration: Yes (CIBERBBN). Competitividad. Type: International. Funding: 156.453 €. Funding agency: CYTED. Duration: 2015-2017 Funding: 5.000 €. Duration: 2014-2017

2015 CIBERNED Annual Report / 119 601 GROUP Onofre Combarros Pascual

Onofre Combarros Antonio García Javier Riancho-Zarrabeitia Group PI and Full Professor of Clinical Neurophysiology ASP Neurology MIR Neurology Isabel González Aramburu Eloy Rodríguez Rodríguez Jon Infante Neurology specialist and IFIMAV Neurology ASP Group co-PI, ASP and Assistant postMIR Fellow Professor of Neurology María Ruiz Soto Andrés González-Mandly MICINN Technician José Berciano Radiology ASP Professor of Neurology and Pascual Sánchez-Juan Associate Professor Emeritus Andrea González Suaréz Neurology ASP Neurologist specialist and IDIVAL María Teresa Berciano Fellow Coro Sánchez-Quintana Full Professor of Cell Biology CIBERNED Lab Technician Miguel A. Lafarga Iñigo Casafont Full Professor of Cell Biology Olga Tapia Martínez Assistant Professor Postdoctoral researcher, IDIVAL Jorge Mata Garrido Marta de la Fuente FPI-MINECO PhD student María J. Sedano CIBERNED part-time Neurology ASP Administrative Assistant José Ignacio Mateo Neurology ASP María Sierra Elena Gallardo Neurology Specialist and Area Specialist Physician [ASP] José M. Polo López-Albo IFIMAV Fellow and Assistant Professor of Associate Professor of Neurology Radiology José L. Vázquez-Higuera Ana Pozueta Neurology ASP Psychologist, FIS/IFIMAV Fellow

Contact details

Servicio de Neurología Hospital Universitario Marqués de Valdecilla Universidad de Cantabria 39008 Santander (Spain) [email protected]

2015 CIBERNED Annual Report / 120 Summary

Introduction: We have redefined nosology in two strains of Charcot-Marie-Tooth (CMT) disease with As noted in previous Annual Reports, the individual heterozygous mutations in the NEFL research activity of our group has a clinical gene, either E397K or N98S, which now should component (led by Prof. Onofre Combarros), be integrated into the DI-CMT form. and a basic one (led by Profs. Miguel Lafarga and María Teresa Berciano). The latter is set in We have reported the first study of the the Department of Anatomy and Cell Biology, neurophysiological register of the T reflex in Faculty of Medicine, University of Cantabria. CMT1A, establishing that it can be a simple and convenient diagnostic method, particularly On the occasion of his retirement and after applicable to children with this neuropathy. appropriate approval by the CIBERNED Steering Committee, Prof. José Berciano (Group PI until In two review papers, we provide the role of March 1, 2015) becomes Associate Professor imaging studies in the diagnosis of inflammatory Emeritus. and hereditary neuropathies. On the clinical side, our research activity has Our Group continues its participation into the focused on the following aspects: European Projects IGOS. We maintain a close collaboration with Prof. Albena Jordanova • Clinical-molecular correlations in ataxias and (Neurogenetic Laboratory, University of hereditary neuropathies. Our group consists Antwerp, Belgium), with whom we are redefining of two EU Concerted projects: EUROSCA, the CMT2G genotype. RISCA and AFM/CMT1A. Hereditary ataxias: • Analysis of lower limb amyotrophy in CMT by magnetic resonance imaging. We have continued with the European study Natural History of SCAs, which is part of the • Clinical-molecular correlations in familial EUROSCA Project (see details in http://www. Parkinson’s disease. ataxia-study-group.net/html/studies/eurosca). Also, as part of EUROSCA activities, we have • Risk gene polymorphisms in sporadic continued with the prospective follow-up of Alzheimer’s disease. patients with autosomal dominant ataxia included in the RISCA Project. We have • Prionopathies. participated in a longitudinal follow up study of a large cohort of patients with SCA types 1, 2, 3 On the basic research side, it has focused on and 6 (see list of references). the following points: Parkinson’s disease: • The Cajal body and the nuclear fibrillar centers of the nucleolus as markers of We have shown that serum uric acid levels neuronal activity. are not associated with the risk of developing dementia in Parkinson’s disease. • Organization and dynamics of nuclear compartments of DNA injury/repair in In two international collaborative studies, we neurons. have investigated the temporal profile of non- motor symptoms in idiopathic and LRRK2 G2019S • Cellular Basis of the neuroprotective effect mutation-associated Parkinson’s disease (PD). of retinoids in a mouse model of ALS. By transcriptomic analysis in blood samples, we have identified 13 candidate genes in sporadic Peripheral neuropathies: cases of parkinsonism, in asymptomatic carriers of the G2019S mutation, and controls. We have demonstrated the pathogenic relevance of lesions in the proximal nerve trunks, Within our activity as members of the EMSA-SG particularly those affecting the spinal nerves, in (www.emsa-sg.org/), we have participated in a prospective study of six consecutive patients the study that discards the pathogenic role of with Guillain-Barré syndrome (GBS) in early the COQ2 mutation in multiple systems atrophy stage (days 1 to 9 of symptom onset). This notion (MSA). applies to both demyelinating and axonal forms. Our contributions to the knowledge In collaboration with the Department of of SGB have been collected in a book (see Neurology Purpam of Toulouse Hospital Publications 2015). (INSERM), project to identify clinical and

2015 CIBERNED Annual Report / 121 preclinical biomarkers (multimodal MRI) in within the European project JPND DEMTEST. LRRK2-PD is ongoing. Cellular Neurobiology: Alzheimer’s disease: Studies have focused on three research lines: Our group is a founding member of the Dementia Genetics Spanish Consortium (DEGESCO). Effect of bexarotene in a mouse model of As a result of this collaboration, sponsored ALS (SOD1-G93A transgenic mouse). by CIBERNED, this year we have reported the association between MAPT gene variants and Nuclear compartmentalization and dynamics various neurodegenerative diseases in a sample • of foci of DNA injury/repair in sensory ganglion of 11,572 subjects. neurons.

In the research line focused on Alzheimer’s Effect acetylated survival motor neuron disease biomarkers we are carrying out FDG- • factor (SMN) in the biogenesis of spicesomal and PiB-PET analysis on: 1) AD patients, 2) subjects snRNP and Cajal bodies assembly. with mild cognitive impairment and 3) healthy controls. We have an article in preparation to With respect to the first line, we have shown that describe our clinical experience in the use of •treatment with bexarotene, an agonist of retinoid PET amyloid and we have collaborated with receptors, improves survival of mice, increases our data in international consortia that have the expression of RXR and reduces proteostasis resulted in a double publication in JAMA dysfunction in motor neurons. Regarding the describing the prevalence of brain amyloid neuronal response to DNA damage, we have deposits in healthy subjects and in subjects with analyzed the dynamics of RNA polymerase cognitive impairment II, which governs the transcription of genes encoding proteins and the involvement of the Prionopathies: ubiquitin-proteasome pathway in DNA repair. Finally, we have shown that human SMN is This year we have published an analysis of acetylated in lysine K119, a process mediated genome-wide association (GWAs) with the by the CBP acetyltransferase. In addition, the largest sample of patients enrolled to date study of the SMN interactome has allowed (1,543 samples from patients with sporadic us to show that its acetylation regulates the Creutzfeldt-Jakob disease, from 7 European molecular assembly of spliceosomal snRNPs countries and Australia, together with 4,203 and Cajal bodies (manuscript in preparation). controls). This study is part of the collaboration

Keywords

Ataxia, SCA, Charcot-Marie-Tooth disease, Parkinson’s disease, Alzheimer’s disease, Creutzfeldt- Jakob disease, Cajal nuclear bodies, oculopharyngeal dystrophy

Publications

• Jun G, Ibrahim-Verbaas CA, Vronskaya M, Lambert JC, Chung J, Naj AC et al. A novel Alzheimer disease locus located near the gene encoding tau protein.Molecular psychiatry. 2016;21(1): 108-117. Epub 2015. • Carr AS, Pelayo-Negro AL, Evans MR, Laurà M, Blake J, Stancanelli C et al. A study of the neuropathy associated with transthyretin amyloidosis (ATTR) in the UK.Journal of neurology, neurosurgery, and psychiatry. 2015. [Epub ahead of print]. • Alcolea D., Martinez-Lage P., Sanchez-Juan P., Olazaran J., Antunez C., Izagirre A. et al. Amyloid precursor protein metabolism and inflammation markers in preclinical Alzheimer disease. Neurology. 2015;85(7):626-633. • Pelayo-Negro AL, Carr AS, Laura M, Skorupinska M, Reilly MM. An observational study of asymmetry in CMT1A.Journal of neurology, neurosurgery, and psychiatry. 2015;86(5): 589-90 .

2015 CIBERNED Annual Report / 122 Program 2 Group: Onofre Combarros Pascual

• Dols-Icardo O, Nebot I, Gorostidi A, Ortega-Cubero S, Hernández I, Rojas-García R et al. Analysis of the CHCHD10 gene in patients with frontotemporal dementia and amyotrophic lateral sclerosis from Spain.Brain : a journal of neurology. 2015;138(Pt 12): e400. . • Sierra M, Carnicella S, Strafella AP, Bichon A, Lhommée E, Castrioto A et al. Apathy and Impulse Control Disorders: Yin & Yang of Dopamine Dependent Behaviors.Journal of Parkinson's disease. 2015;5(3): 625-36. • Dols-Icardo O., Iborra O., Valdivia J., Pastor P., Ruiz A., de Munain A.L. et al. Assessing the role of TUBA4A gene in frontotemporal degeneration. Neurobiology of Aging. 2015. [Epub ahead of print]. • Romero A.M., Palanca A., Ruiz-Soto M., Llorca J., Marin M.P., Renau-Piqueras J. et al. Chronic Alcohol Exposure Decreases 53BP1 Protein Levels Leading to a Defective DNA Repair in Cultured Primary Cortical Neurons. Neurotoxicity Research. 2015. [Epub ahead of print]. • International Genomics of Alzheimer's Disease Consortium (IGAP), Lleó Bisa Alberto, Combarros Pascual Onofre, Bullido M. Convergent genetic and expression data implicate immunity in Alzheimer's disease.Alzheimer's & dementia : the journal of the Alzheimer's Association. 2015;11(6): 658-71. • Ferrer-Mayorga G., Alvarez-Diaz S., Valle N., De Las Rivas J., Mendes M., Barderas R. et al. Cystatin D locates in the nucleus at sites of active transcription and modulates gene and protein expression. Journal of Biological Chemistry. 2015;290(44):26533-26548. • Casafont I., Palanca A., Lafarga V., Mata-Garrido J., Berciano M.T., Lafarga M. Dynamic Behavior of the RNA Polymerase II and the Ubiquitin Proteasome System During the Neuronal DNA Damage Response to Ionizing Radiation. Molecular Neurobiology. 2015;:1-10. • Berciano J., Gallardo E., Orizaola P., de Lucas E.M., Garcia A., Pelayo-Negro A.L. et al. Early axonal Guillain-Barré syndrome with normal peripheral conduction: Imaging evidence for changes in proximal nerve segments. Heart. 2015. [Epub ahead of print]. • Berciano J. Early Guillain-Barré syndrome with normal peripheral conduction: which is the pathological hallmark?. Clinical neurology and neurosurgery. 2015;137: 11 . • García A, Pelayo-Negro AL, Álvarez-Paradelo S, Antolín FM, Berciano J. Electromyographic tendon reflex recording: An accurate and comfortable method for diagnosis of charcot-marie- tooth disease type 1a.Muscle & nerve. 2015;52(1): 39-44. • Infante J, Prieto C, Sierra M, Sánchez-Juan P, González-Aramburu I, Sánchez-Quintana C et al. Identification of candidate genes for Parkinson's disease through blood transcriptome analysis in LRRK2-G2019S carriers, idiopathic cases, and controls.Neurobiology of aging. 2015;36(2): 1105-9. • Riancho J., Navasa J.M., Sedano-Tous M.J., Polo J.M. Indirect carotid-cavernous fistula successfully treated with carotid compression. Medicina Clinica. 2015. [Epub ahead of print]. • Pastor P, Moreno F, Clarimón J, Ruiz A, Combarros O, Calero M et al. MAPT H1 Haplotype is Associated with Late-Onset Alzheimer's Disease Risk in APOEɛ4 Noncarriers: Results from the Dementia Genetics Spanish Consortium.Journal of Alzheimer's disease : JAD. 2015;49(2): 343- 52. • Carr A.S., Polke J.M., Wilson J., Pelayo-Negro A.L., Laura M., Nanji T. et al. MFN2 deletion of exons 7 and 8: Founder mutation in the UK population. Journal of the Peripheral Nervous System. 2015;20(2):67-71. • Barbagallo G., Sierra-Pena M., Nemmi F., Traon A.P.-L., Meissner W.G., Rascol O. et al. Multimodal MRI assessment of nigro-striatal pathway in multiple system atrophy and Parkinson disease. Movement Disorders. 2015. [Epub ahead of print]. • Gazulla J., Berciano J. Multiple-system atrophy. The New England journal of medicine. 2015;372(14):1374-. [Epub ahead of print]. • Berciano J., Garcia A., Peeters K., Gallardo E., De Vriendt E., Pelayo-Negro A.L. et al. NEFL E396K mutation is associated with a novel dominant intermediate Charcot–Marie–Tooth disease phenotype. Journal of Neurology. 2015;262(5):1289-1300.

2015 CIBERNED Annual Report / 123 • Berciano J., Peeters K., Garcia A., Lopez-Alburquerque T., Gallardo E., Hernandez-Fabian A. et al. NEFL N98S mutation: another cause of dominant intermediate Charcot–Marie–Tooth disease with heterogeneous early-onset phenotype. Journal of Neurology. 2016;263(2):361-369. • Riancho J, Ruiz-Soto M, Berciano MT, Berciano J, Lafarga M. Neuroprotective Effect of Bexarotene in the SOD1(G93A) Mouse Model of Amyotrophic Lateral Sclerosis.Frontiers in cellular neuroscience. ;9: 250. • Baltanas F.C., Valero J., Alonso J.R., Berciano M.T., Lafarga M. Nuclear signs of pre- neurodegeneration. Methods in Molecular Biology. 2015;1254:43-54. • Banzo I, Jiménez-Bonilla JF, Martínez-Rodríguez I, Quirce R, de Arcocha-Torres M, Bravo-Ferrer Z et al. Patterns of ¹¹C-PIB cerebral retention in mild cognitive impairment patients. Revista espanola de medicina nuclear e imagen molecular. 2015. [Epub ahead of print]. • Linnemann C., Tezenas du Montcel S., Rakowicz M., Schmitz-Hubsch T., Szymanski S., Berciano J. et al. Peripheral Neuropathy in Spinocerebellar Ataxia Type 1, 2, 3, and 6. Cerebellum. 2015. [Epub ahead of print]. • Ossenkoppele R, Jansen WJ, Rabinovici GD, Knol DL, van der Flier WM, van Berckel BN et al. Prevalence of amyloid PET positivity in dementia syndromes: a meta-analysis.JAMA. 2015;313(19):1924-38. • Jansen WJ, Ossenkoppele R, Knol DL, Tijms BM, Scheltens P, Verhey FR et al. Prevalence of cerebral amyloid pathology in persons without dementia: a meta-analysis.JAMA. 2015;313(19): 1924-38. • Alcolea D., Vilaplana E., Pegueroles J., Montal V., Sanchez-Juan P., Gonzalez-Suarez A. et al. Relationship between cortical thickness and cerebrospinal fluid YKL-40 in predementia stages of Alzheimer's disease. Neurobiology of Aging. 2015;36(6):2018-2023. • Berciano J, García A. Sural-sparing in Guillain-Barré syndrome: Does it mean lack of histopathological changes?. Clinical neurophysiology : official journal of the International Federation of Clinical Neurophysiology. 2015. [Epub ahead of print]. • Carr A.S., Pelayo-Negro A.L., Jaunmuktane Z., Scalco R.S., Hutt D., Evans M.R.B. et al. Transthyretin V122I amyloidosis with clinical and histological evidence of amyloid neuropathy and myopathy. Neuromuscular Disorders. 2015;25(6):511-515. • Gallardo E., Noto Y.-I., Simon N.G. Ultrasound in the diagnosis of peripheral neuropathy: Structure meets function in the neuromuscular clinic. Journal of Neurology, Neurosurgery and Psychiatry. 2015;86(10):1066-1074. • Sanchez-Juan P., Bishop M.T., Kovacs G.G., Calero M., Aulchenko Y.S., Ladogana A. et al. A genome wide association study links glutamate receptor pathway to sporadic Creutzfeldt- Jakob disease risk. PLoS ONE. 2015;10(4). [Epub ahead of print]. • Infante J., Prieto C., Sierra M., Sanchez-Juan P., Gonzalez-Aramburu I., Sanchez-Quintana C. et al. Comparative blood transcriptome analysis in idiopathic and LRRK2 G2019S-associated Parkinson's disease. Neurobiology of Aging. 2015. [Epub ahead of print]. • Jacobi H., du Montcel S.T., Bauer P., Giunti P., Cook A., Labrum R. et al. Long-term disease progression in spinocerebellar ataxia types 1, 2, 3, and 6: A longitudinal cohort study. The Lancet Neurology. 2015;14(11):1101-1108. • Garcia A., Pelayo-Negro A.L., Alvarez-Paradelo S., Antolin F.M., Berciano J. Reply. Muscle and Nerve. 2015;52(6):1140-1141. • Riancho J., Berciano M.T., Ruiz-Soto M., Berciano J., Landreth G., Lafarga M. Retinoids and motor neuron disease: Potential role in amyotrophic lateral sclerosis. Journal of the Neurological Sciences. 2016;360:115-120. Epub 2015. • Gallardo E., Sedano M.J., Orizaola P., Sanchez-Juan P., Gonzalez-Suarez A., Garcia A. et al. Spinal nerve involvement in early Guillain-Barré syndrome: A clinico-electrophysiological, ultrasonographic and pathological study. Clinical Neurophysiology. 2015;126(4):810-819. • Riancho J., Jimenez-Lopez Y., Marco-de Lucas E., Berciano J. Sudden onset of facial diplegia and aphagia. Revista Clinica Espanola. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 124 Program 2 Group: Onofre Combarros Pascual

• Riancho J., Gonzalo I., Ruiz-Soto M., Berciano J. Why do motor neurons degenerate? Actualization in the pathogenesis of amyotrophic lateral sclerosis. Neurologia. 2015. [Epub ahead of print]. • Berciano J. Guillain-Barré syndrome. Saarbrücken: Lambert Academic Publishing. (pp. 178), 2015.ISBN: 978-3-659-71134-3 • Berciano J, Gallardo E. Charcot-Marie-Tooth disease. En: Imaging in neurodegenerative disorders. (pp. 437-60), 2015. Saba L (eds). Oxford: Oxford University Press. ISBN: 978-0-19- 967161-8. • Pont-Sunyer C., Hotter A., Gaig C., Seppi K., Compta Y., Katzenschlager R. et al. The onset of nonmotor symptoms in parkinson's disease (the onset pd study). Movement Disorders. 2015;30(2):229-237. • Real J., Galindo G., Galvan L., Lafarga M.A., Rodrigo M.D., Ortega M. Use of oral bisphosphonates in primary prevention of fractures in postmenopausal women:A population-based cohort study. PLoS ONE. 2015;10(4): e0118178. • Querol L, Rojas-García R, Diaz-Manera J, Barcena J, Pardo J, Ortega-Moreno A et al. Rituximab in treatment-resistant CIDP with antibodies against paranodal proteins.Neurology(R) neuroimmunology & neuroinflammation. 2015;2(5): e149.

Research projects

• Code: PI12/02288. • Code: CIBERNED 2014/02. Title: Estudio multimodal de biomarcadores Title: Mecanismos epigenéticos de enfermedad de Alzheimer en deterioro implicados en la etiología y progresión cognitivo postoperatorio. de las demencias neurodegenerativas Principal Investigator: Pascual Sánchez- rápidamente progresivas. Juan. Principal Investigator: MIguel Calero. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G114, G509, G503, G601. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: Intramurales. Funding agency: Fondo de Investigaciones Funding agency: CIBERNED. Sanitarias. Funding: 252.074 €. Funding: 36.905 €. Duration: 2015-2017 Duration: 2013-2016 • Code: BFU20011-23983 • Code: FI11/00259. Title: Señalización y reparación del daño Title: Enfermedad de Parkinson causada en el DNA en neuronas. Organización por la mutación G2019S de LRRK2 en estructural, molecular, espacial y temporal Cantabria: aspectos genéticos y estudio de de los focos de lesión/reparación del DNA. biomarcadores en portadores asintomáticos Diferencial del transcriptoma en pacientes de la mutación. y portadores asintomáticos de mutaciones Principal Investigator: María Sierra Pena. en LRRK2. CIBERNED’s collaboration: No. Principal Investigator: Miguel Ángel Lafarga CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: National. Funding agency: Dirección general de Funding agency: Instituto de Salud Carlos III. Investigación, Ministerio de Economía y Funding: 108.000 €. Competitividad Duration: 2011-2015 Duration: 2012-2015 Funding: 139.150 €.

2015 CIBERNED Annual Report / 125 • Code: BFU2014-54754P • Title: Amiotrofia de extremidades inferiores Title: Regulación por acetilación del factor en la enfermedad de Charcot-Marie- de supervivencia de las neuronas motoras. Tooth tipo 1A: estudio longitudinal clínico, Su importancia en la biogénesis de snRNPs neurofisiológico y de resonancia magnética y en el ensamblaje de los cuerpos nucleares Principal Investigator: Ana Lara Pelayo Negro de Cajal. Diferencial del transcriptoma en (Director del proyecto, Jon Infante) pacientes y portadores asintomáticos de CIBERNED’s collaboration: No. mutaciones en LRRK2. CIBERNED groups: Principal Investigator: María Teresa Berciano Funding agency: Fundación Marqués de Blanco y Miguel Ángel Lafarga Coscojuela Valdecilla-IFIMAV CIBERNED’s collaboration: No. Funding: 72.000 €. CIBERNED groups: Duration: 2013-2015 Funding agency: Dirección general de Investigación, MIinisterio de Economía y • Title: Identificación mediante resonancia Competitividad. multimodal de biomarcadores de la fase Participating entities: Departamento de premotora de la enfermedad de Parkinson Anatomía y Biología Celular, Universidad de asociada a la mutación G2019S de LRRK2 Cantabria (proyecto de colaboración con Hospital Duration: 2015-2016. Purpan de Toulouse y con el INSERM, Financiación: 114.950 €. Francia). Principal Investigator: María Sierra Peña • Code: PI11/00228 CIBERNED’s collaboration: No. Title: Selección de genes candidato para la CIBERNED groups: enfermedad de Parkinson idiopática a través Funding agency: Sociedad Española de del análisis diferencial del transcriptoma en Neurología pacientes y portadores asintomáticos de Funding: 3.000 €. mutaciones en LRRK2. Duration: 2013-2014 Principal Investigator: Jon Infante Ceberio CIBERNED’s collaboration: No. • Title: Estudio de la influencia de los factores CIBERNED groups: genéticos reguladores de los niveles séricos Funding agency: Instituto de Salud Carlos III. de Progranulina sobre la penetrancia de Funding: 105.941,55 €. la mutación G2019S de LRRK2 y sobre el Duration: 2012-2014. Prorrogado hasta marzo riesgo de demencia en la enfermedad de 2016 Parkinson. Principal Investigator: Isabel González • Title: Estudio de la influencia de los factores Aramburu (Director del proyecto, Jon genéticos reguladores de los niveles séricos Infante) de Progranulina sobre la penetrancia de CIBERNED’s collaboration: No. la mutación G2019S de LRRK2 y sobre el CIBERNED groups: riesgo de demencia en la enfermedad de Funding agency: FIS (Ayuda predotoral) Parkinson. Funding: 82.500 €. Principal Investigator: Isabel González Duration: 2012-2015 Aramburu (Director del proyecto, Jon Infante) • Title: Biomarcadores de enfermedad de CIBERNED’s collaboration: No. Alzheimer como factores pronóstico en CIBERNED groups: hidrocefalia a presión normal idiopática. Funding agency: FIS (Ayuda predotoral) Principal Investigator: Eloy Manuel Rodríguez Funding: 82.500 €. Rodríguez Duration: 2012-2015 CIBERNED’s collaboration: No. CIBERNED groups: • Code: NVAL 2015/17 Funding agency: Fondo de Investigación Title: Estudio de la utilidad de biomarcadores Sanitaria, Instituto Carlos III. HUMV-IFIMAV y de neuroimagen y olfación en la enfermedad HCUVA de Parkinson asociada a la mutación G2019S Funding: 51.606,5€. de LRRK2 como predictores de conversión a Duration: 2014-2016 enfermedad de Parkinson motora. Principal Investigator: María Sierra Peña • Code: PI11/03028 CIBERNED’s collaboration: No. Title: Proyecto DEMTEST. CIBERNED groups: Principal Investigator: Pascual Sánchez- Funding agency: IDIVAL Juan Funding: 14.000 €. CIBERNED’s collaboration: No. Duration: 2015-2017 CIBERNED groups: Funding agency: FIS (JPND). Funding: 117.370 €. Duration: 2011-2015

2015 CIBERNED Annual Report / 126 Program 2 Group: Onofre Combarros Pascual

• Code: PT13/0010/0024 Title: Plataforma de Biobancos, nodo Hospital "Universitario Marqués de Valdecilla". Principal Investigator: Pascual Sánchez Juan CIBERNED’s collaboration: No. CIBERNED groups: Funding agency: Instituto de Salud Carlos III. Funding: 98.000 € Duration: 2014-2017

PhD dissertations

• Author: Javier Riancho Zarrabeitia. Title: Efecto del Bexaroteno en ratones transgénicos con esclerosis lateral amiotrófica (ELA), estudio histológico y molecular. Date: 27th March 2015. Supervisor: Miguel Ángel Lafarga Coscojuel.

2015 CIBERNED Annual Report / 127 101 GROUP Antonio Cuadrado Pastor

Antonio Cuadrado Pastor Ángel Juan García Yagüe Marta Pajares Cabetas Principal investigator and Full Postdoctoral fellow PhD student Professor María Isabel Escoll Ruth Blanco García Isabel Lastres Becker Guerrero PhD student Assitant Professor Postdoctoral fellow Alba Álvarez Ana Isabel Rojo Sanchís Natalia Robledinos Antón Grade student Postdoctoral fellow PhD student

Contact details

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]

2015 CIBERNED Annual Report / 128 Summary

Despite the promising results obtained in 2) We have developed a new mouse model preclinical animal models of neurodegenerative for Alzheimer combining the expression diseases, neuroprotective pharmacological of APP(V717I) and TAU(P301L) transgenes strategies have consistently failed in its in a wild type genetic background or null clinical translation. We believe that this is for expression of NRF2. In these mice the because animal models merely express the pathology associated with EA develops after proteinopathy (synucleinopathy in Parkinson’s one year of age but is faster in NRF2 deficient disease; amiloidopathy and tauopathy in mice. These mice represent a new tool for the Alzheimer’s disease), only reflect terminal and study of NRF2 as a new therapeutic target. marginal aspects of the disease and do not meet the factors risk and comorbidities found 3) The studies on NRF2 regulation in response in patients. The transcription factor NRF2 is to WNT signaling have led us to describe a master regulator of genes that provide a multiprotein complex that includes Axin, cytoprotection against oxidative, inflammatory GSK-3 and β-TrCP involved in maintaining and proteotoxic stress. Its activity decreases antioxidant basal metabolism in the brain. with age, in animal models is associated with Given the importance of the WNT pathway the risk factors of neurodegenerative diseases in the maintenance and differentiation of and, in addition, there are associations between neurogenic niches, we have initiated a study functional gene haplotypes and risk of Parkinson on the role of NRF2 in the development of or Alzheimer. Therefore, we are studying the neural progenitors of the dentate gyrus (SGZ) pharmacological regulation of NRF2 in the brain in the Alzheimer mouse model mentioned as a possible therapeutic target for Parkinson’s above. and Alzheimer’s. During 2015 we have focused our work on three lines of action: 4) We have continued with the development of a new mouse model of PD deficiency 1) Development of a new mouse model carrying NRF2 with the expression of of tauopathy based on estareotaxic human α-synuclein. In line with our line of delivery of an adeno-associated vector to technology transfer, we have worked with hippocampus that expresses mutant human companies of the biopharmaceutical sector protein TAU(P301L). We have described a to validate new compounds that activate communication between damaged neurons NRF2 in preclinical models of Parkinson’s and microglia aimed at repairing the brain disease. We have also participated in the parenchyma that is based on the release development and intellectual protection of of the chemokine fractalkin by the neuron a hybrid bifunctional compound made of and the activation of a signaling pathway melatonin and sulforaphane that induces leading to microglial activation of NRF2. The neuroprotection against oxidative stress. most relevant results of this study in mice have been validated in postmortem samples from patients with AD and progressive supranuclear palsy.

Keywords

Oxidative stress, neuroinflammation, proteinopathy, neuroprotection, NRF2, WNT, GSK3, TAU

Publications

• Prieto P., Rosales-Mendoza C.E., Terron V., Toledano V., Cuadrado A., Lopez-Collazo E. et al. Activation of autophagy in macrophages by pro-resolving lipid mediators. Autophagy. 2015;11(10):1729-1744.

• Freitas AE, Egea J, Buendía I, Navarro E, Rada P, Cuadrado A et al. Agmatine induces Nrf2 and protects against corticosterone effects in hippocampal neuronal cell line. Molecular neurobiology. 2015;51(3): 1504-19.

2015 CIBERNED Annual Report / 129 • Freitas A.E., Egea J., Buendia I., Gomez-Rangel V., Parada E., Navarro E. et al. Agmatine, by Improving Neuroplasticity Markers and Inducing Nrf2, Prevents Corticosterone-Induced Depressive-Like Behavior in Mice. Molecular Neurobiology. 2015. [Epub ahead of print]. • Schmidt HH, Stocker R, Vollbracht C, Paulsen G, Riley D, Daiber A et al. Antioxidants in Translational Medicine.Antioxidants & redox signaling. 2015;23(14): 1130-43. • Frijhoff J., Winyard P.G., Zarkovic N., Davies S.S., Stocker R., Cheng D. et al. Clinical Relevance of Biomarkers of Oxidative Stress. Antioxidants and Redox Signaling. 2015;23(14):1144-1170. • Pilar Valdecantos M., Prieto-Hontoria P.L., Pardo V., Modol T., Santamaria B., Weber M. et al. Essential role of Nrf2 in the protective effect of lipoic acid against lipoapoptosis in hepatocytes. Free Radical Biology and Medicine. 2015;84:263-278. • Fittschen M., Lastres-Becker I., Halbach M.V., Damrath E., Gispert S., Azizov M. et al. Genetic ablation of ataxin-2 increases several global translation factors in their transcript abundance but decreases translation rate. Neurogenetics. 2015;16(3):181-192. • Egea J., Buendia I., Parada E., Navarro E., Rada P., Cuadrado A. et al. Melatonin-sulforaphane hybrid ITH12674 induces neuroprotection in oxidative stress conditions by a ‘drug-prodrug’ mechanism of action. British Journal of Pharmacology. 2015;172(7):1807-1821. • Delgado-Buenrostro NL, Medina-Reyes EI, Lastres-Becker I, Freyre-Fonseca V, Ji Z, Hernández- Pando R et al. Nrf2 protects the lung against inflammation induced by titanium dioxide nanoparticles: A positive regulator role of Nrf2 on cytokine release.Environmental toxicology. 2015;30(7):-. • Dao V.T.-V., Casas A.I., Maghzal G.J., Seredenina T., Kaludercic N., Robledinos-Anton N. et al. Pharmacology and Clinical Drug Candidates in Redox Medicine. Antioxidants and Redox Signaling. 2015;23(14):1113-1129. • Casas A.I., Dao V.T.-V., Daiber A., Maghzal G.J., Di Lisa F., Kaludercic N. et al. Reactive Oxygen- Related Diseases: Therapeutic Targets and Emerging Clinical Indications. Antioxidants and Redox Signaling. 2015;23(14):1171-1185. • Pajares M., Jimenez-Moreno N., Dias I.H.K., Debelec B., Vucetic M., Fladmark K.E. et al. Redox control of protein degradation. Redox Biology. 2015;6:409-420. • Gonzalez-Rodriguez A., Santamaria B., Mas-Gutierrez J.A., Rada P., Fernandez-Millan E., Pardo V. et al. Resveratrol treatment restores peripheral insulin sensitivity in diabetic mice in a sirt1- independent manner. Molecular Nutrition and Food Research. 2015;59(8):1431-1442. • Cuadrado A. Structural and functional characterization of NRF2 degradation by glycogen synthase kinase 3/β-TrCP. Free Radical Biology and Medicine. 2015. [Epub ahead of print]. • Rada P, Rojo AI, Offergeld A, Feng GJ, Velasco-Martín JP, González-Sancho JM et al. WNT- 3A regulates an Axin1/NRF2 complex that regulates antioxidant metabolism in hepatocytes. Antioxidants & redox signaling. 2015;22(7): 555-71.

Research projects

• Code: SAF2013-43271-R. • Code: 2015/02. Title: Papel de NRF2 como modulador Title: Metabolismo oxidoreductor y antioxidante de la neuroinflamacion en la enfermedad de Parkinson: validacion de enfermedad de Alzheimer. nuevas dianas terapeuticas y nuevos Principal Investigator: Antonio Cuadrado biomarcadores. Pastor. Principal Investigator: José Luis Labandeira. CIBERNED’s collaboration: No. CIBERNED´s collaboration: Yes. CIBERNED groups: CIBERNED groups: G101, G202, G208, G203. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: National. Funding agency: Ministerio de Economía y Funding agency: CIBERNED. Competitividad. Funding: 240.000 €. Funding: 393.250 €. Duration: 2015-2017 Duration: 2014-2016

2015 CIBERNED Annual Report / 130 Program 2 Group: Antonio Cuadrado Pastor

• Code: COEN2008. Title: WNT signaling: biomarker and target evaluation in Alzheimer’s disease. Principal Investigator: Antonio Cuadrado Pastor. CIBERNED’s collaboration: No. CIBERNED groups: Other CIBER’s collaboration: No. Type: International. Funding agency: Instituto de Salud Carlos III. Funding: 616.300 €. Duration: 2014-2015

2015 CIBERNED Annual Report / 131 102 GROUP Isabel Fariñas Gómez

Isabel Fariñas Salomé Sirerol Piquer Ana Domingo Muelas Full Professor Cell Biology. Postdoctoral fellow PhD student Principal investigator Alex Bizy Raquel Montalbán Francisco Pérez Sánchez Postdoctoral fellow PhD student Scientific staff María Pimentel Santillana María José Palop Benlloch Martina Kirstein Postdoctoral fellow Technician Scientific staff Carmen Ovejero Benito Fabrice Durupt Sacramento Rodríguez Postdoctoral fellow Technician Ferrón Scientific staff Beatriz Martí Prado Elba Barberà Ferragud PhD student Technician Ana Pérez Villalba Postdoctoral fellow Germán Belenguer Sánchez José Manuel Morante PhD student Postdoctoral fellow

Contact details

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]

2015 CIBERNED Annual Report / 132 Summary

Our laboratory studies two topics related to the neurogenic niche of the subependymal neurodegeneration and cell therapy: zone and is required for the maintenance of the NSCs, possibly through the regulation of The study of cellular and molecular alterations dopamine bioavailability. To do this, we have 1) that underlie degeneration of dopaminergic been collaborating with Drs. Toledo (IBiS, neurons. Sevilla, CB06/05/0027) and Moratalla (Cajal Institute, Madrid; CB06/05/0055) in order to The study of neural stem cell (NSC) self- better understand the effects of dopamine 2) renewal. In recent years, we have also in adult NSCs. We have also observed an been working in combining both aspects by interaction between neurogenesis and aging in studying the regulation of adult neurogenesis models of accelerated senescence in rodents in parkinsonism and during aging. in collaboration with Dr. Pallás (University of Barcelona; CB06/05/0024). Finally, in the context In 2015, we have been analyzing the effect of of the intramural 2013/09 collaborative project dopamine and the synaptic regulator alpha- with teams led by Drs. Fernández Chacón synuclein in the regulation of adult NSCs, both (IBiS, Sevilla; CB06/05/0606) and Lucas Lozano in physiological situations and in experimental (CBM, Madrid, CB06 / 05/0062), we have been models of parkinsonism. In this line, we have studying adult neurogenesis that occurs in the characterized the phenotype of a null mutant subependymal zone of mice deficient in the for alpha-synuclein and shown that this synaptic synaptic regulator CSP-alpha and in mice with regulator is located in the terminals innervating abnormal levels CPEBs.

Keywords

Adult neurogenesis and cell therapy, neural stem cells, neurogenic niches, neurotrophic factors, mouse models of neurodegenerative disease, Parkinson´s disease

Publications

• Rubio A., Belles M., Belenguer G., Vidueira S., Farinas I., Nacher J. Characterization and isolation of immature neurons of the adult mouse piriform cortex. Developmental Neurobiology. 2015. [Epub ahead of print]. • Jones J, Estirado A, Redondo C, Pacheco-Torres J, Sirerol-Piquer MS, Garcia-Verdugo JM et al. Mesenchymal stem cells improve motor functions and decrease neurodegeneration in ataxic mice.Molecular therapy : the journal of the American Society of Gene Therapy. 2015;23(1):-. • Andreu Z., Khan M.A., Gonzalez-Gomez P., Negueruela S., Hortiguela R., San Emeterio J. et al. The cyclin-dependent kinase inhibitor p27<sup>kip1</sup> regulates radial stem cell quiescence and neurogenesis in the adult hippocampus. Stem Cells. 2015;33(1):219-229. • Coppiello G., Collantes M., Sirerol-Piquer M.S., Vandenwijngaert S., Schoors S., Swinnen M. et al. Meox2/Tcf15 heterodimers program the heart capillary endothelium for cardiac fatty acid uptake. Circulation. 2015;131(9):815-826. • Soriano-Cantón R, Perez-Villalba A, Morante-Redolat JM, Marqués-Torrejón MÁ, Pallás M, Pérez- Sánchez F et al. Regulation of the p19(Arf)/p53 pathway by histone acetylation underlies neural stem cell behavior in senescence-prone SAMP8 mice.Aging cell. 2015;14(3): 453-62.

2015 CIBERNED Annual Report / 133 Research projects

• Code: RD12/0019. Other CIBER’s collaboration: No. Title: RETIC de Terapia Celular. Type: International. Principal Investigator: Coordinador de red: Funding agency: Conselleria de Educacion José María Moraleda. de la Generalitat Valenciana. CIBERNED’s collaboration: Yes. Funding: 400.000 €. CIBERNED groups: G102, G301, G113, G208, Duration: 2015 G105, G607. Other CIBER’s collaboration: No. • Code: Convenio Botin-UV. Type: National. Title: Estudio de células madre en el ámbito Funding agency: Fondo de Investigaciones de las investigaciones básicas en terapia Sanitarias. celular. Funding: 296.700 €. Principal Investigator: Isabel Farinas. Duration: 2013-2016 CIBERNED’s collaboration: No. CIBERNED groups: • Code: PROMETEOII/2013/020. Other CIBER’s collaboration: No. Title: Efectos del microambiente vascular en Type: National. las células madre del cerebro adulto. Funding agency: Fundacion Botín-Banco Principal Investigator: Isabel Farinas. Santander. CIBERNED’s collaboration: No. Funding: 625.000 €. CIBERNED groups: Duration: 2014-2018

2015 CIBERNED Annual Report / 134 606 GROUP Rafael Fernández-Chacón

Rafael Fernández-Chacón Macarena Cabrera Serrano Josif Mircheski Associate Professor. Principal PhD student Postdoctoral fellow Investigator Fabiola Mavillard Saborido Leonardo Gómez Sánchez Carmen Paradas López Researcher PhD student Postdoctoral fellow Gloria Cantero Nieto Marina Valenzuela Villatoro Emilia Servián Postdoctoral fellow PhD student Researcher Ismael Valladares Millán Masters student

Contact details

Instituto de Biomedicina de Sevilla (IBiS) Hospital Universitario Virgen del Rocío. CSIC. Universidad de Sevilla Departamento de Fisiología Médica y Biofísica. CIBERNED 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

2015 CIBERNED Annual Report / 135 Summary

Our group studies the molecular mechanisms our observations indicate that such an underlying the normal function of nerve alteration in neurogenesis is not secondary to terminals and the synaptic dysfunctions related neurodegeneration. These observations have with neurodegeneration. Neurotransmitter unveiled as yet unknown functions for CSP-α release requires a tight coupling between beyond its previously described synaptic roles. synaptic vesicle exocytosis and endocytosis. We Our laboratory is currently investigating the have examined the time course of endocytosis molecular mechanisms that link CSP-α and at mouse motor nerve and separated two neurogenesis. Our unpublished results might sequential phases of endocytosis taking place help to find out novel key contributions of CSP-α during the stimulation train: early and late for the maintenance of neuronal integrity. endocytosis. Freshly released synaptic vesicle proteins are preferentially retrieved during the Furthermore, our scientific activity within early phase while synaptic vesicle proteins pre- this project has been reinforced by the existing at the plasma membrane before the collaboration with the groups led by Dr. Isabel stimulation are preferentially retrieved during Fariñas and Dr. José J. Lucas. Our investigations the late phase (J Physiol. 593:2867-88 (2015)). on neurogenesis at the hippocampal subgranular zone have been extended, in In addition, our group is studying the response of collaboration with Dr. Fariñas’s group, to the the brain to neurodegeneration in genetically study of neurogenesis at the subventricular zone modified mice. Knock-out mice lacking the to clarify if our observations reflect a general synaptic vesicle molecular co-chaperone role of CSP-α in neurogenesis. In collaboration Cysteine String Protein-α (CSP-α) suffer from with Dr. Lucas’s group we have advanced the presynaptic degeneration of GABAergic electrophysiological analysis of the synaptic terminals. These terminals regulate adult function in new genetically modified mice with neurogenesis, a remarkable phenomenon behavioral phenotypes that could be useful to consisting in the integration of newborn understand molecular alterations underlying neurons in the adult central nervous system. psychiatric disorders in humans. On the other We have investigated in those mice the hand, we maintain our collaboration with relationship between both, neurogenesis and Dr. Carmen Paradas’s group to investigate neurodegeneration, and we have found that the molecular mechanisms underlying the the initial stages of adult neurogenesis are physiopathology of a novel familial limb-girdle hyperactivated in CSP-α KO mice. Interestingly, muscular dystrophy.

Keywords

Synapse, synaptic vesicle cycle, motorneurons, hippocampus, neurogenesis, neuronal ceroid lipofuscinosis, genetically modified mice

Publications

• Cabrera-Serrano M, Fabian VA, Boutilier J, Wise C, Faiz F, Lamont PJ et al. Adult onset distal and proximal myopathy with complete ophthalmoplegia associated with a novel de novo p.(Leu1877Pro) mutation in MYH2.Clinical genetics. 2015;88(6): 573-8. • Linares-Clemente P., Rozas J.L., Mircheski J., Garcia-Junco-Clemente P., Martinez-Lopez J.A., Nieto-Gonzalez J.L. et al. Different dynamin blockers interfere with distinct phases of synaptic endocytosis during stimulation in motoneurones. Journal of Physiology. 2015;593(13):2867- 2888. • Cabrera-Serrano M., Junckerstorff R.C., Atkinson V., Sivadorai P., Allcock R.J., Lamont P. et al. Novel CHKB mutation expands the megaconial muscular dystrophy phenotype. Muscle and Nerve. 2015;51(1):140-143. • Camacho A., Esteban J., Paradas C. Report by the Spanish Foundation for the Brain on the social impact of amyotrophic lateral sclerosis and other neuromuscular disorders. Neurologia. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 136 Program 2 Group: Rafael Fernández-Chacón

• Nieto-Gonzalez J.L., Holm M.M., Vardya I., Christensen T., Wiborg O., Jensen K.. Presynaptic plasticity as a hallmark of rat stress susceptibility and antidepressant response. PLoS ONE. 2015;10(3): e0119993. • Martin-Rodriguez J.F., Munoz-Bravo J.L., Ibanez-Costa A., Fernandez-Maza L., Balcerzyk M., Leal-Campanario R. et al. Molecular Characterization of Growth Hormone-producing Tumors in the GC Rat Model of Acromegaly. Scientific Reports. 2015;5: 16298.

Research projects

• Code: P12-CTS-2232. CIBERNED groups: Title: Análisis genético y funcional de Co- Other CIBER’s collaboration: No. Chaperones sinápticos en el mantenimiento Type: International. y la degeneracion sináptica. Funding agency: Ministerio de Economía y Principal Investigator: Rafael Fernández- Competitividad. Chacón. Funding: 280.000 €. CIBERNED’s collaboration: No. Duration: 2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: PI-0017- 2014. Type: International. Title: Estudio terapéutico preclínico en un Funding agency: Consejería de Economía, modelo Murino Knock-In de distrofia muscular Innovación, Ciencia y Empleo, Junta de asociado a una mutación humana en el Andalucía. Gen Poglut1. Funding: 268.853 €. Principal Investigator: José Luis Nieto Duration: 2015 González. CIBERNED’s collaboration: No. • Code: BFU2013-47493-P. CIBERNED groups: Title: Mecanismos moleculares y celulares Other CIBER’s collaboration: No. de la respuesta cerebral y neuronal a la Type: CC.AA. degeneración presináptica. Funding agency: Fundación Pública Principal Investigator: Rafael Fernández- Andaluza Progreso y Salud. Chacón. Funding: 36.961 €. CIBERNED’s collaboration: No. Duration: 2015

2015 CIBERNED Annual Report / 137 303 GROUP Javier Fernández Ruiz

Javier Fernández Ruiz Onintza Sagredo Ezkioga María Gómez Cañas Full Professor - Principal Assistant Professor PhD student Investigator Concepción García García Sara Valdeolivas Rojas José Antonio Ramos Postdoctoral researcher PhD student Atance Full Professor Mª Ruth Pazos Rodríguez Francisco Espejo Porras Postdoctoral researcher PhD student Mariluz Hernández Gálvez Associate Professor Carmen Rodríguez Cueto María Ceprián Costoso Postdoctoral researcher PhD student Eva de Lago Femia Assistant Professor Valentina Satta Laura García Toscano PhD student Postdoctoral visiting reseacher María Gómez Ruiz Assistant Professor Cristina Palomo Garo Yolanda García Movellán Administrative Assistant PhD student

Contact details

Departamento de Bioquímica y Biología Molecular III Facultad de Medicina, Universidad Complutense de Madrid Avenida Complutense s/n Pabellón IV, 4ª planta, laboratorios 9, 11 y 12 28040 Madrid (Spain) Tel.: +34 91 394 14 50 Fax: +34 91 394 16 91 [email protected] [email protected]

2015 CIBERNED Annual Report / 138 Summary

The work during 2015 has been concentrated of inflammation in this disease, by comparing in the study of the neuroprotective different experimental models and including potential of cannabinoids in several chronic studies in mice deficient for this receptor and neurodegenerative disorders with motor also LRRK2 mutant mice, a genetic model symptoms, and, in particular, in the identification of Parkinson’s disease. This study has been of cellular and molecular mechanisms that conducted within a project funded by the underlie these effects. Michael J. Fox Foundation which has finalized in 2015. In the case of Huntington’s disease, the work has continued the collaboration with In the case of amyotrophic lateral sclerosis, the the companies GW Pharmaceuticals and work has again addressed the identification Vivacell Biotechnology-Spain in relation with of the changes that the disease provokes in the evaluation of the neuroprotective effects the endocannabinoid system and completed of different phytocannabinoids (∆9-THC, the evaluation of phytocannabinoids cannabidiol, cannabigerol) in experimental and derivatives of the companies GW models of this disease, in particular R6/2 mice. Pharmaceuticals and Vivacell Biotechnology, During this last year, we have progressed in using SOD-1 mutant mice and also TDP- those studies addressed to determine the 43 transgenic mice, which also serves for molecular and cellular mechanisms underlying the experimental study of frontotemporal these effects using in vitro models of this disease dementia. In the case of Vivacell Biotechnology, (Q7 versus Q111 cells), in which were studying these studies are being carried out within a the Nrf-2 signaling as the key mechanism. Lastly, project funded in the 2014 call of the RETOS- we are preparing for publication the clinical trial COLABORACION program of the MINECO. conducted in the “Hospital Ramón y Cajal” in We have also continued the biochemical and Madrid in patients using Sativex®, trial in which pharmacological (with Sativex) studies initiated our group has participated in the analysis of in 2013 in dogs with degenerative myelopathy, different biomarkers in collaboration with other an ALS-like disorder also produced by SOD-1 CIBERNED groups. mutations, in which, during 2015, we have concentrated on demonstrating the existence In the case of Parkinson’s disease, the work has of changes in endocannabinoid signaling in the also continued the collaboration with the two spinal cord of dogs affected by this disease. companies, in the case of GW Pharmaceuticals in investigating the advantages of the Lastly, in the case of spinocerebellar ataxias, combination of cannabidiol and ∆9-THCV for the the work has been concentrated in finishing symptomatic and neuroprotective treatment the biochemical and histopathological studies of this disease, and, in the case of Vivacell addressed to demonstrate the occurrence of Biotechnology, in determining the potential an endocannabinoid dysregulation in different of cannabigerol-quinones (they are ligands of CNS areas during the course of the disease, using PPARs) and cannabidiol-quinones (CB2 and SCA-3 transgenic mice, and we are carrying out PPAR agonists) in experimental models of this pharmacological studies addressed to correct disease. We are particularly interested in the such dysregulation and to elicit neuroprotective role of CB2 receptors in relation with the control effects.

Keywords

Cannabinoids, CB1 receptors, CB2 receptors, neuroprotection, Huntington’s disease, Parkinson’s disease, amyotrophic lateral sclerosis/frontotemporal dementia, spinocerebellar ataxias

2015 CIBERNED Annual Report / 139 Publications

• Feliú A, Moreno-Martet M, Mecha M, Carrillo-Salinas FJ, de Lago E, Fernández-Ruiz J et al. A Sativex(®) -like combination of phytocannabinoids as a disease-modifying therapy in a viral model of multiple sclerosis.British journal of pharmacology. 2015;172(14): 3579-95. • Erdozain AM, Rubio M, Meana JJ, Fernández-Ruiz J, Callado LF. Altered CB1 receptor coupling to G-proteins in the post-mortem caudate nucleus and cerebellum of alcoholic subjects. Journal of psychopharmacology (Oxford, England). 2015;29(11): 1137-45 . • García C, Palomo-Garo C, Gómez-Gálvez Y, Fernández-Ruiz J. Cannabinoid-dopamine interactions in the physiology and physiopathology of the basal ganglia.British journal of pharmacology. 2015. [Epub ahead of print]. • Fernández-Ruiz J, Moro MA, Martínez-Orgado J. Cannabinoids in Neurodegenerative Disorders and Stroke/Brain Trauma: From Preclinical Models to Clinical Applications.Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics. 2015;12(4): 793-806. • Espejo-Porras F, Piscitelli F, Verde R, Ramos JA, Di Marzo V, de Lago E et al. Changes in the endocannabinoid signaling system in CNS structures of TDP-43 transgenic mice: relevance for a neuroprotective therapy in TDP-43-related disorders.Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology. 2015;10(2): 233-44. • de Lago E., Moreno-Martet M., Espejo-Porras F., Fernandez-Ruiz J. Endocannabinoids and amyotrophic lateral sclerosis. Cannabinoids in Neurologic and Mental Disease. 2015;:99-123. • Fernandez-Ruiz J., Romero J. Endocannabinoids and Neurodegenerative disorders: Parkinson’s disease, Huntington’s chorea, Alzheimer’s disease, and others. Handbook of Experimental Pharmacology. 2015;231:233-259. • Vazquez C., Tolon R.M., Pazos M.R., Moreno M., Koester E.C., Cravatt B.F. et al. Endocannabinoids regulate the activity of astrocytic hemichannels and the microglial response against an injury: In vivo studies. Neurobiology of Disease. 2015;79:41-50. • Valdeolivas S, Navarrete C, Cantarero I, Bellido ML, Muñoz E, Sagredo O. Neuroprotective properties of cannabigerol in Huntington’s disease: studies in R6/2 mice and 3-nitropropionate- lesioned mice.Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics. 2015;12(1): 185-99. • Gómez-Gálvez Y, Palomo-Garo C, Fernández-Ruiz J, García C. Potential of the cannabinoid CB(2) receptor as a pharmacological target against inflammation in Parkinson’s disease. Progress in neuro-psychopharmacology & biological psychiatry. 2016;64. Epub 2015. • de Salas-Quiroga A, Díaz-Alonso J, García-Rincón D, Remmers F, Vega D, Gómez-Cañas M et al. Prenatal exposure to cannabinoids evokes long-lasting functional alterations by targeting CB1 receptors on developing cortical neurons.Proceedings of the National Academy of Sciences of the United States of America. 2015;112(44): 13693-8. • Morales P., Blasco-Benito S., Andradas C., Gomez-Canas M., Flores J.M., Goya P. et al. Selective, nontoxic CB2 cannabinoid o-quinone with in vivo activity against triple-negative breast cancer. Journal of Medicinal Chemistry. 2015;58(5):2256-2264. • Ragusa G, Gómez-Cañas M, Morales P, Hurst DP, Deligia F, Pazos R et al. Synthesis, pharmacological evaluation and docking studies of pyrrole structure-based CB2 receptor antagonists. European journal of medicinal chemistry. 2015;101: 651-67. • Moreno-Martet M, Feliú A, Espejo-Porras F, Mecha M, Carrillo-Salinas FJ, Fernández-Ruiz J et al. The disease-modifying effects of a Sativex-like combination of phytocannabinoids in mice with experimental autoimmune encephalomyelitis are preferentially due to Δ(9)- tetrahydrocannabinol acting through CB1 receptors.Multiple sclerosis and related disorders. 2015;4(6): 505-11. • Erdozain AM, Rubio M, Valdizan EM, Pazos A, Meana JJ, Fernández-Ruiz J et al. The endocannabinoid system is altered in the post-mortem prefrontal cortex of alcoholic subjects. Addiction biology. 2015;20(4): 773-83. • Garcia M.C., Cinquina V., Palomo-Garo C., Rabano A., Fernandez-Ruiz J. Identification of CB2 receptors in human nigral neurons that degenerate in Parkinson’s disease. Neuroscience Letters. 2015;587:1-4.

2015 CIBERNED Annual Report / 140 Program 2 Group Javier Fernández Ruiz

Research projects

• Code: GR3/14. • Code: GW2015. Title: Programa de Financiación Grupos de Title: Preclinical development of Investigación. Cannabinoides 950344. phytocannabinoid-based therapies for Principal Investigator: Javier Fernández Ruiz, the treatment of disease progression in Manuel Guzmán Pastor. amyotrophic lateral sclerosis/frontotemporal CIBERNED’s collaboration: Yes. dementia using TDP-43 transgenic mice. CIBERNED groups: G303, G305. Principal Investigator: Javier Fernández Ruiz, Other CIBER’s collaboration: No. Eva de Lago Femia. Type: Intramurales. CIBERNED’s collaboration: No. Funding agency: Universidad Complutense CIBERNED groups: de Madrid - Banco Santander. Other CIBER’s collaboration: No. Funding: 5.614,34 €. Type: International. Duration: 2014-2015 Funding agency: GW Research Ltd. (UK). Funding: 108.846 €. • Code: GW2013. Duration: 2015-2017 Title: Studies with phytocannabinoids as disease modifying agents in different • Code: CIBERNED 2013/05. neurodegenerative disorders. Title: Identificación y caracterización Principal Investigator: Javier Fernández Ruiz. molecular de subpoblaciones de receptores CIBERNED’s collaboration: No. cannabinoides en poliglutaminopatias. CIBERNED groups: Principal Investigator: Manuel Guzmán Other CIBER’s collaboration: No. Pastor. Type: Private. CIBERNED’s collaboration: Yes. Funding agency: GW Pharmaceuticals, Ltd. CIBERNED groups: G305, G201, G303, G304. Funding: 46.000 €. Other CIBER’s collaboration: No. Duration: 2013-2015 Type: International. Funding agency: CIBERNED. • Code: PI12/00192. Funding: 200.000 €. Title: Sinergia terapéutica entre Cannabidiol Duration: 2013-2015 e Hipotermia sobre el daño cerebral y extracerebral secundario a asfixia • Code: SAF2012-39173. perinatal en un modelo de cerdo neonatal. Title: El sistema cannabinoide como diana Mecanismos implicados. para una terapia neuroprotectora en la Principal Investigator: Mª Ruth Pazos esclerosis lateral amiotrófica. Rodríguez. Principal Investigator: Javier Fernández Ruiz. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: Internacional. Funding agency: Ministerio de Economía y Funding agency: Ministerio de Economía Competitividad. y Competitividad. Dirección Gral. de Funding: 68.365 €. Investigación y Gestión Plan Nacional I+D+i. Duration: 2013-2015 Funding: 220.300 €. Duration: 2013-2015 • Code: MJFOXF. Title: Cannabinoid CB2 receptors as a • Code: RTC-2014-1877-1. new target for the treatment of disease Title: Desarrollo preclínico de nuevos progression in Parkinson’s disease: studies in cannabinoides para el tratamiento LRRK2-transgenic mice. de esclerodermia y esclerosis lateral Principal Investigator: Javier Fernández Ruiz/ amiotrófica. Mª Concepción García García. Principal Investigator: Javier Fernández Ruiz. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: International. Funding agency: Michael J. Fox Foundation. Funding agency: Retos-Colaboración del Funding: 107.781 €. Programa Estatal de Investigación. Ministerio Duration: 2012-2015 Economía y Competitividad. Funding: 330.212 €. Duration: 2015-2016

2015 CIBERNED Annual Report / 141 • Code: FIS PIE13/00040. • Code: S2010/BMD-2308. Title: Modulation of cellular micro RNAs as Title: Neurofarmacología del sistema a therapeutic strategy for he cure of HIV endocannabinoide: del laboratorio a la infection. clínica. Principal Investigator: Santiago Moreno. Principal Investigator: Manuel Guzmán CIBERNED’s collaboration: No. Pastor. CIBERNED groups: CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G305, G303, G304. Type: International. Other CIBER’s collaboration: Yes Funding agency: Instituto de Salud Carlos (CIBERSAM). III. Estrategia de Salud 2013. Proyectos Type: CC.AA. Integrados de Excelencia en los Institutos de Funding agency: Comunidad de Madrid. Investigación Acreditados. Funding: 922.975 €. Funding: 605.000 €. Duration: 2012-2015 Duration: 2014-2016

PhD dissertations

• Author: Carmen Aurora Rodríguez Cueto. Title: Desregulación del sistema endocannabinoide en las ataxias espinocerebelosas. Date: 3rd June de 2015. Supervisor: Javier Fernández Ruiz.

2015 CIBERNED Annual Report / 142 103 GROUP José Manuel Fuentes Rodríguez

José Manuel Fuentes Ignacio Casado Naranjo Elisabet Uribe Carretero Rodríguez Head of Department PhD student Full Professor-Principal Investigator Sokhna Maryama Seydina Guadalupe Martínez Rosa Ana González Polo Yakhine DIOP Chacón Researcher PhD student Technician

Mireia Niso Santano Mario Rodríguez Arribas María Pura Luceño Postdoctoral fellow PhD student Technician

Contact details

Grupo PARK 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]

2015 CIBERNED Annual Report / 143 Summary

Our efforts during 2015 have continued showing as a key regulator in macroautophagic focusing on the study of basic molecular response. mechanisms involved in the pathogenesis of Parkinson’s disease, autophagy regulation and Our group has also started to screening potential role of PARK genes derived proteins in both neuroprotective in natural and mediated processes. Moreover we have also continued compounds by a change in the cell autophagic our collaboration and service delivery to activity. In this sense we have published a review Extremadura Health Service in molecular that reflects the current state on the subject in diagnosis of Parkinson’s disease. relation with the potential use in the treatment of neurodegenerative diseases. Following the work we carried out since 2009 in collaboration with the group of Dr. Adolfo Furthermore, we have initiated in 2015 a López de Munain (CIBERNED), have continued Cooperative project (PI2015/03) with the groups to characterize the basal macroautophagy of Dr. Perez Tur and Dr. Perez Castillo (with the deregulation in fibroblasts from Parkinson participation of Dr. Lopez de Munain) where patients carriers of pathogenic G2019S mutation we will study differential metabolic profiles in in LRRK2. Fibroblasts from individuals with G2019S Parkinson’s disease. LRRK2 mutation presents levels of autophagy basally higher than those observed in fibroblasts Finally we have followed interested in the from individuals without the mutation, this flare study of general mechanisms of regulation is accompanied by an increased sensitivity of of autophagy, this part of our business is still the cells to toxic substances, such as MPP+ , carried out in collaboration with the laboratory regarding Parkinson ‘s disease . of Dr. Guido Kroemer of Apoptosis, Cancer & Immunity Laboratory, INSERM Cordeliers Finally we are moving in functional studies using Research Center, Paris, Francia. fibroblasts from individuals with the mutation of LRRK2 R1441G. In this case we have also seen Regarding our collaboration with the an accelerated basal autophagy, however Extremadura Health Service should be noted no higher degradation than fibroblasts from that we have signed four agreements to provide individuals without this mutation. In both models services in relation to the molecular diagnosis we begin the analysis of the expression of of Parkinson’s. It has been increased to twenty actyltransferase and deacetylase enzymes and the number of products in our portfolio of the degree of acetylation of proteins related to services, and participate in research training for autophagy, since this posttranslational marking is healthcare professionals.

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

Publications

• Casado-Naranjo I, Romero Sevilla R, Portilla Cuenca JC, Duque de San Juan B, Calle Escobar ML, Fernández Pereira L et al. Association between subclinical carotid atherosclerosis, hyperhomocysteinaemia and mild cognitive impairment.Acta neurologica Scandinavica. 2015. [Epub ahead of print]. • Casado Naranjo I., Portilla Cuenca J.C., Duque De San Juan B., Garcia A.F., Sevilla R.R., Serrano Cabrera A. et al. Association of vascular factors and amnestic mild cognitive impairment: A comprehensive approach. Journal of Alzheimer’s Disease. 2015;44(2):695-704. • Galluzzi L, Bravo-San Pedro JM, Vitale I, Aaronson SA, Abrams JM, Adam D et al. Essential versus accessory aspects of cell death: recommendations of the NCCD 2015.Cell death and differentiation. 2015;22(1):-. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1). Epub 2015.

2015 CIBERNED Annual Report / 144 Program 2 Group: José Manuel Fuentes Rodríguez

• Rodríguez-Arribas M, Pizarro-Estrella E, Gómez-Sánchez R, Yakhine-Diop SM, Gragera-Hidalgo A, Cristo A et al. IFDOTMETER: A New Software Application for Automated Immunofluorescence Analysis.Journal of laboratory automation. 2015. [Epub ahead of print]. • González-Polo RA, Pizarro-Estrella E, Yakhine-Diop SM, Rodríguez-Arribas M, Gómez-Sánchez R, Pedro JM et al. Is the Modulation of Autophagy the Future in the Treatment of Neurodegenerative Diseases?. Current topics in medicinal chemistry. 2015;15(21): 2152-74. • Enot DP, Niso-Santano M, Durand S, Chery A, Pietrocola F, Vacchelli E et al. Metabolomic analyses reveal that anti-aging metabolites are depleted by palmitate but increased by oleate in vivo.Cell cycle (Georgetown, Tex.). 2015;14(15): 2399-407. • Heřmanová I, Arruabarrena-Aristorena A, Vališ K, Nůsková H, Alberich-Jorda M, Fišer K et al. Pharmacological inhibition of fatty-acid oxidation synergistically enhances the effect of l-asparaginase in childhood ALL cells.Leukemia. 2015. [Epub ahead of print]. • Gómez-Sánchez R, Pizarro-Estrella E, Yakhine-Diop SM, Rodríguez-Arribas M, Bravo-San Pedro JM, Fuentes JM et al. Routine Western blot to check autophagic flux: cautions and recommendations.Analytical biochemistry. 2015;477: 13-20. • Pietrocola F, Lachkar S, Enot DP, Niso-Santano M, Bravo-San Pedro JM, Sica V et al. Spermidine induces autophagy by inhibiting the acetyltransferase EP300.Cell death and differentiation. 2015;22(3): 509-16. • Niso-Santano M, Malik SA, Pietrocola F, Bravo-San Pedro JM, Mariño G, Cianfanelli V et al. Unsaturated fatty acids induce non-canonical autophagy.The EMBO journal. 2015;34(8): 1025- 41. • Control of Autophagy in Parkinson’s Disease.Rosa A González-Polo, Rubén Gómez-Sánchez, Elisa Pizarro-Estrella, Sokhna MS Yakhine-Diop, Mario Rodríguez-Arribas, José M Fuentes. En: Toxicity and Autophagy in Neurodegenerative Disorders.(pp. 91-122), 2015. • Association of vascular factors and amnestic mild cognitive impairment: A comprehensive approach.Naranjoa, I.C. , Cuenca, J.C.P., Juan De, B.D.S., García, A.F., Sevilla, R.R., Cabrer, A.S., Hijón, C.C., Chala, S.R., Fuentes, J.M., Moreno, J.M.R. . En: Handbook of Depression in Alzheimer’s Disease.(pp. 237-246), 2015. • Toxicity and autophagy in neurodegenerative disorders.Fuentes J.M .2015.

Research projects

• Code: PPGRU15L1. • Code: GR15045. Title: Ayuda básica a grupos de investigación Title: Ayudas para el fortalecimiento de los 2015. Programa propio de investigación. grupos de investigación de Extremadura. Principal Investigator: José Manuel Fuentes Principal Investigator: José Manuel Fuentes Rodríguez. Rodríguez. CIBERNED’s collaborator: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: International. Funding agency: Universidad de Funding agency: Gobierno de Extremadura. Extremadura. Funding: 3.082,74 €. Funding: 64.640,36 €. Duration: 2015 Duration: 2015-2017

2015 CIBERNED Annual Report / 145 • Code: PI14/00170. • Code: UNEX13-1E-1721. Title: Búsqueda de nuevas dianas Title: Laboratorio de Ensayos no clínicos BPL. terapéuticas en la enfermedad de Parkinson. Principal Investigator: Luis García Marín. Análisis de los sistemas de reciclaje celular y CIBERNED’s collaboration: No. estado mitocondrial en células procedentes CIBERNED groups: de pacientes. Other CIBER’s collaboration: No. Principal Investigator: Rosa Ana González Type: International. Polo. Funding agency: Ministerio de Economía y CIBERNED’s collaboration: No. Competitividad. Ayudas a infraestructuras CIBERNED groups: y equipamiento científico-técnico Other CIBER’s collaboration: No. subprograma estatal de infrestructuras Type: National. científicas y equipamiento. Funding agency: Instituto de Salud Carlos Funding: 137.445 €. III. Subdirección General de Evaluación y Duration: 2015 Fomento de la Investigación. Funding: 110.715 €. • Code: 2015/03. Duration: 2015-2017 Title: Perfiles metabólicos diferenciales en enfermedad de Parkinson. • Code: PI12/02280. Principal Investigator: José Manuel Fuentes Title: Acetilación, autofagia y Enfermedad Rodríguez. de Parkinson. CIBERNED´’s collaboration: Yes. Principal Investigator: José Manuel Fuentes CIBERNED groups: G103, G609, G110, G209. Rodríguez. Other CIBER’s collaboration: No. CIBERNED´’s collaboration: No. Type: International. CIBERNED groups: Funding agency: CIBERNED. Other CIBER’s collaboration: No. Funding: 210.000 €. Type: International. Duration: 2015 Funding agency: Ministerio de Economía y Competitividad. Instituto de Salud Carlos III. Fondo de Investigación Sanitaria. Funding: 116.765 €. Duration: 2013-2015

PhD dissertations

• Author: Pedro Jesús Labrador Gómez. Title: Perfil bioquímico del riesgo cardiovascular en población extremeña y su relación con la asistencia nefrológica. Date: 11th Decembre 2015. Supervisor: José Manuel Fuentes Rodríguez.

2015 CIBERNED Annual Report / 146 113 GROUP José Manuel García Verdugo

José Manuel García Vicente Herranz Pérez María Durán Moreno Verdugo Postdoctoral fellow PhD student Full professor - Principal investigator Sara García Gil-Perotín Susana González Granero Postdoctoral fellow Tecnician Juan Antonio Barcia Albacar Arantxa Cebrián Silla Research associate PhD student

José Miguel Soria López Research associate

Contact details

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]

2015 CIBERNED Annual Report / 147 Summary

Adult neurogenesis in mammals remains as our stem cells are capable of surviving even under main research line because of its regenerative prolonged hypoxia. potential in diseases of the nervous system. In the past year we have focused some of our studies Another aspect involved in the organization of on the human brain. We previously described neurogenic niches during development is the a migratory stream towards our olfactory bulbs establishment of planar polarity of ependymal in infants and, on the one hand, a second cells. We have recently studied the direct migration to the ventral prefrontal cortex. At involvement in this process of mechanosensory this moment, we are studying a migration, not Pkd1 and Pkd2 proteins, present in the cilia described so far, to the dorsal prefrontal cortex. of radial glia cells, the embryonic precursors The organization of these streams of migrating of ependymal cells in the lateral ventricles. cells, their temporal evolution, and the molecular Mutation of these proteins causes alterations markers that identify these cells as migrating in the organization of cilia of ependymal neuroblasts, make them strong candidates for cells and, as a result, deficits in cerebrospinal explaining structural and cognitive alterations fluid circulation in the mouse brain and during the development of the young brain. hydrocephalus. There are significant differences at the levels of In addition to the lateral ventricles of the brain, adult neurogenesis between different species we are also interested in the study of other of vertebrates. This has led us to perform a ventricular compartments. At the level of comparative study between higher vertebrates, the third ventricle, we performed a study on which reflects a progressive decrease in adult tanycyte populations, where we described new neurogenesis, especially in the human brain. subtypes based on the basal bodies of cilia and The possible cause of this decrease could the presence of vimentin. be attributed to the progressive thickening, during evolution, of the cerebral cortex, and In our interest in the knowledge of diseases to the great distances that new neurons need affecting the human brain, we sought to to travel in order to integrate into their final the study of animal models of Alzheimer’s destinations. However, although it seems that disease. Currently we are working with various there is a low rate of new neuronal addition molecules, such as genistein, which seems to in the human brain, there are indicators have a beneficial effect in terms of image (PET), which support the idea that production of cognitive function, and the size of amyloid new oligodendrocytes that contribute to plaques. In this regard, we plan to start clinical myelination process remains relatively high. In trials using this molecule. this regard, we have conducted a study which confirms that even if the neurogenesis in the Finally, we are offering a microscopy platform, aged mouse brain is disrupted, the formation of both with optical and electron microcopy new oligodendrocytes is maintained. Related equipment, and the possibility to establish to the above, we have also observed that correlations between them (the so-called chronic hypoxia affects the organization of correlative microscopy). We are working with neurogenic niches in the mouse. However, cell cultures, tissues, as well as in animal models, although neurogenesis and oligodendrogenesis which is bringing us a broader view of the brain are affected, the most undifferentiated neural organization and its function.

Keywords

Neurogenesis, stem cells, subventricular zone, third ventricle, oligodendrocytes, Alzheimer’s disease

Publications

• Sanchez P., Infante A., de Eguino G.R., Fuentes-Maestre J.A., Garcia-Verdugo J.M., Rodriguez C.I. Age-Related Lipid Metabolic Signature in Human LMNA-Lipodystrophic Stem Cell-Derived Adipocytes. The Journal of clinical endocrinology and metabolism. 2015;100(7):E964-E973. • Paredes M.F., Sorrells S.F., Garcia-Verdugo J.M., Alvarez-Buylla A. Brain size and limits to adult neurogenesis. Journal of Comparative Neurology. 2016;524(3):646-664. Epub 2015.

2015 CIBERNED Annual Report / 148 Program 2 Group: José Manuel García Verdugo

• Jimenez-Xarrie E., Davila M., Gil-Perotin S., Jurado-Rodriguez A., Candiota A.P., Delgado- Mederos R. et al. In vivo and ex vivo magnetic resonance spectroscopy of the infarct and the subventricular zone in experimental stroke. Journal of Cerebral Blood Flow and Metabolism. 2015;35(5):828-834. • Ohata S., Herranz-Perez V., Nakatani J., Boletta A., Garcia-Verdugo J.M., Alvarez-Buylla A. Mechanosensory genes Pkd1 and Pkd2 contribute to the planar polarization of brain ventricular epithelium. Journal of Neuroscience. 2015;35(31):11153-11168. • Garcia-Morales V., Montero F., Gonzalez-Forero D., Rodriguez-Bey G., Gomez-Perez L., Medialdea-Wandossell M.J. et al. Membrane-Derived Phospholipids Control Synaptic Neurotransmission and Plasticity. PLoS Biology. 2015;13(5):-. [Epub ahead of print]. • Coppiello G., Collantes M., Sirerol-Piquer M.S., Vandenwijngaert S., Schoors S., Swinnen M. et al. Meox2/Tcf15 heterodimers program the heart capillary endothelium for cardiac fatty acid uptake. Circulation. 2015;131(9):815-826. • Iglesias-García O, Baumgartner S, Macrí-Pellizzeri L, Rodriguez-Madoz JR, Abizanda G, Guruceaga E et al. Neuregulin-1β induces mature ventricular cardiac differentiation from induced pluripotent stem cells contributing to cardiac tissue repair.Stem cells and development. 2015;24(4): 484-96. • Paradells S, Rocamonde B, Llinares C, Herranz-Pérez V, Jimenez M, Garcia-Verdugo JM et al. Neurotoxic effects of ochratoxin A on the subventricular zone of adult mouse brain.Journal of applied toxicology : JAT. 2015;35(7): 737-51. • Ballester-Lurbe B, González-Granero S, Mocholí E, Poch E, García-Manzanares M, Dierssen M et al. RhoE deficiency alters postnatal subventricular zone development and the number of calbindin-expressing neurons in the olfactory bulb of mouse.Brain structure & function. 2015;220(6): 3113-30. • Macri-Pellizzeri L., Pelacho B., Sancho A., Iglesias-Garcia O., Simon-Yarza A.M., Soriano-Navarro M. et al. Substrate stiffness and composition specifically direct differentiation of induced pluripotent stem cells. Tissue Engineering - Part A. 2015;21(9-10):1633-1641. • Capilla-Gonzalez V., Herranz-Perez V., Garcia-Verdugo J.M. The aged brain: Genesis and fate of residual progenitor cells in the subventricular zone. Frontiers in Cellular Neuroscience. 2015;9(September). [Epub ahead of print]. • Alfaro-Cervello C., Soriano-Navarro M., Ramirez M., Bernet L., Banaclocha M.M., Cano R. et al. Ultrastructural pathology of anaplastic and grade II ependymomas reveals distinctive ciliary structures - Electron microscopy redux. Ultrastructural Pathology. 2015;39(1):23-29. • Agirre X., Castellano G., Pascual M., Heath S., Kulis M., Segura V. et al. Whole-epigenome analysis in multiple myeloma reveals DNA hypermethylation of B cell-specific enhancers. Genome Research. 2015;25(4):478-487. • Jones J, Estirado A, Redondo C, Pacheco-Torres J, Sirerol-Piquer MS, Garcia-Verdugo JM et al. Mesenchymal stem cells improve motor functions and decrease neurodegeneration in ataxic mice.Molecular therapy : the journal of the American Society of Gene Therapy. 2015;23(1): 130-8.

Research projects

• Code: 111111. Code: 277990-2. Title: Use of ACA-generated stem cells for Title: Multipotent Adult Progenitor Cells to the treatment of Parkinson’s disease. treat Stroke II (Strokemap II). Principal Investigator: Zorica Becker Kojic. Principal Investigator: Catherine Verfaillie. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: European. Type: European. Funding agency: Unión Europea - H2020. Funding agency: European Comission. Funding: N.D. Funding: N.D. Duration: 2015 Duration: 2012-2015

2015 CIBERNED Annual Report / 149 • Code: SEP-210212921. • Code: 2015/01-1. Title: Use of ACA-generated stem cells for Title: Terapia génica dirigida a neuregulinas cure of neural degenerative disease like para el tratamiento de enfermedades Cerebral Ischemia. degenerativas de las motoneuronas. Principal Investigator: Zorica Becker Kojic. Principal Investigator: Xavier Navarro. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G607, G113, G406, G407. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: European. Type: Intramurales. Funding agency: Unión Europea - H2020. Funding agency: Instituto de Salud Carlos III, Funding: N.D. CIBERNED. Duration: 2015 Funding: 240.000 €. Duration: 2015-2017 • Code: Ayudas a la investigación Ignacio Hernando Larramendi 2014. Protocolo de • Code: RD12/0019. actuación. Title: RETIC de Terapia Celular. Title: Aplicación de la resonancia Principal Investigator: Coordinador de red: magnética nuclear con tensor de difusión José María Moraleda. y la tractografia cerebral en ictus para la CIBERNED’s collaboration: Yes. valoracion del daño cerebral. CIBERNED groups: G102, G301, G113, G208, Principal Investigator: José Manuel García G105, G607. Verdugo. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: Funding agency: Fondo de Investigaciones Other CIBER’s collaboration: No. Sanitarias. Type: Private. Funding: 296.700 €. Funding agency: Fundación Mapfre. Duration: 2013-2016 Funding: 15.000 €. Duration: 2015 • Code: 479/C/2012. Title: Efecto de la migración de células • Code: PROMETEOII/2014/075. madre mediante campos magnéticos en la Title: Celulas madre adultas: Aplicaciones recuperación del infarto cerebral. en terapia regenerativa. Principal Investigator: Juan Sahuquillo Barris. Principal Investigator: José Manuel García CIBERNED’s collaboration: No. Verdugo. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: CC.AA. Other CIBER’s collaboration: No. Funding agency: Fundació la Marató de Type: CC.AA. TV3. Funding agency: Conselleria de Cultura, Funding: 305.675 €. Educación y Deportes, Generalitat Duration: 2013-2016 Valenciana. Funding: 56.065 €. • Code: CB06/05/1131. Duration: 2014-2017 Title: CIBERNED. Principal Investigator: José Manuel García • Code: SAF2012-33683. Verdugo. Title: Análisis del proceso de mielinización en CIBERNED’s collaboration: No. el estriado: efectos del EGF E IGF-1. CIBERNED groups: Principal Investigator: José Manuel García Other CIBER’s collaboration: No. Verdugo. Type: National. CIBERNED’s collaboration: No. Funding agency: Instituto de Salud Carlos III. CIBERNED groups: Funding: 710.381,01 €. Other CIBER’s collaboration: No. Duration: 2007-2015 Type: National. Funding agency: Ministerio de Economía y Competitividad. Funding: 105.300 €. Duration: 2013-2015

2015 CIBERNED Annual Report / 150 104 GROUP José González Castaño

José González Castaño Raúl Sánchez Lanzas Teresa Bermejo Pastor Principal investigator PhD student Lab technician

Contact details

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]

2015 CIBERNED Annual Report / 151 Summary

Alpha-synuclein is a small protein (140 aa) levels or in the rates of degradation of those implicated in the pathophysiology of Parkinson’s selective CMA protein substrates. These results disease (PD). The degradation of alpha- indicate that the Lamp-2A protein seems to synuclein is a critical step of its proteotasis and be dispensable for protein degradation by regulates the levels and balance between the lysosome. AS a consequence, either other native and aggregated forms of the protein. lysosomal proteins can compensate for the lack Three pathways have been implicated in its of expression of Lamp-2 in the CMA pathway or degradation, the proteasome, the chaperon this pathway is not very relevant in proteostasis, mediated autophagy and macroautophagy. and the proteasome and macro-autophagy pathways are the main pathways responsible We have approached the study of the role of proteostasis in the cell. of the CMA pathway in alpha-synuclein degradation, reported to be mediated During last year we have also continued the by the lysosomal receptor Lamp-2A, in a study of the pathway of degradation of the lymphoblastoid cell line derived from a patient different DJ-1 point mutants that have been with Danon disease. Danon disease is a X-linked described as responsible of familiar autosomal genetic disease characterized by myopathy, recessive forms of PD. Similar to the well cardiomyopathy and intellectual disability and characterized L166P point mutant that we is due to mutations or deletions of the LAMP2 have previously studied, there are several DJ-1 gene. Accordingly, in those patients there is point mutants whose increased instability in a lack of expression of the different Lamp-2 the cell is due, at least in part, to proteasomal proteins generated by alternative splicing, degradation. Nevertheless, we are at present including Lamp-2A. The degradation of several studying the relevance of alternative pathways selective substrates of the CMA pathway, in their degradation. Both lines of research fit including alpha-synuclein, have been studied with our commitment to study the role of protein both in control and Lamp2-defficient cells. The degradation in neurodegenerative disease, results obtained show no significant changes especially in connection with PD pathology. either in the steady-state protein and mRNA

Keywords

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

Research projects

• Code: BM1307. • Code: S2010_BMD. Title: 2013 COST action proteostasis. Title: Redes de senalización y vías efectoras Principal Investigator: Manuel S. Rodríguez, en modelos celulares y animales de Rosa Barrios (Spain). enfermedades neurodegenerativas. CIBERNED’s collaboration: No. Principal Investigator: José González CIBERNED groups: Castaño. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: International. CIBERNED groups: G111, G401, G104, G307, Funding agency: UE. G409, G412. Funding: 13.500 €. Other CIBER’s collaboration: Yes (CIBERER). Duration: 2014-2017 Type: CC.AA. Funding agency: Comunidad de Madrid. Funding: 483.000 €. Duration: 2012-2016

2015 CIBERNED Annual Report / 152 305 GROUP Manuel Guzmán Pastor

Manuel Guzmán Pastor Javier Díaz Alonso Daniel García Rincón Full Professor-Principal Investigator Postdoctoral fellow PhD student

Ismael Galve Roperh Anna Paola Chiarlone Juan Paraíso Luna Associate Professor. Principal Co- Postdoctoral fellow PhD student Investigator Raquel Bajo Grañeras Irene Berenice Maroto Guillermo Velasco Díez Postdoctoral fellow Martínez Associate Professor PhD student Adán de Salas Quiroga Cristina Blázquez Ortiz PhD student José Aguareles Gorines Associate Professor PhD student Andrea Ruiz Calvo Luigi Bellocchio PhD student Eva Resel Mariné Postdoctoral fellow Lab manager

Elena García Taboada Lab technician

Contact details

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]

2015 CIBERNED Annual Report / 153 Summary

Our research focuses globally on the study of the 2) A remarkable and dorsolaterally-selective molecular and cellular mechanisms underlying down-regulation of the CB1 receptor has the control of neural cell physiopathology by been documented in the basal ganglia of the endocannabinoid system. Thus, in 2015 we Huntington’s disease patients and animal have kept on studying how this system tunes models, and, of interest, this loss of CB1 neural progenitor proliferation, differentiation receptors seems to reflect, at least in part, and survival. the neuron-damage pattern characteristic of the disease. Specifically, in the context of our CIBERNED program, we have evaluated the role of the 3) This loss of CB1 receptors occurs at early endocannabinoid system in Huntington’s stages of Huntington’s disease and prior to disease. As a matter of fact, Huntington’s the appearance of overt clinical symptoms, disease constitutes, in our opinion, the best neurodegeneration and bulk changes in currently available model disease to assess the other neurochemical parameters. pathophysiological relevance and therapeutic potential of the endocannabinoid system in In 2015, our studies on the endocannabinoid neurodegenerative diseases. This is due to system in Huntington´s disease have aimed several reasons: at defining (1) whether stimulation of the endocannabinoid system in Huntington’s 1) CB1 is the most abundant G protein-coupled disease models promotes neuroprotection and receptor in the brain; specifically, it is highly therefore delays the onset and/or attenuates expressed in the basal ganglia at synapses the progression of the pathology, (2) whether established by neurons containing GABA the alterations of CB1 cannabinoid receptors [especially medium-sized spiny neurons observed in MSNs and/or corticostriatal terminals (MSNs), the cells that primarily degenerate at early stages of the disease are involved in in Huntington’s disease] or glutamate the pathogenesis of Huntington’s disease, and (especially corticostriatal projecting neurons, (3) whether the induction of CB2 cannabinoid which critically control MSN function) as receptors in reactive microglial cells modulates transmitters, and play a key role in the control the excitotoxicity processes associated with of motor behaviour (one of the processes Huntington’s disease . We aim at unravelling that is most characteristically affected in the molecular and cellular bases as well as the Huntington’s disease). potential clinical relevance of those processes.

Keywords

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

Publications

• Cardinal P., Bellocchio L., Guzman-Quevedo O., Andre C., Clark S., Elie M. et al. Cannabinoid Type 1 (CB1) Receptors on Sim1-Expressing Neurons Regulate Energy Expenditure in Male Mice. Endocrinology. 2015;156(2):411-418. • Abrams D.I., Guzman M. Cannabis in cancer care. Clinical pharmacology and therapeutics. 2015;97(6):575-586. • Díaz-Alonso J, Aguado T, de Salas-Quiroga A, Ortega Z, Guzmán M, Galve-Roperh I. CB1 Cannabinoid Receptor-Dependent Activation of mTORC1/Pax6 Signaling Drives Tbr2 Expression and Basal Progenitor Expansion in the Developing Mouse Cortex.Cerebral cortex (New York, N.Y. : 1991). 2015;25(9): 2395-408. • Pietropaolo S., Bellocchio L., Ruiz-Calvo A., Cabanas M., Du Z., Guzman M. et al. Chronic cannabinoid receptor stimulation selectively prevents motor impairments in a mouse model of Huntington’s disease. Neuropharmacology. 2015;89:368-374.

2015 CIBERNED Annual Report / 154 Program 2 Group: Manuel Guzmán Pastor

• Oddi D, Subashi E, Middei S, Bellocchio L, Lemaire-Mayo V, Guzmán M et al. Early social enrichment rescues adult behavioral and brain abnormalities in a mouse model of fragile X syndrome.Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology. 2015;40(5): 1113-22. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1). Epub 2015. • Salazar M, Lorente M, García-Taboada E, Pérez Gómez E, Dávila D, Zúñiga-García P et al. Loss of Tribbles pseudokinase-3 promotes Akt-driven tumorigenesis via FOXO inactivation.Cell death and differentiation. 2015;22(1): 131-44 . • McKinnon C., Goold R., Andre R., Devoy A., Ortega Z., Moonga J. et al. Prion-mediated neurodegeneration is associated with early impairment of the ubiquitin–proteasome system. Acta Neuropathologica. 2015;:1-15. • Blázquez C, Chiarlone A, Bellocchio L, Resel E, Pruunsild P, García-Rincón D et al. The CB₁ cannabinoid receptor signals striatal neuroprotection via a PI3K/Akt/mTORC1/BDNF pathway. Cell death and differentiation. 2015;22(10): 1618-29. • Martin R., Bajo-Graneras R., Moratalla R., Perea G., Araque A. Circuit-specific signaling in astrocyte-neuron networks in basal ganglia pathways. Science. 2015;349(6249):730-734. • de Salas-Quiroga A, Díaz-Alonso J, García-Rincón D, Remmers F, Vega D, Gómez-Cañas M et al. Prenatal exposure to cannabinoids evokes long-lasting functional alterations by targeting CB1 receptors on developing cortical neurons.Proceedings of the National Academy of Sciences of the United States of America. 2015;112(44): 13693-8. • Perez-Gomez E., Andradas C., Blasco-Benito S., Caffarel M.M., Garcia-Taboada E., Villa-Morales M. et al. Role of cannabinoid receptor CB<inf>2</inf> in HER2 pro-oncogenic signaling in breast cancer. Journal of the National Cancer Institute. 2015;107(6). [Epub ahead of print].

Research projects

• Code: GR3/14. Code: PI12/02248. Title: Programa de Financiación Grupos de Title: Mecanismos de regulación de la Investigación. Cannabinoides 950344. autofagia en células tumorales: muerte Principal Investigator: Javier Fernández Ruiz, • mediada por autofagia en respuesta a Manuel Guzmán Pastor. fármacos antitumorales y participación en CIBERNED’s collaboration: Yes. el control de la tumorigénesis. CIBERNED groups: G303, G305. Principal Investigator: Guillermo Velasco Other CIBER’s collaboration: No. Díez. Type: Intramurales. CIBERNED’s collaboration: No. Funding agency: UCM - Banco Santander. CIBERNED groups: Funding: 5.614,34 €. Other CIBER’s collaboration: No. Duration: 2014-2015 Type: National. Funding agency: Ministerio de Economía y Code: PI12/00919. Competitividad-Instituto de Salud Carlos III- • Title: Papel del receptor CB1 cannabinoide Fondo de Investigaciones Sanitarias. en alteraciones de la neurogénesis cortical: Funding: 191.500 €. Implicaciones en excitabibilidad neuronal y Duration: 2013-2015 función corticoespinal. Principal Investigator: Ismael Galve Roperh. CIBERNED’s collaboration: No. CIBERNED groups: Other CIBER’s collaboration: No. Type: National. Funding agency: Ministerio de Economía y Competitividad-Instituto de Salud Carlos III- Fondo de Investigaciones Sanitarias. Funding: 147.015 €. Duration: 2013-2015

2015 CIBERNED Annual Report / 155 • Code: CIBERNED 2013/05. • Code: S2011/BMD 2336. Title: Identificación y caracterización Title: Estudio de la biología de células madre molecular de subpoblaciones de receptores neurales para su empleo en terapia celular cannabinoides en poliglutaminopatías. en enfermedades neurodegenerativas. Principal Investigato: Manuel Guzmán Principal Investigator: Rosario Moratalla. Pastor. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G204, G305, G108. CIBERNED groups: G305, G201, G303, G304. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: CC.AA. Type: International. Funding agency: Comunidad Autónoma de Funding agency: CIBERNED. Madrid. Funding: 200.000 €. Funding: 598.000 €. Duration: 2013-2015 Duration: 2012-2015 • Code: SAF2012-35759. • Code: S2010/BMD-2308. Title: Neuroprotección por el receptor Title: Neurofarmacología del sistema CB1 cannabinoide en la enfermedad endocannabinoide: del laboratorio a la de Huntington: relevancia de las vías clínica. corticoestriatales directa e indirecta. Principal Investigator: Manuel Guzmán Principal Investigator: Manuel Guzmán Pastor. Pastor. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G305, G303, G304. CIBERNED groups: Other CIBER’s collaboration:Yes (CIBERSAM). Other CIBER’s collaboration: No. Typ: CC.AA. Type: National. Funding agency: Comunidad de Madrid. Funding agency: Ministerio de Economía y Funding: 922.975 €. Competitividad-Plan Nacional. Duration: 2012-2015 Funding: 290.000 €. Duration: 2013-2015

2015 CIBERNED Annual Report / 156 111 GROUP Teresa Iglesias Vacas

Teresa Iglesias Vacas Andrea Gamir Morralla Aurelio del Pino Beitia Principal Investigator, CSIC Postdoctoral Fellow Master’s student Tenured Scientist Julia Pose Utrilla Raquel López Labiano Lucía García Guerra Master’s student Grade student Postdoctoral Fellow Noelia Sofía de León Reyes Esther Otero Vidal Ana Mª del Puerto del Pino Master’s student Technician Postdoctoral Fellow

Contact details

Instituto de Investigaciones Biomédicas “Alberto Sols” CSIC-UAM y CIBERNED 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]

2015 CIBERNED Annual Report / 157 Summary

Our group has spent recent years studying increasing neuronal viability (Gamir-Morralla & Kidins220 and protein kinase D (PKD) in different López—Menéndez et al, 2015). The application neuropathologies. First we discovered that of peptide Tat-K may be useful in the treatment Kidins220 is crucial for neuronal survival and plays of neuropathologies associated with either a neuroprotective role. This protein undergoes acute (stroke, hypoxia, acute trauma, epilepsy) a drastic down-regulation by proteolysis largely or chronic excitotoxicity (neurodegenerative dependent on the protease calpain under diseases). conditions which induce neuronal death by excitotoxicity as those occurring during On the other hand we have previously found cerebral ischemia. This type of neuronal death that Kidins220 is increased in brain necropsies of is also involved in the development of several patients with Alzheimer´s disease (AD), where neurodegenerative diseases such as Parkinson’s, it accumulates with hyperphosphorylated Huntington’s or Alzheimer´s. We have identified tau, one of the two main neuropathological that the disappearance of Kidins220 is markers of this disease. At present we want to associated with inactivation of pro-survival determine whether these alterations in Kidins220 neuronal pathways, and further characterized are cause or consequence of this dementia, the molecular mechanisms that contribute if they could be at the bases of the disease to reduce Kidins220 under these conditions etiopathology. For this purpose, we will study of neuronal death. To continue these studies the effect of manipulating Kidins220 levels on we decided to identify calpain processing various pathophysiological processes related sites within Kidins220 aminoacidic sequence to the initial steps of neurodegeneration in and design an approach that would avoid AD. In addition, knowledge of the proteolytic such processing as possible neuroprotective mechanisms of proteins such as tau and strategy. We have mapped the major calpain Kidins220 could help to design strategies to cleavage site, and with this information we prevent their pathological accumulation. have developed a new small molecule called Therefore, we aim to clarify the routes and peptide Tat-K with neuroprotective properties. molecular mechanisms involved in modulating This peptide, which prevents Kidins220 cut by Kidins220 proteostasis and trafficking, and some calpain in excitotoxicity, helps to preserve the of the proteins or complexes involved in these levels of this protein and pro-survival signaling processes. pathways associated with it (pERK and pCREB),

Keywords

Kidins220/ARMS, protein kinase D (PKD), neuroprotection, neurotoxicity, neuronal survival, Alzheimer, Parkinson

Publications

• Gamir-Morralla A, López-Menéndez C, Ayuso-Dolado S, Tejeda GS, Montaner J, Rosell A et al. Development of a neuroprotective peptide that preserves survival pathways by preventing Kidins220/ARMS calpain processing induced by excitotoxicity.Cell death & disease. 2015;6: e1939. • Schmieg N, Thomas C, Yabe A, Lynch DS, Iglesias T, Chakravarty P et al. Novel Kidins220/ARMS Splice Isoforms: Potential Specific Regulators of Neuronal and Cardiovascular Development. PloS one. ;10(6): e0129944. • Del Puerto A., Fronzaroli-Molinieres L., Perez-Alvarez M.J., Giraud P., Carlier E., Wandosell F. et al. ATP-P2X7 Receptor Modulates Axon Initial Segment Composition and Function in Physiological Conditions and Brain Injury. Cerebral Cortex. 2015;25(8):2282-2294.

2015 CIBERNED Annual Report / 158 Program 2 Group: Teresa Iglesias Vacas

Research projects

• Code: SAF2014-52737-P. • Code: 2013/07A. Title: Participación y regulación de PKD y Title: Papel de GSK-3 β en las alteraciones KIDINS220 en los procesos y rutas moleculares de los circuitos corticales que ocurren en la de la neurodegeneración. enfermedad de Alzheimer. Principal Investigator: Teresa Iglesias Vacas. Principal Investigator: Teresa Iglesias Vacas. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G504, G401, G403, G111, Other CIBER’s collaboration: No. G307. Type: National. Other CIBER’s collaboration: No. Funding agency: Ministerio de Economía y Type: International. Competitividad. Funding agency: CIBERNED. Funding: 193.600 €. Funding: 316.000 €. Duration: 2015-2017 Duration: 2013-2015 • Code: SAF2011-26233. • Code: S2010_BMD. Title: Participation of PKD and its substrate, Title: Redes de senalización y vías efectoras Kidins220, in the molecular pathways of en modelos celulares y animales de neuronal survival and neurodegeneration. enfermedades neurodegenerativas. Principal Invstigator: Teresa Iglesias Vacas. Principal Investigator: José González CIBERNED’s collaboration: No. Castaño. CIBERNED groups: CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED groups: G111, G401, G104, G307, Type: National. G409, G412. Funding agency: Ministerio de Economía y Other CIBER’s collaboration: Yes (CIBERER). Competitividad. Type: CC.AA. Funding: 205.710 €. Funding agency: Comunidad de Madrid. Duration: 2012-2015 Funding: 483.000 €. Duration: 2012-2016

PhD dissertations

• Author: Andrea Gamir Morralla. Title: Identificación de los mecanismos moleculares y rutas mediante los que Kidins220 participa en procesos de supervivencia neuronal y neurodegeneración. Diseño de posibles estrategias neuroprotectoras. Date: 20th March de 2015. Supervisor: Teresa Iglesias Vacas.

2015 CIBERNED Annual Report / 159 202 GROUP Jaime Kulisevsky Bojarski

Jaime Kulisevsky Bojarsky Javier Pagonabarraga Ramón Fernández de Principal investigator Mora Bobadilla Researcher PhD student Helena Berj Kasem Marco Researcher Berta Pascual Sedano Juan Martín Lahoz Researcher PhD student Antonia Campolongo Perillo Carles Roig Arnall Saúl Martínez Horta Researcher Researcher PhD student

Carmen García Sánchez Fabián Arenas Ríos Jesús Pérez Pérez Researcher Researcher PhD student

Alexandre Gironell Carreró María Victoria Sosti Sosa Researcher Researcher

Contact details

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]

2015 CIBERNED Annual Report / 160 Summary

Directed by Jaime Kulisevsky MD, PhD EP avanzada: (Movement Disorders Unit, Sant Pau Hospital, Biomedical Research Institute), we focus on • Erradicación de Helicobacter Pylori en research of Parkinson’s Disease (PD) and other pacientes con levodopa intraduodenal vs. Movement Disorders. During 2015 we initiated oral: efectos en estado motor, cognición y new projects and different objectives have calidad de vida. been achieved: • Análisis del estado nutricional de pacientes con levodopa intraduodenal. Genetics: • Estudio prospectivo de aspectos cognitivos y calidad de vida en EP y estimulación • Research on the influence of Mendelian cerebral profunda subtalámica bilateral. genes for sporadic forms of PD (Spataro et al, Mol Hum Genet 2015). “Musicoterapia” y biofeedback: • Participation in national multicenter project: study of MAPT H1 haplotype gene in • Double-blind study using biofeedback neurodegenerative diseases, including PD training Interfaces® Think about memory (Pastor et al, J Alz Dis 2015). and attention in Attention Deficit and • Study of the relationship of variations in the Hyperactivity Disorder (ADHD). number of copies (CNV) of the 17q21.31 • Double-blind study of music therapy and chromosomal region with the risk of cognitive stimulation on gait disorders in PD. sinucleopathies (Cervera-Carles et al, Am J Med Genet 2015). Huntington’s disease (HD): PD and cognition: • Study of prevalence of neuropsychiatric disorders in asymptomatic and in de novo • Continuation of the longitudinal study of PD individuals. patients <5 years of disease: motor, cognitive, • Development of cognitive assessment emotional and behavioral predictors of instruments “Cognitive Phenotype Working cognitive impairment. Group” of the European Huntington’s • FIS PI14/02058: neurophysiological markers Disease Network (EHDN). in blood and progression of cognitive • Participation in the task force to design new impairment in PD. functional assessment scale Furst 2.0. • Validation of cognitive (PD-CRS) and Collaboration with the UB in the project functional scales (PD-CFRS) with the new • “The nuclear lamina in HD: its role in diagnostic criteria for mild cognitive pathophysiology and therapeutic impairment (MCI) in PD (paper under applications” (Grant Ramon Areces). review). Collaboration with the UB in the project • Development of an alternative version of • “Overcoming Depression in Huntington’s the PD-CRS for re-test in clinical trials (paper Disease: Cdk5 as a potential biomarker and under review). pharmacological target”. • Validation of cognitive scales (PD-CRS) • Longitudinal changes of functional and (paper in preparation) and functional structural connectivity in pre-symptomatic (PD-CFRS) (published in J Neur Sci) in patients (FIS PI14/00834). neurodegenerative diseases different of PD. • “Phenotype and haplotypes associated with • Psychometric comparative study of different intermediate alleles of HD gene (htt)”. (FIS) cognitive scales for PD. • Prospective analysis of nutritional status of • Broadcasting, translation and validation of patients with pre-motor and motor HD. neuropsychological PD scales in international web platform (Www.movementscales. com). Tremor: • Design of DuoCog Project: randomized, double-blind crossover study comparing • Development of online system for immediate vs. intraduodenal levodopa in neurophysiological study of tremor by cognition and mood in PD patients. Smartphone. Application for grant with the company SEIDOR and the University of Vic. • Characterization of different tremors in PD. Apathy and behavior in PD: • Drug efficacy study of ethosuximide in essential tremor. • FIS PI15/00962: functional anatomical correlates of structural abnormalities on apathy, hallucinations and cognitive impairment.

2015 CIBERNED Annual Report / 161 Dystonia: Translational research:

• Spanish draft consensus on “Evidence and • Transgenic rat model of Huntington (BACHD): myths about the use of botulinum toxin in longitudinal study of motor, emotional and dystonia and spasticity”. cognitive disorders. • Model of hemi parkinsonism in rats: study of transcriptional striatal changes with different Ataxia-premutation tremor associated with doses of levodopa. Fragile X syndrome (FXTAS):

• Assistance and research in collaboration with the Genetics Department of Hospital Clínic (Barcelona) and the Catalan Association X Fragile (ACXF).

Keywords

Parkinson, cognition, mild cognitive impairment, Huntington, dystonia- FxTAS, essential tremor - animal models

Publications

• Puigdemont D., Portella M.J., Perez-Egea R., Molet J., Gironell A., de Diego-Adelino J. et al. A randomized double-blind crossover trial of deep brain stimulation of the subcallosal cingulate gyrus in patients with treatment-resistant depression: A pilot study of relapse prevention. Journal of Psychiatry and Neuroscience. 2015;40(4):224-231. • Gomez-Anson B., Roman E., De Bobadilla R.F., Pires-Encuentra P., Diaz-Manera J., Nuez F. et al. Alterations in cerebral white matter and neuropsychology in patients with cirrhosis and falls. PLoS ONE. 2015;10(3). [Epub ahead of print]. • Pagonabarraga J., Kulisevsky J., Strafella A.P., Krack P. Apathy in Parkinson’s disease: Clinical features, neural substrates, diagnosis, and treatment. The Lancet Neurology. 2015;14(5):518- 531. • Krack P., Pagonabarraga J., Strafella A.P., Kulisevsky J. Apathy: Who cares?. The Lancet Neurology. 2015;14(5):465-. [Epub ahead of print]. • Marti-Fabregas J., Delgado-Mederos R., Crespo J., Pena E., Marin R., Jimenez-Xarrie E. et al. Circulating endothelial progenitor cells and the risk of vascular events after ischemic stroke. PLoS ONE. 2015;10(4). [Epub ahead of print]. • Odin P., Ray Chaudhuri K., Slevin J.T., Volkmann J., Dietrichs E., Martinez-Martin P. et al. Collective physician perspectives on non-oral medication approaches for the management of clinically relevant unresolved issues in Parkinson’s disease: Consensus from an international survey and discussion program. Parkinsonism and Related Disorders. 2015;21(10):1133-1144. • Perez-Perez J., Pagonabarraga J., Martinez-Horta S., Fernandez-Bobadilla R., Sierra S., Pascual- Sedano B. et al. Head-to-Head Comparison of the Neuropsychiatric Effect of Dopamine Agonists in Parkinson’s Disease: A Prospective, Cross-Sectional Study in Non-demented Patients. Drugs and Aging. 2015;32(5):401-407. • Marin-Lahoz J., Gironell A. Linking Essential Tremor to the Cerebellum: Neurochemical Evidence. Cerebellum. 2015. [Epub ahead of print]. • Pagonabarraga J., Martinez-Horta S., Fernandez de Bobadilla R., Perez J., Ribosa-Nogue R., Marin J. et al. Minor hallucinations occur in drug-naive Parkinson’s disease patients, even from the premotor phase. Movement Disorders. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 162 Program 2 Group: Jaime Kulisevsky Bojarski

• Martinez-Horta S., Perez-Perez J., van Duijn E., Fernandez-Bobadilla R., Carceller M., Pagonabarraga J. et al. Neuropsychiatric symptoms are very common in premanifest and early stage HD. Parkinsonism and Related Disorders. 2015. [Epub ahead of print]. • Ruzafa-Valiente E., Fernandez-Bobadilla R., Garcia-Sanchez C., Pagonabarraga J., Martinez- Horta S., Kulisevsky J. Parkinson’s Disease - Cognitive Functional Rating Scale across different conditions and degrees of cognitive impairment. Journal of the Neurological Sciences. 2016;361:66-71. Epub 2015. • Trenkwalder C., Chaudhuri K.R., Martinez-Martin P., Rascol O., Ehret R., Valis M. et al. Prolonged- release oxycodone-naloxone for treatment of severe pain in patients with Parkinson’s disease (PANDA): A double-blind, randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(12):1161-1170. • Perez-Perez J., Martinez-Horta S., Pagonabarraga J., Carceller M., Kulisevsky J. Rasagiline for the treatment of parkinsonism in Huntington’s disease. Parkinsonism and Related Disorders. 2015;21(3):340-342. • Gironell A, Ribosa-Nogué R, Gich I, Marin-Lahoz J, Pascual-Sedano B. Severity stages in essential tremor: a long-term retrospective study using the glass scale.Tremor and other hyperkinetic movements (New York, N.Y.). ;5: 299. • Riba J., Valle M., Sampedro F., Rodriguez-Pujadas A., Martinez-Horta S., Kulisevsky J. et al. Telling true from false: Cannabis users show increased susceptibility to false memories. Molecular Psychiatry. 2015;20(6):772-777. • Cattaneo G, Calabria M, Marne P, Gironell A, Abutalebi J, Costa A. The role of executive control in bilingual language production: A study with Parkinson’s disease individuals.Neuropsychologia. 2015;66: 99-110. • Pagonabarraga J., Pinol G., Cardozo A., Sanz P., Puente V., Otermin P. et al. Transdermal rotigotine improves sleep fragmentation in parkinson’s disease: Results of the multicenter, prospective SLEEP-FRAM study. Parkinson’s Disease. 2015;2015. [Epub ahead of print]. • Lovestone S, Boada M, Dubois B, Hüll M, Rinne JO, Huppertz HJ et al. A phase II trial of tideglusib in Alzheimer’s disease.Journal of Alzheimer’s disease : JAD. 2015;45(1): 75-88. • Cervera-Carles L., Pagonabarraga J., Pascual-Sedano B., Pastor P., Campolongo A., Fortea J. et al. Copy number variation analysis of the 17q21.31 region and its role in neurodegenerative diseases. American Journal of Medical Genetics, Part B: Neuropsychiatric Genetics. 2015. [Epub ahead of print]. • Poewe W., Seppi K., Fitzer-Attas C.J., Wenning G.K., Gilman S., Low P.A. et al. Efficacy of rasagiline in patients with the parkinsonian variant of multiple system atrophy: A randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(2):145-152. • Buongiorno M, Antonelli F, Cámara A, Puente V, de Fabregues-Nebot O, Hernandez-Vara J et al. Long-term response to continuous duodenal infusion of levodopa/carbidopa gel in patients with advanced Parkinson disease: The Barcelona registry.Parkinsonism & related disorders. 2015;21(8): 871-6. • Pastor P, Moreno F, Clarimón J, Ruiz A, Combarros O, Calero M et al. MAPT H1 Haplotype is Associated with Late-Onset Alzheimer’s Disease Risk in APOEε4 Noncarriers: Results from the Dementia Genetics Spanish Consortium.Journal of Alzheimer’s disease : JAD. 2015;49(2): 343- 52. • Hor H., Francescatto L., Bartesaghi L., Ortega-Cubero S., Kousi M., Lorenzo-Betancor O. et al. Missense mutations in TENM4, a regulator of axon guidance and central myelination, cause essential tremor. Human Molecular Genetics. 2015;24(20):5677-5686.

2015 CIBERNED Annual Report / 163 Research projects

• Code: 2010 SGR 1203. • Code: PI14/02058. Title: AGAUR SGR 2014. Title: Marcadores en sangre y neurofisiológicos Principal Investigator: Jaime Kulisevsky. de la progresión del deterioro cognitivo en CIBERNED’s collaboration: No. la enfermedad de Parkinson. CIBERNED groups: Principal Investigator: Javier Pagona Other CIBER’s collaboration: No. Barraga. Type: Otros. CIBERNED’s collaboration: No. Funding agency: Agència de Gestio d'Ajuts CIBERNED groups: Universitaris i de Recerca. Other CIBER’s collaboration: No. Funding: 21.000 €. Type: International. Duration: 2014-2016 Funding agency: Ministerio de Economía y Competitividad. • Code: 2014 SGR 1203. Funding: 65.945 €. Title: Grupo de Trastornos del Movimiento Duration: 2015 Hospital de la Santa Creu i Sant Pau. Principal Investigator: Jaime Kulisevsky. • Code: 112610. CIBERNED’s collaboration: No. Title: Early implementation of music therapy CIBERNED groups: in rehabilitation of Aphasic patients after Other CIBER’s collaboration: No. acute adquired brain injury /Efectivity of Type: International. music therapy in the rehabilitation of acute Funding agency: Agència de Gestio d'Ajuts aphasic patients. Universitaris i de Recerca (AGAUR). Principal Investigator: Jaime Kulisevsky. Funding: 21.000 €. CIBERNED’s collaboration: No. Duration: 2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: PI12/03005. Type: Private. Title: Desarrollo, validación y estudio de Funding agency: Fundació La Marató de la sensibilidad al cambio de una versión TV3. alternativa de la Parkinson's Disease - Funding: 105.091,25 €. Cognitive Rating Scale. Duration: 2012-2015 Principal Investigator: Jaime Kulisevsy CIBERNED’s collaboration: No. • Code: Telemarato 2011 Lesiones medulares CIBERNED groups: y cerebrales. Other CIBER’s collaboration: No. Title: Efectividad de la musicoterapia en Type: National. la rehabilitación en pacientes afásicos Funding agency: Ministerio de Economía y agudos. Competitividad. Principal Investigator: Carmen García Funding: 28.435 €. Sánchez. Duration: 2013-2015 CIBERNED’s collaboration: No. CIBERNED groups: • Code: MIA 2012-05. Other CIBER’s collaboration: No. Title: Contrato MIA 2012. Type: National. Principal Investigator: Jesús Pérez. Funding agency: Fundacio La Marato de CIBERNED’s collaboration: No. TV3. CIBERNED groups: Funding: 105.091,25 €. Other CIBER’s collaboration: No. Duration: 2012-2015 Type: Private. Funding agency: Fundación Privada Hospital • Code: La lámina nuclear en la enfermedad de la Santa Creu i Sant Pau. de Huntington: papel en la fisiopatología y Funding: 45.222,91 €. aplicaciones terapéuticas. Duration: 2015 Title: La lámina nuclear en la enfermedad de Huntington: papel en la fisiopatología y • Code: CM13/00284. aplicaciones terapéuticas. Title: Contratos Rio Hortega 2013. Principal Investigator: Esther Pérez Navarro. Principal Investigator: Jesús Pérez. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G202, G301. CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: International. Funding agency: Fundación Ramón Areces. Funding agency: Ministerio de Economía y Funding: 121.440 €. Competitividad. Duration: 2015 Funding: 53.732 €. Duration: 2015

2015 CIBERNED Annual Report / 164 Program 2 Group: Jaime Kulisevsky Bojarski

• Code: 484/U/2014 MARATO. • Code: 477/U/2014 MARATO. Title: Blood-based and neurophysiological Title: Prediction of apathy and impulse markers of cognitive deterioration and control disorders in Parkinson Disease based dementia in Parkinson's Disease. on feedback related negativity. Principal Investigator: Javier Principal Investigator: Jaime Kulisevsky. Pagonabarraga. CIBERNE’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: National. Funding agency: Fundació La Marató de Funding agency: Fundació La Marató de TV3. TV3. Funding: 199.607,5 €. Funding: 199.486,25 €. Duration: 2015-2017 Duration: 2015-2017 • Code: 2015/02. • Code: 477. Title: Metabolismo oxidoreductor y Title: Prediction of apathy and impulse enfermedad de Parkinson: validación control disorders in Parkinson's disease based de nuevas dianas terapéuticas y nuevos on the Feedback Related Negativity (FRN), biomarcadores. a neurophysiological marker of incentive Principal Investigator: José Luis Labandeira. processing. CIBERNED’s collaboration: Yes. Principal Investigator: Jaime Kulisevsky. CIBERNED groups: G101, G202, G208, G203. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED: Type: National. Other CIBER’s collaboration: No. Funding agency: CIBERNED. Type: CC.AA. Funding: 240.000 €. Funding agency: Fundació la Marató de Duration: 2015-2017 TV3. Funding: 199.607,5 €. Duration: 2015

2015 CIBERNED Annual Report / 165 208 GROUP José Luis Labandeira García

José Luis Labandeira Ana I. Rodríguez Pérez Antonio Domínguez Meijide García Associate professor Postdoctoral researcher Full professor Carmen Díaz Ruiz Mª Alicia Costa Besada María J. Guerra Seijas Associate professor Predoctoral researcher Full professor Begoña Villar Cheda María García Garrote Ramón Soto Otero Associate professor Predoctoral researcher Full professor Pablo Garrido Gil Iria Novoa Pérez Estefanía Méndez Álvarez Postdoctoral researcher Specialist Technician Full professor Rita Valenzuela Limiñana Pilar Aldrey García Jannette Rodríguez Pallares Predoctoral researcher Specialist Technician Associate professor Juan A. Parga Martín José A. Trillo Franco Ana M. Muñoz Patiño Postdoctoral researcher Specialist Technician Associate professor Mª Cristina Gianzo Quintela Specialist Technician Contact details

Center for Research in Molecular Medicine and Chronic Diseases (CIMUS) 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]

2015 CIBERNED Annual Report / 166 Summary

In 2015, we have investigated mechanisms 63:466-82; Neuroscientist 21:616-29). Additional involved in the higher dopaminergic mechanisms related with aging were analyzed vulnerability with aging. First, we have to know their possible role in aging-related collaborated in the design of more adequate higher dopaminergic vulnerability, and their animal models of Parkinson´s disease (PD; J possible manipulation for neuroprotective NeurochemPMID:26500044). Using in vitro and therapy in PD: the brain renin-angiotensin in vivo models, we have also investigated system(Biochem Pharmacol 96:131-42), sirtuins the current mechanisms responsible for the (Oncotarget 6:26675-89), and CB1 y CB2 conflicting results between experimental and receptors (Brain Struct Funct 220:2721-38). We clinical studies on the beneficial effects of also investigated new mechanisms involved menopausal hormonal replacement therapy, in dyskinesias induced by L-dopa therapy in particularly in PD; we investigated the window animal models of PD in order to develop more of opportunity or critical period hypothesis and effective therapies (Mol Neurobiol 52:1408- possible mechanisms involved (Neurobiol Aging 20; Mol Neurobiol PMID:26452359). Finally, we 36:11904-208). have also collaborated in the design of new multitarget inhibitors of monoamine oxidase We also studied the role of Rho kinase (ROCK) B and cholinesterases for PD (J Med Chem in the aging-related higher dopaminergic 58:5561-78; Eur J Med Chem 89:98-105). An vulnerability (J Gerontol A Biol Sci Med Sci. important part of these studies were undertaken PMID:26503374) as well as the role of ROCK in collaboration with other groups of Ciberned in neuroinflammation and dopaminergic (Dr. Lanciego, Drs Canela/R. Franco, Drs. Lopez- degeneration, and the possible use of ROCK Barneo/Toledo) and CiberObn (Dieguez/ inhibitors as neuroprotectors for PD (Glia Beiroa).

Keywords

Aging, angiotensin, dyskinesias, neurodegeneration, neuroinflammation, neuroprotection, Parkinson.

Publications

• Muñoz A, Corrêa CL, Villar-Cheda B, Costa-Besada MA, Labandeira-Garcia JL. Aging-related Increase in Rho Kinase Activity in the Nigral Region Is Counteracted by Physical Exercise.The journals of gerontology. Series A, Biological sciences and medical sciences. 2015. [Epub ahead of print]. • Munoz-Manchado A.B., Villadiego J., Romo-Madero S., Suarez-Luna N., Bermejo-Navas A., Rodriguez-Gomez J.A. et al. Chronic and progressive Parkinson’s disease MPTP model in adult and aged mice. Journal of Neurochemistry. 2015. [Epub ahead of print]. • Franco R, Casadó-Anguera V, Muñoz A, Petrovic M, Navarro G, Moreno E et al. Hints on the Lateralization of Dopamine Binding to D₁ Receptors in Rat Striatum.Molecular neurobiology. 2015. [Epub ahead of print]. • Rodriguez-Perez A.I., Borrajo A., Rodriguez-Pallares J., Guerra M.J., Labandeira-Garcia J.L. Interaction between NADPH-oxidase and Rho-kinase in angiotensin II-induced microglial activation. GLIA. 2015;63(3):466-482. • Diaz-Ruiz C., Rodriguez-Perez A.I., Beiroa D., Rodriguez-Pallares J., Labandeira-Garcia J.L. Reciprocal regulation between sirtuin-1 and angiotensin-II in the substantia nigra: Implications for aging and neurodegeneration. Oncotarget. 2015;6(29):26675-26689. • Labandeira-Garcia JL, Rodríguez-Perez AI, Villar-Cheda B, Borrajo A, Dominguez-Meijide A, Guerra MJ. Rho Kinase and Dopaminergic Degeneration: A Promising Therapeutic Target for Parkinson’s Disease.The Neuroscientist : a review journal bringing neurobiology, neurology and psychiatry. 2015;21(6): 616-29. • Farina R., Pisani L., Catto M., Nicolotti O., Gadaleta D., Denora N. et al. Structure-Based Design and Optimization of Multitarget-Directed 2 H -Chromen-2-one Derivatives as Potent Inhibitors of Monoamine Oxidase B and Cholinesterases. Journal of Medicinal Chemistry. 2015;58(14):5561- 5578.

2015 CIBERNED Annual Report / 167 • Rodriguez-Perez AI, Borrajo A, Valenzuela R, Lanciego JL, Labandeira-Garcia JL. Critical period for dopaminergic neuroprotection by hormonal replacement in menopausal rats.Neurobiology of aging. 2015;36(2): 1194-208. • Sierra S, Luquin N, Rico AJ, Gómez-Bautista V, Roda E, Dopeso-Reyes IG et al. Detection of cannabinoid receptors CB1 and CB2 within basal ganglia output neurons in macaques: changes following experimental parkinsonism.Brain structure & function. 2015;220(5): 2721-38. • Martinez-Pinilla E., Rodriguez-Perez A.I., Navarro G., Aguinaga D., Moreno E., Lanciego J.L. et al. Dopamine D2 and angiotensin II type 1 receptors form functional heteromers in rat striatum. Biochemical Pharmacology. 2015;96(2):131-142.

Research projects

• Code: 2015-CE009. • Code: PI 2015/02. Title: Financiación línea de investigación Title: Validación de nuevas dianas en enf. neuromuscuylares. Trastornos del terapéuticas y nuevos biomarcadores en movimiento. enfermedad de Parkinson. Principal Investigator: José luis Labandeira. Principal Investigator: José Luis Labandeira. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: National. Funding agency: Fundación de La Luz. Funding agency: CIBERNED. Funding: 31.000 €. Funding: 240.000 €. Duration: 2015-2016 Duration: 2015-2017 • Code: R2014/50. • Code: RD12/0019. Title: Red Gallega de Terapia Celular. Title: RETIC de Terapia Celular. Principal Investigator: Clara Álvarez (IP Nodo Principal Investigator: Coordinador de red: J.L. Labandeira). José María Moraleda. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G102, G301, G113, G208, Other CIBER’s collaboration: Yes (CIBERBBN, G105, G607. CIBEROBN). Other CIBER’s collaboration: No. Type: CC.AA. Type: National. Funding agency: Conselleria de Educación. Funding agency: Fondo de Investigaciones Xunta Galicia. Sanitarias. Funding: 120.000 €. Funding: 296.700 €. Duration: 2014-2015 Duration: 2013-2016 • Code: 2015/02. • Code: GRC2014/002. Title: Metabolismo oxidoreductor y Title: Consolidación y estructuración de enfermedad de Parkinson: validación grupos de referencia competitiva. de nuevas dianas terapéuticas y nuevos Principal Investigator: José Luis Labandeira. biomarcadores. CIBERNED’s collaboration: No. Principal Investigator: José Luis Labandeira. CIBERNED groups: CIBERNED’s collaboration: Yes. Other CIBER’s collaboration: No. CIBERNED group: G101, G202, G208, G203. Type: International. Other CIBER’s collaboration: No. Funding agency: Conselleria de Educación. Type: National. Xunta de Galicia. Funding agency: CIBERNED. Funding: 320.000 €. Funding: 240.000 €. Duration: 2015-2018 Duration: 2015-2017

PhD dissertations

• Author: Antonio Domínguez Meijide. Title: Interacción entre el sistema dopaminérgico nigro-estriatal y el sistema renina-angiotensina local. Implicaciones en la enfermedad de Parkinson. Date: 18th Decembre 2015. Supervisor: José Luis Labandeira García.

2015 CIBERNED Annual Report / 168 203 GROUP José Luis Lanciego Pérez

José Luis Lanciego Iria María González- Diego Sucunza Guibert Principal investigator Dopeso Reyes PhD student PhD Gloria González- Elvira Roda Recalde Aseguinolaza Eva Martínez-Pinilla Lab technician Research PhD

Alberto José Rico Martín José Diego Pignataro PhD PhD student

Contact details

Fundación para la Investigación Médica Aplicada (FIMA) Centro de Investigación Médica Aplicada (CIMA) Departamento de Neurociencias Laboratorio de Neuroanatomía 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]

2015 CIBERNED Annual Report / 169 Summary

Studies on heteromers made of G protein- vectors for modifying brain circuits. It is expected coupled receptors (GPCRs): that these genetic manipulations of brain circuits will ultimately sustain a direct therapeutic effect. It might be argued that GPCRs have a natural By using relevant animal models of Parkinson’s tendency to form heteromeric complexes disease, we have designed different types of that represent novel molecular entities adeno-associated (AAVs) and lentiviral vectors with properties distinct from those of each that are currently being explored within the component receptor, when considered following specific projects: separately. GPCR heteromers represent novel targets for pharmacological developments • Reconstruction of the nigrostriatal pathway in the field of Parkinson’s disease. At present, in animals showing a severe parkinsonian our main activities focus on GPCR heteromers syndrome by using Rheb-carrying AAVs. made of dopaminergic receptors (D1 and D2), • Selective elimination of hyperactive basal as well as those formed by endocannabinoid ganglia circuits with pseudotyped lentiviruses, receptors (CB1, CB2 and GPR55). strategy known as “immunotoxin-mediated tract-targeting”. Gene therapy for Parkinson’s disease: • Direct in situ cellular reprogramming using AAVs coding for a combination of 3 The field of gene therapy has recently witnessed transcription factors. a numer of major conceptual changes. Instead • Selective clearance of alpha-synuclein of using viral vectors for the delivery of a given aggregates by means of pseudotyped therapeutic gene, the current tendency in the lentiviruses carrying the gene coding for field relies on the use of gene-carrying viral beta-glucocerebrosidase.

Keywords

GPCR, dopamine, cannabis, basal ganglia, gene therapy, adeno-associated viral vectors, lentiviruses

Publications

• Rodriguez-Perez AI, Borrajo A, Valenzuela R, Lanciego JL, Labandeira-Garcia JL. Critical period for dopaminergic neuroprotection by hormonal replacement in menopausal rats.Neurobiology of aging. 2015;36(2): 1194-208. • Sierra S, Luquin N, Rico AJ, Gómez-Bautista V, Roda E, Dopeso-Reyes IG et al. Detection of cannabinoid receptors CB1 and CB2 within basal ganglia output neurons in macaques: changes following experimental parkinsonism.Brain structure & function. 2015;220(5): 2721-38. • Deng Y., Lanciego J., Goff L.K.-L., Coulon P., Salin P., Kachidian P. et al. Differential organization of cortical inputs to striatal projection neurons of the matrix compartment in rats. Frontiers in Systems Neuroscience. 2015;9(APR). [Epub ahead of print]. • Martinez-Pinilla E., Rodriguez-Perez A.I., Navarro G., Aguinaga D., Moreno E., Lanciego J.L. et al. Dopamine D2 and angiotensin II type 1 receptors form functional heteromers in rat striatum. Biochemical Pharmacology. 2015;96(2):131-142. • Blesa J., Lanciego J.L., Obeso J.A. Editorial: Parkinson’s disease: Cell vulnerability and disease progression. Frontiers in Neuroanatomy. 2015;9(September). [Epub ahead of print]. • Franco R, Casadó-Anguera V, Muñoz A, Petrovic M, Navarro G, Moreno E et al. Hints on the Lateralization of Dopamine Binding to D₁ Receptors in Rat Striatum.Molecular neurobiology. 2015. [Epub ahead of print]. • Garcia-Gutierrez M.S., Navarrete F., Aracil A., Bartoll A., Martinez-Gras I., Lanciego J.L. et al. Increased vulnerability to ethanol consumption in adolescent maternal separated mice. Addiction Biology. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 170 Program 2 Group: José Luis Lanciego Pérez

• Aguirre M.E.E., Zelaya M.V., de Gordoa J.S.R., Tunon M.T., Lanciego J.L. Midbrain catecholaminergic neurons co-express α-synuclein and tau in progressive supranuclear palsy. Frontiers in Neuroanatomy. 2015;9(MAR). [Epub ahead of print]. • Liberia T., Blasco-Ibanez J.M., Nacher J., Varea E., Lanciego J.L., Crespo C. Synaptic connectivity of the cholinergic axons in the olfactory bulb of the cynomolgus monkey. Frontiers in Neuroanatomy. 2015;9(MAR). [Epub ahead of print]. • Gomez-Vallejo V., Ugarte A., Garcia-Barroso C., Cuadrado-Tejedor M., Szczupak B., Dopeso- Reyes I.G. et al. Pharmacokinetic investigation of sildenafil using positron emission tomography and determination of its effect on cerebrospinal fluid cGMP levels. Journal of Neurochemistry. 2016;136(2):403-415. Epub 2015.

Research projects

• Code: BFU2012-37907. CIBERNED’s collaboration: Yes. Title: Coexpresión de receptores de CIBERNED groups: G101, G202, G208, G203. adenosina 2A y de endocannabinoides 1 Other CIBER’s collaboration: No. y 2 en los núcleos de salida de los ganglios Type: National. basales en el modelo de enfermedad de Funding agency: CIBERNED. Parkinson en monos. Funding: 240.000 €. Principal Investigator: José Luis Lanciego. Duration: 2015-2017 CIBERNED’s collaboration: No. CIBERNED groups: Code: COEN-PATHFINDER. Other CIBER’s collaboration: No. • Title: In vivo neuronal cell reprogramming for Type: International. a new regenerative approach in Parkinson's Funding agency: Ministerio de Economía y disease. Competitividad. Principal Investigator: José Luis Lanciego. Funding: 127.050 €. CIBERNED’s collaboration: No. Duratio: 2013-2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: 20141330. Type: International. Title: Cannabinoid receptor heteromeric Funding agency: Instituto de Salud Carlos III- complexes as therapeutic targets in CIBERNED. Parkinson's disease. Funding: 385.500 €. Principal Investigator: Rafael Franco Duration: 2013-2015 Fernández. CIBERNED’s collaboration: Yes. • Code: 340527 ERC Advanced. CIBERNED groups: G201, G203. Title: New Experimental therapeutic Other CIBER’s collaboration: No. approaches for Parkinson's disease by direct Type: Private. DA neuronal reprogramming. Funding agency: Fundació La Maratoóde Principal Investigator: José Luis Lanciego TV3. (Co-IP). Funding: 150.000 €. CIBERNED’s collaboration: No. Duration: 2015-2017 CIBERNED groups: Other CIBER’s collaboration: No. • Code: 2015/02. Type: International. Title: Metabolismo oxidoreductor y Funding agency: European Research enfermedad de Parkinson: validación Council (ERC Advanced Grants). de nuevas dianas terapéuticas y nuevos Funding: 2.415.767 €. biomarcadores. Duration: 2014-2018 Principal Investigator: José Luis Labandeira.

2015 CIBERNED Annual Report / 171 105 GROUP José López Barneo

José López Barneo Ivette López López Pablo Mir Rivera Principal investigator Technician MD

Candela Caballero Erazo José Antonio Rodríguez Patricia González MD Gómez Rodríguez Postdoctoral fellow Postdoctoral fellow Francisco Javier Villadiego Luque Juan José Toledo Aral Patricia Ortega Sáenz Postdoctoral fellow Researcher Researcher

Gloria Paula García Flores Lin Gao Pilar Gómez Garre Tecnician Researcher Researcher

Helia Sarmiento Soto María del Carmen Ricardo Pardal Redondo Technician Fernández-Aguera Researcher Rodríguez Ignacio Arias Mayenco PhD student Victoria Bonilla Henao Technician Technician Nela Suárez Luna Technician Xavier d’Anglemont de Tassigny Postdoctoral fellow Contact details

Hospital Universitario Virgen del Rocío Universidad de Sevilla Avenida Manuel Siurot s/n 41013 Sevilla (Spain) Tel.: +34 95 592 30 00 Fax: +34 95 592 31 01 [email protected]

2015 CIBERNED Annual Report / 172 Summary

In 2015, our group has developed three research Cell therapy in PD: lines related to CIBERNED: In continuation with the transplantations Pathogenesis of Parkinson’s disease (PD): of rat carotid body (CB) and CB-derived neurospheres on parkinsonian mice, we have We are currently studying the early stages performedd in collabration with Dr. Juan José of neurodegenerative diseases, such as Toledo-Aral, transplants of human CB on mice Parkinson’s disease (PD), using three transgenic to study the trophic effect exerted by GDNF on animal models carrying deletions in distinct the nigrostriatal neurons. Indeed, these human mitochondrial complexes (I, II and III). CB grafts have shown that human CB (as much as in experimental models) provides a trophic We have developed several animal models action on the nigrostriatal pathway. Moreover, that are now in the characterization step to i) various criterias (i.e. age and sex of the human advance in the understanding of the role of donors) have been evaluated on the trophic endogenous neurotrophic glial derived factor action exerted by the transplanted CB tissue. (GDNF) on the maintenance of the nigrostriatal In this regard, age is represents a limiting factor pathway, ii) study the difference at the that reduces the therapeutic efficacy of carotid molecular level of the intracellular mechanisms tissue, and we are now further studying the leading to the synthesis and secretion of GDNF molecular mechanisms involved in this aspect. in the striatum, and iii) investigate methods to In any case, the neurotrophic actions on the increase the intrastriatal release of GDNF. For this, nigrostriatal pathway by the human CB and we are also setting up a cell sorting protocol to the human CB-derived neurospheres are under compare genomic and proteomic patterns of writing for publication. the striatal parvalbumin neurons (GDNF+) from the same neurons located in the nearby cortex Effect of hypoxia on the neurogenic niches in (negative for GDNF). This will help to understand the central and peripheral nervous system: the specific regulation of endogenous GDNF synthesis. As part of this research project, we are investigating the effects of chronic and In parallel to the animal models, we are intermittent hypoxia on the neurogenic niche. continuing the study on genetic alterations We have developed and characterized two related to PD and movement disorders. models of chronic and intermittent hypoxia in Additionally, we are monitoring the putative rodent animals. These models are now in use cortical activity changes in PD patients by using in combination with several state-of-the-art transcranial magnetic stimulation. techniques to analyze the changes occuring in the neurogenic niche at both the central (subventricular zone SVZ) and peripheral (CB) levels.

Keywords

Hypoxia, ion channels, carotid body, movement disorders, Parkinson’s disease

Publications

• Martin-Rodriguez J.F., Ruiz-Rodriguez M.A., Palomar F.J., Caceres-Redondo M.T., Vargas L., Porcacchia P. et al. Aberrant cortical associative plasticity associated with severe adult Tourette syndrome. Movement Disorders. 2015;30(3):431-435. • Fernandez-Santiago R., Carballo-Carbajal I., Castellano G., Torrent R., Richaud Y., Sanchez- Danes A. et al. Aberrant epigenome in iPSC-derived dopaminergic neurons from Parkinson’s disease patients. EMBO Molecular Medicine. 2015. [Epub ahead of print]. • Galan-Cobo A., Ramirez-Lorca R., Toledo-Aral J.J., Echevarria M. Aquaporin-1 plays important role in proliferation by affecting cell cycle progression. Journal of Cellular Physiology. 2016;231(1):243-256. Epub 2015.

2015 CIBERNED Annual Report / 173 • Lopez-Barneo J., Macias D., Platero-Luengo A., Ortega-Saenz P., Pardal R. Carotid body oxygen sensing and adaptation to hypoxia. Pflugers Archiv European Journal of Physiology. 2016;468(1):59-70. Epub 2015. • Labrador-Garrido A, Cejudo-Guillén M, Daturpalli S, Leal MM, Klippstein R, De Genst EJ et al. Chaperome screening leads to identification of Grp94/Gp96 and FKBP4/52 as modulators of the α-synuclein-elicited immune response.FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 2016;30(2):-. Epub 2015. • Odin P., Ray Chaudhuri K., Slevin J.T., Volkmann J., Dietrichs E., Martinez-Martin P. et al. Collective physician perspectives on non-oral medication approaches for the management of clinically relevant unresolved issues in Parkinson’s disease: Consensus from an international survey and discussion program. Parkinsonism and Related Disorders. 2015;21(10):1133-1144. • Wasserman K, Kisaka T, Luehrs RE, Bates ML, Kumar VH, Lopez-Barneo J et al. Commentaries on Viewpoint: Why do some patients stop breathing after taking narcotics? Ventilatory chemosensitivity as a predictor of opioid-induced respiratory depression.Journal of applied physiology (Bethesda, Md. : 1985). 2015;119(4): 423-5. • Poewe W., Seppi K., Fitzer-Attas C.J., Wenning G.K., Gilman S., Low P.A. et al. Efficacy of rasagiline in patients with the parkinsonian variant of multiple system atrophy: A randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(2):145-152. • Huertas-Fernandez I., Gomez-Garre P., Madruga-Garrido M., Bernal-Bernal I., Bonilla-Toribio M., Martin-Rodriguez J.F. et al. GDNF gene is associated with tourette syndrome in a family study. Movement Disorders. 2015. [Epub ahead of print]. • De Tassigny X.D., Pascual A., Lopez-Barneo J. GDNF-based therapies, GDNF-producing interneurons, and trophic support of the dopaminergic nigrostriatal pathway. Implications for parkinson’s disease. Frontiers in Neuroanatomy. 2015;9(FEB):1-15. • González-Rodríguez P, Falcón D, Castro MJ, Ureña J, López-Barneo J, Castellano A. Hypoxic induction of T-type Ca(2+) channels in rat cardiac myocytes: role of HIF-1α and RhoA/ROCK signalling.The Journal of physiology. 2015;593(21): 4729-45. • Oropesa-Ruiz J.M., Huertas-Fernandez I., Jesus S., Caceres-Redondo M.T., Vargas-Gonzalez L., Carrillo F. et al. Low serum uric acid levels in progressive supranuclear palsy. Movement Disorders. 2015. [Epub ahead of print]. • Huertas-Fernandez I., Garcia-Gomez F.J., Garcia-Solis D., Benitez-Rivero S., Marin-Oyaga V.A., Jesus S. et al. Machine learning models for the differential diagnosis of vascular parkinsonism and Parkinson’s disease using [123I]FP-CIT SPECT. European Journal of Nuclear Medicine and Molecular Imaging. 2015;42(1):112-119. • Ortega-Saenz P., Villadiego J., Pardal R., Toledo-Aral J.J., Lopez-Barneo J. Neurotrophic Properties, Chemosensory Responses and Neurogenic Niche of the Human Carotid Body. Advances in Experimental Medicine and Biology. 2015;860:139-152. • Ganos C., Erro R., Mir P., Lang A.E., Bhatia K.P., Vidailhet M. On the long-term outcome of orthostatic tremor. Parkinsonism and Related Disorders. 2015;21(10):1290-1291. • Fernandez-Aguera M.C., Gao L., Gonzalez-Rodriguez P., Pintado C.O., Arias-Mayenco I., Garcia-Flores P. et al. Oxygen Sensing by Arterial Chemoreceptors Depends on Mitochondrial Complex i Signaling. Cell Metabolism. 2015;22(5):825-837. • Lopez-Barneo J, Gonzalez-Rodriguez P, Gao L, Fernandez-Aguera MC, Pardal R, Ortega-Saenz P. OXYGEN SENSING BY THE CAROTID BODY: MECHANISMS AND ROLE IN ADAPTATION TO HYPOXIA.American journal of physiology. Cell physiology. 2016. Epub 2015. • López-Barneo J, Ortega-Sáenz P, González-Rodríguez P, Fernández-Agüera MC, Macías D, Pardal R et al. Oxygen-sensing by arterial chemoreceptors: Mechanisms and medical translation.Molecular aspects of medicine. ;47-48:-. Epub 2015. • Albanese A., Abbruzzese G., Dressler D., Duzynski W., Khatkova S., Marti M.J. et al. Practical guidance for CD management involving treatment of botulinum toxin: a consensus statement. Journal of Neurology. 2015;262(10):2201-2213. • Diaz-Castro B., Pardal R., Garcia-Flores P., Sobrino V., Duran R., Piruat J.I. et al. Resistance of glia- like central and peripheral neural stem cells to genetically induced mitochondrial dysfunction - Differential effects on neurogenesis. EMBO Reports. 2015;16(11):1511-1519.

2015 CIBERNED Annual Report / 174 Program 2 Group: José López Barneo

• Ganos C., Maugest L., Apartis E., Gasca-Salas C., Caceres-Redondo M.T., Erro R. et al. The long-term outcome of orthostatic tremor. Journal of Neurology, Neurosurgery and Psychiatry. 2015. [Epub ahead of print]. • Gómez-Caravaca MT, Cáceres-Redondo MT, Huertas-Fernández I, Vargas-González L, Carrillo F, Carballo M et al. The use of botulinum toxin in the treatment of sialorrhea in parkinsonian disorders.Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology. 2015;36(2): 275-9. • Munoz-Manchado A.B., Villadiego J., Romo-Madero S., Suarez-Luna N., Bermejo-Navas A., Rodriguez-Gomez J.A. et al. Chronic and progressive Parkinson’s disease MPTP model in adult and aged mice. Journal of Neurochemistry. 2015. [Epub ahead of print].

Research projects

• Code: Fundación Mutua Madrilena 13. Funding agency: Instituto de Salud Carlos III Title: Estudio de asociación de los genes Ministerio de Sanidad y Consumo. relacionados con plasticidad neuronal y Funding: 121.605 €. distonia primaria. Duration: 2014-2016 Principal Investigator: Pablo Mir Rivera. CIBERNED’s collaboration: No. • Code: SAF2012-33816. CIBERNED groups: Title: Caracterización de los mecanismos Other CIBER’s collaboration: No. intrínsecos implicados en el mantenimiento Type: National. de las neuronas de la SNPC. Papel de Funding agency: Fundación Mutua GDNF y de genes modulados por el Madrilena. envejecimiento. Funding: 33.000 €. Principal Investigator: Alberto Pascual Duration: 2013-2015 Bravo. CIBERNED’s collaboration: No. • Code: 278367. CIBERNED groups: Title: EMTICS (European Multicentre Tics in Other CIBER’s collaboration: No. Children Studies). Type: International. Principal Investigator: Pablo Mir Rivera. Funding agency: Ministerio de Economía y CIBERNED’s collaboration: No. Competitividad- FEDER. CIBERNED groups: Funding: 187.200 €. Other CIBER’s collaboration: No. Duration: 2013-2015 Type: European. Funding agency: Séptimo Programa Marco. • Code: PIE 2012 Excelencia (P12-CTS-2739 Comisión Europea. FP7-HEALTH-2011. MO). Funding: 100.533,6 €. Title: Terapia celular en la enfermedad de Duration: 2011-2016 Parkinson. Principal Investigator: Juan José Toledo • Code: FIS 12 (PI12/02574). Aral. Title: Uso de cuerpo carotideo en terapia CIBERNED’s collaboration: No. celular en la enfermedad de Parkinson. CIBERNED groups: Principal Investigator: Juan José Toledo Other CIBER’s collaboration: No. Aral. Type: National. CIBERNED’s collaboration: No. Funding agency: Consejeria de Economía, CIBERNED groups: Innovación, Ciencia y Empleo. Other CIBER’s collaboration: No. Funding: 191.875 €. Type: National. Duration: 2014-2017 Funding agency: Instituto de Salud Carlos III Ministerio de Sanidad y Consumo. • Code: SAF2013-48535-P. Funding: 109.505 €. Title: Fisiopatología de celulas madre Duration: 2013-2015 derivadas de la cresta neural. Principal Investigator: Ricardo Pardal. • Code: FIS 13 (PI13/01461). CIBERNED’s collaboration: No. Title: Plasticidad neuronal en las discinesias CIBERNED groups: inducidas por levodopa en la enfermedad Other CIBER’s collaboration: No. de Parkinson. Type: National. Principal Investigator: Pablo Mir Rivera. Funding agency: Ministerio de Economía y CIBERNED’s collaboration: No. Competitividad. CIBERNED groups: Funding: 250.000 €. Other CIBER’s collaboration: No. Duration: 2014-2016 Type: National.

2015 CIBERNED Annual Report / 175 • Code: Axontherapix (Prorroga). • Code: Fundación Marcelino Botín (Nuevo Title: Axontherapix. convenio). Principal Investigator: José López Barneo. Title: Sensibilidad al oxígeno y CIBERNED’s collaboration: No. neurodegeneracion. CIBERNED groups: Principal Investigator: José López Barneo. Other CIBER’s collaboration: Yes (Redes de CIBERNED’s collaboration: No. Terapia Celular). CIBERNED groups: Type: Nacional. Other CIBER’s collaboration: No. Funding agency: Axontherapix. Type: International. Funding: 251.217,5 €. Fundin agency: Fundación Marcelino Botin. Duration: 2015-2016 Funding: 650.000 €. Duration: 2013-2016 • Code: RD12/0019. Title: RETIC de Terapia Celular. • Code: ERC (FP7-260964). Principal Investigator: Coordinador de red: Title: Physiology of the adult carotid body José María Moraleda. stem cell niche. CIBERNED’s collaboration: Yes. Principal Investigator: Ricardo Pardal. CIBERNED groups: G102, G301, G113, G208, CIBERNED’s collaboration: No. G105, G607. CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: European. Funding agency: Fondo de Investigaciones Funding agency: Comisión de la Comunidad Sanitarias. Europea. Funding: 296.700 €. Funding: 1.476.000 €. Duration: 2013-2016 Duration: 2010-2015 • Code: SAF2012/39343. Title: Sensibilidad al oxígeno y neurodegeneracion. Principal Investigator: José López Barneo. CIBERNED’s collaboration: No. CIBERNED groups: Other CIBER’s collaboration: No. Type: National. Funding agency: Ministerio de Economía y Competividad. Funding: 400.000 €. Duration: 2013-2015

PhD dissertations

• Author: María del Carmen Fernández- Aguera Rodríguez. Title: Mecanismos moleculares de la sensibilidad aguda al oxígeno en los quimioreceptores arteriales: función del complejo i mitocondrial. Date: 17th July de 2015. Supervisor: José López Barneo.

2015 CIBERNED Annual Report / 176 609 GROUP Adolfo López de Munain

Adolfo López de Munain M. Pilar Camaño Haritz Jiménez Urbieta Juanjo Poza Aldea Principal investigator González PhD student PhD PhD student María Cruz Rodríguez Jaione Lacalle Prieto Patrícia de la Riva Juez Oroz Leire Casas Fraile PhD Neurologist Co Principal investigator PhD student José Félix Martí Masso Tatiana Rodríguez Ana Aiastui Pujana Manuel Delgado PhD Chinchilla PhD Alvarado PhD student PhD student Pablo Martínez - Lage Garazi Aldanondo Alvarez Javier Ruiz Martínez Aristizabal Roberto Fernández Torrón Bachelor degree PhD PhD student PhD student Alba Mateos Mateos Amets Sáenz Peña Miren Karmele Arnaiz Belén Gago Calderón Ayerdi PhD Pérez PhD PhD student Office manager Andone Sistiaga Federico García Fermín Moreno Izco Berrondo Myriam Barandiarán Bragado PhD student PhD Amillano PhD PhD student Neia Naldaiz Gastezi Ivan Toral Ojeda Francisco Javier Gil Bea PhD student PhD student Alberto Bergareche PhD Yarza Irene Navalpotro Gómez Ainara Vallejo PhD student María Goicoechea PhD student Illarramendi Bianchi PhD Cristina Caballero PhD Nahikari Pastoriza Polo Martínez Technician Miren Zulaica Ijurco Researcher Ana Gorostidi Pagola Bachelor degree PhD

Contact details

Instituto de Investigación Biodonostia Pº Dr. Beguiristain s/n 20014 San Sebastián (Spain) Leyre Curto Arzac Tel.: +34 94 300 62 95 [email protected]

2015 CIBERNED Annual Report / 177 Summary

The main research lines are: using in vivo PET and post-mortem studies. On the other hand, the study of iPS derived from • Impulse control disorders (ICD) and dyskinesia: genetic forms (patients and asymptomatic Physiopathology, risk factors and therapeutic carriers of the LRRK2 R1441G mutation) and approaches in these complications derived sporadic cases of PD will allow to deepen from chronic dopaminergic treatment in in the relationship between glial cells, Parkinson’s disease. We focus on clinical and neuroinflammation and neuronal death in experimental research in animal models with both sporadic and genetic forms of PD. several approaches comprising behavioural, neuropsychological, molecular, genetic and On the oher side, the neuromuscular group is image studies (PET using different radiotracers composed by neurologists, neuropsychologists and multimodal MRI). A multicentre clinical and biologists who develop a comprehensive trial on ICD coordinated by this group is also approach to neuromuscular pathology under development. (including motor neuron disease, spinal atrophy, polyneuropathies, endplate diseases and • Dementia and mild cognitive impairment muscular diseases) both in basic and clinical (MCI) in PD. This line is focused on the care. The group is also interested in the role identification of biomarkers of MCI that might of muscle metabolic control and its influence be used in the early diagnosis of dementia in on the development of central phenomena PD as well as in the implementation of early associated with aging (aging brain and therapeutic interventions. In this regard, proteinopathies) cognitive stimulation is also one of the lines of study at this moment. The approach to this Current research lines of the group are focused line of research includes multimodal MRI, PET, on the pathophysiology of the main forms of electrophysiological and molecular studies muscular dystrophy (myotonic dystrophy, limb in CSF and plasma as well as proteomic and girdle dystrophy type 2A, facioscapulohumeral epigenetic studies. dystrophy and Duchenne dystrophy), and on abnormal muscle homeostasis in aging and • Genetic and premotor biomarkers of PD. This brain neurodegeneration. line is focused on the study of the phenotype of patients with the LRRK2 R1441G mutation (frequent in the Baque country) and of • Myotonic dystrophy: study of the pathways asymptomatic carriers of this mutation that determine the risk of oncogenicity and aiming to know pre-motor biomarkers of brain disorders. PD including clinical, molecular, (CSF and plasma) and image (DATSCAN and fMRI) • Duchenne dystrophy: study of alterations studies as well as their evolution. in calcium homeostasis and its rescue with compounds targeting ryanodine receptor • Familiar esencial tremor. This line comprises stability. clinical, neurophysiológical, genetic (exoming and genetic linkage analysis), and • Facioscapulohumeral dystrophy: study image (MRI) studies. The objective is to study of genomic and epigenetic mechanisms genetic, molecular functional abnormalities that regulate the phenotypic expression in in families with different clinical phenotypes. scapuloperoneales syndromes.

• Inflammation in PD. It comprises studies in • Limb Girdle Dystrophy type 2 A: study of animal models of parkinsonism and in iPS the influence of calpain 3 in proliferation, derived from patients with PD. The aim is to differentiation and replacement of muscle study the relationship between inflammation satellite cells. and dopaminergic death in animal models

Keywords

Parkinson disease, Dyskinesias, impulse control disorder, dementia, cognitive impairment, LRRK2, neuroinflammation, muscular dystrophies, frontotemporal dementia, calpain, progranulin, biomarkers, neurogenetics

2015 CIBERNED Annual Report / 178 Program 2 Group: Adolfo López de Munain

Publications

• Irizar H., Goni J., Alzualde A., Castillo-Trivino T., Olascoaga J., de Munain A.L. et al. Age gene expression and coexpression progressive signatures in peripheral blood leukocytes. Experimental Gerontology. 2015;72:50-56. • Toledo J.B., Zetterberg H., Van Harten A.C., Glodzik L., Martinez-Lage P., Bocchio-Chiavetto L. et al. Alzheimer’s disease cerebrospinal fluid biomarker in cognitively normal subjects. Brain. 2015;138(9):2701-2715. • Alcolea D., Martinez-Lage P., Sanchez-Juan P., Olazaran J., Antunez C., Izagirre A. et al. Amyloid precursor protein metabolism and inflammation markers in preclinical Alzheimer disease. Neurology. 2015;85(7):626-633. • Dols-Icardo O, Nebot I, Gorostidi A, Ortega-Cubero S, Hernández I, Rojas-García R et al. Analysis of the CHCHD10 gene in patients with frontotemporal dementia and amyotrophic lateral sclerosis from Spain.Brain : a journal of neurology. 2015;138(Pt 12): e400. • Martinez-Martin P., Chaudhuri K.R., Rojo-Abuin J.M., Rodriguez-Blazquez C., Alvarez-Sanchez M., Arakaki T. et al. Assessing the non-motor symptoms of Parkinson’s disease: MDS-UPDRS and NMS Scale. European Journal of Neurology. 2015;22(1):37-43. • Dols-Icardo O., Iborra O., Valdivia J., Pastor P., Ruiz A., de Munain A.L. et al. Assessing the role of TUBA4A gene in frontotemporal degeneration. Neurobiology of Aging. 2015. [Epub ahead of print]. • Abete I, Arriola L, Etxezarreta N, Mozo I, Moreno-Iribas C, Amiano P et al. Association between different obesity measures and the risk of stroke in the EPIC Spanish cohort.European journal of nutrition. 2015;54(3): 365-75. • Kalia LV, Lang AE, Hazrati LN, Fujioka S, Wszolek ZK, Dickson DW et al. Clinical correlations with Lewy body pathology in LRRK2-related Parkinson disease.JAMA neurology. 2015;72(1): 100-5. • de la Riva P., Martinez-Zabaleta M.T., Pardo E., Sampron N., Mondragon-Rezola E., Arruti Gonzalez M. et al. Congenital Spinal Malformation and Stroke: Aneurysmal Dilatations and Bilateral Rotational Vertebral Artery Occlusion. Journal of Stroke and Cerebrovascular Diseases. 2015. [Epub ahead of print]. • Jaka O, Casas-Fraile L, López de Munain A, Sáenz A. Costamere proteins and their involvement in myopathic processes.Expert reviews in molecular medicine. 2015;17: e12. • Ugarte A, Gil-Bea F, García-Barroso C, Cedazo-Minguez Á, Ramírez MJ, Franco R et al. Decreased levels of guanosine 3’, 5’-monophosphate (cGMP) in cerebrospinal fluid (CSF) are associated with cognitive decline and amyloid pathology in Alzheimer’s disease.Neuropathology and applied neurobiology. 2015;41(4): 471-82. • Nalls M.A., McLean C.Y., Rick J., Eberly S., Hutten S.J., Gwinn K. et al. Diagnosis of Parkinson’s disease on the basis of clinical and genetic classification: A population-based modelling study. The Lancet Neurology. 2015;14(10):1002-1009. • van den Boogaard ML, Jfl Lemmers R, Camaño P, van der Vliet PJ, Voermans N, van Engelen BG et al. Double SMCHD1 variants in FSHD2: the synergistic effect of two SMCHD1 variants on D4Z4 hypomethylation and disease penetrance in FSHD2.European journal of human genetics : EJHG. 2015. [Epub ahead of print]. • Jiménez-Urbieta H, Gago B, de la Riva P, Delgado-Alvarado M, Marin C, Rodriguez-Oroz MC. Dyskinesias and impulse control disorders in Parkinson’s disease: From pathogenesis to potential therapeutic approaches.Neuroscience and biobehavioral reviews. 2015;56: 294-314. • Abete I., Gomez-Uriz A.M., Mansego M.L., De Arce A., Goyenechea E., Blazquez V. et al. Epigenetic changes in the methylation patterns of KCNQ1 and WT1 after a weight loss intervention program in obese stroke patients. Current Neurovascular Research. 2015;12(4):321- 333. • Mar J, Soto-Gordoa M, Arrospide A, Moreno-Izco F, Martínez-Lage P. Fitting the epidemiology and neuropathology of the early stages of Alzheimer’s disease to prevent dementia.Alzheimer’s research & therapy. ;7(1):2. • Marin C, Bonastre M, Mengod G, Cortés R, Rodríguez-Oroz MC. From unilateral to bilateral parkinsonism: Effects of lateralization on dyskinesias and associated molecular mechanisms. Neuropharmacology. 2015;97: 365-75.

2015 CIBERNED Annual Report / 179 • Ruiz-Martinez J, Krebs CE, Makarov V, Gorostidi A, Martí-Massó JF, Paisán-Ruiz C. GIGYF2 mutation in late-onset Parkinson’s disease with cognitive impairment.Journal of human genetics. 2015;60(10): 637-40. • Gutiérrez-Rivas E, Illa I, Pascual-Pascual SI, Pérez-López J, Vílchez-Padilla JJ, Bautista-Lorite J et al. Guidelines for monitoring late-onset Pompe disease.Revista de neurologia. 2015;60(7): 321-8. • Agalliu I, San Luciano M, Mirelman A, Giladi N, Waro B, Aasly J et al. Higher frequency of certain cancers in LRRK2 G2019S mutation carriers with Parkinson disease: a pooled analysis. JAMA neurology. 2015;72(1): 58-65. • Irizar H., Munoz-Culla M., Saenz-Cuesta M., Osorio-Querejeta I., Sepulveda L., Castillo-Trivino T. et al. Identification of ncRNAs as potential therapeutic targets in multiple sclerosis through differential ncRNA - mRNA network analysis. BMC Genomics. 2015;16(1. [Epub ahead of print]. • Alquezar C, Esteras N, de la Encarnación A, Moreno F, López de Munain A, Martín-Requero Á. Increasing progranulin levels and blockade of the ERK1/2 pathway: upstream and downstream strategies for the treatment of progranulin deficient frontotemporal dementia.European neuropsychopharmacology : the journal of the European College of Neuropsychopharmacology. 2015;25(3): 386-403. • Lemmers RJ, Goeman JJ, van der Vliet PJ, van Nieuwenhuizen MP, Balog J, Vos-Versteeg M et al. Inter-individual differences in CpG methylation at D4Z4 correlate with clinical variability in FSHD1 and FSHD2.Human molecular genetics. 2015;24(3): 659-69. • Martí Massó JF. Javier Urcola Echeverría (1937-2014).Neurologia (Barcelona, Spain). 2015;30(2): 135-6. • Batini C., Hallast P., Zadik D., Delser P.M., Benazzo A., Ghirotto S. et al. Large-scale recent expansion of European patrilineages shown by population resequencing. Nature Communications. 2015;6. [Epub ahead of print]. • Samuel M, Rodriguez-Oroz M, Antonini A, Brotchie JM, Ray Chaudhuri K, Brown RG et al. Management of impulse control disorders in Parkinson’s disease: Controversies and future approaches.Movement disorders : official journal of the Movement Disorder Society. 2015;30(2): 150-9. • Molinet-Dronda F, Gago B, Quiroga-Varela A, Juri C, Collantes M, Delgado M et al. Monoaminergic PET imaging and histopathological correlation in unilateral and bilateral 6-hydroxydopamine lesioned rat models of Parkinson’s disease: a longitudinal in-vivo study. Neurobiology of disease. 2015;77: 165-72. • Mateos-Aierdi AJ, Goicoechea M, Aiastui A, Fernández-Torrón R, Garcia-Puga M, Matheu A et al. Muscle wasting in myotonic dystrophies: a model of premature aging.Frontiers in aging neuroscience. ;7: 125. • Masso J.F.M., Zarranz J.J., Otaegui D., De Munain A.L. Neurogenetic Disorders in the Basque Population. Annals of Human Genetics. 2015;79(1):57-75. • Gomez-Uriz A.M., Milagro F.I., Mansego M.L., Cordero P., Abete I., De Arce A. et al. Obesity and ischemic stroke modulate the methylation levels of KCNQ1 in white blood cells. Human Molecular Genetics. 2015;24(5):1432-1440. • Delgado-Alvarado M, de la Riva P, Jiménez-Urbieta H, Gago B, Gabilondo A, Bornstein B et al. Parkinsonism, cognitive deficit and behavioural disturbance caused by a novel mutation in the polymerase gamma gene.Journal of the neurological sciences. 2015;350(1-2): 93-7. • Gallo V, Brayne C, Forsgren L, Barker RA, Petersson J, Hansson O et al. Parkinson’s Disease Case Ascertainment in the EPIC Cohort: The NeuroEPIC4PD Study.Neuro-degenerative diseases. ;15(6): 331-8. • Bastide M.F., Meissner W.G., Picconi B., Fasano S., Fernagut P.-O., Feyder M. et al. Pathophysiology of L-dopa-induced motor and non-motor complications in Parkinson’s disease. Progress in Neurobiology. 2015;132:96-168.

2015 CIBERNED Annual Report / 180 Program 2 Group: Adolfo López de Munain

• Alquézar C, de la Encarnación A, Moreno F, López de Munain A, Martín-Requero Á. Progranulin deficiency induces overactivation of WNT5A expression via TNF-α/NF-κB pathway in peripheral cells from frontotemporal dementia-linked granulin mutation carriers.Journal of psychiatry & neuroscience : JPN. 2015;41(1): 150131. • Soto-Gordoa M, Arrospide A, Moreno-Izco F, Martínez-Lage P, Castilla I, Mar J. Projecting Burden of Dementia in Spain, 2010-2050: Impact of Modifying Risk Factors.Journal of Alzheimer’s disease : JAD. 2015;48(3): 721-30. • Fernandez-Torron R., De Munain A.L., Camano P., Garcia-Bragado F. Rapidly reversible winging scapula. Arthritis and Rheumatology. 2015;67(9):2502-. [Epub ahead of print]. • Arbizu J, García-Ribas G, Carrió I, Garrastachu P, Martínez-Lage P, Molinuevo JL. Recommendations for the use of PET imaging biomarkers in the diagnosis of neurodegenerative conditions associated with dementia: SEMNIM and SEN consensus.Revista espanola de medicina nuclear e imagen molecular. ;34(5): 303-13. • Ruiz J.R., Gil-Bea F., Bustamante-Ara N., Rodriguez-Romo G., Fiuza-Luces C., Serra-Rexach J.A. et al. Resistance training does not have an effect on cognition or related serum biomarkers in nonagenarians: A randomized controlled trial. International Journal of Sports Medicine. 2015;36(1):54-60. • Bergareche A, Bednarz M, Sánchez E, Krebs CE, Ruiz-Martinez J, De La Riva P et al. SCN4A pore mutation pathogenetically contributes to autosomal dominant essential tremor and may increase susceptibility to epilepsy.Human molecular genetics. 2015;24(24): 7111-20. • Gasca-Salas C., Clavero P., Garcia-Garcia D., Obeso J.A., Rodriguez-Oroz M.C. Significance of visual hallucinations and cerebral hypometabolism in the risk of dementia in Parkinson’s disease patients with mild cognitive impairment. Human Brain Mapping. 2015. [Epub ahead of print]. • Sánchez E, Bergareche A, Krebs CE, Gorostidi A, Makarov V, Ruiz-Martinez J et al. SORT1 Mutation Resulting in Sortilin Deficiency and p75(NTR) Upregulation in a Family With Essential Tremor.ASN neuro. ;7(4). [Epub ahead of print]. • Gutiérrez-Rivas E, Bautista J, Vílchez JJ, Muelas N, Díaz-Manera J, Illa I et al. Targeted screening for the detection of Pompe disease in patients with unclassified limb-girdle muscular dystrophy or asymptomatic hyperCKemia using dried blood: A Spanish cohort.Neuromuscular disorders : NMD. 2015;25(7): 548-53. • Ganos C., Maugest L., Apartis E., Gasca-Salas C., Caceres-Redondo M.T., Erro R. et al. The long-term outcome of orthostatic tremor. Journal of Neurology, Neurosurgery and Psychiatry. 2015. [Epub ahead of print]. • Rodriguez-Oroz MC, Gago B, Clavero P, Delgado-Alvarado M, Garcia-Garcia D, Jimenez- Urbieta H. The relationship between atrophy and hypometabolism: is it regionally dependent in dementias?. Current neurology and neuroscience reports. 2015;15(7): 44. • Mauri Llerda JA, Suller Marti A, de la Peña Mayor P, Martínez Ferri M, Poza Aldea JJ, Gomez Alonso J et al. The Spanish Society of Neurology’s official clinical practice guidelines for epilepsy. Special considerations in epilepsy: comorbidities, women of childbearing age, and elderly patients.Neurologia (Barcelona, Spain). 2015;30(8): 510-7. • Bocchetta M, Galluzzi S, Kehoe PG, Aguera E, Bernabei R, Bullock R et al. The use of biomarkers for the etiologic diagnosis of MCI in Europe: an EADC survey.Alzheimer’s & dementia : the journal of the Alzheimer’s Association. 2015;11(2): 195-206. • Hallast P, Batini C, Zadik D, Maisano Delser P, Wetton JH, Arroyo-Pardo E et al. The Y-chromosome tree bursts into leaf: 13,000 high-confidence SNPs covering the majority of known clades. Molecular biology and evolution. 2015;32(3): 661-73. • Castillo-Trivino T., Lopetegui I., Alarcon-Duque J.A., Lopez De Munain A., Olascoaga J. Ventricular tachycardia on chronic fingolimod treatment for multiple sclerosis. Journal of Neurology, Neurosurgery and Psychiatry. 2015;86(8):931-932. • Bergareche A., Rodriguez-Oroz M.C., Estanga A., Gorostidi A., Lopez de Munain A., Castillo- Trivino T. et al. DAT imaging and clinical biomarkers in relatives at genetic risk for LRRK2R1441G Parkinson’s disease. Movement Disorders. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 181 • Díaz-Manera J, Alejaldre A, González L, Olivé M, Gómez-Andrés D, Muelas N et al. Muscle imaging in muscle dystrophies produced by mutations in the EMD and LMNA genes.Neuromuscular disorders : NMD. 2015. [Epub ahead of print].

Research projects

• Code: FMM14-005. • Code: PI14/00763. Title: Testado de nuevas sustancias con Title: Deterioro cognitivo en la enfermedad potencial terapéutico in vitro e in vivo para de Parkinson: correlato histológico del el tratamiento de la distrofia muscular de hipometabolismo y la atrofia cerebral en un Duchenne. modelo animal y efecto de la estimulación Principal Investigator: Ainara Vallejo. cognitiva en pacientes. CIBERNED’s collaboration: No. Principal Investigator: Maria Cruz Rodríguez CIBERNED groups: Oroz. Other CIBER’s collaborarion: No. CIBERNED’s collaboration: No. Type: National. CIBERNED groups: Funding agency: Fundación Mutua Other CIBER’s collaboration: No. Madrilena. Type: International. Funding: 35.000 €. Funding agency: Instituto de Salud Carlos III. Duration: 2014-2015 Funding: 72.000 €. Duration: 2015 • Code: DPPE14/001. Title: Testado de nuevas sustancias con • Code: UE/2014/CAPN3_CA. potencial terapéutico in vitro e in vivo para Title: Targeting calcium handling proteins in el tratamiento de la distrofia muscular de LGMD2A muscular dystrophy. Duchenne. Principal Investigator: Ainara Vallejo. Principal Investigator: Ainara Vallejo. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: International. Funding agency: Association Française Funding agency: Asociación Duchenne Contre Les Myopathies. Parent Project España. Funding: 80.000 €. Funding: 50.000 €. Duration: 2015-2016 Duration: 2014-2016 • Code: PI14/02129. • Code: GV-2011111074. Title: Modulación farmacológica del Title: Estudio de biomarcadores en la fase receptor de rianodina en la distrofia muscular temprana de la demencia en la enfermedad de Duchenne. de Parkinson. Principal Investigator: Ainara Vallejo. Principal Investigator: María Cruz Rodríguez CIBERNED’s collaboration: No. Oroz. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: International. Other CIBER’s collaboration: No. Funding agency: Instituto de Salud Carlos III Type: International. - ISCIII. Funding agency: Gobierno Vasco. Funding: 91.500 €. Funding: 52.250 €. Duration: 2015 Duration: 2013-2015 • Code: PI14/00436. • Code: DFG14/025. Title: Estudio de la capacidad regenerativa Title: Trastorno de impulsividad en la de los progenitores musculares derivados del enfermedad de Parkinson: fisiopatología e ipss del pacientes con distrofia de cinturas impacto lingüístico. tipo 2ª. Estudio in vitro y en un modelo murino Principal Investigator: María Cruz Rodríguez de dano tisular. Oroz. Principal Investigator: Adolfo López De CIBERNED’s collaboration: No. Munain. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: International. Other CIBER’s collaboration: No. Funding agency: Diputación Foral de Type: National. Guipúzcoa. Funding agency: Instituto de Salud Carlos III Funding: 56.658 €. - ISCIII. Duration: 2015 Funding: 131.000 €. Duration: 2015-2017

2015 CIBERNED Annual Report / 182 Program 2 Group: Adolfo López de Munain

• Code: RTC-2015-3750-1. • Code: MINECO13/803. Title: Desarrollo de agentes terapéuticos Título: Programa Ramón y Cajal. basados en ácidos nucléicos para el Principal Investigator: Ainara Vallejo. tratamiento de enfermedades neuromotoras CIBERNED’s collaboration: No. y neuromusculares. CIBERNED groups: Principal Investigator: Rubén Lépez Vales. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: International. CIBERNED groups: G607, G609. Funding agency: Ministerio de Economía y Other CIBER’s collaboration: No. Competitividad. Type: National. Funding: 258.600 €. Funding agency: Ministerio de Economía y Duration: 2015 Competitividad. Funding: 204.315 €. Duration: 2015-2018 • Code: 2015/03. Title: Perfiles metabólicos diferenciales en enfermedad de Parkinson. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED’s collaboration: Yes. CIBERNED groups: G103, G609, G110, G209. Other CIBER´s collaboration: No. Type: International. Funding agency: CIBERNED. Funding: 210.000 €. Duration: 2015

PhD dissertations

• Author: Ainara Estanga Alustiza. • Author: Jon Toledo Atucha. Title: Alteración cognitiva asociada a la Title: Correlación clínico-electrofisiológica mutación R1441G en el gen lrrk2: estudio de la actividad oscilatoria del núcleo neuropsicológico en pacientes de Parkinson subtalámico en la enfermedad de y portadores asintomáticos. Parkinson. Date: 18th Decembre 2015. Date: 26th Novembre 2015. Supervisor: María Cruz Rodríguez Oroz. Supervidor: María Cruz Rodríguez Oroz.

2015 CIBERNED Annual Report / 183 306 GROUP José Javier Lucas Lozano

José Javier Lucas Lozano Marta Fernández Nogales Elena Llorente Martín Principal Investigator PhD student Undergrad student

Ainara Elorza Peregrina Alberto Parras Rodríguez Miriam Lucas Santamaría Postdoctoral fellow PhD student Technician

Jorge Rubén Cabrera Ivó Hernández Hernández María Santos Galindo Heredia PhD student Technician Postdoctoral fellow

Contact details

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

2015 CIBERNED Annual Report / 184 Summary

Huntington’s disease is an autosomal dominant appearance that, however, did not correlate neurodegenerative disease characterized by with an increase in nuclear indentations involuntary movements, psychiatric symptoms (Fernández-Nogales et al, submitted). Thus, and dementia. It is caused by an expansion TNR detection could be useful as marker of tau of the trinucleotide CAG located in exon 1 of imbalance. the huntingtin gene. In our lab we study the molecular basis of HD through the analysis of Splicing factors as SRSF6 are often act on cell models and in vivo (by generating and functionally related genes. We have studied characterizing transgenic models that could the levels and isoforms of MAP2, another mimic as well as revert the disease) and through microtubule-associated protein closely related studies on human tissue from patients. to tau, functionally as well as phylogenetically. We found a marked decrease in its levels and By using human brain post-mortem tissue from HD an imbalance between high and low molecular individuals we could demonstrate an imbalance weight isoforms in HD. This splicing can be in the tau isoforms with 3 or 4 Microtubule Binging performed by SRSF6 according to our results in Domains and discover a new histopathological cellular models. Furthermore, MAP2 is restrained hallmark (the Tau Nuclear Rods or TNRs) in this to the cell body and doesn’t fill the dendritic disease, consisting on the presence of tau in compartment and it is besides found in nuclear nuclear indentations. The use of transgenic indentations (Cabrera and Lucas, submitted). animals with varying levels of tau allowed us to observe how a moderate decrease in this protein The tau and MAP2 splicing alterations found in in HD model mice was able to attenuate the HD have opened a new research line in which disease phenotype. Regarding the mechanism we are performing the newest techniques: we underlying tau imbalance in HD, we have have performed RNAseq to characterize the discovered that the splicing factor SRSF6, that splicing alterations in HD to identify which events was known to modulate tau exon 10 splicing are altered in HD as well as in presymptomatic and to bind CAG repeats, is altered at levels animal models to then explore which events and phosphorylation in HD and is sequestered could contribute to pathogenesis. into the mutant huntingtin inclusion bodies (Nat On the other hand, the characterization of Med. 2014, 20(8):881-5). This discovery relates model of genetic or pharmacological inhibition HD with tauopathies, which include Alzheimer’s of GSK-3 has allowed us to observe an HD-like disease, thus broadening the possible therapies phenotype with motor deficits and neuronal applicable to HD to those generated for apoptosis (EMBO J., 2007, 26(11):2743-54) tauopathies. mediated by Fas/NFAT (J Clin Invest. 2010, 120(7):2432-45; PLoS One, 2013, 8(8):e70952). In order to characterize if tau alteration is We have also described how GSK3 levels enough for TNR appearance, we have tested are reduced in HD patients and models and a mouse model of frontotemporal dementia demonstrated its involvement in pathogenesis, with parkinsonism associated to chromosome as HD mouse models with a mild overexpression 17, a disease caused by altered tau. We could of GSK-3 show an improvement of their HD-like observe, in addition to the characteristic phenotype (Hum Mol Genet 2015, 24(17):5040- tau overexpression in these animals, TNR 52).

Keywords

Huntington’s disease, transgenic mice, GSK-3, Tau, MAP2, SRSF6, splicing

Publications

• Fernández-Nogales M, Hernández F, Miguez A, Alberch J, Ginés S, Pérez-Navarro E et al. Decreased glycogen synthase kinase-3 levels and activity contribute to Huntington's disease. Human molecular genetics. 2015;24(17): 5040-52. • Ettcheto M, Junyent F, de Lemos L, Pallas M, Folch J, Beas-Zarate C et al. Mice Lacking Functional Fas Death Receptors Are Protected from Kainic Acid-Induced Apoptosis in the Hippocampus. Molecular neurobiology. 2015;52(1): 120-9.

2015 CIBERNED Annual Report / 185 Research projects

• Code: NEURO-MIR. • Code: SAF2012-34177. Title: microRNA as novel therapeutic targets Title: Convergencia de alteraciones génicas, and disease biomarkers in Alzheimer's transcripcionales y traduccionales en la Disease, Frontotemporal dementia and enfermedad de Huntington. Amyotrophic lateral sclerosis (NEURO-MIR). Principal Investigator: José J. Lucas. Principal Investigator: José J. Lucas. CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type International. Funding agency: Ministerio de Economía y Funding agency: iniciativa COEN. Competitividad, Plan Nacional de I+D+I. Funding: 200.000 €. Funding: 330000 €. Duration: 2013-2015 Duration: 2013-2015

PhD dissertations

• Author: Marta Fernández Nogales. Title: Papel de GSK3 y tau en la patogénesis de la enfermedad de Huntington. Date: 14th May 2015. Supervisor: José Javier Lucas Lozano.

2015 CIBERNED Annual Report / 186 304 GROUP María Ángeles Mena

María Ángeles Mena Patricia Trigo Carolina Ruiz Principal Investigador Neurophychologist PhD student

Justo García de Yébenes María José Casarejos Marian Fernández Neurologist Associate Researcher PhD student

Guillermo García Ribas Juan Perucho Andrea Mohedano Neurologist Postdoctoral Fellow Nurse

José Luis López Sendón Conceição Bettencourt Ana Gómez Neurologist Postdoctoral Fellow Technician

Juan García Caldentey Raquel Ros María Paz Muñoz Neurologist Postdoctoral Fellow Technician María Ángeles Robles Administrative assintant

Contact details

Hospital Ramón y Cajal Carretera de Colmenar km 9,1 28034 Madrid(Spain) Tel.: +34 91 336 88 21 Fax: +34 91 336 90 16 [email protected] [email protected]

2015 CIBERNED Annual Report / 187 Summary

In 2015 our group has maintained the research in cell models) of the pathogenic mechanism neurodegenerative disorders mainly dementias and the neuroprotective treatments in these like Alzheimer disease, Parkinson’s disease, diseases. Progressive supranuclear palsy, Huntington’s We continued the study of the amyloidogenic disease, dystonias, ataxias and paraparexias. role of volatile anesthetics and the role of Our work centred in: biomarkers in human cerebrospinal fluid in dementia, the systems of cellular processing - Clinical and molecular characterization of of abnormal proteins in models of Parkinson neurodegenerative diseases: Identification of disease and the role of cannabinoids in new genes and mutations implicated in these Huntington’s disease. diseases. We continued in collaboration with others In the second half of 2015 we have initiated a groups, Spanish, Portuguese, Dutch and English. new line of work, the development of digital We participated in the identification of new multicentric registries for the investigation of genes using the whole-exome sequencing in neurological diseases and for the evaluation patients with ataxia and PSP diseases. of movement disorders. We have designed a digital registry for Parkinson´s disease that is -Clinical studies on neuroprotection compounds going to be implemented in the next few weeks in these diseases. This is the case of the clinical by the Federation of associations of patients trial of SATIVEX drug in Hungtinton patients. with Parkinson’s disease and one method for the quantitative analysis of dyskinesias of the - Study in experimental models (transgenic and head.

Keywords

Neurodegeneratives diseases, neurogenetics, cell cultures, transgenics mouses, neuroprotection

Publications

• Perucho J., Gonzalo-Gobernado R., Bazan E., Casarejos M.J., Jimenez-Escrig A., Asensio M.J. et al. Optimal excitation and emission wavelengths to analyze amino acids and optimize neurotransmitters quantification using precolumn OPA-derivatization by HPLC. Amino Acids. 2015;47(5):963-973.

Research projects

• Code: CIBERNED 2013/05. • Code: S2010/BMD-2308. Title: Identificación y caracterización Title: Neurofarmacología del sistema molecular de subpoblaciones de receptores endocannabinoide: del laboratorio a la cannabinoides en poliglutaminopatias. clínica. Principal Investigator: Manuel Guzmán Principal Investigator: Manuel Guzmán Pastor. Pastor. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: Yes. CIBERNED groups: G305, G201, G303, G304. CIBERNED groups: G305, G303, G304. Other CIBER’s collaboration: No. Other CIBER’s collaboration: Yes Type: International. (CIBERSAM). Funding agency: CIBERNED. Type: CC.AA. Funding: 200.000 €. Funding agency: Comunidad de Madrid. Duration: 2013-2015 Funding: 922.975 €. Duration: 2012-2015

2015 CIBERNED Annual Report / 188 204 GROUP Rosario Moratalla Villalba

Rosario Moratalla Noelia Granado Martínez Irene Ruiz De Diego CSIC Tenured Scientist-Principal PhD PhD Investigator Luz María Suárez Beatriz Pro Oscar Solís Castrejón Postdoctoral fellow Technician PhD student Patricia García Sanz Emilia Rubio Rubio Guillermo Bueno Gil Postdoctoral fellowl Technician PhD student

Contact details

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

2015 CIBERNED Annual Report / 189 Summary

In this period, we have studied the structural Biol Psychiatry). L-DOPA activates the cAMP- and synaptic plasticity in the striatum involved dependent signaling cascade through the Golf in dyskinesia induced by L-DOPA treatment protein, which couples D1 receptor to adenylyl and the role of dopamine and the intracellular cyclase to activate the cAMP. We have shown cAMP-dependent signaling cascade through that Golf is modulated by dopamine and by D1 the DREAM and Golf proteins. To achieve these receptor stimulation, since dopamine absence goals, we are using genetic and chemical upregulates Golf and increasing dopamine models of Parkinson’s disease in mice and levels decrease its expression. We have also chronic treatment with L-DOPA. The genetic shown that dyskinesias are inversely related to model we use is the aphakia mouse model the expression of Golf (Ruiz-DeDiego et al, 2015, and the chemical model is generated by the Mov Disord). In addition, we have also shown unilateral 6-OHDA striatal lesion, which is the that nitric oxide inhibitors decrease dyskinesias most used model in the field of dyskinesia. We and the associated molecular changes, without also use BAC-transgenic mice expressing the modifying the therapeutic effect of L-DOPA green fluorescent protein under the D2 receptor (Solis et al, 2015, Neurobiol Dis). promoter and the red tomato protein under the D1 receptor promoter to identify the two During this year, we have also shown by striatal neuronal populations. We have shown electrophysiological experiments conducted that dopaminergic denervation reduces the in collaboration with Dr. Araque, that neurons number of dendritic spines in the two neuronal in the striatum are homotypically coactivated, populations, in the striatonigral neurons, which i.e. D1-neurons with D1-neurons and D2 facilitate movement, and in the striatopallidal with D2, through specific subpopulations of neurons, which decrease it. However, L-DOPA astrocytes. That is, the activation of a particular treatment, which induces dyskinesias, increases subpopulation of astrocytes specifically the number of dendritic spines only in the stimulates neurons D1 or D2 neurons but they striatopallidal neurons, although these neurons never do it heterotypically (Martin et al, 2015, do not change their firing frequency (Suarez et Science) and this coactivation occurs via al, 2014, Biol Psychiatry). endocannabinoids. In relation to dopaminergic toxicity exerted by On the other hand, in collaboration with Dr. amphetamine derivates we have unequivocally J.R. Naranjo, we have seen that mice lacking shown that meth kills dopaminergic neurons (Ares- the calcium binding protein DREAM have Santos et al, 2014, Neuropsychopharmacology) more dyskinesia and that the dominant active which would increase the risk of developing DREAM mice have fewer dyskinesias and also Parkinson’s disease (Moratalla et al, 2015, Prog have lower expression of the gene changes Neurobiol). In summary, these results have activated by L-DOPA (Ruiz-DeDiego et al, 2015, important implications in Parkinson’s disease.

Keywords

Parkinson’s disease, dyskinesia, synaptic plasticity, neurotoxicity, methamphetamine

Publications

• Ruiz-Dediego I., Mellstrom B., Vallejo M., Naranjo J.R., Moratalla R. Activation of DREAM (Downstream Regulatory Element Antagonistic Modulator), a calcium-binding protein, reduces L-DOPA-induced dyskinesias in mice. Biological Psychiatry. 2015;77(2):95-105. • Moratalla R, Khairnar A, Simola N, Granado N, García-Montes JR, Porceddu PF et al. Amphetamine-related drugs neurotoxicity in humans and in experimental animals: Main mechanisms.Progress in neurobiology. 2015. [Epub ahead of print]. • Martin R., Bajo-Graneras R., Moratalla R., Perea G., Araque A. Circuit-specific signaling in astrocyte-neuron networks in basal ganglia pathways. Science. 2015;349(6249):730-734. • Solís O, Garcia-Montes JR, González-Granillo A, Xu M, Moratalla R. Dopamine D3 Receptor Modulates l-DOPA-Induced Dyskinesia by Targeting D1 Receptor-Mediated Striatal Signaling. Cerebral cortex (New York, N.Y. : 1991). 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 190 Program 2 Group: Rosario Moratalla Villalba

• Ruiz-Dediego I., Naranjo J.R., Herve D., Moratalla R. Dopaminergic regulation of olfactory type G-protein α subunit expression in the striatum. Movement Disorders. 2015. [Epub ahead of print]. • Carmena A, Granado N, Ares-Santos S, Alberquilla S, Tizabi Y, Moratalla R. Methamphetamine- induced toxicity in indusium griseum of mice is associated with astro- and microgliosis. Neurotoxicity research. 2015;27(3): 209-16. • Solis O., Espadas I., Del-Bel E.A., Moratalla R. Nitric oxide synthase inhibition decreases l-DOPA- induced dyskinesia and the expression of striatal molecular markers in Pitx3-/- aphakia mice. Neurobiology of Disease. 2015;73:49-59. • Vergaño-Vera E, Díaz-Guerra E, Rodríguez-Traver E, Méndez-Gómez HR, Solís Ó, Pignatelli J et al. Nurr1 blocks the mitogenic effect of FGF-2 and EGF, inducing olfactory bulb neural stem cells to adopt dopaminergic and dopaminergic-GABAergic neuronal phenotypes.Developmental neurobiology. 2015;75(8): 823-41. • Rodriguez-Traver E., Solis O., Diaz-Guerra E., Ortiz O., Vergano-Vera E., Mendez-Gomez H.R. et al. Role of Nurr1 in the Generation and Differentiation of Dopaminergic Neurons from Stem Cells. Neurotoxicity Research. 2015:1-18. • Castro-Hernández J, Afonso-Oramas D, Cruz-Muros I, Salas-Hernández J, Barroso-Chinea P, Moratalla R et al. Prolonged treatment with pramipexole promotes physical interaction of striatal dopamine D3 autoreceptors with dopamine transporters to reduce dopamine uptake. Neurobiology of disease. 2015;74: 325-35.

Research projects

• Code: Cajal Blue Brain Project. • Code: PNSD. Title: Cajal Blue Brain Project. International Title: Caracterización de la función de DREAM Blue Brain Proyect. en la neurotoxicidad de la metanfetamina y Principal Investigator: Javier de Felipe. otros psicoestimulantes. CIBERNED’s collaboration: Yes. Principal Investigator: R Moratalla. CIBERNED groups: G204, G403. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: International. Other CIBER’s collaboration: No. Funding agency: Ecole Polytechnique Type: International. Federale de Lausanne (Suiza), IBM, Funding agency: Ministerio de Sanidad Universidad Politécnica de Madrid, Consejo Servicios Sociales e Igualdad. Plan Nacional Superior de Investigaciones Cientificas, Sobre Drogas. España. Funding: 120.800 €. Funding: N.D. Duration: 2013-2015 Duration: 2009-2019 • Code: SECITI nº 065/2013. • Code: PCIN-2015-098. Title: Efecto de la estimulación magnética Title: Development of a new in vivo transcraneal repetitiva (EMTr) sobre la radiotracer for alpha-synuclein. terapia dopaminérgica y las discinesias en Principal Investigator: Mireille Dumoilin. modelos experimentales y en pacientes con CIBERNED’s collaboration: No. enfermedad de Parkinson avanzada. CIBERNED groups: Principal Investigator: Rosario Moratalla. Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: International. CIBERNED groups: Funding agency: Unión Europea, ERANET Other CIBER’s collaboration: No. Euronanomed, Ministerio de Economia y Type: International. Competitividad. Funding agency: Secretaria de Ciencia, Funding: 110.000 €. Tecnología e Innovación del Distrito Federal, Duration: 2015-2018 México. Funding: 138.320 €. Duration: 2014-2016

2015 CIBERNED Annual Report / 191 • Code: PI2013/01. CIBERNED’s collaboration: No. Title: Propiedades emergentes de la CIBERNED groups: relación neurona-glia que subyacen a Other CIBER’s collaboration: No. neurodegeneración y demencia en la Type: National. enfermedad de Alzheimer. Funding agency: Ministerio de Economía y Principal Investigator: Ignacio Torres- Competitividad. Alemán. Funding: 399.299 €. CIBERNED’s collaboration: Yes. Duration: 2014-2016 CIBERNED groups: G415, G413, G204, G409, G108, G411. • Code: S2011/BMD 2336. Other CIBER’s collaboration: No. Title: Estudio de la biología de células madre Type: National. neurales para su empleo en terapia celular Funding agency: CIBERNED. en enfermedades neurodegenerativas. Funding: 350.000 €. Principal Investigator: Rosario Moratalla. Duration: 2013-2015 CIBERNED’s collaboration: Yes. CIBERNED groups: G204, G305, G108. • Code: SAF2013-48532-R. Other CIBER’s collaboration: No. Title: Mecanismos moleculares responsables Type: CC.AA. de la plasticidad sináptica y remodelación Funding agency: Comunidad Autónoma estructural de las neuronas estriatales de de Madrid. proyección en ratones disquinéticos. Funding: 598.000 €. Principal Investigator: Rosario Moratalla. Duration: 2012-2015

PhD dissertations

• Author: Isabel Espadas Villanueva. Title: Dopamine signaling is essential in cognitive process and motor performance. Date: 22th Octobre 2015. Supervisor: Rosario Moratalla Villalba.

2015 CIBERNED Annual Report / 192 604 GROUP Pura Muñoz-Cánoves

Pura Muñoz Cánoves Yacine Kharraz Mónica Zamora Villafaina ICREA Research Professor-Princi- Postdoctoral Fellow Postdoctoral Fellow pal Investigator Vera Lukesova Jessica Segales Dalmau Antonio L. Serrano Sánchez Technician Postdoctoral Fellow Senior Researcher Laura Ortet Diana Sobral Mesquita Eusebio Perdiguero Postdoctoral Fellow Pre-doctoral Fellow Santamaría Senior Researcher Patrizia Pessina Begoña Ampudia Carrasco Postdoctoral Fellow Animal Facility Technician Laura García Prat Postdoctoral Fellow Marina Raya Beatriz de Lucas Moreno Labmanager Pre-doctoral Fellow Susana Gutarra Díaz Technician Vanessa Ruíz-Bonilla Postdoctoral Fellow Mercè Jardí Ripoll Technician

Contact details

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

2015 CIBERNED Annual Report / 193 Summary

We have identified a new regulatory role for Fibrosis is a hallmark of DMD and aggravates p38 MAPK in myogenesis, whereby this kinase disease progression, being also an obstacle regulates the transition of muscle stem cells from for therapies. In addition to muscle resident the proliferation to differentiation stages, via a fibroblasts, we have shown that endothelial and transcriptional and epigenetic mechanism. This muscle cells undergo a fibrogenic conversion will help understand the molecular mechanisms at advanced stages of the disease. This cellular underlying the formation of skeletal muscle. interconversion depends on TGFbeta. This Regulation of skeletal myogenesis by p38 MAPK mechanisms will help in the understanding signaling during myogenesis, at the different of the cellular and molecular basis of fibrosis stages of the in vivo regeneration process : development in DMD, and will open doors for activation, proliferation, differentiation and therapeutic intervention. return to quiescence (self-renewal). We have shown that muscle stem cells from We have identified new roles for p38 in controlling individuals of advanced age (geriatric age) macrophage phenotypic transitions necessary switch reversible quiescence into senescence, for appropriate muscle stem cell–dependent due to derepression of p16INK4a. We tissue repair. This novel regulatory mechanism demonstrated that p16INK4a silencing in may have implications in chronic inflammatory geriatric satellite cells restores quiescence and degenerative diseases. muscle regenerative functions. This may provide a basis for stem cell rejuvenation in sarcopenic We have identified new cellular origins for muscles. fibrosis in Duchenne Muscular Dystrophy (DMD).

Keywords

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

Publications

• Lopez-Arribillaga E., Rodilla V., Pellegrinet L., Guiu J., Iglesias M., Roman A.C. et al. Bmi1 regulates murine intestinal stem cell proliferation and self-renewal downstream of Notch. Development (Cambridge). 2015;142(1):41-50. • Sousa-Victor P., Garcia-Prat L., Munoz-Canoves P. Dual mTORC1/C2 inhibitors: Gerosuppressors with potential anti-aging effect. Oncotarget. 2015;6(27):23052-23054. • Segales J., Perdiguero E., Munoz-Canoves P. Epigenetic control of adult skeletal muscle stem cell functions. FEBS Journal. 2015;282(9):1571-1588. • Pessina P., Kharraz Y., Jardi M., Fukada S.-I., Serrano A.L., Perdiguero E. et al. Fibrogenic Cell Plasticity Blunts Tissue Regeneration and Aggravates Muscular Dystrophy. Stem Cell Reports. 2015;4(6):1046-1060. • Munoz-Canoves P., Serrano A.L. Macrophages decide between regeneration and fibrosis in muscle. Trends in Endocrinology and Metabolism. 2015;26(9):449-450. • Sousa-Victor P., Garcia-Prat L., Serrano A.L., Perdiguero E., Munoz-Canoves P. Muscle stem cell aging: Regulation and rejuvenation. Trends in Endocrinology and Metabolism. 2015;26(6):287- 296. • Guerra J., Ferrer B., Giralt M., Comes G., Carrasco J., Molinero A. et al. Muscular interleukin-6 differentially regulates skeletal muscle adaptation to high-fat diet in a sex-dependent manner. Cytokine. 2015;74(1):145-151. • Zamora M., Pardo R., Villena J.A. Pharmacological induction of mitochondrial biogenesis as a therapeutic strategy for the treatment of type 2 diabetes. Biochemical Pharmacology. 2015;98(1):16-28. • Artandi SE, Blau HM, de Haan G, Geiger H, Goodell MA, Jones L et al. Stem Cells and Aging: What’s Next?. Cell stem cell. 2015;16(6): 578-81 .

2015 CIBERNED Annual Report / 194 Program 2 Group: Pura Muñoz Cánoves

• Brack A.S., Munoz-Canoves P. The ins and outs of muscle stem cell aging. Skeletal Muscle. 2016. Epub 2015. • Garcia-Prat L., Martinez-Vicente M., Perdiguero E., Ortet L., Rodriguez-Ubreva J., Rebollo E. et al. Autophagy maintains stemness by preventing senescence. Nature. 2016;529(7584):37-42. Epub 2015. • Aso E, Serrano AL, Muñoz-Cánoves P, Ferrer I. Fibrinogen-Derived γ377-395 Peptide Improves Cognitive Performance and Reduces Amyloid-β Deposition, without Altering Inflammation, in AβPP/PS1 Mice.Journal of Alzheimer’s disease : JAD. 2015;47(2): 403-12.

Research projects

• Code: AGAUR SGR 102. • Principal Investigator: Pura Muñoz-Cánoves. Title: Biología Celular. CIBERNED’s collaboration: No. Principal Investigator: Pura Muñoz-Cánoves. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED: Type: National. Other CIBER’s collaboration: No. Funding agency: Ministerio de Economía y Type: CC.AA. Competitividad. Funding agency: AGAUR (Generalitat de Funding: 250.000 €. Catalunya) “Grup de Recerca Consolidat”. Duration: 2013-2015 Funding: 48.800 €. Duration: 2015-2017 Code: Inter-Cibers. Título: Molecular links between diabetes and Code: La Marató-TV3. neurodegenerative disorders. Title: Novel approaches for degenerative Principal Investigator: Ángel Raya. • muscular dystrophies therapies. • CIBERNED’s collaboration: Yes. Principal Investigator: Pura Muñoz-Cánoves. CIBERNED groups: G409, G604. CIBERNED’s collaboration: No. Other CIBER’s collaboration: Yes CIBERNED groups: (Metabolismo). Other CIBER’s collaboration: No. Type: National. Type: National. Funding agency: Instituto de Salud Carlos III. Funding agency: La Marató-TV3. Funding: 660.000 €. Funding: 120.000 €. Duration: 2015 Duration: 2013-2015 Code: SAF2012-38547. Title: Regulation and dysregulation of skeletal muscle regeneration and growth.

PhD dissertations

• Author: Laura García Prats. Title: Mechanisms controlling stem cell self- renewal. Date: 14th Octobre 2015. Supervisor: Pura Muñoz Cánoves.

2015 CIBERNED Annual Report / 195 307 GROUP José Ramón Naranjo Orovio

José Ramón Naranjo Alberto Rábano M. Paz González Principal Investigator Researcher CIBERNED Assistant researcher

Britt Mellström Javier Morón Oset Xosé Manuel Dopazo CIBERNED Senior Researcher Student CIBERNED Assistant researcher

Juan Casado Elena Higuera Postdoctoral Fellow. JAE-DOC Master’s student

Contact details

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

2015 CIBERNED Annual Report / 196 Summary

We investigate the nuclear components of L-DOPA-induced dyskinesias. activity- and Ca2+-dependent transcriptional responses in neurons to: We foresee the Ca2+-dependent transcriptional repressor DREAM as an active/central 1) Understand the molecular determinants component of several different nucleoprotein of downstream events that are responsible complexes that mediate specifically in the for plastic changes in synaptic functionality various transcriptional cascades triggered by during neurodegeneration. membrane depolarization in neurons that are essential in neuronal plasticity and synaptic 2) To develop tools to expose early markers of dysfunction. Work in progress analyzes the role the neurodegenerative process. of DREAM in the regulation of transcription in cell and animal models of NDDs to better 3) To design small molecules to intervene understand early changes in the transcriptome in physiological and pathological and epigenome and to explore new venues for output processes including learning therapeutic intervention oriented to boost early and memory, motor coordination and endogenous neuroprotective mechanisms. neurodegeneration. Furthermore, previous work in our group has shown that many biological activities of DREAM Early compensatory changes in transcriptional depend on specific protein-protein interactions. programs and altered neuronal calcium In this regard, we have been using NAPPA homeostasis are common features of many protein arrays to further expand our knowledge neurodegenerative pathologies including of the DREAM interactome. Work in progress Alzheimer’s (AD), Down syndrome (DS) and investigates in depth some of these new Huntington’s disease (HD). DREAM (DRE interactions, especially those between DREAM Antagonist Modulator), a Ca2+-dependent and key components of the unfolded protein transcriptional repressor, also known as calsenilin response pathway. because its interaction with presenilins, has an important role in neurodegenerative diseases Finally, we have developed in vitro and in vivo (NDD) through the control of Ca2+ homeostasis. assays to screen for small molecules able to Importantly, changes in DREAM levels are bind to and modify DREAM biological activity. found in several NDD mouse models, including The goal is to identify potent inhibitors of the AD, DS and HD. Genetic experiments show DREAM activity in vivo that could represent that this could be part of a neuroprotective new therapeutic venues for the treatment of mechanism. In addition, we have found that NDD patients. DREAM is important in the development of

Keywords

Calcium pathologies, DREAM, Huntington’s, Alzheimer’s, dyskinesias, UPR, DREAM inhibitors

Publications

2015 CIBERNED Annual Report / 197 • Pisa D., Alonso R., Juarranz A., Rabano A., Carrasco L. Direct visualization of fungal infection in brains from patients with Alzheimer’s disease. Journal of Alzheimer’s Disease. 2015;43(2):613- 624. • Garcia M.C., Cinquina V., Palomo-Garo C., Rabano A., Fernandez-Ruiz J. Identification of CB2 receptors in human nigral neurons that degenerate in Parkinson’s disease. Neuroscience Letters. 2015;587:1-4. • Neant I., Mellstrom B., Gonzalez P., Naranjo J.R., Moreau M., Leclerc C. Kcnip1 a Ca²+- dependent transcriptional repressor regulates the size of the neural plate in Xenopus. Biochimica et biophysica acta. 2015;1853(9):2077-2085. • Llorens-Martin M., Rabano A., Avila J. The ever-changing morphology of hippocampal granule neurons in physiology and pathology. Frontiers in Neuroscience. 2016;9(JAN). Epub 2015. • Ruiz-Dediego I., Mellstrom B., Vallejo M., Naranjo J.R., Moratalla R. Activation of DREAM (Downstream Regulatory Element Antagonistic Modulator), a calcium-binding protein, reduces L-DOPA-induced dyskinesias in mice. Biological Psychiatry. 2015;77(2):95-105. • Ruiz-Dediego I., Naranjo J.R., Herve D., Moratalla R. Dopaminergic regulation of olfactory type G-protein α subunit expression in the striatum. Movement Disorders. 2015. [Epub ahead of print]. • Sánchez-Ferro Á, Rábano A, Catalán MJ, Rodríguez-Valcárcel FC, Fernández Díez S, Herreros- Rodríguez J et al. In vivo gastric detection of α-synuclein inclusions in Parkinson’s disease. Movement disorders : official journal of the Movement Disorder Society. 2015;30(4): 517-24.

Research projects

• Code: 2013/07A. • Code: S2010_BMD. Title: Papel de GSK-3 β en las alteraciones Title: Redes de señalización y vías efectoras de los circuitos corticales que ocurren en la en modelos celulares y animales de enfermedad de Alzheimer. enfermedades neurodegenerativas. Principal Investigator: Teresa Iglesias Vacas. Principal Investigator: José González CIBERNED’s collaboration: Yes. Castaño. CIBERNED groups: G504, G401, G403, G111, CIBERNED’s collaboration: Yes. G307. CIBERNED groups: G111, G401, G104, G307, Other CIBER’s collaboration: No. G409, G412. Type: International. Other CIBER’s collaboration: Yes (CIBERER). Funding agency: CIBERNED. Type: CC.AA. Funding: 316.000 €. funding agency: Comunidad de Madrid. Duration: 2013-2015 Funding: 483.000 €. Duration: 2012-2016

2015 CIBERNED Annual Report / 198 607 GROUP Xavier Navarro Acebes

Xavier Navarro Jordi Bruna Escuer Natalia De la Oliva Eva Santos Nogueira Acebes Assistant Professor Muñoz PhD Student Full Professor-Principal PhD Student Investigator Roser Velasco Daniel Santos Rojas Fargas Isaac Francos PhD Student Esther Udina Bonet Assistant Professor Quijorna Associate Professor- PhD Studentl Jordi Badia Researcher Stefano Cobianchi Casahuja Postdoctoral Fellow Francisco González Researcher Rubèn López Vales Pérez Associate Professor- Jaume del Valle PhD Student Jessica Jaramillo Researcher Macià Rodríguez Postdoctoral Fellow Víctor Manuel Technician Caty Casas Louzao López Álvarez Associate Professor- Joaquim Hernández PhD Student Israel Blasco Ruano Researcher Martín Technician Postdoctoral Fellow Clara López Serrano Assumpció Bosch PhD Student Marta Morell Merino Mireia Herrando Orduña Associate Professor- Grabulosa Renzo Mancuso Technician Researcher Postdoctoral Fellow PhD Student Guillermo García Jesús Amo Aparicio Anna Martínez Alías PhD Student Muriana Ramon y Cajal PhD Student Researcher Ariadna Arbat Plana PhD Student David Romeo Jordi Cuadras Mas Guitart Emeritous Professor- Marina Coll Miró PhD Student Researcher PhD Studentl

Contact details

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]

2015 CIBERNED Annual Report / 199 Summary

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

Keywords

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

Publications

• Cutrone A., Del Valle J., Santos D., Badia J., Filippeschi C., Micera S. et al. A three-dimensional self-opening intraneural peripheral interface (SELINE). Journal of Neural Engineering. 2015;12(1). [Epub ahead of print]. • Thwaite R., Pages G., Chillon M., Bosch A. AAVrh.10 immunogenicity in mice and humans. Relevance of antibody cross-reactivity in human gene therapy. Gene Therapy. 2015;22(2):196- 201. • Santos-Nogueira E., Lopez-Serrano C., Hernandez J., Lago N., Astudillo A.M., Balsinde J. et al. Activation of lysophosphatidic acid receptor type 1 contributes to pathophysiology of spinal cord injury. Journal of Neuroscience. 2015;35(28):10224-102035. • Arbat-Plana A., Torres-Espin A., Navarro X., Udina E. Activity dependent therapies modulate the spinal changes that motoneurons suffer after a peripheral nerve injury. Experimental Neurology. 2015;263:293-305. • Porquet D., Andres-Benito P., Grinan-Ferre C., Camins A., Ferrer I., Canudas A.M. et al. Amyloid and tau pathology of familial Alzheimer’s disease APP/PS1 mouse model in a senescence phenotype background (SAMP8). Age. 2015;37(1). [Epub ahead of print].

2015 CIBERNED Annual Report / 200 Program 2 Group: Xavier Navarro Acebes

• Mancuso R., Navarro X. Amyotrophic lateral sclerosis: Current perspectives from basic research to the clinic. Progress in Neurobiology. 2015;133:1-26. • Del Valle J., De La Oliva N., Muller M., Stieglitz T., Navarro X. Biocompatibility evaluation of parylene C and polyimide as substrates for peripheral nerve interfaces. International IEEE/EMBS Conference on Neural Engineering, NER. 2015;2015-July:442-445. • Simo M., Vaquero L., Ripolles P., Jove J., Fuentes R., Cardenal F. et al. Brain damage following prophylactic cranial irradiation in lung cancer survivors. Brain Imaging and Behavior. 2015. [Epub ahead of print]. • Peluffo H., Solari-Saquieres P., Negro-Demontel M.L., Francos-Quijorna I., Navarro X., Lopez- Vales R. et al. CD300f immunoreceptor contributes to peripheral nerve regeneration by the modulation of macrophage inflammatory phenotype. Journal of Neuroinflammation. 2015;12(1). [Epub ahead of print]. • Casals-Diaz L., Casas C., Navarro X. Changes of voltage-gated sodium channels in sensory nerve regeneration and neuropathic pain models. Restorative Neurology and Neuroscience. 2015;33(3):321-334. • Meyer C, Stenberg L, Gonzalez-Perez F, Wrobel S, Ronchi G, Udina E et al. Chitosan-film enhanced chitosan nerve guides for long-distance regeneration of peripheral nerves.Biomaterials. 2016;76. Epub 2015. • Maciejasz P., Badia J., Souquet G., Cvancara P., Picq C., Stieglitz T. et al. Decreasing stimulation charge by delaying the discharge phase - Comparison of efficacy for various stimulation waveforms. International IEEE/EMBS Conference on Neural Engineering, NER. 2015;2015- July:402-405. • Maciejasz P., Badia J., Boretius T., Andreu D., Stieglitz T., Jensen W. et al. Delaying discharge after the stimulus significantly decreases muscle activation thresholds with small impact on the selectivity: an in vivo study using TIME. Medical and Biological Engineering and Computing. 2015;53(4):371-379. • López-Álvarez VM, Modol L, Navarro X, Cobianchi S. Early increasing-intensity treadmill exercise reduces neuropathic pain by preventing nociceptor collateral sprouting and disruption of chloride cotransporters homeostasis after peripheral nerve injury.Pain. 2015;156(9): 1812-25. • Navarro X. Functional evaluation of peripheral nerve regeneration and target reinnervation in animal models: A critical overview. European Journal of Neuroscience. 2015. [Epub ahead of print]. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016; 12(1). Epub 2015. • Torres-Espín A, Redondo-Castro E, Hernandez J, Navarro X. Immunosuppression of allogenic mesenchymal stem cells transplantation after spinal cord injury improves graft survival and beneficial outcomes.Journal of neurotrauma. 2015;32(6): 367-80. • Reginensi D., Carulla P., Nocentini S., Seira O., Serra-Picamal X., Torres-Espin A. et al. Increased migration of olfactory ensheathing cells secreting the Nogo receptor ectodomain over inhibitory substrates and lesioned spinal cord. Cellular and Molecular Life Sciences. 2015;72(14):2719- 2737. • Alvarez-Berdugo D., Rofes L., Farre R., Casamitjana J.F., Enrique A., Chamizo J. et al. Localization and expression of TRPV1 and TRPA1 in the human oropharynx and larynx. Neurogastroenterology and Motility. 2016;28(1):91-100. Epub 2015. • Casas C., Isus L., Herrando-Grabulosa M., Mancuso F.M., Borras E., Sabido E. et al. Network- based proteomic approaches reveal the neurodegenerative, neuroprotective and pain- related mechanisms involved after retrograde axonal damage. Scientific Reports. 2015;5. [Epub ahead of print]. • Modol L., Santos D., Cobianchi S., Gonzalez-Perez F., Lopez-Alvarez V., Navarro X. NKCC1 activation is required for myelinated sensory neurons regeneration through JNK-dependent pathway. Journal of Neuroscience. 2015;35(19):7414-7427.

2015 CIBERNED Annual Report / 201 • Velasco R., Videla S., Villoria J., Ortiz E., Navarro X., Bruna J. Reliability and accuracy of quantitative sensory testing for oxaliplatin-induced neurotoxicity. Acta Neurologica Scandinavica. 2015;131(5):282-289. • Massó A, Sánchez A, Gimenez-Llort L, Lizcano JM, Cañete M, García B et al. Secreted and Transmembrane αKlotho Isoforms Have Different Spatio-Temporal Profiles in the Brain during Aging and Alzheimer’s Disease Progression.PloS one. ;10(11): e0143623. • Badia J, Raspopovic S, Carpaneto J, Micera S, Navarro X. Spatial and Functional Selectivity of Peripheral Nerve Signal Recording With the Transversal Intrafascicular Multichannel Electrode (TIME).IEEE transactions on neural systems and rehabilitation engineering : a publication of the IEEE Engineering in Medicine and Biology Society. 2016;24(1). Epub 2015. • Gonzalez-Perez F., Ale A., Santos D., Barwig C., Freier T., Navarro X. et al. Substratum preferences of motor and sensory neurons in postnatal and adult rats. European Journal of Neuroscience. 2015. [Epub ahead of print]. • Ale A., Bruna J., Herrando M., Navarro X., Udina E. Toxic Effects of Bortezomib on Primary Sensory Neurons and Schwann Cells of Adult Mice. Neurotoxicity Research. 2015;27(4):430-440. • Gonzalez-Perez F, Cobianchi S, Geuna S, Barwig C, Freier T, Udina E et al. Tubulization with chitosan guides for the repair of long gap peripheral nerve injury in the rat.Microsurgery. 2015;35(4): 300-8.

Research projects

• Code: PI15/01303. Other CIBER’s collaboration: No. Title: Estudio exploratorio sobre la Type: International. etiopatogenia, biomarcadores de riesgo y Funding agency: International Foundation mecanismos implicados en la regeneración for Research in Paraplegia. de la neuropatía inducida por platinos. Funding: 125.000 €. Principal Investigator: Jordi Bruna. Duration: 2014-2016 CIBERNED’s collaboration: No. CIBERNED groups: • Code: SAF2013-48431-R. Other CIBER’s collaboration: No. Title: Participación de los receptores del Type: National. aácido lisofosfatídico en la fisiopatología de Funding agency: Instituto de Salud Carlos III. la lesión medular. Funding: 86.515 €. Principal Investigator: Rubèn Lopez-Vales. Duration: 2015-2018 CIBERNED’s collaboration: No. CIBERNED groups: • Code: SAF 2014-59701. Other CIBER’s collaboration: No. Title: Reprogramación neuronal para Type: International. promover los mecanismos endogenos de Funding agency: Ministerio de Economía y neuroprotección usando biología sintética Competitividad. en un modelo de degeneración retrógrada Funding: 157.300 €. de motoneuronas. Duration: 2014-2017 Principal Investigator: Caty Casas. CIBERNED’s collaboration: No. • Code: TV32014-2810. CIBERNED groups: Title: Terapia génica dirigida a neuregulinas Other CIBER’s collaboration: No. para el tratamiento de la esclerosis lateral Type: National. amiotrófica. Funding agency: Ministerio de Economía y Principal Investigator: Xavier Navarro. Competitividad. CIBERNED’s collaboration: No. Funding: 108.900 €. CIBERNED groups: Duration: 2015-2018 Other CIBER’s collaboration: No. Type: CC.AA. • Code: IRP Research Grant P148. Funding agency: Fundació La Marató-TV3. Title: Role of pro-resolving lipid mediators in Funding: 199.682 €. SCI. Duration: 2015-2017 Principal Investigator: Rubèn Lopez-Vales. CIBERNED’s collaboration: No. CIBERNED groups:

2015 CIBERNED Annual Report / 202 Program 2 Group: Xavier Navarro Acebes

• Code: FP7-602547. • Code: FP7-NMP4-SL-2012-280778. Title: Natural sensory feedback for phantom Title: Micro and nano engineered bidirectional limb pain modulation and therapy carbon interfaces for advanced peripheral (EPIONE). nervous system prosthetic and hybrid bionics Principal Investigator: Navarro Acebes, (MERIDIAN). Xavier. Principal Investigator: Navarro Acebes, CIBERNED’s collaboration: No. Xavier. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: International. Other CIBER’s collaboration: No. Funding agency: Project FP7-602547. Type: International. Funding: 199.840 €. Funding agency: European Commission. Duration: 2013-2017 Funding: 310.000 €. Duration: 2012-2015 • Code: RTC-2015-3750-1. Title: Desarrollo de agentes terapéuticos • Code: FP7-278612. basados en ácidos nucleicos para el Title: Biohybrid templates for peripheral nerve tratamiento de enfermedades neuromotoras regeneration (BIOHYBRID). y neuromusculares. Principal Investigator: Navarro Acebes, Principal Investigator: Rubèn López Vales. Xavier. CIBERNED’s collaboration: Yes. CIBERNED’s collaboration: No. CIBERNED groups: G607, G609. CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: European. Funding agency: Ministerio de Economía y Funding agency: European Commission. Competitividad. Funding: 360.000 €. Funding: 204.315 €. Duration: 2011-2015 Duration: 2015-2018 • Code: FP7-611687. • Code: 2015/01-1. Title: Neurocontrolled bidirectional artificial Title: Terapia génica dirigida a neuregulinas upper limb and hand prosthesis (NEBIAS). para el tratamiento de enfermedades Principal Investigator: Navarro Acebes, degenerativas de las motoneuronas. Xavier. Principal Investigator: Xavier Navarro. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G607, G113, G406, G407. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: Intramurales. Funding agency: European Commission. Funding agency: Instituto de Salud Carlos III, Funding: 517.424 €. CIBERNED. Duration: 2013-2017 Funding: 240.000 €. Duration: 2015-2017 • Code: RTC-2015-3731-1. Title: Optimización y desarrollo de un agente • Code: RD12/0019. terapéutico basado en ácidos nucleicos Title: RETIC de Terapia Celular. para el tratamiento de la enfermedad de Principal Investigator: Coordinador de red: Huntington. Jose Maria Moraleda. Principal Investigator: Assumpció Bosch CIBERNED’s collaboration: Yes. (G607)/Esther Pérez (G301). CIBERNED groups: G102, G301, G113, G208, CIBERNED’s collaboration: Yes. G105, G607. CIBERNED groups: G607, G301. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: National. Type: International. Fundig agency: Fondo de Investigaciones Funding agency: Ministerio de Economía y Sanitarias. Competitividad. Funding: 296.700 €. Funding: 1.582.767,92 €. Duration: 2013-2016 Duration: 2015-2017

2015 CIBERNED Annual Report / 203 PhD dissertations

• Author: Gemma Pagès Pi. • Author: Marina Coll Miró. Title: Intrathecal administration of AAVrh10 Title: Therapeutic role of IL-37 after injury to coding for ß-glucuronidase corrects the nervous system. biochemical and histological hallmarks of Date: 16th Novembre 2015. mucopolysaccharidosis type VII mice and Supervisor: Rubén López Vales. improves behavior and survival. Date: 10th July 2015. Supervisor: Assumpció Bosch Merino.

• Author: Marta Simó Parra. Title: Effects of chemotherapy and cranial radiation in brain structures and cognitive functions of lung cancer patients. Date: 4th May 2015. Supervisor: Jordi Bruna Escuer.

2015 CIBERNED Annual Report / 204 205 GROUP José Ángel Obeso Inchausti

José Ángel Obeso Cristina Calvo González Ignacio Obeso Martín Inchausti Administrative assistant PhD Principal investigator Francisco Javier Blesa de Lydia Vela Desojo María Ledia Fernández los Mozos PhD Hernández PhD Research staff Fernando Alonso Frech Stéphanie Etienne PhD Inés Trigo Damas PhD student Research staff Raúl Martínez Fernández Francisco Molinet Dronda PhD student PhD student

Contact details

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

2015 CIBERNED Annual Report / 205 Summary

Dr. Obeso heads the movement disorders define these mechanisms. lab located at the HM Puerta del Sur hospital in Móstoles (Madrid) and it is mainly focused Evolution and treatment of the disease on the study of Parkinson´s disease (PD) from a multidisciplinary approach. The principal HIFU (High Intensity Focused Ultrasound) is a novel objective is the study of the onset and technique which allows performing controlled progression of the disease using basic and and focused brain lesions without surgery. Thus, clinical experiments aiming to define the mobility and mortality and economic costs are etiopathogenesis of the disease, optimizing greatly reduced compared to current surgical the diagnosis and advancing in the treatment treatments. This technique has demonstrated of neurodegenerative and neuropsiquiatric efficacy and safety in the performance of diseases. Main research fields are: thalamotomies aimed to cease essential tremor and PD tremor. At CINAC, the team is aimed Onset of the disease: compensatory mechanisms to use this technique not only to improve the and selective vulnerability symptoms, but also to intervene in the evolution of the disease. Also, at CINAC it is being tested So far, treatments for PD have not been a novel hypothesis which postulates that successful and one of the reasons is because nigrostriatal degeneration might be caused by diagnosis tends to be delayed respect to the cortical alterations. Thus, this approach could onset of the neurodegenerative process. At imply a revolutionary step at the therapeutical CINAC, patients are treated with magnetic approach, since it gives the possibility to exert stimulation (TMS) and neuroimaging directed to a cortical effect on the symptoms and the define the onset of the disease. Interestingly, PD evolution of the disease. Also, in the recent is principally characterized by the progressive years, it has been also demonstrated the degeneration of dopaminergic neurons of relevance of the risk factors and mechanisms the substantia nigra pars compacta (SNc), involved in the cognitive deficit associated with which are crucial during learning and habit the disease. At CINAC, clinical studies (TMS, acquisition. At CINAC, studies with rodents using cognitive rehabilitation) directed to define electrophysiology, optogenetics and calcium these factors and the development of new in vivo microscopy based on continuous and therapies which could predict or delay their simultaneous execution of tasks are aimed to progression are performed. examine the role of these neurons during these processes. Additionally, since the dopaminergic All studies mentioned above are supplemented loss is previous to the diagnosis it is assumed with neuroimaging studies using positron that there might be a series of compensatory emission topography (PET) and magnetic mechanisms which delay the rise of the resonance (MR) which help to determine symptoms. At CINAC, in vivo neuronal activity potential biomarkers of the disease. recordings in PD animals models are used to

Keywords

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

Publications

2015 CIBERNED Annual Report / 206 Program 2 Group: José Ángel Obeso Inchausti

• Perez T., Tijero B., Gabilondo I., Luna A., Llorens V., Berganzo K. et al. Cardiocirculatory manifestations in Parkinson’s disease patients without orthostatic hypotension. Journal of Human Hypertension. 2015;29(10):604-609. • Benito-León J, Louis ED, Puertas-Martín V, Romero JP, Matarazzo M, Molina-Arjona JA et al. Cognitive and neuropsychiatric features of orthostatic tremor: A case-control comparison. Journal of the neurological sciences. 2016;361. Epub 2015. • Garcia-Ruiz PJ, Ayerbe J, Vela Desojo L, Feliz CE, Del Val Fernandez J, Obeso Inchausti José Ángel. Deep brain stimulation for pantothenate kinase-associated neurodegeneration.Case reports in neurological medicine. ;2015: 1513-6. • Nachev P., Lopez-Sosa F., Gonzalez-Rosa J.J., Galarza A., Avecillas J., Pineda-Pardo J.A. et al. Dynamic risk control by human nucleus accumbens. Brain. 2015;138(12):3496-3502. • Kohl S., Aggeli K., Obeso I., Speekenbrink M., Limousin P., Kuhn J. et al. In Parkinson’s disease pallidal deep brain stimulation speeds up response initiation but has no effect on reactive inhibition. Journal of Neurology. 2015;262(7):1741-1750. • Cuesta P., Garces P., Castellanos N.P., Lopez M.E., Aurtenetxe S., Bajo R. et al. Influence of the APOE ε4 allele and mild cognitive impairment diagnosis in the disruption of the MEG resting state functional connectivity in sources space. Journal of Alzheimer’s Disease. 2015;44(2):493- 505. • Morales-Chacon L.M., Alfredo Sanchez Catasus C., Minou Baez Martin M., Rodriguez Rojas R., Lorigados Pedre L., Estupinan Diaz B. Multimodal imaging in nonlesional medically intractable focal epilepsy. Frontiers in bioscience (Elite edition). 2015;7:42-57. • Canuet L., Pusil S., Lopez M.E., Bajo R., Pineda-Pardo J.A., Cuesta P. et al. Network disruption and cerebrospinal fluid amyloid-beta and phospho-tau levels in mild cognitive impairment. Journal of Neuroscience. 2015;35(28):10325-10330. • Mellone M., Stanic J., Hernandez L.F., Iglesias E., Zianni E., Longhi A. et al. NMDA receptor gluN2A/ gluN2B subunit ratio as synaptic trait of levodopa-induced dyskinesias: From experimental models to patients. Frontiers in Cellular Neuroscience. 2015;9(JULY). [Epub ahead of print]. • Bermejo-Pareja F., Contador I., Trincado R., Lora D., Sanchez-Ferro A., Mitchell A.J. et al. Prognostic Significance of Mild Cognitive Impairment Subtypes for Dementia and Mortality: Data from the NEDICES Cohort. Journal of Alzheimer’s Disease. 2016;50(3):719-731. Epub 2015. • Giancardo L., Sanchez-Ferro A., Butterworth I., Mendoza C.S., Hooker J.M. Psychomotor Impairment Detection via Finger Interactions with a Computer Keyboard During Natural Typing. Scientific Reports. 2015;5. [Epub ahead of print]. • Abi-Jaoude E, Segura B, Obeso I, Cho SS, Houle S, Lang AE et al. Similar striatal D2/D3 dopamine receptor availability in adults with Tourette syndrome compared with healthy controls: A [(11) C]-(+)-PHNO and [(11) C]raclopride positron emission tomography imaging study.Human brain mapping. 2015;36(7). • Sánchez-Catasús CA, Cabrera-Gomez J, Almaguer Melián W, Bosch Bayard J, Rodríguez Rojas R, Valdes-Sosa P et al. The number of optic neuritis attacks is a potential confounder when comparing patients with NMO vs. controls by voxel-based neuroimaging analysis.Acta radiologica (Stockholm, Sweden : 1987). 2015. [Epub ahead of print]. • Hallett M., Obeso J. Where does chorea come from? Cortical excitability findings challenge classic pathophysiological concepts. Movement Disorders. 2015;30(2):169-170. • Blesa J., Lanciego J.L., Obeso J.A. Editorial: Parkinson’s disease: Cell vulnerability and disease progression. Frontiers in Neuroanatomy. 2015;9(September). [Epub ahead of print]. • Molinet-Dronda F, Gago B, Quiroga-Varela A, Juri C, Collantes M, Delgado M et al. Monoaminergic PET imaging and histopathological correlation in unilateral and bilateral 6-hydroxydopamine lesioned rat models of Parkinson’s disease: a longitudinal in-vivo study. Neurobiology of disease. 2015;77: 165-72. • Gasca-Salas C., Clavero P., Garcia-Garcia D., Obeso J.A., Rodriguez-Oroz M.C. Significance of visual hallucinations and cerebral hypometabolism in the risk of dementia in Parkinson’s disease patients with mild cognitive impairment. Human Brain Mapping. 2015. [Epub ahead of print].

2015 CIBERNED Annual Report / 207 • Ganos C., Maugest L., Apartis E., Gasca-Salas C., Caceres-Redondo M.T., Erro R. et al. The long-term outcome of orthostatic tremor. Journal of Neurology, Neurosurgery and Psychiatry. 2015. [Epub ahead of print]. • Parkinson’s disease and Other Movement Disorders.Olanow CW, Schapira AE, Obeso JA. En: Harrison’s Principles of Medicine, 19th Edition. 2015.

Research projects

• Code: SAF2012-40216-C02-01. • Code: MJFF Staff Initiated 2013. Title: Progresión del déficit dopaminérgico Title: Prion-like dissemination of synuclein y mecanismos extra-estriatales en la pathology: a non-human primate study. enfermedad de Parkinson: compensación Principal Investigator: Erwan Bezard inicial vs progresión secundaria. (Universite de Bordeaux, France). Principal Investigator: José Ángel Obeso CIBERNED’s collaboration: Yes. Inchausti. CIBERNED groups: G109, G205. CIBERNED’s collaboration: No. Other CIBER’s collaboration: No. CIBERNED groups: Type: International. Other CIBER’s collaboration: No. Funding agency: Michael J. Fox Foundation. Type: International. Funding: 374.375 €. Funding agency: Ministerio de Economía y Duration: 2013-2016 Competitividad. Funding: 128.700 €. Duration: 2013-2016 • Code: 2014/06. Title: Inicio y progresión de la enfermedad de Parkinson: papel de la activación glial. Principal Investigator: María Cruz Rodríguez Oroz. CIBERNED’s collaboration: Yes. CIBERNED groups: G206, G404, G205. Other CIBER’s collaboration: No. Type: International. Funding agency: Funding: 280.000 €. Duration: 2015

2015 CIBERNED Annual Report / 208 110 GROUP Ana María Pérez Castillo

Ana Pérez-Castillo Elena Giné Isabel Barriuso Ortega Principal investigator PhD Bachelor degree

Ángel Santos Montes Sandra Alonso Gil Rocío Prado Calvo Full professor Technician Technician

José A. Morales García Marina Sanz San Cristóbal PhD Technician

Contact details

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]

2015 CIBERNED Annual Report / 209 Summary

In the past year we have continued in neurogenesis, using neurospheres isolated analyzing the effects of PDE7 inhibition on the from the subventricular zone of adult rats. Our neurodegeneration and neuroinflammation results show that some of these compounds processes that takes place in animal models are able to stimulate early neurogenesis and of neurodegenerative diseases, specifically neuronal maturation in these neurosphere Parkinson’s disease (PD) and progressive cultures and that these effects are probably multiple sclerosis. Besides using chemical mediated via serotonergic and melatonergic inhibitors of this enzyme, this year we have stimulation . In addition, we have demonstrated shown that lentiviral-mediated delivery of that the PDE7 inhibitor S14 is able to induce PDE7B shRNA conferred marked in vitro and in endogenous neuroregenerative processes vivo neuroprotection against 6-OHDA and LPS toward a dopaminergic phenotype. Our results toxicity in dopaminergic neurons, and preserved show a population of actively dividing cells, motor function involving the dopamine system which give rise to new neurons in the SNpc of in mouse. These results corroborate our previous hemiparkinsonian rats in those animals treated data and provide an unequivocally validation with this compound. These data identify of PDE7B enzyme as a valuable new target for S14 as a novel regulator of dopaminergic therapeutic development in the treatment of neuron generation. Finally, we have shown a PD. relationship between the transcription factor C/EBPβ and complement component 3 (C3) We have also initiated a collaboration with Dr. in neuroinflammation and excitotoxic process, Rodriguez at the Medicinal Chemistry Institute which underlie many neurodegenerative (CSIC) to analyze the effects of a new family of disorders. compounds (pinolines, derived from melatonin)

Keywords

Neuroprotection, neuroinflammation, neurogenesis, Parkinson, PDE7, C/EBPβ, C3

Publications

• Prati F, De Simone A, Armirotti A, Summa M, Pizzirani D, Scarpelli R et al. 3,4-Dihydro-1,3,5- triazin-2(1H)-ones as the First Dual BACE-1/GSK-3β Fragment Hits against Alzheimer’s Disease. ACS chemical neuroscience. 2015;6(10): 1665-82. • Hernandez-Encinas E, Aguilar-Morante D, Cortes-Canteli M, Morales-Garcia JA, Gine E, Santos A et al. CCAAT/enhancer binding protein β directly regulates the expression of the complement component 3 gene in neural cells: implications for the pro-inflammatory effects of this transcription factor.Journal of neuroinflammation. 2015;12: 14. • Aguilar-Morante D, Morales-Garcia JA, Santos A, Perez-Castillo A. CCAAT/enhancer binding protein β induces motility and invasion of glioblastoma cells through transcriptional regulation of the calcium binding protein S100A4.Oncotarget. 2015;6(6): 4369-84. • Prati F, De Simone A, Bisignano P, Armirotti A, Summa M, Pizzirani D et al. Multitarget drug discovery for Alzheimer’s disease: triazinones as BACE-1 and GSK-3β inhibitors.Angewandte Chemie (International ed. in English). 2015;54(5): 1578-82. • de la Fuente Revenga M, Pérez C, Morales-García JA, Alonso-Gil S, Pérez-Castillo A, Caignard DH et al. Neurogenic Potential Assessment and Pharmacological Characterization of 6-Methoxy- 1,2,3,4-tetrahydro-β-carboline (Pinoline) and Melatonin-Pinoline Hybrids.ACS chemical neuroscience. 2015;6(5): 800-10. • De La Fuente Revenga M., Fernandez-Saez N., Herrera-Arozamena C., Morales-Garcia J.A., Alonso-Gil S., Perez-Castillo A. et al. Novel N -Acetyl Bioisosteres of Melatonin: Melatonergic Receptor Pharmacology, Physicochemical Studies, and Phenotypic Assessment of Their Neurogenic Potential. Journal of Medicinal Chemistry. 2015;58(12):4998-5014. • Mestre L, Redondo M, Carrillo-Salinas FJ, Morales-García JA, Alonso-Gil S, Pérez-Castillo A et al. PDE7 inhibitor TC3.6 ameliorates symptomatology in a model of primary progressive multiple sclerosis.British journal of pharmacology. 2015;172(17): 4277-90.

2015 CIBERNED Annual Report / 210 Program 2 Group: Ana María Pérez Castillo

• Morales-Garcia J.A., Alonso-Gil S., Gil C., Martinez A., Santos A., Perez-Castillo A. Phosphodiesterase 7 inhibition induces dopaminergic neurogenesis in hemiparkinsonian rats. Stem Cells Translational Medicine. 2015;4(6):564-575. • Morales-Garcia JA, Aguilar-Morante D, Hernandez-Encinas E, Alonso-Gil S, Gil C, Martinez A et al. Silencing phosphodiesterase 7B gene by lentiviral-shRNA interference attenuates neurodegeneration and motor deficits in hemiparkinsonian mice.Neurobiology of aging. 2015;36(2): 1160-73.

Research projects

• Code: INNOGRAPH H2020-PHC-2014-two- Other CIBER’s collaboration: No. stage. Type: Internacional. Title: INNOvative in vitro Point-of-Care Funding agency: Unión Europea. GRAPHene Technologies for Clinical Funding: N.D. Applications in Neurodegenerative Duration: 2015 Diseases. Principal Investigator: Emmanuel Ifeachor. • Code: Functional validation in animal and CIBERNED’s collaboration: No. cellular models of genetic determinants of CIBERNED groups: diseases and ageing processes. Other CIBER’s collaboration: No. Title: Identification of novel pharmacotherapy Type: International. in epilepsy disorders. A multimodal Funding agency: Unión Europea. investigation (Patho-physiology and therapy Funding: ND. of epilepsy and epileptiform disorders). Duration: 2015 Principal Investigator: Ana Pérez Castillo. CIBERNED’s collaboration: No. • Code: 2014 Target validation. CIBERNED groups: Title: Effects of new agonists of melatonin Other CIBER’s collaboration: No. receptors on neurodegeneration, Type: International. inflammation, and neuroregeneration in Funding agency: Unión Europea. Parkinson’s disease. Funding: N.D. Principal Investigator: Ana Pérez Castillo. Duration: 2015 CIBERNED’s collaboration: No. CIBERNED groups: • Code: Human Brain Project (HBP Other CIBER’s collaboration: No. Neurorobotics Toolkit, Robot Designer, Type: International. The Virtual Environment Designer, and the Funding agency: Michael J. Fox Foundation. Closed-Loop Engine. Funding: N.D. Title: Virtual robotic environments, agents, Duration: 2015-2016 sensory and motor systems. Principal Investigator: Janusz Bedkowski. • Code: 2014 Therapeutic Pipeline Program. CIBERNED’s collaboration: No. Title: Preclinical study of co-administration CIBERNED groups: of S14, a selective PDE7 inhibitor, and Other CIBER’s collaboration: No. subactive doses of L-DOPA as potential add- Type: International. on treatment for Parkinson. Funding agency: Unión Europea. Principal Investigator: Ana Pérez Castillo. Funding: N.D. CIBERNED’s collaboration: No. Duration: 2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: Marie Skłodowska-Curie Actions. Type: International. H2020-MSCA-ITN-2014. Funding agency: Michael J. Fox Foundation. Title: Training and Research in epilepsy Funding: N.D. disease. Duration: 2015 Principal Investigator: Ana Pérez Castillo. CIBERNED’s collaboration: No. • Code: FP7-ICT-2013-10. CIBERNED groups: Title: Early Diagnosis of Parkinson Disease Other CIBER’s collaboration: No. and Prediction of Disease Progression with Type: International. Computer-based Models. Funding agency: Unión Europea. Principal Investigator: University of Plymouth. Funding: N.D. CIBERNED’s collaboration: No. Duration: 2015 CIBERNED groups:

2015 CIBERNED Annual Report / 211 • Code: Marie Skłodowska-Curie Actions. • Code: 2015/03. H2020-MSCA-ITN-2015. Title: Perfiles metabólicos diferenciales en Title: Neuroregenerative drug discovery. enfermedad de Parkinson. Principal Investigator: María Laura Principal Investigator: José Manuel Fuentes Bolognesi. Rodríguez. CIBERNED’s collaboration: No. CIBERNED’s collaboration: Yes. CIBERNED groups: CIBERNED groups: G103, G609, G110, G209. Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: European. Type: International. Funding agency: Unión Europea. Funding agency: CIBERNED. Funding: N.D. Funding: 210.000 €. Duration: 2015-2017 Duration: 2015 • Code: SAF2014. • Code: SAF2010-16365. Title: Papel del factor de transcripción Title: Assessment of novel PPARγ activators CEBPβ en la enfermedad de Parkinson: and GSK-3β and PDE7 inhibitors as Identificación y caracterización de los genes possible therapeutic agents for the mediadores de su acción. treatment of Parkinson Disease and other Principal Investigator: Ana Pérez Castillo. neurodegenerative disorders. CIBERNED’s collaboration: No. Principal Investigator: Ana Pérez Castillo. CIBERNED groups: CIBERNED’s collaboration: No. Other CIBERs’s collaboration: No. CIBERNED groups: Type: National. Other CIBER’s collaboration: No. Funding agency: Ministerio de Economía y Type: National. Competitividad. Funding agency: Ministerio de Economía y Funding: 100.000 €. Competitividad. Duration: 2015-2017 Funding: 266.200 €. Duration: 2011-2015

2015 CIBERNED Annual Report / 212 209 GROUP Jordi Pérez Tur

Jordi Pérez Tur Fernando Cardona Serrate Concepción Rubio Granero Principal Investigator Advanced Technical Junior researcher

Contact details

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]

2015 CIBERNED Annual Report / 213 Summary

During this year, we have focused primarily but could also contribute to other neurological on the study of genetic factors related to the phenotypes. disease spectrum that ranges from Amyotrophic Lateral Sclerosis (ALS) to Frontotemporal As for other projects in 2015, we have completed Dementia (FTD) exploiting synergies with other the study of genetic factors related to dementia groups through CIBERNED’s DEGESCO. Thus, we in Parkinson’s disease, the characterization of have contributed to the analysis of the impact DNA variants related to the appearance of a that changes in TUB4A and CHCHD10 have in familial form of primary lateral sclerosis as well the Spanish population suffering from FTD or ALS- as in the definition of the mutation that causes a FTD respectively. We have also characterized familial form of progressive supranuclear palsy. the appearance of an expansion in C9ORF72 We have also continued with the functional in a family in which the ALS appears with other characterization of variants that may be neurological diagnoses such as schizophrenia related to the risk for Parkinson’s and variants or mental retardation, reinforcing the notion that modify the expression of genes related to that expansions in this gene may be associated this disease disease. not only with neurodegeneration in ALS, or FTD,

Keywords

Alzheimer, Parkinson, Neurodegeneration, Neurogenetics, ALS

Publications

• PUBLICATIONS 2015 Dols-Icardo O., Iborra O., Valdivia J., Pastor P., Ruiz A., de Munain A.L. et al. Assessing the role of TUBA4A gene in frontotemporal degeneration. Neurobiology of Aging. 2015. [Epub ahead of print]. • Vazquez-Costa J.F., Beltran E., Sopena P., Sabater A., Cardona F., Vilchez J.J. et al. Clinical and neuroimaging characterization of two C9orf72-positive siblings with amyotrophic lateral sclerosis and schizophrenia. Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration. 2015; 1-4. • Dols-Icardo O, Nebot I, Gorostidi A, Ortega-Cubero S, Hernández I, Rojas-García R et al. Analysis of the CHCHD10 gene in patients with frontotemporal dementia and amyotrophic

Research projects

• Code: 2015/03. Title: Perfiles metabólicos diferenciales en enfermedad de Parkinson. Principal Investigator: José Manuel Fuentes Rodríguez. CIBERNED’s collaboration: Yes. CIBERNED groups: G103, G609, G110, G209. Other CIBER’s collaboration: No. Type: International. Funding agency: CIBERNED. Funding: 210.000 €. Duration: 2015

2015 CIBERNED Annual Report / 214 114 GROUP José Antonio del Río Fernández

José Antonio del Río Ariadna Pérez Balaguer Ágata Mata Rodríguez Fernández Postdoctoral fellow PhD student Principal Investigator Arnau Hervera Laura Urrea Zazurca Rosalina Gavín Marín Postdoctoral fellow PhD student Lecturer Andreu Matamoros Anglès Miriam Segura Feliú Vanessa Gil Fernández PhD student Lab technician Postdoctoral fellow

Contact details

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]

2015 CIBERNED Annual Report / 215 Summary

Our research interests are focused on four F. Wandosell from CIDERNED. These studies aspects of Neurodegeneration and cell were published in Molecular Neurobiology. therapy: Next experiments will continue in this line but also will expand to Parkinson´s disease. Our Cell therapy and pharmacological treatment hypothesis is that PrPc is a cross-link protein to potentiate axon regeneration in lesioned between different neurodegenerative diseases central nervous system: presenting taupathy. In addition, our group In the last years we focused to genetically collaborates in some “omics” studies of specific modify olfactory ensheathing cells (OECs) to markers in rapid progressive dementia (e.g., be able to increase their survival and migration CJD) in collaboration with I. Ferrer. in the lesioned spinal cord. Thus, results were recently published in Cellular and Molecular Development of new lab on a chip devices for Life Science. In the experiments OECs were neurobiological research: genetically modified to overexpress the We recently developed a new device able ectodomain of Nogo Receptor being able to to reproduce the neuromuscular junction in a increase their migration in inhibitory substrates single chip (published in RSC advances 2014). in vitro and in contused spinal cord in vivo. This In addition, in 2015 we also developed a study was developed in collaboration of X. specific device to allow for axonal lesions that Navarro and F. Wandosell groups of CIBERNED, mimic Wallerian-like degeneration, molecular and X. Trepat and J. Samitier from CIBERBBN. gradient generation for migrating neurons and in silico 3D modelization, or neurodegenerative Neurodegenerative diseases, pTau and PrPc: diseases: Parkinson´s disease). The axotomy chip We determined the role of a natural neural was published in RSC advances in collaboration cellular prion protein protein (PrPc) in the with J. Samitier (CIBERBBN). evolution of the Alzheimer´s disease (published in Molecular Neurobiology). Results point PrPc, epilepsy and prionopathies: to PrPc as key neuroprotective factor in In 2015 we determined the role of the cellular Alzheimer´s disease. In fact, when present Tau prion protein in epilepsy by using 6 mice phosphorylation by ADDLs is impaired. However models in a collaborative project between in Braak stages III, PrPc down-regulates by 4 laboratories in collaboration with I. Ferrer unknown reasons and Tau phosphorylation (CIBERNED). Our data published in Scientific increases. These experiments were developed Reports, demonstrate that the absence of PrPc in vitro, in mouse models of AD as well in post- induce a susceptible phenotype for epilepsy, mortem human samples. Experiments were one of the hallmarks of several prionopathies. developed in collaboration with I. Ferrer and

Keywords

p-Tau, amyloid, cellular prion protein, axon regeneration, cell therapy

Publications

• Tong Z., Segura-Feliu M., Seira O., Homs-Corbera A., Del Rio J.A., Samitier J. A microfluidic neuronal platform for neuron axotomy and controlled regenerative studies. RSC Advances. 2015;5(90):73457-73466. • Vilches S., Vergara C., Nicolas O., Mata A., Del Rio J.A., Gavin R. Domain-Specific Activation of Death-Associated Intracellular Signalling Cascades by the Cellular Prion Protein in Neuroblastoma Cells. Molecular Neurobiology. 2015. [Epub ahead of print]. • Reginensi D., Carulla P., Nocentini S., Seira O., Serra-Picamal X., Torres-Espin A. et al. Increased migration of olfactory ensheathing cells secreting the Nogo receptor ectodomain over inhibitory substrates and lesioned spinal cord. Cellular and Molecular Life Sciences. 2015;72(14):2719- 2737. • Carulla P., Llorens F., Matamoros-Angles A., Aguilar-Calvo P., Espinosa J.C., Gavin R. et al. Involvement of PrP C in kainate-induced excitotoxicity in several mouse strains. Scientific Reports. 2015;5. [Epub ahead of print].

2015 CIBERNED Annual Report / 216 Program 2 Group: José Antonio del Río Fernández

• Vergara C, Ordóñez-Gutiérrez L, Wandosell F, Ferrer I, del Río JA, Gavín R. Role of PrP(C) Expression in Tau Protein Levels and Phosphorylation in Alzheimer’s Disease Evolution.Molecular neurobiology. 2015;51(3): 1206-20. • Llorens F, Zafar S, Ansoleaga B, Shafiq M, Blanco R, Carmona M et al. Subtype and regional regulation of prion biomarkers in sporadic Creutzfeldt-Jakob disease.Neuropathology and applied neurobiology. 2015;41(5): 631-45.

Research projects

• Code: SRI-NEUROMEMs. Other CIBER’s collaboration: No. Title: Neuromems. Type: Private. Principal Investigator: José Antonio del Río Funding agency: Fundació Marato TV3. CIBERNED’s collaboration: No. Funding: 195.646,25 €. CIBERNED groups: Duration: 2015-2017 Other CIBER’s collaboration: No. Type: Private. • Code: CIBERNED 2014/02. Funding agency: Obra Social La Caixa. Title: Mecanismos epigenéticos Funding: 25.000 €. implicados en la etiología y progresión Duration: 2014-2015 de las demencias neurodegenerativas rápidamente progresivas. • Code: 392/U/2014. Principal Investigator: Miguel Calero. Title: Role of the cellular prion protein as CIBERNED’s collaboration: Yes. “cross-talk” protein between alpha-syn/ CIBERNED groups: G114, G509, G503, G601. LRRK2 and p-Tau in sporadic and familiar Other CIBER’s collaboration: No. Parkinson´s disease. Type: Intramurales. Principal Investigator: José Antonio del Funding agency: CIBERNED. Río (Fundacio Institut de Bioenginyeria de Funding: 252.074 €. Catalunya). Duration: 2015-2017 CIBERNED’s collaboration: Yes. CIBERNED groups: G109, G114. • Code: BFU2012-32617. Other CIBER’s collaboration: No. Title: Nuevas funciones de PLEXIND1/SEMA3E, Type: Private. PRPC y las proteínas asociadas a la mielina Funding agency: Fundació la Marató TV3. durante el desarrollo de la corteza cerebral Funding: 195.646 €. de roedores y en neurodegeneración. Duration: 2014-2017 Principal Investigator: José Antonio del Río. CIBERNED’s collaboration: No. • Code: Marató TV3-6. CIBERNED groups: Title: Paper de la PrPc com a “cross-talk Other CIBER’s collaboration: No. protein” entre alpha-sinucleina/LRRK2 i Type: National. p-Tau en la malaltia de Parkinson de tipus Funding agency: Ministerio de Economía y esporàdic i/o familiar. Competitividad. Principal Investigator: José Antonio del Río. Funding: 304.200 €. CIBERNED’s collaboration: No. Duration: 2013-2015 CIBERNED groups:

PhD dissertations

• Author: Diego Reginensi Espinoza. • Author: Cristina Vergara Paños. Title: Modulación de las propiedades Title: Papel regulador de la proteina priónica migratorias de las células de la glia celular en la enfermedad de Alzheimer y envolvente olfatoria. uso de gamma-péptidos como potenciales Date: 5th May 2015. agentes terapéuticos. Superviso: José Antonio Del Río Fernández. Date: 21th April 2015. Supervisor: José Antonio Del Río Fernández.

2015 CIBERNED Annual Report / 217 206 GROUP Manuel Rodríguez Díaz

Manuel Rodríguez Díaz Alberto Sánchez Fernández Fernando Isidro Montón Full Professor. Principal Research staff Álvarez Investigator Research staff Ingrid Morales Pérez Tomás González Hernández Postdoctoral fellow Jesús Norellis Lorenzo Brito Research staff Associate group researcher Clara Rodríguez Sabaté Magdalena Sabaté Bel PhD student Research staff

Contact details

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]

2015 CIBERNED Annual Report / 218 Summary

An animal model was developed to study the basal ganglia circuits. Data analysis with Multiple main axonal and glial mechanisms involved in Linear Regression showed relevant non-linear the retrograde degeneration of dopaminergic components in the basal ganglia interactions. neurons in Parkinson’s disease. This model Thus, a new non-linear method based on the induces an apoptotic degeneration of striatal Multiple Correspondence Analysis was also dopaminergic terminals accompanied by applied to basal ganglia interactions. This is a a selective reactive astrogliosis and not by multivariate method which may identify massive microgliosis. This model allows the study of interacting networks with an “unsupervised” the action of reactive astrocytes on the data-driven procedure. This method generates metabolization of debris degenerated by a self-organizing clustering of centers according the dopaminergic degeneration, on the re- to their interactions which is similar to that innervation of the denervated striatal areas, generated by Independent Component and on the protection of dopaminergic Analysis, but which may be particularly suitable nigrostriatal neurons from a dying back axonal when non-linear interactions are being studied. degeneration. This model may be particularly The Multiple Correspondence Analysis isolated useful to study the functional efficiency of six networks within the cortico-subcortical motor astrocytes generated from iPS cells obtained loop of basal ganglia. The activity of these from parkinsonian patients. Our group is networks is now being studied in Parkinson’s collaborating in the iPS project with other groups disease. belonging to CIBERNED who facilitate patient samples and the differentiation of astrocytic The membrane transporter involved in the cells. dopamine uptake by dopaminergic neurons is a relevant factor for the vulnerability of By using a procedure developed to identify mesencephalic dopaminergic neurons in and to get representative BOLD-signals of the Parkinson’s disease. We have studied the main basal ganglia from magnetic resonance possible role of D3 dopamine receptors on images, the main interactions of these centers the dopamine transporter activity in mice and were studied in the human brain. The partial cell cultures. D3 agonists decrease dopamine correlation methods showed a circular cortico- uptake, changing the integration of the subcortical interaction which was globally transporter in the cytoplasmatic membrane similar in humans to that previously found in of the dopaminergic synaptic terminals, and animals with other methods. This procedure was the phosforylation and ubiquitinization of the used to analyze the functional connectivity of dopamine transporter. the intralaminar thalamic centers with the main

Keywords

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

Publications

• Rodriguez M, Rodriguez-Sabate C, Morales I, Sanchez A, Sabate M. Parkinson’s disease as a result of aging.Aging cell. 2015;14(3): 293-308. • Castro-Hernández J, Afonso-Oramas D, Cruz-Muros I, Salas-Hernández J, Barroso-Chinea P, Moratalla R et al. Prolonged treatment with pramipexole promotes physical interaction of striatal dopamine D3 autoreceptors with dopamine transporters to reduce dopamine uptake. Neurobiology of disease. 2015;74: 325-35. • Trenkwalder C., Chaudhuri K.R., Martinez-Martin P., Rascol O., Ehret R., Valis M. et al. Prolonged- release oxycodone-naloxone for treatment of severe pain in patients with Parkinson’s disease (PANDA): A double-blind, randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(12):1161-1170. • Morales I., Sanchez A., Rodriguez-Sabate C., Rodriguez M. The degeneration of dopaminergic synapses in Parkinson’s disease: A selective animal model. Behavioural Brain Research. 2015;289:19-28.

2015 CIBERNED Annual Report / 219 • Rodriguez-Sabate C, Llanos C, Morales I, Garcia-Alvarez R, Sabate M, Rodriguez M. The functional connectivity of intralaminar thalamic nuclei in the human basal ganglia.Human brain mapping. 2015;36(4): 1335-47.

Research projects

• Code: 2014/06. Title: Inicio y progresión de la enfermedad de Parkinson: papel de la activación glial. Principal Investigator: María Cruz Rodríguez Oroz. CIBERNED’s collaboration: Yes. CIBERNED groups: G206, G404, G205. Other CIBER’s collaboration: No. Type: International. Funding agency: N.D. Funding: 280.000 €. Duration: 2015

2015 CIBERNED Annual Report / 220 207 GROUP Eduard Tolosa Sarró

Eduard Tolosa Sarró Alejandro Iranzo de Riquer Carme Junqué Plaja Principal Investigator Consultant Full Profesor

María José Martí Carles Gaig Dolores Vilas Doménech Specialist PhD student Consultant Mario Ezquerra Trabalón Judith Navarro Francesc Valldeoriola i Researcher PhD student Serra Consultant Yaroslau Compta Hinrjy Mercè Bonastre García Specialist Lab Technician Esteban Muñoz Martínez Consultant Rubén Fernández-Santiago Manuel Fernández Sánchez Researcher Lab Technician Joan Santamaría Cano Senior Consultant Mónica Serradell Eroles PhD student Josep Valls-Solé Senior Consultant

Contact details

Hospital Clinic de Barcelona Universidad de Barcelona Villarroel 171 08036 Barcelona, Catalunya (Spain) Tel.: +34 93 227 57 85 Fax: +34 93 227 57 83 [email protected]

2015 CIBERNED Annual Report / 221 Summary

In continuation of our studies on peripheral observation underlying cognitive deterioriation synuclien as a diagnostic tissue marker for and apathy in this condition (Mov disord 2015; Parkinson disease (PD). We have reported Hum Brain Mapp 2015). negative results for synuclein aggregates in the maleolar region in PD (J Neurol Transm In the field of experimental therapeutics 2014) and different patterns of alpha-synuclein we have coordinated the first report of the inmunoreactivity in the gut of PD (Neurosci Barcelona Registry on intrayeyunal infusions of Letters 2015). In the framework of studies on levodopa (Parkinsonism Related Disord 2015), premotor PD we have reported different and participated and coauthored trials on patterns of premotor symptoms in PD, the rasagiline in multisystem atrophy (Lancet Neurol ONSET PD study (Mov Disorders 2015) , that 2015) and rotigotine on non motor symptoms olfactory dysfunction predicts early transition of PD (Europ J Neurol 2015). to a Lewy body disorder in idiopathic RBD ( a premotor synucleinopahty) (Neurology 2015), We have consolidated the line of research on and minimal presence of RBD in asymptomatic cerebrospinal fluid and imaging biomarkers carriers of the LRRK2 mutation (PLOSone in Parkinson’s disease related dementia (J 2015). We have investigated the expression Neurol 2015). Particularly we have completed levels of PD-associated microRNAs in serum the longitudinal analyses of cerebrospinal samples from patients with idiopathic rapid- fluid levels of tau and amyloid-beta proteins eye-movement (REM) sleep behavior disorder as well as molecular imaging of amyloid-beta (IRBD), a condition that eventually evolves into and tau (by means of the PET-tracer FDDNP) a synucleinopathy such as PD. We found that showing that these might be predictors of the microRNA miR-19b is down-regulated in Parkinson’s dementia. We have also finished patients with RBD and antedates the diagnosis the recruitment of atypical and unclassifiable of PD and dementia with Lewy bodies (DLB) Parkinsonism cases as part of the project on the by 5 years, suggesting its potential role as early potential usefulness of the FDDNP PET probe biomarker for motor conversion (Annals Neurol as an in vivo marker of tau pathology. Lastly, 2015). a Catalan network of MSA has been launched and several different types of tissue samples are Imaging studies in PD have continued in the now being collected as part of an MSA-specific search for the neural substrate of nonmotor biorepository. symptoms and we have reported novel imaging

Keywords

Neuroimaging and cognition, Biological, Diagnostic pre-motor, non-motor symptoms, motor complications, genetic variants, Sleep disturbances, experimental therapy

Publications

• Marques-Iturria I., Scholtens L.H., Garolera M., Pueyo R., Garcia-Garcia I., Gonzalez-Tartiere P. et al. Affected connectivity organization of the reward system structure in obesity. NeuroImage. 2015;111:100-106. • Aldecoa I., Navarro-Otano J., Stefanova N., Sprenger F.S., Seppi K., Poewe W. et al. Alpha- synuclein immunoreactivity patterns in the enteric nervous system. Neuroscience Letters. 2015;602:145-149. • Vilas D., Iranzo A., Pont-Sunyer C., Serradell M., Gaig C., Santamaria J. et al. Brainstem raphe and substantia nigra echogenicity in idiopathic REM sleep behavior disorder with comorbid depression. Journal of Neurology. 2015;262(7):1665-1672. • Sierra-Rio A., Balasa M., Olives J., Antonell A., Iranzo A., Castellvi M. et al. Cerebrospinal fluid biomarkers predict clinical evolution in patients with subjective cognitive decline and mild cognitive impairment. Neurodegenerative Diseases. 2016;16(1-2):69-76. Epub 2015. • Baggio H.-C., Segura B., Sala-Llonch R., Marti M.-J., Valldeoriola F., Compta Y. et al. Cognitive impairment and resting-state network connectivity in Parkinson’s disease. Human Brain Mapping. 2015;36(1):199-212.

2015 CIBERNED Annual Report / 222 Program 2 Group: Eduard Tolosa Sarró

• Odin P., Ray Chaudhuri K., Slevin J.T., Volkmann J., Dietrichs E., Martinez-Martin P. et al. Collective physician perspectives on non-oral medication approaches for the management of clinically relevant unresolved issues in Parkinson’s disease: Consensus from an international survey and discussion program. Parkinsonism and Related Disorders. 2015;21(10):1133-1144. • Simuni T., Caspell-Garcia C., Coffey C., Chahine L.M., Lasch S., Oertel W.H. et al. Correlates of excessive daytime sleepiness in de novo Parkinson’s disease: A case control study. Movement Disorders. 2015;30(10):1371-1381. • Navarro-Otano J, Casanova-Mollà J, Morales M, Valls-Solé J, Tolosa E. Cutaneous autonomic denervation in Parkinson’s disease.Journal of neural transmission (Vienna, Austria : 1996). 2015;122(8): 1149-55. • Urbizu A., Canet-Pons J., Munoz-Marmol A.M., Aldecoa I., Lopez M.T., Compta Y. et al. Cystatin C is differentially involved in multiple system atrophy phenotypes. Neuropathology and Applied Neurobiology. 2015;41(4):507-519. • Bergareche A., Rodriguez-Oroz M.C., Estanga A., Gorostidi A., Lopez de Munain A., Castillo- Trivino T. et al. DAT imaging and clinical biomarkers in relatives at genetic risk for LRRK2R1441G Parkinson’s disease. Movement Disorders. 2015. [Epub ahead of print]. • Cabib C, Llufriu S, Casanova-Molla J, Saiz A, Valls-Solé J. Defective sensorimotor integration in preparation for reaction time tasks in patients with multiple sclerosis.Journal of neurophysiology. 2015;113(5): 1462-9. • Coelho M, Marti MJ, Sampaio C, Ferreira JJ, Valldeoriola F, Rosa MM et al. Dementia and severity of parkinsonism determines the handicap of patients in late-stage Parkinson’s disease: the Barcelona-Lisbon cohort.European journal of neurology. 2015;22(2): 305-12. • Pont-Sunyer C., Bressman S., Raymond D., Glickman A., Tolosa E., Saunders-Pullman R. Disclosure of research results in genetic studies of Parkinson’s disease caused by LRRK2 mutations. Movement Disorders. 2015;30(7):904-908. • Veciana M., Becerra J.L., Fossas P., Muriana D., Sansa G., Santamarina E. et al. EEG extreme delta brush: An ictal pattern in patients with anti-NMDA receptor encephalitis. Epilepsy and Behavior. 2015;49:280-285. • Antonini A., Bauer L., Dohin E., Oertel W.H., Rascol O., Reichmann H. et al. Effects of rotigotine transdermal patch in patients with Parkinson’s disease presenting with non-motor symptoms - results of a double-blind, randomized, placebo-controlled trial. European Journal of Neurology. 2015;22(10):1400-1407. • Poewe W., Seppi K., Fitzer-Attas C.J., Wenning G.K., Gilman S., Low P.A. et al. Efficacy of rasagiline in patients with the parkinsonian variant of multiple system atrophy: A randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(2):145-152. • Sprenger FS, Stefanova N, Gelpi E, Seppi K, Navarro-Otano J, Offner F et al. Enteric nervous system α-synuclein immunoreactivity in idiopathic REM sleep behavior disorder.Neurology. 2015;85(20): 1761-8. • Fernandez-Santiago R., Ezquerra M. Epigenetic research of neurodegenerative disorders using patient iPSC-based models. Stem Cells International. 2015;2016. [Epub ahead of print]. • Ferini-Strambi L, Oertel W, Dauvilliers Y, Postuma RB, Marelli S, Iranzo A et al. Erratum to: Autonomic symptoms in idiopathic REM behavior disorder: a multicentre case-control study. Journal of neurology. 2015;262(1): 249-50. • Ferreira J.J., Colosimo C., Bhidayasiri R., Marti M.J., Maisonobe P., Om S. Factors influencing secondary non-response to botulinum toxin type A injections in cervical dystonia. Parkinsonism and Related Disorders. 2015;21(2):111-115. • Lees A.J., Tolosa E., Olanow C.W. Four pioneers of L-dopa treatment: Arvid Carlsson, Oleh Hornykiewicz, George Cotzias, and Melvin Yahr. Movement Disorders. 2015;30(1):19-36. • Antelmi E., Ferri R., Iranzo A., Arnulf I., Dauvilliers Y., Bhatia K.P. et al. From state dissociation to status dissociatus. Sleep Medicine Reviews. 2016;28:1-13. Epub 2015.

2015 CIBERNED Annual Report / 223 • Marin C, Bonastre M, Mengod G, Cortés R, Rodríguez-Oroz MC. From unilateral to bilateral parkinsonism: Effects of lateralization on dyskinesias and associated molecular mechanisms. Neuropharmacology. 2015;97: 365-75. • Garcia-Garcia I., Jurado M.A., Garolera M., Marques-Iturria I., Horstmann A., Segura B. et al. Functional network centrality in obesity: A resting-state and task fMRI study. Psychiatry Research - Neuroimaging. 2015;233(3):331-338. • Sala-Llonch R, Palacios EM, Junqué C, Bargalló N, Vendrell P. Functional networks and structural connectivity of visuospatial and visuoperceptual working memory.Frontiers in human neuroscience. ;9: 340. • Guerreiro R., Escott-Price V., Darwent L., Parkkinen L., Ansorge O., Hernandez D.G. et al. Genome-wide analysis of genetic correlation in dementia with Lewy bodies, Parkinson’s and Alzheimer’s diseases. Neurobiology of Aging. 2015. [Epub ahead of print]. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1). Epub 2015. • Ninerola-Baizan A., Rojas S., Bonastre M., Tudela R., Lomena F., Pavia J. et al. In vivo evaluation of the dopaminergic neurotransmission system using [123I]FP-CIT SPECT in 6-OHDA lesioned rats. Contrast Media and Molecular Imaging. 2015;10(1):67-73. • Buongiorno M, Antonelli F, Cámara A, Puente V, de Fabregues-Nebot O, Hernandez-Vara J et al. Long-term response to continuous duodenal infusion of levodopa/carbidopa gel in patients with advanced Parkinson disease: The Barcelona registry.Parkinsonism & related disorders. 2015;21(8): 871-6. • Fernandez-Santiago R., Iranzo A., Gaig C., Serradell M., Fernandez M., Tolosa E. et al. MicroRNA association with synucleinopathy conversion in rapid eye movement behavior disorder. Annals of Neurology. 2015;77(5):895-901. • Hor H., Francescatto L., Bartesaghi L., Ortega-Cubero S., Kousi M., Lorenzo-Betancor O. et al. Missense mutations in TENM4, a regulator of axon guidance and central myelination, cause essential tremor. Human Molecular Genetics. 2015;24(20):5677-5686. • Vidal-Pineiro D., Martin-Trias P., Falcon C., Bargallo N., Clemente I.C., Valls-Sole J. et al. Neurochemical modulation in posteromedial default-mode network cortex induced by transcranial magnetic stimulation. Brain Stimulation. 2015;8(5):937-944. • Fabbri M., Leodori G., Fernandes R.M., Bhidayasiri R., Marti M.J., Colosimo C. et al. Neutralizing Antibody and Botulinum Toxin Therapy: A Systematic Review and Meta-analysis. Neurotoxicity Research. 2016;29(1):105-117. Epub 2015. • Mahlknecht P., Iranzo A., Hogl B., Frauscher B., Muller C., Santamaria J. et al. Olfactory dysfunction predicts early transition to a Lewy body disease in idiopathic RBD. Neurology. 2015;84(7):654-658. • Tolosa E., Vilas D. Peripheral synuclein tissue markers: A step closer to Parkinson’s disease diagnosis. Brain. 2015;138(8):2120-2122. • Aguirre-Mardones C., Iranzo A., Vilas D., Serradell M., Gaig C., Santamaria J. et al. Prevalence and timeline of nonmotor symptoms in idiopathic rapid eye movement sleep behavior disorder. Journal of Neurology. 2015;262(6):1568-1578. • Trenkwalder C., Chaudhuri K.R., Martinez-Martin P., Rascol O., Ehret R., Valis M. et al. Prolonged- release oxycodone-naloxone for treatment of severe pain in patients with Parkinson’s disease (PANDA): A double-blind, randomised, placebo-controlled trial. The Lancet Neurology. 2015;14(12):1161-1170. • Guaita M., Melia U., Vallverdu M., Caminal P., Vilaseca I., Montserrat J.M. et al. Regularity of cardiac rhythm as a marker of sleepiness in sleep disordered breathing. PLoS ONE. 2015;10(4). [Epub ahead of print]. • Sala-Llonch R, Bartrés-Faz D, Junqué C. Reorganization of brain networks in aging: a review of functional connectivity studies.Frontiers in psychology. ;6: 663.

2015 CIBERNED Annual Report / 224 Program 2 Group: Eduard Tolosa Sarró

• Tell-Martí G, Puig-Butille JA, Potrony M, Badenas C, Milà M, Malvehy J et al. Reply.Annals of neurology. 2016;79(1). Epub 2015. • Baggio H.C., Segura B., Garrido-Millan J.L., Marti M.-J., Compta Y., Valldeoriola F. et al. Resting- state frontostriatal functional connectivity in Parkinson’s disease-related apathy. Movement Disorders. 2015;30(5):671-679. • Baggio H.C., Segura B., Junque C. Resting-State Functional Brain Networks in Parkinson’s Disease. CNS Neuroscience and Therapeutics. 2015;21(10):793-801. • Baggio H.C., Segura B., Junque C., De Reus M.A., Sala-Llonch R., Van Den Heuvel M.P. Rich club organization and cognitive performance in healthy older participants. Journal of Cognitive Neuroscience. 2015;27(9):1801-1810. • Postuma R.B., Iranzo A., Hogl B., Arnulf I., Ferini-Strambi L., Manni R. et al. Risk factors for neurodegeneration in idiopathic rapid eye movement sleep behavior disorder: A multicenter study. Annals of Neurology. 2015;77(5):830-839. • Tercero-Uribe A, Gaig C, Iranzo A, Gómez JB, Graus F, Santamaria J. Small fast rhythmic eye movements (SFREM) misdiagnosed as frontal seizures.Clinical neurophysiology : official journal of the International Federation of Clinical Neurophysiology. 2016;127(1). Epub 2015. • Guaita M., Salamero M., Vilaseca I., Iranzo A., Montserrat J.M., Gaig C. et al. The Barcelona sleepiness index: A new instrument to assess excessive daytime sleepiness in sleep disordered breathing. Journal of Clinical Sleep Medicine. 2015;11(11):1289-1298. • Tell-Marti G., Puig-Butille J.A., Potrony M., Badenas C., Mila M., Malvehy J. et al. The MC1R melanoma risk variant p.R160W is associated with Parkinson disease. Annals of Neurology. 2015;77(5):889-894. • Ortega-Cubero S, Lorenzo-Betancor O, Lorenzo E, Agúndez JA, Jiménez-Jiménez FJ, Ross OA et al. TREM2 R47H variant and risk of essential tremor: a cross-sectional international multicenter study.Parkinsonism & related disorders. 2015;21(3): 306-9. • Matos N, Gaig C, Santamaria J, Iranzo A. Tricks to rapidly terminate episodes of cataplexy in narcolepsy.Sleep medicine. 2015. [Epub ahead of print]. • Fernandez-Santiago R., Carballo-Carbajal I., Castellano G., Torrent R., Richaud Y., Sanchez- Danes A. et al. Aberrant epigenome in iPSC-derived dopaminergic neurons from Parkinson’s disease patients. EMBO Molecular Medicine. 2015. [Epub ahead of print]. • Vilas D., Ispierto L., Alvarez R., Pont-Sunyer C., Marti M.J., Valldeoriola F. et al. Clinical and imaging markers in premotor LRRK2 G2019S mutation carriers. Parkinsonism and Related Disorders. 2015;21(10):1170-1176. • Kalia LV, Lang AE, Hazrati LN, Fujioka S, Wszolek ZK, Dickson DW et al. Clinical correlations with Lewy body pathology in LRRK2-related Parkinson disease.JAMA neurology. 2015;72(1): 100-5. • Grau-Rivera O., Gelpi E., Nos C., Gaig C., Ferrer I., Saiz A. et al. Clinicopathological Correlations and Concomitant Pathologies in Rapidly Progressive Dementia: A Brain Bank Series. Neurodegenerative Diseases. 2015;15(6):350-360. • Compta Y, Valente T, Saura J, Segura B, Iranzo Á, Serradell M et al. Correlates of cerebrospinal fluid levels of oligomeric- and total-α-synuclein in premotor, motor and dementia stages of Parkinson’s disease.Journal of neurology. 2015;262(2): 294-306. • Nalls M.A., McLean C.Y., Rick J., Eberly S., Hutten S.J., Gwinn K. et al. Diagnosis of Parkinson’s disease on the basis of clinical and genetic classification: A population-based modelling study. The Lancet Neurology. 2015;14(10):1002-1009. • Albanese A., Abbruzzese G., Dressler D., Duzynski W., Khatkova S., Marti M.J. et al. Practical guidance for CD management involving treatment of botulinum toxin: a consensus statement. Journal of Neurology. 2015;262(10):2201-2213. • Pont-Sunyer C., Iranzo A., Gaig C., Fernandez-Arcos A., Vilas D., Valldeoriola F. et al. Sleep disorders in parkinsonian and nonparkinsonian LRRK2 mutation carriers. PLoS ONE. 2015;10(7). [Epub ahead of print].

2015 CIBERNED Annual Report / 225 • Grau-Rivera O., Sanchez-Valle R., Bargallo N., Llado A., Gaig C., Nos C. et al. Sporadic MM2- thalamic+cortical Creutzfeldt-Jakob disease: Utility of diffusion tensor imaging in the detection of cortical involvement in vivo. Neuropathology. 2015. [Epub ahead of print]. • Pont-Sunyer C., Hotter A., Gaig C., Seppi K., Compta Y., Katzenschlager R. et al. The onset of nonmotor symptoms in parkinson’s disease (the onset pd study). Movement Disorders. 2015;30(2):229-237.

Research projects

• Code: 2014\FTPGB-LAGUNA. CIBERNED groups: G408, G301, G207, G410, Title: Estudio del perfil transcripcional en G109. subgrupos de pacientes prodrómicos con Other CIBER’s collaboration: No. enfermedad de Parkinson. Type: National. Principal Investigator: Ariadna Laguna Funding agency: Instituto de Salud Carlos III. Tuset. Funding: 300.000 €. CIBERNED’s collaboration: Yes. Duration: 2013-2015 CIBERNED groups: G109, G207. Other CIBER’s collaboration: No. • Code: 00000. Type: National. Title: The Parkinson's Progression Markers Funding agency: Fundación Tatiana Pérez Initiative (PPMI). de Guzmán el Bueno. Principal Investigator: Eduard Tolosa. Funding: 44.000 €. CIBERNED’s collaboration: No. Duration: 2014-2017 CIBERNED groups: Other CIBER’s collaboration: No. • Code: LRRK2 Resource RFA 2013. Type: International. Title: Alpha-synuclein and other biomarkers Funding agency: The Michael J. Fox in biological samples of LRKK2 PD. Foundation for Parkinson's Research. Principal Investigator: Eduard Tolosa. Funding: 1.830.210 €. CIBERNED’s collaboration: Yes. Duration: 2013-2018 CIBERNED groups: G410, G207. Other CIBER’s collaboration: No. • Code: COURAGE - PD. Type: International. Title: Comprehensive unbiased risk factor Funding agency: Michael J. Fox Foundation. assessment for genetics and environment in Funding: 60.000 €. Parkinson‘s disease. Duration: 2014-2016 Principal Investigator: Thomas Gasser. CIBERNED’s collaboration: No. • Code: PI2013_08-1. CIBERNED groups: Title: La dinámica mitocondrial y mitofagia Other CIBER’s collaboration: No. como dianas terapéuticas en las Type: International. enfermedades de Parkinson y Huntington. Funding agency: JPND. Principal Investigator: Eduard Soriano. Funding: 4.678.945 €. CIBERNED’s collaboration: Yes. Duration: 2013-2016

PhD dissertations

• Author: Roser Sala Llonch. Title: Multimodal MRI study of human brain connectivity: cognitive networks. Date: 13th January 2015. Supervisor: Carmen Junqué Plaja.

2015 CIBERNED Annual Report / 226 108 GROUP Carlos Vicario Abejón

Carlos Vicario Abejón Inmaculada Crespo Galán Mª José Román CSIC Senior Scientist and Principal Postdoctoral Fellow Technician Investigator Çagla Defterali Carlos Omar Oueslati Eva Díaz Guerra PhD Student Morales Postdoctoral Fellow Master Student Eva Rodríguez Traver Vanesa Nieto Estévez PhD Student Inmaculada Luque Molina Postdoctoral Fellow Master Student

Contact details

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

2015 CIBERNED Annual Report / 227 Summary

We have obtained induced pluripotent stem graphene with neurons, astrocytes and with cells (iPS cells or iPSCs) by reprogramming adult neurogenesis (Defterali et al., 2015). These fibroblasts from Alzheimer´s disease patients are the first published results from the analysis of carrying the epsilon 3 and epsilon 4 alleles of the effects of graphene on adult neural stem APOE gene or a mutation in the Presenilin gene. cells (NSCs), neurons and glia in vivo. The iPS cells grow as flat colonies, express the pluripotency markers OCT4, SOX2, NANOG, We have studied the role of Tbr1 and Nurr1 TRA-1-60 and SSEA-4, and give rise to embryoid transcription factors (Méndez-Gómez et al., bodies which generate the three body lineages 2011; Vergaño-Vera et al., 2014) in neurogenesis, (ectoderm, endoderm and mesoderm). We and neuronal migration and survival in the adult have confirmed that these iPSCs differentiate brain. With this aim, we have used strategies of into neurons and astrocytes. The first results gain- and loss-of function of these transcription of this project were presented during the factors in vivo as well as cell transplantation. III International Meeting on Research and The first results have been recently reported Innovation on Neurodegenerative Diseases/9º (Rodríguez-Traver et al. 2015, Neurotoxicity CIBERNED Scientific Forum (Díaz-Guerra et Research). al.,). Thanks to this achievement, we are participating in an European consortium, which The function of insulin-like growth factor-I has presented a grant application to the call receptor (IGF-IR) has been investigated in a Innovative Medicine Initiative H2020-JTI-IMI2- mouse line in which the levels of this receptor 2015-05. Our project received a favourable can be drastically reduced by Cre recombinase evaluation in the first stage of the call, and expression in dividing NSCs and progenitors. thus we are preparing the full proposal to be Deletion of IGF-IR reduces self-renewal and presented in stage two. proliferation of postnatal-adult hipocampal stem cells. We have published an article in the journal Biomaterials reporting the biocompatibility of

Keywords

Alzheimer´s disease, Parkinson´s disease, neurogenesis, iPS cells, NSCs, transcription factors, growth factors

Publications

• Defterali C., Verdejo R., Peponi L., Martin E.D., Martinez-Murillo R., Lopez-Manchado M.A. et al. Thermally reduced graphene is a permissive material for neurons and astrocytes and de novo neurogenesis in the adult olfactory bulb in vivo. Biomaterials. 2016;82:84-93. Epub 2015. • Vergaño-Vera E, Díaz-Guerra E, Rodríguez-Traver E, Méndez-Gómez HR, Solís Ó, Pignatelli J et al. Nurr1 blocks the mitogenic effect of FGF-2 and EGF, inducing olfactory bulb neural stem cells to adopt dopaminergic and dopaminergic-GABAergic neuronal phenotypes.Developmental neurobiology. 2015;75(8): 823-41. • Rodriguez-Traver E., Solis O., Diaz-Guerra E., Ortiz O., Vergano-Vera E., Mendez-Gomez H.R. et al. Role of Nurr1 in the Generation and Differentiation of Dopaminergic Neurons from Stem Cells. Neurotoxicity Research. 2015. 1-18.

2015 CIBERNED Annual Report / 228 Program 2 Group: Carlos Vicario Abejón

Research projects

• Code: SAF2013-47596-R. CIBERNED’s collaboration: Yes. Title: Formación de neuronas en el ser CIBERNED groups: G415, G413, G204, G409, humano y ratón adultos por activación de G108, G411. la reprogramación celular y la neurogénesis Other CIBER’s collaboration: No. endógena. Type: National. Principal Investigator: Carlos Vicario Funding agency: CIBERNED. Abejón. Funding: 350.000 €. CIBERNED’s collaboration: No. Duration: 2013-2015 CIBERNED groups: Other CIBER’s collaboration: No. • Code: S2011/BMD 2336. Type: National. Title: Estudio de la biología de células madre Funding agency: Ministerio de Economía y neurales para su empleo en terapia celular Competitividad. en enfermedades neurodegenerativas. Funding: 145.200 €. Principal Investigator: Rosario Moratalla. Duration: 2014-2016 CIBERNED’s collaboration: Yes. CIBERNED groups: G204, G305, G108. • Code: PI2013/01. Other CIBER’s collaboration: No. Title: Propiedades emergentes de la Type: CC.AA. relación neurona-glia que subyacen a Funding agency: Comunidad Autónoma de neurodegeneración y demencia en la Madrid. enfermedad de Alzheimer. Funding: 598.000 €. Principal Investigator: Ignacio Torres- Duration: 2012-2015 Alemán.

PhD dissertations

• Author: Çala Defterali. Title: Sudying the neurogenic potential of local progenitors in the adult olfactory bulb and their biocompatibility with graphene. Date: 3rd July de 2015. Supervisor: Carlos Vicario Abejón.

2015 CIBERNED Annual Report / 229 109 GROUP Miquel Vila Bover

Miquel Vila Bover Iria Carballo Carbajal Annabelle Parent ICREA Research Professor-Princi- Postdoctoral fellow Technician pal Investigator Ainhoa Plaza Zabala Thais Cuadros Arasa Celine Perier Postdoctoral fellow Technician Ramón y Cajal Researcher Ariadna Laguna Tuset Beatriz Rodríguez Galván Jordi Bové Badell Postdoctoral fellow Technician Miguel Servet Researcher Sandra Franco Iborra Jordi Romero Giménez Marta Martínez Vicente Predoctoral fellow Technician Miguel Servet Researcher Albert Torra Talavera Oriol de Fàbregues-Boixar Predoctoral fellow Nebot Clinician collaborator

Contact details

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

2015 CIBERNED Annual Report / 230 Summary

In 2015 we have continued our studies on the pathological alterations in the epigenetic pathogenicity of α-synuclein and its potential profile of dopaminergic neurons derived from role on neuron dysfunction/death in the context patients with Parkinson’s disease. These cells Parkinson’s disease. We have also continued were obtained by reprogramming of skin cells with the development of novel therapeutic into induced pluripotent stem cells and their tools to interfere with α-synuclein-mediated subsequent differentiation into dopaminergic neurotoxicity. Moreover, in collaboration with neurons. other groups of the CIBERNED, we have identified

Keywords

Parkinson’s disease, α-synuclein, epigenetics, induced pluripotent stem cells

Publications

• Fernandez-Santiago R., Carballo-Carbajal I., Castellano G., Torrent R., Richaud Y., Sanchez- Danes A. et al. Aberrant epigenome in iPSC-derived dopaminergic neurons from Parkinson’s disease patients. EMBO Molecular Medicine. 2015. [Epub ahead of print]. • Dehay B., Vila M., Bezard E., Brundin P., Kordower J.H. Alpha-synuclein propagation: New insights from animal models. Movement Disorders. 2015. [Epub ahead of print]. • Martinez-Vicente M. Autophagy in neurodegenerative diseases: From pathogenic dysfunction to therapeutic modulation. Seminars in Cell and Developmental Biology. 2015;40:115-126. • Garcia-Prat L., Martinez-Vicente M., Perdiguero E., Ortet L., Rodriguez-Ubreva J., Rebollo E. et al. Autophagy maintains stemness by preventing senescence. Nature. 2016;529(7584):37-42. Epub 2015. • Vilas D., Ispierto L., Alvarez R., Pont-Sunyer C., Marti M.J., Valldeoriola F. et al. Clinical and imaging markers in premotor LRRK2 G2019S mutation carriers. Parkinsonism and Related Disorders. 2015;21(10):1170-1176. • Laguna A., Schintu N., Nobre A., Alvarsson A., Volakakis N., Jacobsen J.K. et al. Dopaminergic control of autophagic-lysosomal function implicates Lmx1b in Parkinson’s disease. Nature Neuroscience. 2015;18(6):826-835. • Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12(1):-. Epub 2015. • Franco-Iborra S., Perier C. Neurodegeneration: The Size Takes It All. Current Biology. 2015;25(18):R797-R800. • Pont-Sunyer C., Iranzo A., Gaig C., Fernandez-Arcos A., Vilas D., Valldeoriola F. et al. Sleep disorders in parkinsonian and nonparkinsonian LRRK2 mutation carriers. PLoS ONE. 2015;10(7):-. [Epub ahead of print]. • Dehay B., Bourdenx M., Gorry P., Przedborski S., Vila M., Hunot S. et al. Targeting α-synuclein for treatment of Parkinson’s disease: Mechanistic and therapeutic considerations. The Lancet Neurology. 2015;14(8):855-866. • Pont-Sunyer C., Hotter A., Gaig C., Seppi K., Compta Y., Katzenschlager R. et al. The onset of nonmotor symptoms in parkinson’s disease (the onset pd study). Movement Disorders. 2015;30(2):229-237. • Franco-Iborra S, Vila M, Perier C. The Parkinson Disease Mitochondrial Hypothesis: Where Are We at?. The Neuroscientist : a review journal bringing neurobiology, neurology and psychiatry. 2015. [Epub ahead of print]. • Mechanisms and prevention of cell death.Miquel Vila, Celine Perier. En: Parkinson’s disease & movement disorders (6th edition). (pp. 17-27), 2015.Joseph Jankovic, Eduardo Tolosa (eds.).

2015 CIBERNED Annual Report / 231 Research projects

• Code: 2014 SGR 1609. Type: International. Title: Apoptosi i Neurodegeneracio (GRC). Funding agency: Fondo de Investigación Principal Investigator: Joan Comella Sanitaria. Carnicé. Funding: 110.715 €. CIBERNED’s collaboration: Yes. Duration: 2014-2017 CIBERNED groups: G109, G413. Other CIBER’s collaboration: No. • Code: SAF2013-43158-R. Type: International. Title: Disfunción mitocondrial en la Funding agency: AGAUR. enfermedad de Parkinson: papel de la Funding: 30.000 €. dinámica mitocondrial. Duration: 2014-2016 Principal Investigator: Celine Perier. CIBERNED’s collaboration: No. • Code: MOUSEMODPARKINSONUK_PUKIIG. CIBERNED groups: Title: Generation of humanized mouse Other CIBER’s collaboration: No. models to study the role of neuromelanin in Type: National. Parkinson disease. Funding agency: Ministerio de Economía y Principal Investigator: Miquel Vila Bover. Competitividad. CIBERNED’s collaboration: No. Funding: 114.950 €. CIBERNED groups: Duration: 2014-2016 Other CIBER’s collaboration: No. Type: International. • Code: PI13/01897. Funding agency: Parkinson's UK (UK). Title: Rol de la neuromelanina y alfa- Funding: 4.3362,06 €. sinucleina en el inicio y extensión de la Duration: 2015 enfermedad de Parkinson: potencial diagnóstico y terapéutico en nuevos • Code: 2014\FTPGB-LAGUNA. modelos pre-clínicos. Title: Estudio del perfil transcripcional en Principal Investigator: Miquel Vila Bover. subgrupos de pacientes prodrómicos con CIBERNED’s collaboration: No. enfermedad de Parkinson. CIBERNED groups: Principal Investigator: Ariadna Laguna Other CIBER’s collaboration: No. Tuset. Type: National. CIBERNED’s collaboration: Yes. Funding agency: Fondo de Investigaciones CIBERNED groups: G109, G207. Sanitarias-Instituto de Salud Carlos III. Other CIBER’s collaboration: No. Funding: 142.780 €. Type: National. Duration: 2014-2016 Funding agency: Fundación Tatiana Pérez de Guzmán el Bueno. • Code: Fundació la Marató de TV3-Malalties Funding: 44.000 €. del cor (320/U/2015). Duration: 2014-2017 Title: New molecular functions of apoptotic genes in cardiac development and stress. • Code: MJFF_TARGET.ADV-2015 (Grant ID: Principal Investigator: Daniel Sanchis Morales 11580). (IRBLLEIDA, Universitat de Lleida). Title: Targeting the transcriptional regulator CIBERNED’s collaboration: No. of autophagy LMX1B for Parkinson's disease CIBERNED groups: therapeutic development. Other CIBER’s collaboration: No. Principal Investigator: Ariadna Laguna Type: Private. Tuset. Funding agency: Fundació la Marató de CIBERNED’s collaboration: No. TV3. CIBERNED groups: Funding: 191.656 €. Other CIBER’s collaboration: No. Duration: 2015-2018 Type: International. Funding agency: The Michael J. Fox • Code: 392/U/2014. Foundation for Parkinson's Research (USA). Title: Role of the cellular prion protein as Funding: 99.402 €. “cross-talk” protein between alpha-syn/ Duration: 2015-2016 LRRK2 and p-Tau in sporadic and familiar Parkinson´s disease. • Code: PI14/01529. Principal Investigator: José Antonio del Title: Rol patogénico de la disfunción Río (Fundació Institut de Bioenginyeria de autofágica/lisosomal en la enfermedad de Catalunya). Parkinson: diagnóstico y nuevas terapias CIBERNED’s collaborator: Yes. pre-clínicas. CIBERNED groups: G109, G114. Principal Investigator: Marta Martínez Other CIBER’s collaboration: No. Vicente. Type: Private. CIBERNED’s collaboration: No. Funding agency: Fundació la Marató de CIBERNED groups: TV3. Other CIBER’s collaboration: Yes (CIBER- Funding: 195.646 €. BBN). Duration: 2014-2017

2015 CIBERNED Annual Report / 232 Program 2 Group: Miquel Vila Bover

• Code: IPT-2012-1208-300000. • Code: MJFF TPP2014. Title: Suppression of alpha-synuclein Title: Pharmacokinetic and expression and PDE10 as a new therapeutic pharmacodynamic characterization of a strategy to Parkinson disease and novel therapy to silence selectively. improvement of cognitive dysfunctions. Principal Investigator: Raquel Revilla (nLife Principal Investigator: Andrés Montefeltro Therapeutics). (nLife Therapeutics). CIBERNED’s collaboration: No. CIBERNED’s collaboration: No. CIBERNED groups: CIBERNED groups: Other CIBER’s collaboration: No. Other CIBER’s collaboration: No. Type: International. Type: European. Funding agency: Michael J. Fox Foundation. Funding agency: European Regional Funding: 353.694 €. Development Fund, INNPACTO program. Duration: 2014-2016 Funding: 255.059 €. Duration: 2012-2015 Code: MJFF Staff Initiated 2013. • Title: Prion-like dissemination of synuclein • Code: Marató TV3 (120531). pathology: a non-human primate study. Title: Regenerating dopaminergic neurons in Principal Investigator: Erwan Bezard Parkinson’s disease via cell-fusion-mediated (Universite de Bordeaux, France). reprogramming. CIBERNED’s collaboration: Yes. Principal Investigator: María Pia Cosma CIBERNED groups: G109, G205. (CRG, Barcelona). Other CIBER’s collaboration: No. CIBERNED’s collaboration: No. Type: International. CIBERNED groups: Funding agency: Michael J. Fox Foundation. Other CIBER’s collaboration: No. Funging: 374.375 €. Type: Private. Duration: 2013-2016 Funding agency: Fundació la Marató TV3. Funding: 300.000 €. Code: RTC-2014-2812-1. Duration: 2013-2015 Title: Nuevas estrategias terapéuticas para • • el tratamiento de sinocleunopatias. Code: PI2013_08-1. Principal Investigator: Raquel Revilla (nLife Title: La dinámica mitocondrial y mitofagia Therapeutics). como dianas terapéuticas en las CIBERNED’s collaboration: No. enfermedades de Parkinson y Huntington. CIBERNED groups: Principal Investigator: Eduardo Soriano. Other CIBER’s collaboration: No. CIBERNED’s collaboration: Yes. Type: National. CIBERNED groups: G408, G301, G207, G410, Funding agency: Ministerio de Economía y G109. Competitividad. Other CIBE’s collaboration: No. Funding: 1.971.513 €. Type: National. Duration: 2014-2016 Funding agency: Instituto de Salud Carlos III. Funding: 300.000 €. Duration: 2013-2015

2015 CIBERNED Annual Report / 233 Cooperative research 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 Centre’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 six calls for projects in total. The first three (2010, 2011, and 2013) has already developed fifteen completed projects and a total accumulated budget allocation of 4,980,100 €.

These projects from the first three calls are listed below:

First call

Nº Code Title Coordinator 1 PI2010/07 Reelin and GSK3 as therapeutic targets in Alzheimer’s Ávila de Grado, Jesús disease

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

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

4 PI2010/06 Neuroprotection in Huntington’s disease García de Yébenes Prous, Justo Generating a dopaminergic neuronal model from Tolosa Sarró, Eduardo 5 PI2010/05 induced pluripotent stem cells from patients with Parkinson’s disease associated to mutations in the LRRK2 gene

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

Second call

Nº Code Title Coordinator 7 PI2011/01 The Nrf2 transcription factor as a new therapeutic Cuadrado Pastor, Antonio target for Parkinson’s disease

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

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

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

2015 CIBERNED Annual Report / 237 Third call

Nº Code Title Coordinator 11 PI2013/01 Emergent properties of the neuron-glia relationship Torres Alemán, Ignacio underlying neurodegeneration and dementia in Alzheimer’s disease

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

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

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

Synaptic degeneration and dysregulation 15 PI2013/09 of adult neurogenesis in murine models of Fernández Chacón, neurodegeneration Rafael

Those fifteen already completed projects have actively involved 54 CIBERNED research groups overall, in addition to some others external ones, representing 85.7% of the existing groups.

The third call for CIBERNED cooperative projects ended in December 2015. As stated in the call remit, the groups involved detailed 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 third call:

• Project 11: Emerging properties of neuron-glia relationship underlying neurodegeneration and dementia in Alzheimer’s disease

Principal Investigator Institution

Torres Alemán, Ignacio CIBERNED, Cajal Institute CSIC, Madrid Vicario Abejón, Carlos CIBERNED, Cajal Institute CSIC, Madrid Vitorica Ferrández, Javier CIBERNED, University of Sevilla Comella Carnicé, Joan X. Vall d’Hebron University Hospital, Barcelona Gutiérrez Pérez, Antonia University of Málaga Moratalla Villaba, Rosario Associated group, CIBERNED, Cajal Institute CSIC, Madrid Boada Rovira, Mercé Associated group, Fundación ACE, Barcelona

In this project, we have determined neuro-inflammatory processes associated to the progression of Alzheimer´s disease (AD). Specifically, we have analyzed adaptive changes in neuron-glia interactions and their toxic impact together with ensuing neuronal demise. The ultimate aim was to identify novel therapeutic targets to address the pervasive neurodegeneration underlying the disease, and consequently, clinical deterioration. We have assessed emerging properties of the relationship between neurons and glial cells taking into account their intense functional crosstalk. Our assumption was that the dyad Abeta/tau might be triggering novel cellular interactions. We have used animal models as well as brain samples from AD patients.

2015 CIBERNED Annual Report / 238 We have obtained the following main results:

1) Glial cells in AD brains evolve differentially. While at advanced stages of the disease microglia degenerates, astroglial cells are resistant to degeneration. It seems that hyperphosphorylated Tau causes glial toxicity in AD.

2) We have modulated the interaction between the phosphatase calcineurin (involved in inflammatory processes) and the transcription factor FOXO (associated to oxidative stress responses) elicited by Abeta. Indeed, we have markedly ameliorated neuronal death by using novel peptide mimetics.

3) We have determined that FAIM-L, a protein involved in neurotoxic processes, is decreased in AD brains and in rodent models of AD. Further, Abeta reduces FAIM-L, a protein required for TNF to protect neurons against death induced by either Abeta or brain supernatants obtained from APP/PS1 mice.

4) In addition, we have developed cellular models of the disease (iPS cells and primary fibroblasts obtained from AD patients) to corroborate findings in the above used models. Firstly, we have performed a descriptive analysis of these cells.

We have completed all major aspects of the initially proposed objectives, as reflected in our main conclusions:

• Microglia and astroglial cells in AD brains display different activation patterns along the disease. We have found important differences between animal models of AD (APP/PS1 mice) and patient´s brains. Hyperphosphorylated Tau appears as the toxic agent causing the microglial degeneration observed in AD brains. • Interactions between calcineurin and FOXO are amenable to pharmacological manipulation through interfering compounds that in this way show a clear therapeutic potential. The compounds provide protection against Abeta-induced cell death in cultured neurons. • FAIM-L levels in AD brains are reduced at advanced stages of the disease. This is also found in animal models (APP/PS1 mice). In addition, FAIM-L appears to play a key role in the neuronal response to TNF, an ambivalent cytokine with beneficial and detrimental actions on neurons. • IGF-I exerts potent synaptic actions both at pre- and post-synaptic sites. • We have derived neurons and astrocytes from iPS cells obtained from AD patients. • Primary fibroblasts from AD patients with different ApoE polymorphisms show altered cholesterol homeostasis and autophagy disturbances.

• Project 12: Mitochondrial dynamics and mitophagy as therapeutic targets in Parkinson’s Disease and Huntington

Principal Investigator Institution

Soriano García, Eduardo CIBERNED, University of Barcelona Vila Bover, Miquel CIBERNED, Vall d’Hebron University Hospital, Barcelona Trullás Oliva, Ramón CIBERNED, IDIBAPS-CSIC , Barcelona Alberch Vié, Jordi CIBERNED, University of Barcelona Tolosa Sarró, Eduardo CIBERNED, Clinic Hospital, Barcelona

2015 CIBERNED Annual Report / 239 Mitochondrial dysfunction is a key process in the pathogenesis of many neurodegenerative diseases, in particular Parkinson’s disease (PD), and Huntington’s disease (HD). In addition to mitochondrial disorders related to cellular respiration and production of ATP, calcium homeostasis and oxidative stress, recent studies suggest a second type of mitochondrial alterations that converge on two essential processes for neuronal viability and physiology: mitochondrial transport and dynamics, and mitochondria recycling through mitophagy. Although some of the genes involved in these processes are known, the exact mechanisms that regulate mitochondrial dynamics and mitophagy in neurons, as well as their involvement in the pathogenesis of PD and HD -especially in early stages- remain unknown.

The objective of this project is to dissect the role of mitochondrial proteins recently identified by the consortium groups as regulators of mitochondrial dynamics and mitophagy processes (the Armcx/Armc10 family, Pentraxin1/NP1 and HTT) and in the pathogenesis of PD and HD. Second, we will determine its potential therapeutic utility in treating these disorders.

To this end, three specific objectives are proposed:

1) To dissect the molecular mechanisms by which the Armcx/Armc10, NP1 and HTT genes regulate mitochondrial dynamics and mitophagy.

2) To characterize the pathological alterations in mitochondrial dynamics and mitophagy in PD and HD (in brains of patients as in cellular models of disease, including fibroblasts and neurons differentiated from patient-derived iPSCs) and their relationship to the above mentioned genes.

3) To investigate whether manipulating the expression of those genes is able to reverse the PD- and HD- associated mitochondrial deficits and restore function/neuronal viability both in vitro (fibroblasts and neurons from patient-derived iPSCs) as in experimental models in vivo.

The results of this project will help to decipher the role of the alterations of mitochondrial dynamics and mitophagy in the pathogenesis of PD and HD, and will be useful in assessing whether Armcx/ Armc10, NP1 and HTT genes may represent new therapeutic targets for the treatment of these diseases.

• Project 13: Identification and molecular characterization of cannabinoid receptors subpopulations in polyglutaminopathies

Principal Investigator Institution

Guzmán Pastor, Manuel CIBERNED, Complutense University of Madrid Fernández Ruiz, Javier CIBERNED, Complutense University of Madrid McCormick, Peter Joseph CIBERNED, University of Barcelona Mena Gómez, Mª Ángeles CIBERNED, Ramón y Cajal University Hospital, Madrid

Understanding the processes of neuron survival is a pivotal issue for characterizing the etiology and progression of neurodegenerative diseases and, therefore, for designing rational therapies for their treatment. In this context, during the last years 4 groups of this proposal have studied how cannabinoid receptors modulate these processes, and what can be the relevance of cannabinoid receptors as potential neuropharmacological targets.

Specifically, the CB1 cannabinoid receptor is the most abundant metabotropic receptor in the mammalian brain, and is highly expressed in the basal ganglia and cerebellum, where it plays a key role in the control of motor activity and coordination, respectively. It has been shown that CB1 receptor expression decreases in the basal ganglia of patients and animal models of Huntington’s disease (HD), the most studied neurodegenerative disease that occurs by triplet

2015 CIBERNED Annual Report / 240 expansion. Although some detailed studies on the protective role of cannabinoid receptors have already been conducted in HD models, we still do not know which precise cannabinoid receptor subpopulations are involved in those neuroprotective effects.

The existence of CB1 receptor subpopulations has been recently suggested by the findings that this receptor has different signaling efficacy in glutamatergic or GABAergic terminals, and that in transfected cells and ex vivo tissues it can form heteromers with other metabotropic receptors. Hence, the global objective of this proposal is to identify and molecularly characterize the subpopulations of cannabinoid receptor (CB1 and CB1-adenosine receptors or CB1-other cannabinoid receptors heteromers) and to define their potential as new targets to achieve neuroprotection in polyglutamine diseases.

This global objective may be divided in the following specific objectives:

1) To study the differential expression of CB1 receptor heteromers and their neuroprotective role in GABAergic and glutamatergic terminals.

2) To study the neuroprotective role of CB1 receptor and CB1 receptor heteromers in mouse models of two polyglutamine diseases (HD and SCA3) as well as in animals in which a neuronal population-selective activation will be achieved by the Designer Receptors Exclusively Activated by Designer Drugs (DREADD) pharmacogenetic technique.

3) To study the possible alterations in the expression of CB1 receptors and CB1 receptor heteromers in post-mortem brain tissue from patients of two polyglutamine diseases (HD and SCA3).

These studies will put together the complementary experiences and skills of the 4 groups of the proposal, and will help to clarify the molecular bases and possible clinical relevance of two important issues: (i) whether the alterations of the cannabinoid system are involved in the pathogenesis of polyglutamine diseases, and (ii) whether the stimulation of cannabinoid signaling promotes neuroprotection and therefore delays the onset and/or attenuates the progression of those diseases.

• Project 14: Role of GSK-3β in cortical circuits alterations occurring in Alzheimer’s disease

Principal Investigator Institution

Iglesias Vacas, Teresa CIBERNED, Institute of Biomedical Resrach CSIC-UAM, Madrid

Ávila de Grado, Jesús CIBERNED, Center of Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid De Felipe Oroquieta, Javier CIBERNED, Cajal Institute CSIC, Center of Biomedical Technology, Madrid Naranjo Orovio, José Ramón CIBERNED, National Center of Biotechnology CSIC, Madrid Lleó Bisa, Alberto CIBERNED, Santa Creu i Sant Pau Hospital, Barcelona

One of the major aims of Alzheimer´s disease (AD) research is to elucidate he relationship between progressive cognitive decline and molecular, physiological and morphological alterations that take place in the cortical circuits. Changes in calcium levels and overstimulation of glycogen synthase kinase β (GSK-3β) are directly related with AD pathogenesis. Therefore, therapies aimed to modulate calcium homeostasis and/or to inhibit GSK-3β applied at initial stages of AD could have a great potential for its treatment.

In the present project, we have analyzed modifications/alterations associated to GSK-3β hyperactivity in the continuum of AD and the relationship with early alterations in calcium homeostasis in order to identify those changes linked to the disease in preclinical or prodromal stages that could help to early diagnosis. For our studies we have used mainly mouse models

2015 CIBERNED Annual Report / 241 overexpressing GSK-3β, overexpressing the calcium regulated protein DREAM, or lacking the later, and brain and cerebrospinal fluid (CSF) samples from control subjects or AD patients at different disease stages.

With this work, we have increased our knowledge on the molecular bases by which GSK-3β participates in AD etiopathogenesis. GSK--3β overexpression in neural precursors leads to an increase in the number of both precursors and mature neurons in the dentate gyrus. Dendritic spines of those newborn neurons, generated by adult neurogenesis, are clearly distinct from those pre-existing mature neurons. On the other hand, microanatomical studies show differential phosphorylation and location of tau in early and late stages of the disease. Finally, a novel CSF marker has been identified and we have mapped a phospho-epitope, whose phosphor-specific antibody could be tested in AD, to establish its temporal pattern of appearance along disease progression. This data could help early diagnosis and allow therapeutic intervention before symptoms of cognitive impairment occur.

• Project 15: Synaptic degeneration and dysregulation of adult neurogenesis in mouse models of neurodegeneration

Principal Investigator Institution

Fernández Chacón, Rafael CIBERNED, University of Sevilla Lucas Lozano, José Javier CIBERNED, Center of Molecular Biology “Severo Ochoa” CSIC-UAM, Madrid Fariñas Gómez, Isabel CIBERNED, University of Valencia

The involvement of synaptic dysfunctions is well established as a hallmark during early phases of neurodegeneration. Nonetheless, synaptic alterations are more difficult to analyze than neuronal vulnerability and therefore there is a lack of information about how synaptic dysfunction contributes to neurodegeneration.

On the other hand, there are recent evidences indicating that synaptic activity modulates adult neurogenesis and this process might contribute to the progression of some symptoms in neurodegeneration and in the potential response of the brain to degenerative insults. The identification in recent years of molecules involved in the release of neurotransmitters related with different neurodegenerative pathologies has opened the possibility of studying the connection between the maintenance of synaptic function and certain diseases such as Huntington’s disease (HD) or Parkinson’s disease (PD). Because of that, our consortium has studied the relationship between synaptic activity and neurodegeneration through the analysis of mouse models bearing genetic alterations of the three following molecules: Cysteine String Protein-alpha, alpha-synuclein and huntingtin.

We have used three genetically modified mouse models: a model of polyglutamine toxicity (R6/1 mice with mutated huntingtin), a mouse model of presynaptic degeneration (CSP-alpha KO mice) y and mouse models with modifications in the synaptic protein alpha-synuclein.

We have obtained interesting results that are currently being further investigated and that will eventually be published. These results indicate that:

The proteins CSP-alpha and alpha-synuclein are involved in neurogenesis. Fernández-Chacon’s 1) group have found alterations in neurogenesis at the SGZ in mice lacking CSP-alpha that could be explained by a non-synaptic role of CSP-alpha in neural stem cells. On the other hand, Fariñas’ group has reported alterations in neurogenesis at the SVZ in KO mice lacking alpha- synuclein as part of a wider study that demonstrate the role of dopaminergic inputs on the regulation of neurogenesis at the SVZ.

2015 CIBERNED Annual Report / 242 Fernández-Chacón’s and Fariñas’ groups keep collaborating to understand the role of alpha- synuclein on the adult neurogenesis at the SGZ and the role of CSP-alpha on the adult neurogenesis at the SVZ. Interestingly, it seems that at the SGZ both proteins are present at neural stem cells from neurospheres. In contrast, at the SVZ both proteins seems to be expressed only at the synaptic terminals.

2) The absence of CSP-alpha does not protect against the neurotoxicity induced by mutated huntingtin in mice. These results are based in a novel mouse line generated in the consortium of R6/1 mice lacking a copy of the CSP-alpha gene. Lucas’ group has carried out behavioral analysis that indicates that the reduction of CSP-alpha does not improve survival or motor performance in R&/1 mice.

3) There is an unexpected alteration of CPEB4 in Huntington disease. Interestingly such an observation from Lucas’ group has led to unravel a new node in autism gene regulation. Fernandez-Chacón’s group has contributed to find alterations in synaptic transmission in transgenic mice over-expressing CPEB4. On the other hand, Fariñas’ results indicate a deficit in adult neurogenesis at the SVZ in the absence of CPEB4. Part of those results are being resubmitted for publication to Cell.

Fourth call

Nº Code Title Coordinator 16 PI2014/06 Onset and progression of Parkinson’s disease: role of Rodríguez Oroz, Mª Cruz glial activation

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

This fourth call for proposals is funded with 532,074 € for two years, and involves the active participation of eight CIBERNED research groups. Below is a brief description of the scientific activity carried out by each of the projects during its first year of implementation:

• Project 16: Onset and progression of Parkinson’s disease: role of glial activation

Principal Investigator Institution

Rodríguez Oroz, Mª Cruz CIBERNED, Biodonostia Research Institute, San Sebastián Matute Almau, Carlos CIBERNED, University of the Basque Country, Bilbao Obeso Inchausti, José A. CIBERNED, Center for Applied Medical Research, Univ. Navarra, Pamplona Rodríguez Día, Manuel CIBERNED, University La Laguna, Tenerife

Parkinson’ disease (PD) is characterized by degeneration of dopaminergic neurons in substantia nigra pars compacta (SNc) and the presence of cytoplasmic inclusions containing alpha- synuclein aggregates (a-syn). In most cases, it is sporadic, but in 5-10% of cases exits a genetic cause, being LRRK2 gene mutations the most frequent. Moreover, GWAS studies also point to a main role of this gene in sporadic PD. The first motor signs of PD appear when dopaminergic loss is around 40-60%, indicating that neurodegeneration precedes diagnosis in several years. In fact, asymptomatic carriers of LRRK2 mutations have dopaminergic depletion years before the diagnosis of PD.

2015 CIBERNED Annual Report / 243 Many mechanisms have been involved in the pathogenesis and progress of PD among which neuroinflammation and glial activation have charged great relevance. Studies in patients with advanced PD and animal models with acute and severe dopaminergic lesion (6-OHDA rat and MPTP monkey) show numerous evidences of an activation of microglia and astrocytes and high levels of proinflammatory cytokines in the SNc and striatum that are associated with dopaminergic death. However, astrocyte can also have a protective role through several mechanisms.

Moreover, it has been hypothesized that mutations in LRRK2 gene are associated with a proinflammatory state in microglial cells. Thus, existing data show a strong interaction between inflammation and dopaminergic death, although neither human studies nor animal models used until this moment allow explaining the role of microglia and astrocytes in PD (initiation/progress or consequence of dopaminergic death). In addition, it is not known whether LRRK2 mutations causing PD induce proinflammatory changes that can drive to dopaminergic degeneration. In this work, we aim to clarify the role of astrocytes and microglia in PD through several complementary approaches. To this aim, we will analyze the relationship between glial cells, neuroinflammation and dopaminergic neurons death regarding sporadic and LRRK mutation PD.

Studies will be performed in cell cultures derived from patients with and without LRRK2 mutation, asymptomatic carriers and in vivo studies in animal models of progressive dopaminergic nigrostriatal lesion as well as in patients and asymptomatic carriers. More specifically, astrocytes (Ast) differentiated from induced pluripotent stem cells (iPS) derived from fibroblasts of sporadic and LRRK2 mutations PD patients and from asymptomatic carriers of these mutations will be generated and characterized. Survival, proliferation, differentiation and migration of implanted Ast-iPS in the striatum of healthy and parkinsonian rats will be studied in vivo to assess its role in the initiation and progression of dopaminergic death. The temporal relationship between the nigrostriatal lesion and microglial activation will be defined in vivo using PET in the rat model overexpressing mutant human a-syn A53T and the progressive MPTP intoxication in monkey. In addition, in asymptomatic LRRK2 mutation carriers and in de novo sporadic PD patients, the relation between striatal dopaminergic denervation and presence of inflammatory changes will also be study by PET.

• Project 17: Epigenetic mechanisms involved in the etiology and progression of rapidly progressive neurodegenerative dementias

Principal Investigator Institution

Calero Lara, Miguel CIBERNED, Carlos III Institute of Health - CIEN Foundation, Madrid Ferrer Abizanda, Isidro CIBERNED, Bellvitge Biomecial research Institute, Barcelona Del Río Fernández, José A. CIBERNED, Catalonian Bioengineering Institute, Barcelona Sánchez Juan, Pascual CIBERNED, Marqués de Valdecilla Foundation, Santander

This project is focused on the study of epigenetic mechanisms involved in the etiology and progression of rapid progressive neurodegenerative dementias (rpNDD) compared to their slow progressive counterparts. The main objectives are:

1) Identification of epigenetic mechanisms that modulate the molecular expression of rpNDD (CJD, rpAD, and rpLBD) in brain tissue.

2) Analysis of the hypermutability of CpG dinucleotides in genes associated with CJD, AD and LBD as a regulatory mechanism of the expression of neurodegenerative dementias.

3) Targeted identification of epigenetic and transcriptomic signatures of differentiated IPSc.

4) Device a sub-classification and definition of rpNDD endophenotypes, as a function of neuropathological and molecular characteristics based on epigenetic signatures.

2015 CIBERNED Annual Report / 244 5) Elucidate whether the observed changes can be used as biomarkers in accessible fluids such as blood and CSF.

This work is a multidisciplinary approach common to different clinical entities with similar presentation, focused on the epigenetic mechanisms involve in the etiology of rpNDD. Currently, the project is analyzing human samples from the Brain Bank of Institute of Neuropathology (Biobank HUB-ICO-IDIBELL, differentiated IPS cells, as well as ADN samples and blood and CSF samples from collections of the participants from several neurodegenerative disorders (CJD, rpAD, rpLBD, typical AD and LBD and age-matched controls).

Following, we describe the most important results, arranged according to the pertaining objectives:

• Objective 1. Identification of epigenetic mechanisms that modulate the molecular expression of rpNDD (CJD, rpAD, and rpLBD) in brain tissue.

Preliminary results on this objective, led by Dr. Isidro Ferrer, suggest that rapid progressive dementias related to neurodegenerative disorders associated with abnormally-conformed protein deposits show: i) an increase on the inflammatory response, and ii) more pronounced changes in molecules involved in protein synthesis.

• Objective 2. Analysis of the hypermutability of CpG dinucleotides in genes associated with CJD, AD and LBD as a regulatory mechanism of the expression of neurodegenerative dementias.

The results of this objective, led by M. Calero, indicates: i) that the hypermutability of CpG dinucleotides may be involved in creating hot spots in the sequence of genes involved in neurodegenerative dementias, and ii) that variability in genes involved into the regulation of this hypermutability of CpG (DNA methyltransferase, cytosine deaminases, repair-enzymes, etc.) may be associated with neurodegenerative disorders.

• Objective 3. Targeted identification of epigenetic and transcriptomic signatures of differentiated IPSc.

During this year, in this objective led by Dr. Jose Antonio del Rio, several key tools have been generated: i) Spherical neural masses (SNMs) have been differentiated from neural induced pluripotent stem cell (IPSc) from a patient with GSS, and ii) The SNMs generated have been characterized and material for subsequent analyses (RNA, DNA, fixed cell cultures) prepared.

• Objective 4. Device a sub-classification and definition of rpNDD endophenotypes as a function of neuropathological and molecular characteristics based on epigenetic signatures.

This objective, led by Dr. Isidro Ferrer, requires collecting all the information within the collaborative project, and therefore, it will be performed at the end of the second year, at the conclusion of the project.

• Objective 5. Elucidate whether the observed changes can be used as biomarkers in accessible fluids such as blood and CSF.

The preliminary results of this objective, led by Dr. Pascual Sanchez Juan, following a comparative study between EA cases and controls has allowed us to select several miRNA as candidate biomarkers, namely miR-9-5p, miR-134 and miR-598. Analysis of the routes involved by using the web page www.mirsystem.cgm has demonstrated that these miRNA are involved in: i) amyloid formation routes, ii) cellular stress signaling, and iii) Huntington disease.

2015 CIBERNED Annual Report / 245 The overall results obtained during the first year of the project are satisfactory, relevant and according to the initial work plan. It is important to remark that this collaborative project could not be carried out by any of the participants single-handedly, as the project takes advantage and potentiates the technical and clinical expertise as well as the methodological resources of the research team, constituted by four CIBERNED groups with established experience in the neurodegeneration field.

Fifth call

Nº Code Title Coordinator 18 PI2015/02 Validation of new therapeutic targets and biomarkers Labandeira García, José in Parkinson’s disease Luis

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

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

This fifth call for projects it is endowed with 345,000 € annually for two years, of 690,000 € overall, for the 11 CIBERNED research groups and 3 external associated groups that participate in them. A summary of the objectives set for the projects approved in the fourth call, initiated during the second half of 2015 is presented below.

• Project 18: Validation of new therapeutic targets and biomarkers in Parkinson’s disease

Principal Investigator Institution

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

Development of new therapeutic strategies which alleviate the exponential growth of neurodegenerative disorders such as Parkinson’s disease (PD) constitute a scientific, medical and social priority challenge of human health. New strategies are necessary to go beyond current symptomatic treatment. Unfortunately, all neuroprotective strategies have failed. Furthermore, the absence of effective therapies against cognitive impairment which go in varying degrees with disease progression is particularly worrying. This challenge will be undertaken in this project from a multidisciplinary perspective and providing innovative strategies to address several factors that could lead to failure of previous studies:

1) A new model of PD induced by α-synuclein in mice and primates will be introduced.

2) In contrast to previous studies based on the administration of antioxidants, two new therapeutic targets focused on major and common mechanism of redox metabolism and neuroinflammation will be tested: inhibition of the pro-inflammatory angiotensin-NADPH-oxidase-Rho-kinase pathway and reinforcing pharmacological activation of the transcription factor Nrf2 as main regulator of antioxidant defense.

3) Alterations observed in murine and primate preclinical models with findings in postmortem brain tissue of patients with PD will be correlated.

2015 CIBERNED Annual Report / 246 4) Advanced Glycation End products (AGEs) will be assessed in patients and controls plasma as potential predictive biomarkers of cognitive impairment in PD. Plasma levels of AGEs and cognitive impairment in PD primate models will be correlated. Finally, the effects of the proposed therapies on AGEs plasma levels will be assessed in mice and primates animal models.

• Project 19: Neuregulin gene therapy aimed at the treatment of motor neuron degeneration in ALS

Investigador principal Institución

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

Neurodegenerative diseases are disabling and fatal disorders characterized by dysfunction and loss of neurons in specific regions of the central nervous system (CNS). In these diseases, neuron death is associated with several pathogenic hallmarks including, among others, protein aggregation, mitochondrial dysfunction, oxidative stress and synaptic defects. However, common molecular mechanisms causing these pathogenic changes in neurodegenerative diseases are largely unclear. Understanding these mechanisms is crucial for designing efficient therapeutic approaches to prevent, delay or reverse neuron and synaptic dysfunction in these disorders.

Importantly, recent genetic and cellular evidences suggest an association of the neurotrophic factor Neuregulin 1 (Nrg1) and its receptor ErbB4 in Charcot-Marie--Tooth and Amyotrophic Lateral Sclerosis (ALS) among 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 in 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 (AD). However, the pathogenic role of Nrg1 and its processing in motoneuron survival is not well understood. Our hypothesis is that Nrg1/ErbB4 signaling could be a relevant molecular pathogenic mechanism underlying neurodegeneration.

Therefore, targeting Nrg1/ErbB signaling may represent a promising novel therapeutic approach for ALS and other neurodegenerative diseases. To investigate the role of Nrg1/ErbB4 system on motoneuron dysfunction and loss in ALS pathogenesis, we propose to examine Nrg1/ErbB4 signaling and its therapeutic targeting in a faithful mutant mouse model of ALS, harboring a point mutation in the Sod1 gene, which leads to motoneuron degeneration and a phenotype mimicking the usual clinic of sporadic and familial ALS.

To this end, this project intends to bring together four established groups with long and complementary expertise in the study of neurodegeneration with the aim to evaluate new advanced therapies for ALS. We propose the following specific aims:

• To characterize the expression and processing of Nrg1 isoforms and ErbB receptors in spinal cord and muscles of SOD1G93A ALS mice and in ALS patient samples.

• To investigate the role of Nrg1--ErbB in neuronal degeneration and neuroinflammation and the potential cellular signaling pathways involved.

• To evaluate gene therapy strategies for modulating the expression of Nrg1 isoforms to increase neuromuscular junction maintenance and motoneuron survival, and thus to improve motor function and survival in SOD1G93A ALS mice.

2015 CIBERNED Annual Report / 247 • To assess the effects of Nrg1 modifications on maintenance and formation of synapses on the cortical and spinal neurons as well as maintenance (or restoration) of neuromuscular junctions in the ALS mouse model.

• Project 20: Differential metabolic profiles in Parkinson’s disease

Principal Investigator Institution

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

Intermediary metabolism and cell signalling pathways have been considered and studied so far as independent entities. Actually, they are the catabolic and anabolic pathways that provide the energy and cell precursors necessary for cell survival. Thus, knowledge of the metabolic changes is essential to integrate and understand the mechanisms leading to neurodegeneration. The determination of differential metabolic profiles of Parkinson Disease may allow the development of a risk and prognosis diagnostic pattern.

The aim of this study is the identification of differential transcriptomic and metabolomic profiles using cellular and animal models. Both, mice that over expressing two major pathogenic mutations in LRRK2 (G2019S and R1441G) and wild type mice, will be used for in vivo studies. A group of animals will be treated with MPTP in order to differentiate the effect of environmental agents either in controls or in animals carrying the aforementioned mutations. In addition, we will use fibroblasts and plasma from healthy individuals, patients with idiopathic Parkinson, asymptomatic carriers and patients with the G2019S and R1441G pathogenic forms. Similarly, to in vivo models, fibroblasts will be treated with MPP+ toxin (MPTP active metabolite). Finally, once identified the most promising targets, we will analyze the effect of modulators/adapters as possible neuroprotective agents in Parkinson’s disease.

This project is the first study which combines transcriptomics and metabolomics with the identification of specific biomarkers in genetic and neurotoxic models and with a simultaneous study of cellular and animal samples from idiopathic Parkinson’s patients, Parkinson’s patients with pathogenic mutations for LRRK2 (G2019S and R1441G) and healthy carriers with the same mutation. The profiles obtained from these studies can be used to identify genomic or metabolomic patterns which could be utilized as predictive and diagnosis markers of Parkinson’s disease and used for the development of potential new neuroprotective drug therapies.

Finally, during the second half of 2015, the sixth call for projects was launched and the newly approved projects are expected to begin activities in the first half of 2016.

CIBERNED cooperative program has so far invested an overall budget of about 6,812,174 €, distributed among the 20 cooperative projects mentioned above. Up to 55 CIBERNED research groups have actively participated in the cooperative program, which represents 90.47% of all groups belonging to CIBERNED during this period, along with a dozen of outside associated groups. In addition, 40 groups (63,49%) have participated in more than one cooperative project and projects 16-20 are currently still active, with the involvement of 19 research groups CIBERNED (35,18%).

2015 CIBERNED Annual Report / 248 By analyzing previous calls, given that, some are completely closed and others just started an estimation of the budget invested can be made by individual project and participating group and project. Thus:

Average budget Average budget Duration Total number Number of Call Budget for project for project and in years of groups projects and year group

1st call (2010) 2.322.100 € 3 40 6 129.005,56 € 19.350,83 € 2nd call (2011) 1.192.000 € 2 26 4 149.000,00 € 22.923,08 € 3rd call (2013) 1.376.000 € 2 24 5 137.600,00 € 28.666,67 € 4th call (2014) 532.074 € 2 8 2 133.018,50 € 33.254,63 € 5th call (2015) 690.000 € 2 11 3 115.000,00 € 31.363,64 € 6th call (2015) 700.000 € 2 To be launched in 2016

2015 CIBERNED Annual Report / 249 International relations 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.

2015 CIBERNED Annual Report / 253 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. The projects involving CIBERNED research groups that have been active during 2015 are described below.

BIOMARKAPD: Biomarkers for Alzheimer’s disease and Parkinson’s disease

Total funding 8,5 M€ (approx.)

Start date June, 2012

Duration 3 years

Coordinator Bengt Winblad, Karolinska Institutet, Stockholm, Sweden

CIBERNED participation Alberto Lleó and Javier Sáez-Valero groups

2015 CIBERNED Annual Report / 254 Project partners:

• Judes Poirier, McGill Heine University Medical • Marzena Zboch, Scinawa, University, Quebec, School Duesseldorf, Wroclaw Medical Canada Germany University, Poland • Bengt Winblad, Karolinska • Lutz Froehlich, Central • Alexandre Mendonca, Institutet, Stockholm, Institute of Mental Health Instituto de Medicina Sweden Mannheim, Germany Molecular, Portugal • Lennart Minthon, Skåne • Thomas Arendt, University • Catarina Resende de University Hospital, Sweden of Leipzig, Germany Oliveira, University of • Henrik Zetterberg, The • Magda Tsolaki, Aristotle Coimbra, Portugal Sahlgrenska Academy, University of Thessaloniki, • Odete da Cruz e Silva, University of Gothenburg, Greece University of Aveiro, Sweden • Elisabeth Kapaki, University Portugal • Gunhild Waldemar, of Athens, Greece • Norbert Zilka, Slovak Rigshospitalet, • Brian Lawlor, Trinity College Academy of Science, Copenhagen Univ Hospital, Dublin, Ireland Slovakia Denmark • Andrea Urbani, IRCCS • Uros Rot, University Medical • Niels Heegard, Statens Foundation Santa Lucia, Centre, Ljubljana, Slovenia Serum Institut, Denmark Italy • José Luis Molinuevo, • Hilkka Soininen, University of • Fabrizio Tagliavini, IRCCS Hospital Clinic, Barcelona, Eastern Finland, Finland Foundation Carlo Besta, Spain • Juha Rinne, Turku University Italy • Alberto Lleo, Santa Hospital, Finland • Giovanni Frisoni, IRCCS Creu i Sant Pau Hospital, • Bruno Dubois, Salpetriere Foundation San Giovanni Barcelona, Spain University Hospital (UPMC - Di Dio Fatebenefratelli, Italy • Joan Gispert, Pasqual University Paris 6), France • Paul Wilmes, University of Maragall Foundation, • Sylvain Lehmann, CHRU de Luxembourg, Luxembourg Barcelona, Spain Montpellier, France • Pieter Jelle Visser, • Javier Sáez-Valero, • Armand Perret-Liaudet, String of Pearls Initiative University Miguel Université Lyon 1, France Neurodegeneration, The Hernández-CSIC, • Jens Wiltfang, Universität Netherlands CIBERNED, Spain Duisburg-Essen, Germany • Marcel Verbeek, Radboud • Christoph Hock, University • Thomas Klockgether, University Medical Centre, of Zurich, Psychiatric German Center for Nijmegen, The Netherlands & University Hospital, Neurodegenerative • Tormod Fladby, Akershus Switzerland Diseases (DZNE), Germany University Hospital, Norway • Panteleimon • Brit Mollenhauer, • Jacek Kuznicki, Giannakopoulos, University Paracelsus-Elena-Klinik + International Institute of of Geneva Hospital, • Georg August University Molecular and Cell Biology Switzerland Göttingen, Germany (IIMCB), Poland • Sermin Genc, Dokuz Eylul • Piotr Lewczuk, • Andrzej Szczudlik, University, Turkey Universitätsklinikum Jagiellonian University • Esen Saka Topcuoglu, Erlangen, Germany College of Medicine (UJ), Hacettepe University, • Markus Otto, University of Poland Turkey Ulm, Germany • Tomasz Gabryelewicz, • Hakan Gurvit, Istanbul • Katrin Marcus, Ruhr IMDiK, Mossakowski University, Turkey University Bochum, Medical Research Centre, Germany Poland • Dieter Willbold, Heinrich-

In the context of the JPND-BIOMARKADPD, we collaborated in the establishment of analytical protocols for biomarker determination in the cerebrospinal fluid (CSF) of Alzheimer’s patients (AD), as well in the validation of α-synuclein as a potential biomarker for Parkinson’s disease. Most of our effort has been focused in researching for new markers for Alzheimer’s disease. There is evidence for the diagnostic potential of soluble forms of the amyloid precursor (APPα and sAPPβ), and we demonstrated the presence of isoforms containing the transmembrane domain (full-length / unprocessed protein) of APP in the CSF forming oligomers and confounding the ELISA analysis. Similar situation is studied for the enzyme β-secretase BACE1 in the CSF. We have demonstrated that complexes of the γ-secretase enzyme presenilin-1 (PS1) are present in CSF and have potential as biomarker for Alzheimer’s disease. In the AD brain, we found a strong depletion in glycoforms of proteins carrier of the HNK-1 glycoepitope. Finally, we demonstrated that assessment in the CSF of the ApoER2 receptor for ApoER2/ApoE served to estimate the functionality of the signaling pathway.

2015 CIBERNED Annual Report / 255 DEMTEST: Biomarker based diagnosis of rapid progressive dementias- optimisation of diagnostic protocols

Total funding 2,2 M€ (approx.)

Start date May, 2012

Duration 3 years

Coordinator Inga Zerr, University Medical Center Göttingen, Germany

CIBERNED participation Pascual Sánchez-Juan (José Berciano group) and Miguel Calero (Jesús de Pedro group)

Project partners:

• Inga Zerr, University • Stefano Ruggieri & • Pascual Sanchez-Juan, Medical Center Göttingen, Maurizio Pocchiari & Anna IFIMAV and CIBERNED, Germany Ladogana, University Marqués de Valdecilla • Carsten Korth, Heinrich “Sapienza”, Rome, Italy University Hospital, Spain Heine University Medical • Pawel Liberski, Medical • Anna-Lena Hammarin, School, Düsseldorf, University of Lodz, Poland Swedish Institute of Germany • Catarina Resende Oliveira, Communicable Disease • Hans Kretzschmar, Ludwig- University of Coimbra, Control, Sweden Maximilians-University, Portugal • Adriano Aguzzi & Herbert Munich, Germany • Eva Mitrová, Slovak Budka, University Hospital • Jean-Louis Laplanche, Medical University Zürich, Switzerland Hopital Lariboisiere AP-HP, Bratislava, Slovakia • John Collinge, University France • Gorazd Bernard Stokin, College London, United • Olivier Andreoletti, UMR- University Psychiatric Kingdom INRA-EVNT, France Hospital, Ljubljana, Slovenia • Robert G. Will, Western • Theodoros Sklaviadis, • Miguel Calero, National General Hospital, United Aristotle University of Institute of Health Carlos III, Kingdom Thessaloniki, Greece Spain

The DEMTEST collaboration allowed us to perform a genome wide association study (GWAS) in prion diseases. Because of that, we published in 2012 an association between MTMR7 gene and susceptibility to new variant Creutzfeldt-Jakob disease (CJD); this study was followed by an attempt of sequencing our top candidate genes where we found a risk variant in PLCXD3 gene. In 2014, we finalized the second part of our analysis; we performed a GWAS and replicated the results with samples from sporadic CJD patients from all partner countries. We have successfully replicated an intronic SNP in GRM8 gene associated to sporadic CJD risk. In collaboration with our JPND partners (ISCIII Miguel Calero), we sequenced the exons of that gene in a large sample of Spanish cases. In parallel, we correlated genomic data with CSF markers in a sample of German patients from the group of Dr Zerr. Results of these analyses were published during 2015.

The other main results from this project were fruit of direct collaborations with the coordinator of the consortium, Dr. Inga Zerr. We carried out studies evaluating new biomarkers for rapidly progressive dementia (desmoplakin) and analysis of clinical data to generate new diagnostic algorithms for other prion disorders like fatal familial insomnia.

2015 CIBERNED Annual Report / 256 COURAGE-PD: Comprehensive unbiased risk factor assessment for genetics and environment in Parkinson‘s disease

Total funding 4,5 M€ (approx.)

Start date January, 2014

Duration 3 years

Coordinator Thomas Gasser, University of Tuebingen, Germany

CIBERNED participation Eduardo Tolosa group

Project partners:

• Thomas Gasser & Rejko Hospital, Norway Portugal Krüger University of • Nicholas Wood, University • Eduardo Tolosa, University Tübingen, Germany College London, United of Barcelona, Spain • Alexis Elbaz, INSERM, Kingdom • Rudi Balling, University of France • Avi Orr-Urtreger, Tel Aviv Luxembourg, Luxembourg • Stefano Goldwurm, Istituti Sourasky Medical Center, • Peter Heutink, VU Clinici di Perfezionamento, Israel University Medical Italy • Joaquim Ferreira, Institute Centre, Amsterdam, The • Mathias Toft, Oslo University of Molecular Medicine, Netherlands

In 2015, COURAGE-PD has mainly be working on the following tasks of the work programme:

Work package 1:

Identification of genetic risk variants. Task 1: Exome sequencing. Task 2: Resequencing of GWAS loci. Task 3: Bioinformatic analysis.

Work package 2:

Genetic susceptibility, protective factors, and gene-gene and gene-environment interactions in PD. Task 1: COURAGE-GEOPD chip development and analysis. Task 4: Improving causal inference for environmental factors in PD: MR, GEI, GGI analyses.

Work package 3:

Task 1 and Task 2: Functional validation of novel genes for monogenic PD and functional characterization of newly identified genetic risk or protective factors. Task 3: Modeling gene-environment interactions (GEI) in vitro. Task 4: Transcriptional analyses of mDA neurons expressing novel genetic causes for PD.

More specifically, the Tolosa group (CIBERNED) has been directly involved in the following activities:

2015 CIBERNED Annual Report / 257 • First, all DNA samples from several selected Spanish families with several PD affected members have been already acquired and preliminary quality control on DNA has been performed. The workflow of the next-generation-sequencing project has been arranged, and genome sequencing has been performed in 2015, including other samples from different European countries. In addition, we provided around 500 samples (PD patients and controls) for the WP2, in order to be analysed with the COURAGE-GEOPD chip, which will cover all variants defined in the screening approach (WP1).

• Second, We recently published a genomewide DNA methylation and a transcriptome studies in induced pluripotent stem cell (iPSC)-derived dopaminergic neurons (DAn generated by cell reprogramming of somatic skin cells from patients with monogenic LRRK2-associated PD (L2PD) or sporadic PD (sPD), and healthy subjects) (Fernández-Santiago et al 2015) by using the Infinium HumanMethylation450 BeadChip. We observed extensive DNA methylation changes in PD DAn, and of RNA expression, which were common in L2PD and sPD. The PD-associated hypermethylation was prominent in gene regulatory regions such as enhancers and was related to the RNA and/or protein downregulation of a network of transcription factors. In the context of COURAGE project, we also have performed a whole genome bysulphite sequencing analysis of the DNA from these samples in order to have a complete knowledge of the epigenetic alterations in this model of PD. Actually, we are analysing this genomic data with two main objectives: a) To expand the previous published study by analysing the possible enrichment in specific chromatin states (Ernst et al., 2011)., as hypermethylation in enhancers of PD b) To search for genomic correlations between differentially methylated loci and genetic top signal loci in published genome wide association analysis, thus searching genetic risk variants that colocalize with hyper/hypo methylated regions. The latter could be used to prioritize the validation of SNP risk found in these GWAS, and in COURAGE project.

Thus, the possibility is now open to model the potential environmental factors found in WP2, in iPSC)-derived dopaminergic neurons.

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.

2015 CIBERNED Annual Report / 258 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, Slovakia • CIBERNED-CIEN Foundation, Spain • Agence Nationale de la Recherche (ANR), France

The overlapping of the COEN group members with those of the JPND will ensure that their complementary objectives progress in close cooperation with each other. This is accomplished through a two-step process, involving expert workshops for the analysis of needs, followed by a call for proposals for collaborative teams between PIs within the participating national Centers of Excellence.

Since 2012, CIBERNED and CIEN Foundation are part of the Oversight Group to be actively involved in the design of COEN future scientific strategy. Dr. Miguel Medina, CIBERNED Deputy Scientific Director and member of the Scientific Advisory Committee of the CIEN Foundation, represents both institutions in the COEN Oversight Group.

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 three approved projects involving CIBERNED have completed during 2015:

2015 CIBERNED Annual Report / 259 • WNT signaling: biomarker and target evaluation in Alzheimer’s disease

Project partners: Antonio Cuadrado (coordinator, CIBERNED, Spain), James Woodgett (Canada) and Simon Lovestone (UK)

This project was established to determine if novel elements of the canonical and non-canonical WNT signaling pathways could be targeted to modify and monitor AD progression. During the two years of duration of this COEN-pathfinder grant, we have been using multidisciplinary approaches, including brain oriented drug screening, a novel proprietary mouse model of AD, new genetic and pharmacological tools for modulation of WNT/GSK3 and WNT/JNK pathways, and human blood samples from AddNeuroMed, DCF and EMIF platforms to identify novel biomarkers of AD progression. The end goal was to explore the impact on AD of novel WNT elements including but not limited to clusterin, DKK1 and NRF2. The final purpose of this grant was the validation of novel therapeutic targets that would be used in a second R&D program with industry partners. This second program was cancelled by COEN.

This project has been conducted essentially according to the initial work plan, as long as experimental results are concerned, or even better in some aspects related to institutional support, additional sources of funding and interaction with industry. Thus, the use of NRF2 as a target in neurodegenerative diseases has been analyzed in preclinical models of PD in collaboration with BIOGEN and we are currently discussing a new collaboration with and SME called EVGEN located at UK.

According to the work plan, the identification of WNT/β-catenin, WNT/PCP and NRF2 modulators in primary high throughput screening with dual reporter assays has been completed. We found that two small molecules identified by this screening inhibit the DKK1/WNT-PCP pathway and activate NRF2. Similarly, the evaluation of the effects of GSK-3 and JNK activation on NRF2 has also been completed. At the mechanistic level, we have characterized the activation of NRF2 by a pathway that involves inhibition of the GSK-3/β-TrCP branch of canonical WNT signaling while JNK/PCP, unexpectedly, does not seem to have a relevant role on NRF2 phosphorylation.

We have completed the characterization of our bigenic APP/Tau and trigenic APP/Tau/Nrf2-KO mice. The role of NRF2 in regulation of autophagy in these mice is now under second review in Autophagy. The characterization of the amyloidopathy and tauopathy has led to two independent studies that are in preparation for publication. These mice are now available for analysis of our selected compounds and for collaboration with industry in preclinical settings. Another rat model has been used based on intraventricular delivery of Aβ in rats. In relation to the potential use of WNT related biomarkers, we have found that DKK isoforms generate unique signatures that are significantly altered in AD patients and are disease relevant when analyzed using pathway bioinformatics. This study has been submitted for publication.

As a result of the COEN collaborations we have presented communications to several meeting that strongly support our view of WNT/GSK3, WNT/JNK and NRF2 as new targets for AD. Very importantly, we have also established collaborations with biopharmaceutical companies that will strengthen our capacity to translate these COEN-pathfinder results to clinical studies.

• In vivo neuronal cell reprogramming for a new regenerative approach in Parkinson’s disease

Project partners: Vania Broccoli (coordinator, Italy), Alexander Dityatev (Germany) and José Luis Lanciego (CIBERNED, Spain)

Neurodegenerative diseases cause a significant burden on the elderly population in Europe. Parkinson’s disease (PD) affects 1.2 million people in Europe and with the increasing life expectancy, this number will rise, putting more pressure on health care. Treatment of PD is only symptomatic, and

2015 CIBERNED Annual Report / 260 therefore, there is an urgent need for more efficient therapies. Degeneration of mesencephalic DA neurons triggers the initial phases of PD, which raised the concept that cell replacement might represent a long-term restorative option for this neuropathology. Indeed, previous studies in PD patients have indicated that cell therapy has the potential to significantly sustain an enduring symptomatic relief. However, these studies suffered from lacking an ideal source of transplantable human DA neurons. We have recently developed a methodology that promotes transdifferentiation of mouse and human fibroblasts into functional induced dopaminergic neurons (iDANs), which display sophisticated neuronal properties including pacemaking firing activity, synaptic integration, and activity-dependent dopamine release. Therefore, iDAN cells offer an unprecedented cellular source with ideal features for cell therapy in PD, since they can be generated from the patients in high amounts. Here, we propose to push forward this technology by elaborating methods for direct in vivo reprogramming to further induce in situ local neuronal transdifferentiation in relevant animal models of PD (mouse and monkey).

• MicroRNA as novel therapeutic targets and disease biomarkers in Alzheimer’s Disease, Frontotemporal dementia and Amyotrophic lateral sclerosis (NEURO-MIR)

Project partners: Jochen Prehn (coordinator, Ireland), Andre Fischer (Germany), Pierre Lau (Flanders, Belgium), José J. Lucas (CIBERNED, Spain)

The NEURO-MIR Consortium aims to explore the role of micro RNAs (miRNAs), molecules capable of controlling gene expression and sensitive to metabolic or environmental stress, in different neurodegenerative diseases (Alzheimer’s disease -AD-, frontotemporal dementia -FTD- and amyotrophic lateral sclerosis -ALS-), which share common disease pathways as the hyperexcitable state, defects in protein quality control and axonal transport defects. Given their unique biological function and in vivo stability, miRNAs show up as one of the most interesting areas for drug and biomarker development, as well as potential key contributors to the disease progression. Pilot data from partners of this Consortium have already shown neuroprotective effects of silencing a miRNA, namely miR-134.

In order to achieve the main aim of this project, 4 Workpackages were proposed, from which we have participated in three:

Workpackage 1: Identify and interrogate the biological function of AD, FTD and ALS-associated miRNAs.

We have generated animals for the immunoprecipitation of Ago2, a protein required to bring together the miRNA and messenger RNA target, in hippocampal samples from control mice, VLW mice that overexpress the microtubule-associated protein tau with 3 mutations found in FTDP-17 (frontotemporal dementia with parkinsonism associated to chromosome 17) patients and Tau-KO mice that don’t express tau (Partner 4). Subsequently, Partner 1 evaluated the miRNA profile in this samples with OpenArray chips. In addition, Partner 4 generated samples from P301S mice that express tau with a mutation that give rise to a more severe phenotype. This samples will be used for validation studies. The results of this study revealed the presence of 155 miRNAs associated to Ago2, an enrichment of miRNAs in VLW animals and a decrease in miRNAs in Tau-KO animals.

Validation of results:

• miR-134, that controls hyperexcitability, has been validated in two AD models: mice that overexpress tau and mice that overexpress GSK-3/tau (generated by Partner 4). An enrichment of its levels has been detected. In addition, it is not detectable in Tau-KO animals. It is overexpressed in AD brain as well.

• miR-22, that controls inflammation, is reduced in GSK-3/tau mice an in AD brain.

2015 CIBERNED Annual Report / 261 Workpackage 2: Deliver a systems biology platform for the modelling of miRNA biological function in neurodegenerative disorders (without Partner 4 participation).

Workpackage 3: Develop strategies for the delivery of antagomiRs/miRNAs as therapeutics.

Antagomir (stable locked nucleic acid-modified oligonucleotides, capable of blocking miRNAs) delivery has been optimized in animal models of neurodegenerative diseases.

Workpackage 4: Test the role of miRNA network changes as biomarker for disease pathogenesis and therapeutic efficacy.

Plasma samples from patients presenting mild or severe Alzheimer’s disease pathology (provided by Partner 4) were analyzed with OpenArray chips. Subsequently, an additional set of samples from control, dementia and severe AD individuals was provided. The results pointed out that:

• 10 miRNAs are overexpressed in dementia, 17 in AD and 4 in both conditions. These miRNAs include miR-139-5p and miR-22.

• 19 miRNAs are downregulated in dementia, 28 in AD and 13 in both conditions.

In addition, tear drops (provided by Partner 4) from AD patients (control, dementia and AD) are being analyzed with OpenArray chips.

Additional results (Partner four):

Given their high expertise in the study of Huntington’s disease (HD), Partner 4 has performed a pilot study in HD animal models. miR-22 levels are reduced in the striatum (main brain structure affected in this disease) of presymptomatic animals and increased in symptomatic animals.

Likewise, Partner 4 has found alterations in MAP2 splicing in HD that could be a consequence of miRNAs generated from intronic sequences and that could silence the expression of their host genes in an autoregulatory loop (Cabrera and Lucas, submitted).

Regarding dementia animal models, Partner 4 has described how tau alteration (levels and/or 4R/3R ratio) is sufficient for the appearance of Tau Nuclear Rods (TNRs), a new histopathological hallmark previously described by the group in HD, in the mouse model of FTDP-17 called P301S (Fernández-Nogales et al, submitted).

Industrial engagements and new collaborations (Partner four).

We have now started a pilot study in Alzheimer disease patients using newly developed biosensors to identify blood purines as biomarkers for different disease stages. Participants: Partner 1, Partner 4 and Sarissa ltd, Coventry, UK (http://www.sarissa-biomedical.com/).

Biotalentum, Hungary: this company has generated human-induced pluripotent stem cell (hiPSC) lines from healthy control individuals as well as from patients suffering from Alzheimer’s disease. It is a partner in a recently submitted grant application to the MSCA-ITN-ETN European Training Networks (coordinated by RCSI).

During 2015, a second call for Pathfinder projects was resolved in December. The approved project involving CIBERNED are described below:

2015 CIBERNED Annual Report / 262 • 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.

2015 CIBERNED Annual Report / 263 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 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.

International Congress for Research and Innovation in Neurodegenerative Diseases (CIIIEN)

During 21st to 23rd September 2015, it was held in Malaga the Third International Congress on Research and Innovation in Neurodegenerative Diseases (CIIIEN), promoted by the Queen Sofia Foundation in collaboration with CIEN Foundation and CIBERNED. The main objective of CIIIEN is providing a forum in which to share progress and information of interest on neurodegenerative diseases among the scientific community.

CIIIEN was created in 2013 and consolidates the merger of the two major scientific conferences on neurodegenerative diseases in general and Alzheimer’s disease in particular, organized in Spain: the XI International Symposium Advances in Alzheimer’s Disease, promoted annually by the Queen Sofia Foundation and CIEN Foundation, and the 9th CIBERNED Scientific Forum, which brought together every year more than the 58 research groups constituting the CIBER in Neurodegenerative Diseases.

Unifying both congresses is a first step in creating a new operating structure in the two main institutions devoted to research on neurological and neurodegenerative diseases in Spain: CIEN Foundation and CIBERNED, both under the Ministry of Economy and Competitiveness through the Institute of Health Carlos III. This new structure seeks greater effectiveness and efficiency in research, promoting the interaction of the different research groups.

This third edition of CIIIEN was once again chaired by Her Majesty Queen Sofia and the scientific program consisted of three plenary sessions and five scientific sessions. Among the invited speakers at the conference can be highlighted some international researchers who are leaders in their field of research such as Zaven Khachaturian, President of the US 2020 Alzheimer’s disease Prevention Campaign; Martin Rossor, Clinical neurologist from the British National Health System, an expert in prevention of dementia, especially Alzheimer’s disease; or David Rubinsztein, Deputy Director of the Medical Research Council in Cambridge, a scientist pioneer in proposing autophagy positive feedback (a process that plays a role in the elimination of toxic proteins abnormally accumulated in the brain of patients) as a potential therapy in neurodegenerative diseases. Rui Costa, a scientist from the Champalimaud Foundation (Portugal) and Angel Cedazo-Minguez, from the Karolinska Institute (Sweden), completed the international presence, focused on the study of the connectivity between different brain areas and the effects of its dysfunction, the first, and pathological mechanisms associated with known risk factors, the second.

The remaining scientific sessions addressed various aspects of frontier research in neurodegenerative diseases. In addition, less common neurodegenerative diseases such as Charcot-Marie-Tooth were also discussed during the congress, as were topics such as neuroimaging techniques, research in glial cells, neuronal plasticity in Huntington’s disease or neuronal signaling in Parkinson’s disease. Moreover, in line with the interest to boost the training of CIBERNED young researchers, the Young Researcher (basic and clinical research) Prizes were awarded during the Congress to Marta Fernández-Nogales and Juan Fortea, respectively, who made presented the studies which have been granted such recognition.

2015 CIBERNED Annual Report / 264 Ultimately, this event establishes in its third edition as a meeting point for the world’s leading experts in neurodegenerative diseases, enabling sharing of knowledge, working methods, new advances and discoveries in a field in which international cooperation between different institutions is becoming increasingly important to obtain optimum results in research.

2015 CIBERNED Annual Report / 265 Scientific productivity and other activities Scientific productivity and other activities

Analysis of CIBERNED scientific productivity in 2015

To complete this section we provide a table of bibliometric indicators for 2014 and 2015. A quick analysis shows a strengthening of the improved scientific production in 2015 (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 (percentage of publications in quartiles 1 and 2, number of publications in high impact journals, average impact factor, international collaborations).

• Table 1. Control panel. It includes publications indexed in PubMed and Scopus.

Indicator 2014 2015 Change (%)

(A) Total number of publications* 571 541 -5,25

(B) Number of publications in quartiles (Q) 1 y 2 477 457 -4,19

(E) Number of publications in 1st decile 183 165 -9,84

(F) Number of publications (1st decile) as percentage of total (A) 32,05 30,50 -4,84

(G) Number of publications in Q1 (includes 1st decile) 337 361 7,12

(H) Number of publications in Q1 as percentage of total (A) 59,02 66,73 13,06

(I) Number of publications in Q1 as percentage of total Q1 and 70,65 78,99 11,81 Q2 (B)

(J) Number of publications in Q2 140 96 -31,43

(K) Number of publications in Q2 as percentage of total (A) 24,52 17,74 -27,63

(L) Number of publications (Q1+Q2) led by CIBERNED groups (1st 248 246 -0,81 author or corresponding author)

(M) Number of publications in Q1 and Q2 led by CIBERNED groups 51,99 53,83 3,53 as percentage of total (B)

(N) Number of publications (Q1 and Q2) not led by CIBERNED 229 211 -7,86 groups (1st author or corresponding author)

(O) Number of publications in Q1 and Q2 not led by CIBERNED 48,01 46,17 -3,83 groups as percentage of total (B)

(P) Number of publications in Q1 and Q2 in which “CIBERNED” 74,42 82,49 10,84 appears in the affiliation (as percentage of total (B))

(Q) Number of publications in Q1 and Q2 led by CIBERNED groups 83,06 89,43 7,66 in which “CIBERNED” appears in the affiliation (as percentage of total (L))

(R) Number of publications in Q1 and Q2 not led by CIBERNED 68,12 74,41 9,23 groups in which “CIBERNED” appears in the affiliation (as percentage of total (N))

(S) Number of publications in Q1 and Q2 with international 197 208 5,58 groups

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

2015 CIBERNED Annual Report / 269 Indicator 2014 2015 Change (%)

(T) Number of publications in Q1 and Q2 with international groups 41,30 45,51 10,20 (S) as percentage of total number of publications (B)

(U) Number of publications in Q1 and Q2 with international groups, 83 77 -7,23 and led by CIBERNED groups

(V) Number of publications in Q1 and Q2 with international groups, 42,13 37,02 -12,14 and led by CIBERNED groups as percentage of all international publications (S)

(W) Number of publications in Q1 and Q2 with other CIBERS 27 24 -11,11

(X) Percentage of publications in Q1 and Q2 with other CIBERS led 55,56 41,67 -25,00 by CIBERNED groups

(Y) Number Percentage of published in Q1 y Q2 1 0 -100,00

(Z) Percentage of Percentage of led by CIBERNED groups 50 50 0,00

(A2) Addition of impact factors of journals (Q1 y Q2) in which 2.787,901 2.797,949 0,36 articles have been published

(B2) Average of impact factors of journals (Q1 y Q2) in which 5,440 5,632 3,53 articles have been published

(C2) Number of publications in journals with impact factor >15 20 20 0,00

Number of publications in Q1 and Q2 with 2 CIBERNED groups 58 40 -31,03

Number of publications in Q1 and Q2 with 3 CIBERNED groups 16 16 0,00

Number of publications in Q1 and Q2 with 4 CIBERNED groups 3 5 66,67

Number of publications in Q1 and Q2 with 5 CIBERNED groups 0 3 300,00

Number of publications in Q1 and Q2 with 6 CIBERNED groups 0 1 100,00

Cooperative projects – Program 1 0 1 100,00

Cooperative projects – Program 2 0 0 0,00

Cooperative projects – Program 3 0 1 100,00

Program 1 cooperative projects 73 70 -4,11

Program 2 cooperative projects 109 156 43,12

Program 3 cooperative projects 39 34 -12,82

Projects with other CIBERs/RETICs/CIEN networks 66 75 13,64

Basic-type projects 162 152 -6,17

Translational-type projects 126 99 -21,43

(A) Total number of publications* 105 103 -1,90

(B) Number of publications in quartiles (Q) 1 y 2 26 39 50,00

2015 CIBERNED Annual Report / 270 Below are some figures summarizing some of the main bibliometric indicators for CIBERNED scientific output during 2015 and its evolution since 2011:

• Figure 1. Percentage of indexed publications CIBERNED 2015

D1 stands for 1st decile.

30,5%

69.5%

Rest D1

• Figure 2. Percentage of indexed publications CIBERNED 2015 distributed by quartiles

QI: publications in the first quartile. Q4: publications in the last quartile. Q2: publications in the second quartile. No IF: publications for which no impact factor Q3: publications in the third quartile. is available.

3,3% 4.6% 7,6%

17,7% 66,7%

Q1 Q2 Q3 Q4 No IF

• Figure 3. Total annual output CIBERNED 2011-2015 (all publications types)

620

603 600 588

580

560 553

540 535 525 520

500

480 2011 2012 2013 2014 2015

2015 CIBERNED Annual Report / 271 • Figure 4. CIBERNED 2011-2015 production by quartiles/1st decile

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

800

700

603 588 600 553 525 535 477 500 467 457 442 422

400

300

183 200 152 156 165 139

100 2011 2012 2013 2014 2015

Number of publications Q1+Q2 D1

• Figure 5. Percentage of publications of excellence (1st decile) CIBERNED 2011-2015

35%

32,0%

29.9% 30% 30.1% 30,5%

25%

24,0%

20% 2011 2012 2013 2014 2015

2015 CIBERNED Annual Report / 272 • Figure 6. Number of CIBERNED 2011-2015 publications within 1st and 2nd quartiles and 1st decile

400 361 351 350 329 337 314 300

250

200 183 165 152 156 150 139 140 116 108 113 96 100

0 2011 2012 2013 2014 2015

D1 Q1 Q2

• Figure 7. Evolution of the impact factor from 2011 to 2015 IF stands for Impact factor.

3500 6 5,626 5,321 5,365 5,44 5,142 3000 2914,551 2820,33 2869,231 5

2575,568 2555,722 2500 4

2000

1500

2 1000

1 500

0 0 2011 2012 2013 2014 2015

Cumulative IF Mean IF

2015 CIBERNED Annual Report / 273 • Figure 8. CIBERNED 2011-2015 publications in high impact journals

2012 10 2011 525 publications 603 publications 17 38 32

2013 6 535 publications

2014 588 publications

8 44

2015 553 publications 17 34

IF > 30

IF 20-30

IF 10-30

2015 CIBERNED Annual Report / 274 • Figure 9. Scientific journals with 5 or more CIBERNED publications in 2015

Movement Disorders 19

PLoS ONE 17

Parkinsonism and Related Disorders 15

Journal of Alzheimer's Disease 14

Frontiers in Neuroanatomy 10

Human Molecular Genetics 10

Molecular Neurobiology 10

Neurobiology of Aging 10

Frontiers in Cellular Neuroscience 9

Journal of Neuroscience 8

Neurologia 8

The Lancet Neurology 8

Journal of Neurology 7

Scientific Reports 7

Journal of Neurochemistry 6

Journal of the Neurological Sciences 6

Annals of Neurology 5

Brain 5

Neurobiology of Disease 5

0510 15 20

2015 CIBERNED Annual Report / 275 • Figure 10. Most frequent thematic categories (WoS-JCR) in CIBERNED 2015 publications

Neurosciences 202 Clinical Neurology 123

Biochemistry Molecular Biology 60

Pharmacology Pharmacy 32

Geriatrics Gerontology 30

Cell Biology 29

Multidisciplinary Sciences 25

Psychiatry 23 Genetics Heredity 18

Endocrinology Metabolism 15

Anatomy Morphology 13

Medicine Research Experimental 12

Pathology 11

Medicine General Internal 8 Chemistry Medicinal 8

Oncology 7 Immunology 7

Gerontology 7

050100 150200 250

2015 CIBERNED Annual Report / 276 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 (Head: Dr. Alberto Rábano).

• Navarra Neurological Tissue Bank (Head: Dr. María Teresa Tuñón).

• 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. José Luis Cantero Llorente)

Events and other activities

CIBERNED has organized or participated in a number of events during the year; particularly noteworthy are the VIII Annual Scientific Forum (held in Málaga on September 21st-23rd) and the Social Forum with the State Reference Center for the Care of people with Alzheimer’s disease (CREA, from its Spanish acronym), and other dementias, as well as other scientific, outreach and international cooperation events.

• IX CIBERNED Scientific Forum

As every year, CIBERNED held in 2015 its IX Scientific Forum. This annual event, which is essential for the proper functioning of CIBERNED, allowed CIBERNED principal investigators, scientists and collaborators meeting to discuss the their research findings, presenting new data, and disseminate the most relevant advances in research on neurodegenerative diseases throughout the last year.

Like the previous year, between the 21stand 23rd of September 2015 on World Alzheimer’s Day that is celebrated every year on September 21, was held in Málaga the Third International Congress for Research and Innovation in Neurodegenerative Diseases (CIIIEN). This Conference joint together the CIBERNED IX Scientific Forum and the 11th annual International Symposium of the CIEN Foundation in collaboration with the Queen Sofia Foundation. A more detailed description of this event is available at the International Relations section of this report.

• III International Symposium on Advances in Alzheimer’s disease social health research

During past June 8 took place the Third International Symposium “Advances in geriatric research in Alzheimer’s disease” at the headquarters of the General Foundation of the University of Salamanca. The University of Salamanca General Foundation, Queen Sofia Foundation, IMSERSO and the State Reference Center on Alzheimer’s promoted the meeting, which benefited from the Presidency of H.M. Queen Sofia.

2015 CIBERNED Annual Report / 277 The Third International Symposium brought together some of the most relevant experts and researchers from the national and international scene with the aim of creating a space for critical thought and discussion. During the meeting, the dissemination of results of recent psychological research was promoted, as well as deepening on ethical and economic aspects related to care and psychosocial research in the context of dementias.

Dr Felix Bermejo, CIBERNED researcher, participated in the Symposium by delivering a lecture on “Geriatric research in Alzheimer’s disease: a promising future”, within the Session IV on Perspectives on Alzheimer’s disease and socio health research.

• Excellence in Neurodegeneration Seminar Series

During 2015 CIBERNED has continued to carry out the series of lectures “Excellence in Neurodegeneration Seminar Series”, offering seminars 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 2015 within this series:

• Speaker: X. William Yang, Brain Research Institute. University of California. Title: Mouse Genetic Dissection of Huntington’s Disease Pathogenesis in vivo.

• Speaker: Jean-Charles Lambert, Institut Pasteur. Title: Genetics of Alzheimer’s dementia. What’s next?

• Speaker: Zaven Kachaturian, Campaign to Prevent Alzheimer’s Disease 2020, Potomac, Maryland. Title: Guidelines for novel conceptual models of dementia: essential features of an “ideal theory” on dementia.

• Speaker: Martin Rossor, NIHR Queen Square Dementia BRU, UCL, London. Title: Responding to the dementia challenge – Is there a window of opportunity with early intervention?

• Speaker: David Rubinsztein, Cambridge Institute for Medical Research. Title: Autophagy and other therapeutic strategies for neurodegenerative diseases.

• Speaker: Rui Costa, Champalimaud Center for the Unknown, Lisboa. Title: Generating and shaping novel actions in health and disease?

• Speaker: Ángel Cedazo Mínguez, Center for Alzheimer Research, Karolinska Institutet, Stockholm. Title: Towards a mechanistic understanding of risk factors for Alzheimer’s disease.

• Official visit to the Champalimaud Foundation in Lisbon

On February 28, 2015, H.M. Queen Sofia visited the Champalimaud Foundation in Lisbon, a Portuguese institution devoted to advanced biomedical research. Accompanied by José Luis Nogueira, Secretary of the Queen Sofia Foundation and Maria Angeles Perez, CIBERNED Manager, the purpose of the visit is to seek new avenues of collaborative research in neurodegeneration.

Received on arrival by Leonor Beleza, chairman of Champalimaud Foundation, Queen Sofia was accompanied on Her visit to the Foundation facilities (internationally renowned for its work on oncology and neuroscience) by the Scientific Director and Nobel Prize Zvi Fuks , Professors Carlo Greco, Rui Costa and Carlos Cordon-Cardo, as well as the Foundation’s Managing Director, Joao Silveira.

2015 CIBERNED Annual Report / 278 On a day in which a series of meetings of scientific content reviewed various research projects, mainly on neursocience, were held, the participating Institutions (Queen Sofia Foundation, CIEN Foundation/CIBERNED, Champalimaud Foundation) focused on identifying joint collaboration opportunities and research to foster the knowledge base in both countries in a field still as unknown as neuroscience.

On behalf of CIBERNED its Scientific Director, Dr. Jesús Ávila de Grado, and his Deputy Scientific Director, Dr. Miguel Medina Padilla, attended the meeting. Dr. Avila intervened by reviewing the scientific activity that the Institution carries out as part of the Alzheimer Project of theQueen Sofia Foundation, which focused mainly on the knowledge of neurodegenerative diseases (with particular emphasis on Alzheimer’s disease), and converges with the priorities and knowledge of the Champalimaud Foundation.

• Dementia Genetics Spanish Consortium (DEGESCO)

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 born in 2013 with the general aim of promoting and strengthen the development of genetic studies in order to understand the genetic architecture of neurodegenerative dementias in the Spanish population through the implementation of collaborative projects and actions among its members. DEGESCO philosophy is inclusive, that is, it is open to the incorporation of new research groups, whether public or private as long as they can prove their capacity and interest to conduct research in genetics of dementia. The stable and long-term scientific relationship between the parties has been established in a Memorandum of Understanding signed by all partners during 2014.

DEGESCO consists of a structure for coordinating sample repositories that share information and processing protocols. Both its scientific, technical quality and ethical compliance with the Biomedical Research Act (Law 14/2007) and Royal Decree on Biobanks (RD 1716/2011) must characterize samples selected by DEGESCO participants. The transfer of samples between consortium members will be in accordance with the requirements of Royal Decree on Biobanks.

During 2015, CIBERNED has continued to participate very actively in the activities of the working group on Genetics of Alzheimer’s disease and dementias (DEGESCO), to continue the Alzheimer’s disease and related dementias in Spain multicenter study, as well as the Spanish representation in major international consortia.

Since its inception in 2013, the consortium has published the following scientific articles:

• Dols-Icardo O, Iborra O, Valdivia J, Pastor P, Ruiz A, de Munain AL et al. Assessing the role of TUBA4A gene in fronto temporal degeneration. Neurobiol Aging. 2016 Feb; 38:215.e13-4.

• Pastor P, Moreno F, Clarimón J, Ruiz A, Combarros O, Calero M et al. MAPT H1 haplotype is associated with late-onset Alzheimer’s disease risk in APOEε4 noncarriers: results from the dementia genetics spanish consortium. J Alzheimers Dis. 2015; 49(2):343-52.

• Thelen M, Razquin C, Hernández I, Gorostidi A, Sánchez-Valle R, Ortega-Cubero S et al. Investigation of the role of rare TREM2 variants in frontotemporal dementia subtypes. Neurobiol Aging. 2014 Nov; 35(11):2657.e13-9.

• Ruiz A, Dols-Icardo O, Bullido MJ, Pastor P, Rodríguez-Rodríguez E, López de Munain A et al. Assessing the role of the TREM2 p.R47H variant as a risk factor for Alzheimer’s disease and frontotemporal dementia. Neurobiol Aging. 2014 Feb; 35(2):444.e1-4.

2015 CIBERNED Annual Report / 279 • Young Investigator Awards

To promote research excellence, CIBERNED awards a prize annually to an emerging scientific involved in clinical or basic research and, on a biennial basis, a prize for an emerging scientist involved in clinical research in neurodegenerative diseases.

In the case of the Young Investigator Award of the Year, candidates must be under 35 years of age, members of a CIBERNED research group and lead author of an article published in a indexed scientific journal during 2015. Regarding the Young Clinical Investigator Award ofthe Year, candidates must be under 40 years, members of a CIBERNED research group and lead author of an article published in a indexed scientific journal during 2014 or 2015.

The call remit was announced by email to all CIBERNED Principal Investigators and through the Center’s website. Once the deadline for applications has passed, seven candidates fulfilling the specifications of both calls in a timely manner were received. For the selection of both Awards, the jury consisted of members of CIBERNED Steering Committee and PIs outside the Committee, which evaluated the scientific quality and impact of the publication as well as the candidate’s personal contribution.

Upon completion of the evaluation process of the applications received, the CIBERNED Scientific Steering agreed to award the 2014 Young Investigator Award to Marta Fernández Nogales for her work “Huntington’s disease is a four-repeat tauopathy with tau nuclear rods”, published in Nature Medicine. 20(8):881-5 (2014).

The Young Clinical Investigator Award of the Year was awarded to Juan Fortea Ormaechea for his work “Cerebrospinal fluid β-amyloid and phospho-tau biomarker interactions affecting brain structure in preclinical Alzheimer disease” published in Annals of Neurolog. 76(2):223-30 (2014).

Other events

• International Symposium 1st School of Biomedicine IBIS-UIMP: cell and molecular biology of neuronal communication in health and disease

The Menéndez Pelayo International University (UIMP) and the Institute of Biomedicine of Seville (IBIS), sponsored among others by CIBERNED, held on December 16-17 the First joint “School of Biomedicine” on “Cell and molecular biology of neuronal communication in health and disease”.

Rafael Fernandez Chacon, principal investigator of CIBERNED, headed the seminar, which focused on issues such as synaptic transmission, neuronal plasticity, neurogenesis and neurodegeneration, and neuropsychiatric disorders, among others. The aim of the meeting was to present the latest developments in these fields of research, and foster dialogue between basic and translational researchers as well as between young scientists and students.

• University of Extremadura (UMEX) Summer course

As part of the XVI edition of the International Summer School of the University of Extremadura, the course “Advances in neurobiology and neurodegenerative diseases” took place in Caceres during July 8-10, 2015. The course was directed by Jose Manuel Fuentes Rodríguez, CIBERNED researcher and member of the Department of Biochemistry and Molecular Biology and Genetics, Faculty of Nursing and Occupational Therapy at the University of Extremadura; and Miguel Medina, CIBERNED Deputy Scientific Director.

2015 CIBERNED Annual Report / 280 The course was aimed at students and Health Sciences professionals and divided into three sections: common aspects of neurodegenerative diseases (neuronal death, autophagy in the central nervous system, oxidative stress, etc.); Alzheimer’s disease; and Parkinson’s disease. The course objective was to disseminate the latest advances in the Nervous System at the molecular and cellular level (with special emphasis on what refers to neurodegenerative disorders), to integrate this knowledge into our understanding, prevention or treatment of neurodegenerative diseases.

2015 CIBERNED Annual Report / 281 Financial report Financial report

Funding

The main source of funding for CIBERNED activities and overall aims comes from the Carlos III Institute of Health, which as promoter and major funding agency, contributes about 71% of the institution.

CIBERNED was created by virtue of the call for stable cooperative research structures dated March 30, 2006 (BOE April 7, 2006). Said call in its paragraph 13 states that the funds granted to the Centers for Biomedical Networked Research (CIBER) may reach a maximum of 80 percent of the budget of the consortium activities.

On March 4, 2015 the Institute of Health Carlos III signs an Order to grant financial support to the CIBER in the subject area of Neurodegenerative Diseases. Such Order sets out in its annex that the Carlos III Institute of Health will transfer the nominative grants reflected there, totaling three million nine hundred twenty three thousand two hundred and forty euros (3.923.240,00 €) for current expenditure, plus one hundred and sixty-three thousand three hundred and eighty euros (163.380,00 €) for capital expenditure.

The Order of 30 March 2006 from the Carlos III Institute of Health, which calls for grants to finance stable structures of cooperative research in Biomedicine and Health Sciences in the framework of the Ingenio 2010 Initiative, Consolider program, CIBER Actions (BOE No. 83, Friday April 7, 2006) provides in Section 13.1 (conditions of the grant):

The support granted to the Centers for Biomedical Research Network (CIBER) may reach a maximum of 80 percent of consortium activities budget. Once created the Center for Biomedical Research Network, your budget will be determined through a specific agreement of funding and goals conducted with the Carlos III Institute of Health.

In determining CIBER’s total budget, is considered among others as ineligible amount, and therefore to be allocated within the non-funded 20%, the expenditure to fund the salaries of the research groups staff who are part of the CIBER and have working relationship with the institution or public center signatory of the agreement.

In CIBERNED Governing Board meeting held in December 2013 the procedure for establishing and calculating the supporting modules system for the contribution of associated institutions was approved.

The contribution of the ISCIII in 2015 totaled to 4.086.620,00 €. The certifications issued following the modules system by the Associated Institutions for CIBERNED reached the figure of 1.021.655,00 €, amount that covers the required 20% co-funding by the Associated Institutions.

2015 CIBERNED Annual Report / 285 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 2015 financial statements were prepared by the Managing Director of the Institution and will be submitted to approval by the Governing Council.

Accounting principles

We have applied all mandatory accounting principles that have a significant impact on these accounts.

Identification of programs

• Financial resources

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 costs Total

Administration, management and coordination 84.798,20€ 244.315,26€ 329.113,46€

Program 1 “Alzheimer’s disease and other degenerative 802.257,00€ 138.959,18€ 941.216,18€ dementias”

Program 2 “Parkinson’s disease and other degenerative 1.163.368,26€ 230.962,69€ 1.394.330,95€ motor disorders””

Provision extra payment / temporary compensations 73.780,56€ - 73.780,56€

Deputy Scientific Director / Transversal plataforms 164.145,67€ 18.887,52€ 183.033,19€

Training / Young Investigator Award / Mobility - 12.909,13€ 12.909,13€

Media service, internationalization, and transfer of - 70.910,45€ 70.910,45€ knowledge

Cooperative projects, 3rd call 561.309,25€ 101.585,00€ 662.894,25€

Cooperative projects, 4th call 77.412,56€ 65.949,60€ 143.362,16€

Cooperative projects, 5th call 13.846,68€ 332,00€ 14.178,68€

2015 CIBERNED Annual Report / 286 Implementation Personnel costs Total

Competitive projects

RTC-2014-1877-1 16.367,05€ 13.245,75€ 29.612,80€

RTC-2015-3289-1 4.617,03€ - 4.617,03€

PI12/00192 - 35.132,68€ 35.132,68€

FJCI-2014-19673 5.100,28€ - 5.100,28€

PIE14/00061 469,74€ - 469,74€

BBVA Research project 10.905,03€ 0,00 10.905,03€

MJFF-JFR - 23.654,17€ 23.654,17€

PI11/03028 - 3.805,96€ 3.805,96€

JPND-COURAGE 30.243,25€ 20.772,71€ 51.015,96€

MS08/0088 10.797,37€ - 10.797,37€

COEN Projects – Pathfinder call 149.393,75€ 144.609,65€ 294.003,40€

E-Rare/ SIRD 14.219,34€ - 14.219,34€

Other projects

Salud 2000 Foundation - 29,40€ 29,40€

Biogen Evaluating the potential for Nrf2 - 588,36€ 588,36€

GW PHARMA 32.182,03€ 7.618,28€ 39.800,31€

SYLENTIS-CIBERNED 9.746,28€ - 9.746,28€

NOSCIRA (Dendria-CENIT / transgenic mice) 37.946,24€ - 37.946,24€

José Castillejo Fellowship - 19.034,00€ 19.034,00€

Associated institutions agreements - - 65.105,21€

Forum - - 76.130,13€

Depreciation of fixed assets - - 127.754,87€

Contributions associated institutions - - 1.021.655,00€

3.262.905,57€ 1.153.301,79€ 5.706.852,57€

The data in the table above show that CIBERNED allocates 2,335,547.13 € to implementing research programs, which represents 49.85% of the ordinary operating expenses (excluding taxes and without taking into account the contribution of associated institutions). Considering the in- kind contribution made by the associated institutions comprising CIBERNED, that percentage goes up to 40.93%.

The amount allocated in 2015 to cover the costs of the internal calls for cooperative projects adds to 820,435.09€, representing 14.38%-17.51% of the overall budget (depending on whether or not the contribution of the associated institutions is considered). The 3rd call allocated budget is significantly higher than the previous two calls because these projects were completed in December 2015. Projects from the 5th call started in October 2015, which explains its low level of budget execution to date.

2015 CIBERNED Annual Report / 287 The sum of both amounts represents the allocation of more than 67% of expenditure directly related to basic, clinical and population-based research, staying in amounts and percentages similar to previous years, without taking into account in these data the contribution of Associated Institutions.

Spending on staff devoted to stable research structures (groups and collaborative projects) represents 57.45% of total spending (not taking into account the assessing of the contribution of the Associated Institutions). Resources for the Administration Department remain one more year similar to previous years ratios, representing 7% of total recurrent expenditure, excluding the value of the contribution of the Associated Institutions.

Depreciation of current assets is set at 2.23% of spending to total recurrent expenditure, due to the ending of the service life of equipment acquired in previous years. This significant and pronounced decrease in recent years can be explained by the acquisition policy carried out by the Institution, investing significantly in the early years to proceed later on to maintenance, renovation and acquisition of complements for the equipment already purchased.

2015 CIBERNED Annual Report / 288 • 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 Autonomous University of Madrid 5 4 102 Fariñas Gómez, Isabel University of Valencia 16 2 103 Fuentes Rodríguez, José University of Extremadura 9 2 Manuel 104 González Castaño, José Autonomous University of Madrid 2 0 105 López Barneo, José University of Sevilla 19 3 106 Ceña Callejo, Valentín University of Castilla-La Mancha 1 3 108 Vicario Abejón, Carlos CSIC 5 3 109 Vila Bover, Miquel Fundació Instituto de Recerca del Hospital 9 2 Universitari Vall d’Hebrón 110 Pérez Castillo, Ana María CSIC 4 2 111 Iglesias Vacas, Teresa CSIC 1 3 113 García Verdugo, José Manuel University of Valencia 5 3 114 del Río Fernández, José Catalonian Institute de Bioengineering 0 1 Antonio 201 Canela Campos,Enric Isidre University of Barcelona 12 3 202 Kulisevsky Bojarski, Jaime Fundació Institut de Recerca del Hospital 9 1 de la Sant Creu i Sant Pau 203 Lanciego Pérez, José Luis Center of Applied Medical Research 5 3 (CIME) 204 Moratalla Villalba, Rosario CSIC 6 2 205 Obeso Inchausti, José Ángel Hospitales de Madrid Foundation 6 2 206 Rodríguez Díaz, Manuel University of La Laguna 2 2 207 Tolosa Sarró, Eduardo Hospital Clinic of Barcelona 11 5 208 Labandeira García, José Luis University of Santiago de Compostela 12 2 209 Pérez Tur, Jordi CSIC 4 1 301 Alberch Vié, Jordi University of Barcelona 31 3 303 Fernández Ruiz, Javier Complutense University of Madrid 12 6 304 Mena Gómez, María Ángeles Ramón y Cajal Hospital, Madrid 2 0 305 Guzmán Pastor, Manuel Complutense University of Madrid 10 4 306 Lucas Lozano, José Javier CSIC 3 4 307 Naranjo Orovio, José Ramón CSIC 3 3 401 Ávila de Grado, Jesús CSIC 9 6 402 Camins Espuny, Antonio University of Barcelona 8 1 403 de Felipe Oroquieta, Javier CSIC 21 3

2015 CIBERNED Annual Report / 289 Group Principal investigator Institution Associated Staff

404 Matute Almau, Carlos University of the Basque Country 17 4 406 Rodríguez Álvarez, José Autonomous University of Barcelona 9 3 407 Sáez Valero, Javier Miguel Hernández University, Elche 7 2 408 Soriano García, Eduardo University of Barcelona 11 2 409 Torres Alemán, Ignacio CSIC 4 4 410 Trullás Oliva, Ramón CSIC 7 3 411 Vitorica Ferrández, University of Sevilla 4 1 Francisco Javier 412 Wandosell Jurado, Francisco CSIC 4 1 413 Comella Carnicé, Joan Xavier Fundació Institut de Recerca del Hospital 12 4 Universitari Vall d’Hebrón 415 Gutíerrez Pérez, Antonia University of Málaga 8 3 502 Bermejo Pareja, Félix Doce de Octubre University Hospital, Madrid 5 2 503 Ferrer Abizanda, Isidro Bellvitge Institute of Biomedical Research, 19 4 Barcelona 504 Lleó Bisa, Alberto Fundació Institut de Recerca del Hospital 15 4 de la Sant Creu i Sant Pau 506 López de Ceballos Lafarga, CSIC 3 1 María 507 Pastor Muñoz, María Center of Applied Medical Research 7 0 Asunción (CIMA) 508 Mengod Los Arcos, CSIC 3 2 Guadalupe 509 de Pedro Cuesta, Jesús Institute of Health Carlos III, Madrid 12 2 510 Bullido Gómez-Heras, María Autonomous University of Madrid 3 2 Jesús 511 Cantero Lorente, José Luis University Pablo de Olavide 8 0 601 Combarros Pascual, Onofre Marqués de Valdecilla Foundation 21 4 604 Muñoz Cánoves, Pura Pompeu Fabra, University 12 4 606 Fernández Chacón, Rafael University of Sevilla 14 4 607 Navarro Acebes, Xavier Autonomous University of Barcelona 25 4 609 López de Munain Arregui, Biodonostia Research Institute 28 4 Adolfo Oficina técnica 0 9 TOTALS 497 152

2015 CIBERNED Annual Report / 290 Attracting and retaining talent is crucial for the future of CIBERNED. In this sense, CIBERNED makes a major effort to incorporate young researchers in the early stages of their career as well as researchers with proven research record of accomplishment. The current socio-economic situation and the need to increase competitiveness make us boost the motivation of our staff and their commitment to our research model. In this strategy, talent management is essential to the leadership of our research as we support and encourage professional and personal growth.

CIBERNED encourages collaboration of its researchers with research groups with international leadership, capable of attracting public and private resource, transferring knowledge and patents to the industrial sector and the clinical practice. It promotes the interplay between basic, clinical and public health researchers. Further supports translational research and innovation. It fosters collaboration between research centers of the Consortium member Institutions and other national and international research institutions.

CIBERNED follows a strict selection process based on professional merit and personal skills to pick the researchers who make up the various research teams. This model of attracting the best talent has formed high-level teams whose talent bring innovation and experience in scientific research and that follow the Aristotlean quote “Intelligence consists not only in the knowledge, but also in the skill to apply the knowledge into practice”.

CIBERNED is equipped with a modern and efficient structure and is aware that acting in a highly competitive field such as research, requires adding value to society through their technical and financial capabilities. Thanks to the daily efforts of our researchers to achieve excellence at all levels of research CIBERNED has positioned itself as a center of international reference in the field of neurodegenerative diseases research.

CIBERNED effectively and efficiently manages and supports research with quality criteria and innovative instruments, offering a professional service and personalized attention oriented towards user satisfaction, working and strengthening the goal of promoting research, innovation and transferring their results at the service of society.

Calls for personnel

CIBERNED has established a systematic procedure approved by the Board of Directors and the Governing Council in order to cover job vacancies. This procedure is in agreement with section 6.2 of ISO 9001:2008. All job announcements are posted on the website of the consortium Institutions and in compliance with the obligations acquired with the various centers, members of CIBERNED Governing Council, and managers of the Institutions are informed of the call contents.

During 2008 it has been implemented through CIBERNED’s intranet an integrated management system which provides Principal Investigators with a quick and easy way of managing calls for personnel. This system 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.

CIBERNED, in keeping with the policy followed in previous years, continues to guide its selective processes in order to get highly qualified staff, whose levels of technical and behavioral skills are suited to the job profiles sought. Knowing well what our strengths and opportunities of the environment are has allowed the incorporation of highly qualified personnel, whose levels of technical and behavioral skills fit the job profiles sought. All CIBERNED selection processes are conducted under criteria of capacity, merit and publicity. During the year 2015, we have posted 5 calls for employment, with 32 positions offered. There have been a total of 694 applicants.

2015 CIBERNED Annual Report / 291 Call Number of Available Number of Applicants/ year calls positions applicants position

2014 5 32 694 21,69 2015 8 47 1.331 28,32

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 47 positions filled-in during 2015, seven people had a PhD degree, twenty four were graduates, and sixteen were advanced technicians.

Category 2014 2015

Doctor (PhD) 11 7 Graduate (BSc, MSc) 15 24 Technician 0 0 Advanced technician 6 16 32 47

CIBERNED welcomes its new workers

The integration of new researchers in CIBERNED is as important as the selection of candidates. Therefore, all workers beginning their employment at CIBERNED are provided with a “Welcome Handbook”. 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.

CIBERNED policy is to ensure all workers health and welfare in all activities undertaken at the workplace. Therefore provides its employees a favorable work environment to health in accordance with the highest standards of reliability, hygiene and safety. From Human Resources ensures the safety of each worker and compliance with the Health and Safety at the workplace protocols.

CIBERNED does not allow any discrimination and works to ensure equal opportunities. This equality of conditions translates into greater opportunities for career development, access to professional training and fair wages. Therefore, the Human Resources Department follows an active policy to avoid any kind of discrimination based on sex and to promote equal opportunities. One example is that wages do not differ between men and women but according to the responsibilities and work of each. These policies are included in “Welcome handbook” and try to introduce a comprehensive concept of equality in the scope of labor activity, covering and aiming at preventing discrimination in all procedures of the organization, such as selection, salary, promotion and training.

2015 CIBERNED Annual Report / 292 • Jobs posted 2014 – 2015

25 24

16 15 15

11 10 7 6 5

0 PhD Graduate Advanced technician 2014 2015

Distribution of research groups personnel during 2015

• CIBERNED personnel by type of contract 2015

During 2015 the distribution of personnel by type of contract was 39 hired permanently and 113 part-time.

Permanent

Part time

113

2015 CIBERNED Annual Report / 293 • CIBERNED personnel 2015

The distribution of personnel according to their degree was 47 Doctors (PhD), 57 graduates, 4 undergraduates, 38 advanced technicians, and 6 administrative assistants.

6 4

38 Administrative

47 Undergraduate PhD

Graduate

Technician 57

• CIBERNED personnel by gender 2015

CIBERNED workforce during 2015 consisted of 113 women and 39 men.

Male

113 Female

• CIBERNED personnel by region 2015

The geographic distribution of personnel of the research teams by Regions was as follows:

Basque Country 8

Canary Islands 2

Galicia 2

Extremadura 2

Valencia 8

Navarra 3

Madrid 63

Catalonia 46

Castile-La Mancha 3

Cantabria 4

Andalusia 11

010203040506070

2015 CIBERNED Annual Report / 294 Training program

In order to promote and develop programs for scientific research training, CIBERNED has promoted or participated during 2015 in a wide variety of training activities including seminars, lectures, courses and conferences.

• From 9th to 13th of March, the fifth edition of the Multidisciplinary Training Course in Neurodegenerative Dementias organized by CIEN Foundation, CIBERNED and UNED was held. This prestigious course has been conducted one more year, for its comprehensive approach to the problem of dementias.

• As part of the Seminar Series “Excellence in Neurodegeneration” organized by the Network Center for Biomedical Research on Neurodegenerative Diseases (CIBERNED), on May 28 took place in Barcelona the conference “Genetics of Alzheimer’s dementia. What’s next?” by Dr. Jean-Charles Lambert, of the Pasteur Institute in Lille (France).

• From June 29 to July 1, 2015 was held in Seville, the XVI Congress of the Spanish Society for Cell Biology (ESCB) under a conference program that covers all possible areas of this discipline. CIBERNED sponsored the session dedicated to Cellular Neurobiology.

• The Workshop “Mitochondrial DNA and neurodegeneration” brought together between September 23-25 an international group of speakers who have made key scientific contributions in the last three years in various aspects of mitochondrial physiology. Jointly organized by EMBO and CIBERNED, the workshop was aimed at students, scientists and professionals in neurodegeneration and molecular research.

• In addition, CIBERNED has participated in the Annual Congress of the Society for Neuroscience in Chicago within the Satellite Symposium of the European Neuroscience Institutes Network (ENINET).

• On the other hand, during 2015 we have promoted continuing education programs for professional and personal development of employees. Among the courses conducted by our Center are the following:

• Animal experimentation category B. • Transfer of knowledge and valuation of biomedical projects. • Statistics. • Advances in neurobiology and valuation of biomedical projects. • Administrative functions in health centers. • Leadership and teamwork organization.

March Fifth edition of the Multidisciplinary XVI Congress of the June 29 Octobre Annual Congress of the 9-13 Training Course in Spanish Society for July03 30 17-21 Society for Neuroscience 01 Neurodegenerative Dementias Cell Biology 05

Conference: “Genetics of May - Workshop Mitochondrial DNA Alzheimer's dementia. Septembre 0228 23-2504 and neurodegeneration What's next?”

2015 CIBERNED Annual Report / 295 Prevention of occupational risks

During 2015, CIBERNED has carried out a series of preventive activities to comply with the legal requirements on safety and occupational health.

Below is the location of groups for which the risk assessment report has been carried out, together with the planning of the corresponding preventive action and information forms of the evaluated work posts:

Province Workplace of groups assessed

Madrid National Center of Microbiology, ISCIII Madrid CIEN Foundaction Madrid Technical University of Madrid Madrid HM CINAC Málaga University of Málaga Sevilla Institute of Biomedicine of Sevill Guipúzcoa Biodonostia Institute Valencia University of Valencia La Coruña University of Santiago de Compostela

As for preventive training relating to Article 19 of the Law on Prevention of Occupational Risks, three classroom occupational hazards courses in research laboratories activities were given, attended by 42 workers.

Regarding health surveillance, 70 specific medical examinations were performed. The following table lists the different types of medical examinations of personnel depending on the risks inherent in the work, according to Article 22 of the Law on Occupational Health and Safety:

Type of medical assessments Number of workers

RM Data Display Screen 11 RML Biological Agents -Data Display Screen -Awkward Postures- 50 Chemical Reagents RM Biological Agents- Screens- Awkward Postures- Ionizing 8 radiations.- Chemicals RML Biological Agents 1

The vaccination campaigns carried out by CIBERNED are specified beolow:

Type of vaccine Serologies Doses

Hepatitis B vaccination campaign 3 20

2015 CIBERNED Annual Report / 296 Quality

During 2015, the recertification of the Quality Management System according to ISO 9001:2008 from the Management department has successfully passed.

Quality Objectives are established annually in order to achieve continuous improvement in procedures and achieve higher levels of customer satisfaction, both external and internal.

The Quality Management System is based on procedures. To do so, operation procedures are constantly reviewed and updated, which is a tool that allows continuous improvement to meet the customers’ requirements, applicable laws and regulations as well as optimizing resources.

The tools used to carry out the review of the Quality Management System are:

• Audit reports, internal and external. • Supplier Evaluation. • Customer complaints, suggestions and information. • Results of customer satisfaction studies. • Evaluation of corrective and preventive actions. • Procedures quality indicators. • Quality objectives. • Internal or external amendments with influence on the Quality System.

Quality policy aims to ensure and optimize procedures related to: orientation to external and internal customers; leadership; staff involvement; procedures-based approach; continuous improvement.

2015 CIBERNED Annual Report / 297 Collaboration agreements

The Network Center for Biomedical Research in Neurodegenerative Diseases (CIBERNED) has not granted during the year 2015 any Agreements aimed at indirectly managing any public service.

In 2015 the following agreements have been kept in force by the institution:

Partner Institutions Description

State Attorney Office - Ministry of Justice Legal assistance

CIEN Fundación - CSIC - CRG Confidenciality agreement related to patent P201130033 “Compuestos para el tratamiento de enfermedades neurodegenerativas”

Teofilo Hernando Institute (UAM) Framework collaboration agreement to promote and strengthen research, teaching, study, and dissemination activities as well as holding events of scientific, academic and cultural interest

CSIC Co-ownership agreement related to patent application “ADN mitocondrial como marcador para el diagnóstico de enfermedades neurodegenerativas”

FIBIO Ramón y Cajal Hospital Patent co-ownership agreement “Medio condicionado de glía como agente neuroprotector”

Miguel Hernández University, Elche Patent co-ownership agreement “Método para determinar la enfermedad de Alzheimer mediante la detección de glicoproteínas portadoras del glicoepitodo HNK-1”

CSIC Co-ownership agreement related to patent application P201231654 “Proteína Viral recombinante SGG2/Y/O complejos binarios SGG2-FNS para su uso en crecimiento y/o regeneración axonal”

ACE Foundation Framework collaboration agreement

CSIC-JOHNSON & JOHNSON Patent co-ownership agreement P201231807 “Método de diagnóstico y/o pronóstico de enfermedades neurodegenerativas”

CSIC-BIO RAD LABORATORIES INC Confidenciality agreement related to patent application P201231807 “Método de diagnóstico y/o pronóstico de enfermedades neurodegenerativas”

CSIC - NEURON BIO Confidenciality agreement related to patent application P201231654 “Proteína Viral recombinante SGG2/Y/O complejos binarios SGG2-FNS para su uso en crecimiento y/o regeneración axonal”

CSIC - CIEN Foundation Co-ownership agreement related to patent application EP13382108.2 “Methods for the prognosis and diagnosis of neurodegenerative diseases”

BIOCROSS - CIEN Foundation Confidenciality agreement related to patent application EP13382108.2 “Methods for the prognosis and diagnosis of neurodegenerative diseases”

Madrid Scientific Park Association to Madrid Scientific Park and office space

Institute for Research in Biomedicine (IRB) Agreement for the transfer of 50% ownership of Andor Microscope

2015 CIBERNED Annual Report / 298 Partner Institutions Description

FIBIO 12 de Octubre Hospital Co-ownership agreement related to patent application PCT/ES2013/070693 “Modelo animal de déficit cognitivo, procedimiento de obtención y aplicaciones”

Miguel Hernández Universiy of Elche - Foundation for Co-ownership agreement related to patent the Promotion of Health and Biomedical Research of application P201130290 “Método de diagnóstico y/o Valencia pronóstico de la enfermedad de alzheimer”

Ottawa Hospital Research Institute-Institut Cochin- E-Rare-2 “Stimulating Intrinsic Repair for DMD (SIRD)” Universite Pierre and Marie Curie

Valencia Technical University (IAESTE )-INJUVE Training foreign students

National Evaluation and Foresight Agency (ANEP) CIBERNED 2015 Scientific evaluation of cooperative projects

Complutense University of Madrid Co-ownership agreement related to patent application “Regulación de receptores CB1 de cannabinoides para la generación de neuronas corticoespinales a partir de células troncales embrionarias murinas”

University of Extremadura Curricular and extracurricular student internships

Janssen Cilag-Neuron Bio-FCIEN-CSIC Collaboration agreement for execution of actions of public-private cooperation framed in the call RETOS COLABORACIÓN 2015

GW Pharma Research project to be developed in the Complutense University of Madrid

ANEP Evaluation of cooperative projects call CIBERNED 2015

CIC biomaGUNE Cooperative Project CIBERNED 2014/06 “Home and progression of Parkinson enferemdad: role of glial activation” Agreement for student placement (Tatiana Rodríguez Chinchilla)

Secretary of State for Universities Ministry of Education, Management training and mobility grants within the Culture and Sports National Program for Promotion of Talent and their Employability from the National Plan of Scientific and Technical Research and Innovation 2013-2016 and other calls for mobility of students and university professors

CSIC- Autonomous University of Madrid Co-ownership agreement related to patent application 201431898 “Compuestos modulares del sensor neuronal de calcio Dream y sus usos terapéuticos”

GW Pharma (UCM) Academic Research Agreement

CIBERNED Associated Institutions IICC Collaboration Agreemnt (Renewal)

CIBER interCIBER Agreement

2015 CIBERNED Annual Report / 299 Acquisitions of fixed assets

The acquisition of assets made during the year are as follows:

Purchase order Equipment description Date Total cost Responsible

5042141659 Rack Inox 5 drawers 19/06/15 2.739,63 € Gómez Isla, Teresa

1142141658 Microenvironmental camera 03/06/15 4.773,45 € Del Río, José Antonio K-FRAME + Magnatic adapter for 4 PL. PETRI 35MM

1142141659 Temperature controller, touch 04/06/15 9.610,43 € Del Río, José Antonio screen, humidity control and manual mixer

2072171660 HP ELITEDESK 800G1 SFF 02/06/15 1.142,91 € Tolosa Sarro, Eduardo

2072171661 Display HP 18-9” mod. ED190i 02/06/15 1.142,91 € Tolosa Sarro, Eduardo LEG VGA/DVI/DP

6062171662 iPAD mini 3 Wi-Fi + cell 128 GB 12/08/15 709,00 € Fernández Chacón, Silver Rafael

1142141663 M-CU-DT-3 Maerzhaeuser 28/08/15 4.452,80 € Del Río, José Antonio desktop controller for 3-axis engine control for motorized sinkers and motorized focus systems. Communication interfaces: RS232C y USB. Connector for Maerzhaeuser joystick or Ergo control system

5042141664 Rack inox 5 drawers 25 boxes 51 24/07/15 1.095,85 € Gómez Isla, Teresa mm

4012141665 Criterion Blotter Wire Electro 31/07/15 1.809,19 € Ávila de Grado, Jesús

4012141666 Powerpack HC, 100-120/220-240 31/07/15 1.809,19 € Ávila de Grado, Jesús V

1032171667 Desktop computer+ keyboard+ 30/09/15 965,00 € Fuentes Rodríguez, José mouse Manuel

1032171668 Display LED 24” Philips 30/09/15 965,00 € Fuentes Rodríguez, José 246V5LHAB Manuel

6062171669 Equipment black side core i5 16/10/15 628,99 € Fernández Chacón, MM5-15 Rafael

6062171632 Display LG 22EN33S-B 21´5” LED 12/01/15 1.003,80 € Fernández Chacón, Rafael 6062171631 Equipment Differo black side 12/01/15 1.003,80 € Fernández Chacón, I7MA5 Rafael

6062171633 Equipment black side core I5 12/01/15 559,00 € Fernández Chacón, MS12 Rafael

4082171636 Display OC E970SWN 05/02/15 7.834,75 € Soriano García, Eduardo

4082171635 Laptop HP PROBOOK 650 G1 05/02/15 7.834,75 € Soriano García, Eduardo

4082171634 PC desktop HP PAVILLON 500- 05/02/15 7.834,75 € Soriano García, Eduardo 315ns + keyboard and mouse

0012171637 NetAPP FAS2552HA 24*900G, 7 05/02/15 19.450,75 € Dirección MODE 8* 10 Gbe+4*Bbe

2015 CIBERNED Annual Report / 300 Purchase order Equipment description Date Total cost Responsible

2032061639 SnapGene Academic 28/01/15 308,28 € Lanciego Pérez, José Luis

2032141640 Freezer 1,852 NO FROST WHITE 08/02/15 954,69 € Lanciego Pérez, José Luis A+

1032141641 Sorvall ST8R refrigerated 02/02/15 4.785,55 € Fuentes Rodríguez, José MicroCentrifuge Manuel

4012141642 Liebherr G5216 freezer 23/02/15 1.327,37 € Ávila de Grado, Jesús

0012171643 Dockingstation/Hi-speed port 18/02/15 383,57 € Dirección rep II

1032141644 Centrifuge rotor microclick 30 x 11/02/15 931,70 € Fuentes Rodríguez, José 2 ml Manuel

4102171645 Synology disk station DS1515+ 24/02/15 1.792,01 € Trullás Oliva, Ramón server NAS SATA 3 GB/s

4102171646 Haard disk Western Digital 3TB 24/02/15 1.792,01 € Trullás Oliva, Ramón SATA III Purple

4012171647 iMac 27 “ 3.2 qc/GT755M 02/03/15 2.272,38 € Ávila de Grado, Jesús

2072141649 Bucket round 750 ML P/rotor TX- 27/02/15 2.566,41 € Tolosa Sarró, Eduardo 750 Set/4

4012141650 Centrifuge Minispin, INCL. Stand. 20/02/15 1.065,71 € Ávila de Grado, Jesús Rotor EU-PLUGGED

1052141651 Termocycler power cord, T100 26/03/15 10.527,00 € López Barneo, José tube support ring

1052141652 S100 Thermal Cycler with dual 26/03/15 10.527,00 € López Barneo, José 48/48 fas reaction module

5112171653 Laptop ACER ES1-P78B 14/04/15 349,00 € Cantero Lorente, José Luis

1142141654 CMR-STG-MHIX2-1-STD- 02/04/15 7.944,26 € Del Río, José Antonio Motorized Stage

3062141656 Cage set mouse cage and 16/04/15 17.644,64 € Lucas Lozano, José Javier accesories

4012141657 ACMENGP072CN 16/04/15 14.698,48 € Ávila de Grado, Jesús

1032141658 CO2Incubator Stericycle sensor 07/05/15 6.534,01 € Fuentes Rodríguez, José TC Manuel

2015 CIBERNED Annual Report / 301 Principal investigator index Principal investigator index

Principal investigator index

Alberch Vié, Jordi Program 2 Page 109 Ávila de Grado, Jesús Program 1 Page 21 Bermejo Pareja, Félix Program 1 Page 24 Bullido Gómez-Heras, Mª Jesús Program 1 Page 29 Camins Espuny, Antonio Program 1 Page 32 Canela Campos, Enric Isidre Program 2 Page 113 Cantero Lorente, José Luis Program 1 Page 36 Ceña Callejo, Valentín Program 2 Page 118 Combarros Pascual, Onofre Program 2 Page 121 Comella Carnicé, Joan Xavier Program 1 Page 39 Cuadrado Pastor, Antonio Program 2 Page 129 Fariñas Gómez, Isabel Program 2 Page 133 de Felipe Oroquieta, Javier Program 1 Page 43 Fernández Chacón, Rafael Program 2 Page 136 Fernández Ruiz, Javier Program 2 Page 139 Ferrer Abizanda, Isidro Program 1 Page 47 Fuentes Rodríguez, José Manuel Program 2 Page 144 García Verdugo, José Manuel Program 2 Page 148 González Castaño, José Program 2 Page 152 Gutiérrez Pérez, Antonia Program 1 Page 53 Guzmán Pastor, Manuel Program 2 Page 154 Iglesias Vacas, Teresa Program 2 Page 158 Kulisevsky Bojarski, Jaime Program 2 Page 161 Labandeira García, José Luis Program 2 Page 167 Lanciego Pérez, José Luis Program 2 Page 171 Lleó Bisa, Alberto Program 1 Page 56 López Barneo, José Program 2 Page 174 López de Ceballos Lafarga, María Program 1 Page 61 López de Munain Arregui, Adolfo Program 2 Page 179 Lucas Lozano, José Javier Program 2 Page 186 Matute Almau, Carlos Program 1 Page 64 Mena Gómez, Mª Ángeles Program 2 Page 189 Mengod Los Arcos, Guadalupe Program 1 Page 68 Moratalla Villalba, Rosario Program 2 Page 191 Muñoz Cánoves, Pura Program 2 Page 195 Naranjo Orovio, José Ramón Program 2 Page 198 Navarro Acebes, Xavier Program 2 Page 201 Obeso Inchausti, José Ángel Program 2 Page 207 Pastor Muñoz, María Asunción Program 1 Page 71 de Pedro Cuesta, Jesús Program 1 Page 75 Pérez Castillo, Ana María Program 2 Page 211 Pérez Tur, Jordi Program 2 Page 215 del Río Fernández, José Antonio Program 2 Page 217 Rodríguez Álvarez, José Program 1 Page 81 Rodríguez Díaz, Manuel Program 2 Page 220 Sáez Valero, Javier Program 1 Page 84 Soriano García, Eduardo Program 1 Page 87 Tolosa Sarró, Eduardo Program 2 Page 223 Torres Alemán, Ignacio Program 1 Page 90 Trullás Oliva, Ramón Program 1 Page 93 Vicario Abejón, Carlos Program 2 Page 229 Vila Bover, Miquel Program 2 Page 232 Vitorica Ferrández, Fco. Javier Program 1 Page 96 Wandosell Jurado, Francisco Program 1 Page 99

2015 CIBERNED Annual Report / 305

2015 Annual Report

MINISTERIO DE ECONOMÍA Y COMPETITIVIDAD

C/ Valderrebollo, 528031 Madrid Tel.: + 34 913 852 200 MINISTERIO MINISTERIO Fax: +34 913 852 118 DE ECONOMÍA DE ECONOMÍA Y COMPETITIVIDAD www.ciberned.es Y COMPETITIVIDAD