Final Report Summary - NEUROSIGN (Development of a Center of Excellence in Neurosignalling)
Executive Summary:
The goal of NeuroSign FP7-REGPOT 2010-1 project was to create a National Centre of Excellence for the study of Neurosignalling during nervous system function and dysfunction, through capacity development of the scientific potential of the Hellenic Pasteur Institute (HPI). HPI hosts three large Laboratories (Molecular Neurobiology and Immunology; Cellular and Molecular Neurobiology; Molecular Genetics) dealing with research in the study of neuromuscular function and neuroimmune interactions, stem cell biology and differentiation as well as the processes of neurodegeneration. Our studies focus primarily on multiple sclerosis, myasthenia gravis, stroke and neurotrauma and extend to a multitude of neurological disorders ranging through Alzheimer’s and Parkinson’s diseases, amyotrophic lateral sclerosis, neuromyelitis optica, schizophrenia, depression, epilepsy, as well as addiction to smoking. Given the broad range and the complementarity of our research topics, a new Department of Neurobiology was established in HPI. NeuroSign aimed at developing this Department into a national Centre of Excellence for the study of neuronal signalling and the consequences of its interruption during nervous system function and dysfunction. The results of Neurosign met successfully with its objectives for achieving excellence in Neurosignalling and are the following:
1. Recruitment of 10 highly-qualified researchers and 3 technicians with expertise in neuroimaging in whole animal level, calcium imaging, electrophysiology, membrane protein expression and crystallization, inflammation, spinal cord injury and stroke models and stem cell biology and transplantation.
2. Establishment of strong collaborations with 12 front-line European research groups (7 in Germany, 1 in UK and 4 in France) of complementary research interests and accomplishment of two-way secondments of research staff for the mutual transfer of knowledge and expertise. The main gain of know how was on in vivo imaging, electrophysiology, membrane protein purification and crystallization, gene transfer, disease modelling and cell transplantation in mice.
3. Acquisition of state-of-the-art scientific equipment. The main instrumentation purchased was a cutting edge technology multiphoton confocal microscope and two Electrophysiology setups. This instrumentation significantly upgraded our research capacities and synchronized us with major European Centers in allowing for advanced light microscopy studies in vivo whole animal imaging and for electrophysiology recordings of human membrane proteins of pharmacological interest expressed in cell lines or Xenopus oocytes. The multiphoton confocal microscope and the specific electrophysiology set ups are unique in the greater Athens area and invaluable to many scientists in the whole region. Neurosign also supported the renovation of the Imaging Unit in order to meet with the special requirements for the best use of the 2-photon microscope. We also purchased a modern FPLC system for protein purification, a nanophotometer, a multi-crystal gamma counter and an automated agarose gel electrophoresis multiparametric system.
4. Dissemination of knowledge. As a Center of Excellence, we put much effort on disseminating the acquired knowledge to both the scientific and the public of general interests. We organized three workshops dealing with experimental models for neurological diseases, injury and repair, live cell imaging and electrophysiology, and an International Conference dealing with the advances in the treatment of neurodegenerative and neuromuscular disorders, which were attended by 1,000 participants. We published 16 research articles (acknowledging Neurosign) in peer-reviewed journals, and participated in 23 International and 14 National conferences or workshops with oral and poster presentations of our scientific results. We organized 20 lectures in HPI with open access in which Neurosign researchers presented their scientific progress and the knowledge gained by their secondments to collaborators and organized an open-day event for school pupils, students and the public of general interest Dissemination of knowledge was also accomplished by the Neurosign website and press releases.
5. Establishment of professional management. We recruited a professional Manager from abroad, who trained a Scientific and Administrative Manager from HPI so as to further strengthen our management skills and our potential to participate in further funding programs. With help from the Managers we improved our research quality, management strategy, dissemination and promotional activities and infrastructure exploitation.
Project Context and Objectives:
The goal of NeuroSign project was to create a Center of Excellence for the study of Neurosignalling during nervous system function and dysfunction, through capacity development of the neuroscience potential of the Hellenic Pasteur Institute (HPI). Our Neuroscience group consists of three labs (Tzartos, Matsas and Probert labs) and conducts research in topical issues ranging from the study of neuromuscular and neuroimmune interactions to neurodevelopment, stem cell biology and the processes of neurodegeneration. Neuroimaging is a key component underlying these studies. Research projects of our group aim at a) gaining basic understanding of the mechanisms involved in neuronal degeneration, protection and regeneration during the development of a number of neurodegenerative and neuromuscular disorders, and b) using this knowledge to identify new targets and strategies for the diagnosis and treatment of such disorders. NeuroSign aimed at further developing our group to foster its integration in the European area of excellence in neuroscience research.
The objectives of the Neurosign Action Plan were:
a) To acquire cutting-edge instrumentation which will allow advanced light microscopy in the whole animal level, electrophysiology for the functional studies of heterologously expressed membrane proteins, fast performance protein purification, accurate measurement of protein concentration, ligand-binding studies and rapid diagnosis system
b) To recruit highly-qualified researchers and technicians with the relevant experience to implement the new technologies
c) To establish collaborations with pioneer European research groups and to promote two-way secondments of research staff for the exchange of know-how in neuroimaging at the whole animal level, electrophysiology, membrane protein purification and crystallization, stem cell biology and transplantation in human disease models and analysis of neurological functional recovery
d) To disseminate the acquired knowledge through organization of Workshops, Conference, open-day events, website, publications, participation in Conferences, and press releases
e) To establish a professional management mechanism to meet the EC requirements.
The Neurosign Action Plan consisted of six work packages (WP 1-6). Listed below are the objectives of Neurosign for each WP.
WP1: Management. Objectives:
1. To set up an effective management for NeuroSign with help from a recruited research Manager, who will ensure the progress of the project towards its planned objectives.
2. To act as the interface between the NeuroSign researchers and the European Commission.
3. To ensure that all actions are performed correctly and within the rules and regulations established by the EC, including financial management, and to ensure that the received funds are correctly distributed and accounted for.
4. To ensure that the work plan and tasks are performed on time, within budget and to the highest quality and create an early warning system.
5. To keep each NeuroSign scientist, the External Advisory Board (EAB) and the Commission, fully informed about the project status, scientific issues, the work planning (adjustments) and all other issues which are important and relevant to the members, in order to obtain maximum transparency; to ensure that all researchers are informed on all important and impacting information that can influence the outcome of the project.
6. To follow up the recommendations of the EAB for further improvement and future development.
WP2: Strategic partnerships. Objectives:
1. To collaborate with 12 top quality research groups of related research interests.
2. To support the transfer of know-how between the collaborating research groups through two-way secondments of research staff.
3. To adopt the state-of-the art methodological approaches of the collaborators in our groups.
WP3: Recruitment of experienced researchers. Objectives:
1. To motivate the recruitment of researchers with highly competitive skills. The recruited researchers should have a proven expertise in the fields of advanced imaging, electrophysiology, membrane protein expression, purification and crystallization, immunology, stem cells and inflammation.
2. To select the candidate researchers on the basis of their research background and relevant experience in the new methods and techniques we wish to adopt correlated to the acquired equipment.
3. To foster the full development of the research skills of incoming researchers in the new working environment.
WP4: Acquisition of equipment. Objectives:
1. To acquire and successfully set up the new state-of-the art equipment. More precisely, the main instrumentation to be purchased is a cutting edge technology multiphoton confocal microscope and two Electrophysiology setups. This instrumentation would significantly upgrade our research capacities and synchronize us with major European Centers in allowing for advanced light microscopy studies in vivo whole animal imaging and for electrophysiology recordings of human membrane proteins of pharmacological interest expressed in cell lines or Xenopus oocytes. The multiphoton confocal microscope and the specific electrophysiology set ups are unique in the greater Athens area and invaluable to many scientists in the whole region. Other purchased instrumentation is a fast performance liquid chamatography system for protein purification, a nanophotometer, a multi-crystal gamma counter and an automated agarose gel electrophoresis multiparametric system.
2. To educate users on the capacities of the new equipment and promote the updating of their knowledge regarding the new equipment. Training on the acquired instrumentation will be based mainly upon mutual secondments of research staff between HPI and 12 European collaborators with relevant expertise.
3. To adopt effectively the new technology in the experimental strategies of our research projects.
WP5: Organization of Workshops and final Conference. Objectives:
1. To communicate our acquired technological expertise and research results with other research groups and spread knowledge among neuroscientists through the organization of 3 workshops and 1 final conference. More specifically, the titles of the 3 Workshops and Final Conference are: 1st Workshop: “Animal models of neurodegeneration and behavioral tests for assessment of motor and cognitive function”; 2nd Workshop: “Functional analysis of CNS-infiltrating immune cells during the development of CNS autoimmunity, injury and repair”; 3rd Workshop: “Advanced Imaging and Electrophysiology in the mammalian CNS”; Final Conference: “Advances in the treatment of neurodegenerative and neuromuscular disorders”.
2. To educate and train interested people in the new technologies introduced to our groups during the practical sections of the workshops.
3. To attract and enforce the participation of private sector in the organized workshops and Final Conference.
WP6: Dissemination. Objectives:
1. To establish a website for Neurosign project, in which the project will be described in full detail. The website will advertise all dissemination activities of our group (Forthcoming Neurosign meetings, Workshops, Conferences, open-day events) and will inform about the acquired infrastructure (new equipment, methodologies, etc). It will also make the publications of our group available to all visitors.
2. To secure dissemination of knowledge acquired by our seconded staff, upon their return, to all other members of our Institute. Monthly meetings within HPI are organized for this reason.
3. To communicate our research results to the scientific community by publications of our research results in peer-reviewed journals and participation of our research staff in International Conferences/Workshops.
4. To communicate our research results to social groups of a wide range of interests by press releases and open-day events.
5. To promote important findings of our research projects in a more effective way and gain a wider recognition as a Centre of Excellence.
6. To advertise our service provision (eg in vivo imaging, electrophysiology, diagnosis, protein expression and crystallization, etc), so as to broaden our customer list.
7. To attract funding from the public and private sector.
Project Results:
The main goal of Neurosign project was to enhance the research potential of the Neurosciences Department of the Hellenic Pasteur Institute (HPI) and establish a Centre of Excellence in Neurosignalling. This was accomplished by a) recruitment of highly experienced researchers, b) establishment of an effective Management, c) acquisition of state-of-the-art equipment, d) two-way secondments between our staff members and EU collaborating labs of top quality, e) dissemination of the acquired knowledge by publications in peer-reviewed journals, website, open-day events, participation in international conferences and workshops, press releases, lectures and the organization of three Workshops and a Final Conference.
Neurosign project was a 3.5 year-funded project (September 2010-February 2014), since it received a 6-month extension. It yielded 27 deliverables and reached 10 milestones. The main results for each corresponding work package follow:
Work package 1: Management (WP leader: Dr Socrates Tzartos)
A. Establishment of an effective Management for Neurosign.
The Management was conducted by the Coordinator (Prof. Socrates Tzartos) who with the help of a recruited very experienced Manager from abroad (Dr Nicole Kerlero De Rosbo) and a recruited Scientific (Dr Marios Zouridakis) and Administrative Manager (Ms Vassiliki Staikou) within HPI organized the actions of the six WP leaders in order to ensure the progress of the project towards its planned objectives. The Scientific Managers and the Administrative Manager were recruited from the start month of the Neurosign project and were in close collaboration with each other. Thus, this management scheme ensured that all actions were performed correctly and within the rules and regulations established by the European Commission, including financial management and that the received funds were correctly distributed to the participating three research groups and between the workpackages. In addition, the Neurosign work plans and tasks were successfully coordinated and performed on time, within budget and to the highest quality. Also, the Neurosign Management has kept the External Advisory Board, the Commission and the three participating group leaders (Drs Tzartos, Matsas and Probert) fully informed about the project status (periodic reporting), scientific issues, adjustments to the work plans, remaining budget and all other important issues relevant to the research group and WP leaders.
B. Establishment of the Steering Committee and External Advisory Board.
One of the first tasks of the Management was to establish the Steering Committee of Neurosign, which consisted of the Coordinator (chair), the 6 WP leaders and by the Managers. Its task was to supervise the progress of NeuroSign, to coordinate the overall work within the project, to discuss major problems and delays and find appropriate solutions, and to assist the Coordinator in ensuring that deliverables and milestones were reached. The Steering Committee met every 6 months and minutes of the meetings were kept as records. Also, an External Advisory Board (EAB) was established consisting of four eminent scientists, namely Prof Jean-Pierre Changeux (College de France and Institut Pasteur, Paris), Prof Dame Louise N. Johnson (R.I.P.) (University of Oxford and Diamond Synchrotron, UK), Prof Melitta Schachner (University of Hamburg, Germany and Rutgers University, New Jersey, USA), and Prof Angela Vincent (Honorary Consultant in Immunology, University of Oxford). Additionally, a Representative of National Authority (GSRT), Dr Sissy Kolyva was included in the EAB. All members of the EAB were informed about the progress of the project through reports sent to them before the Annual Meetings of Neurosign.
C. Organization of the kick-off meeting, 1st, 2nd and 3rd Annual Meetings and six 6-monthly meetings of the Steering Committee.
Before the Annual Meetings, extensive reporting to the members of the EAB was sent together with invitation to these meetings. Dr Greg Ambroziewicz, Coordinator of REGPOT programs attended the final meeting, representing the European Commission. A pre-final meeting also took place the day before the Third Annual Meeting (Monday, February 24th, 2014) between Neurosign Coordinator (Dr S. Tzartos), WP leaders (Drs R. Matsas, L. Probert, E. Taoufik, D. Thomaidou), Managers (Drs N. Kerlero De Rosbo and M. Zouridakis) and REGPOT Coordinator (Dr G. Ambroziewicz). This pre-final meeting took place at the Coordinator’s office at HPI. During this meeting, Neurosign Coordinator, WP leaders and Scientific Manager discussed with Dr Ambroziewicz the results of the Neurosign project in full detail. The success indicators were presented to Dr Ambroziewicz and sustainability of the project was discussed extensively. Afterwards he visited the three Neurosign labs (Tzartos, Matsas and Probert) and was guided to the purchased NeuroSign equipment.
D. Sustainability of recruited experienced researchers and exploitation of the state-of-the-art scientific equipment.
Management of Neurosign successfully coordinated the actions of the correspondent WP leader for recruiting 10 experienced researchers and 4 technicians. Most of the recruited Neurosign persons continued their research after Month 36 of the Neurosign project, since they were funded from other research grants (National and European grants) acquired by the three Neurosign group leaders (Drs S. Tzartos, R. Matsas and L. Probert). However, for 6 of the recruited persons (4 post-docs and 2 technicians), a 6-month extension of their contracts with Neurosign was catalytic (approved by the project officer together with a 6-month extension of the project) in order to get enrolled in other contracts (supported by other grants) beginning in March 2014 (exactly after Month 42 of the Neurosign project). Regarding the newly acquired equipment, this is now adopted in each day research activities of the associated research groups and is available to other researchers from other Academic/Research Institutions. Furthermore, the multi-photon microscope, the crystallization robot and the Electrophysiology equipment will be used for offering service to clients from Academic/Research Institutions or from Pharmaceutical Industry in the near future.
The establishment of a Professional Management within our group and HPI will help us implementing more effectively and successfully future EC grants.
WP1 yielded six deliverables (D1.1: Establishment of the Steering Committee and External Advisory Board; D1.2: Neurosign Brochure; D1.3: Report on the kick-off meeting; D1.4: Report on the First Annual Review; D1.5: Report on the Second Annual Review; D1.6: Report on the Third Annual Review) and reached its milestone (MS1: kick-off meeting) within the scheduled time.
Work package 2: Strategic partnerships (WP leader: Dr Rebecca Matsas)
We established collaborations with 12 top-quality EU laboratories and arranged two-way secondments to gain the know-how in a) advanced imaging (calcium and in vivo imaging), b) stem cells biology, c) electrophysiology, d) membrane protein purification and crystallography, e) spinal cord injury and stroke. In the original proposal we had proposed the establishment of collaborations with seven EU labs but during the first period of the project we added other five EU collaborators (in total: 12 collaborators) of top research quality. The list of the Neurosign collaborators and contact details follows:
NeuroSign list of external collaborators
1. Prof. Wolfram Welte (University of Konstanz, DE; http://strucbio.biologie.uni-konstanz.de/strucbio/index.php?article_id=2&clang=0)
2. Dr Pierre-Jean Corringer (Institut Pasteur, Paris, FR; http://www.eni-net.org/organization/members/dr-pierre-jean-corringer/)
3. Dr Pierre Charneau (Institut Pasteur, Paris, FR; http://www.pasteur.fr/ip/easysite/pasteur/fr)
4. Dr Spencer Shorte (Institut Pasteur, Paris, FR; http://www.openmicroscopy.org/site/about/development-teams/spencer)
5. Dr Tom Mrsic-Flogel (UCL, London, UK; https://iris.ucl.ac.uk/iris/browse/profile?upi=TMRSI56)
6. Prof. Nikolaus Plesnila (LMU Munich Medical School, Munich, DE; http://www.amgenscholars.mcn.uni-muenchen.de/faculty/molecular_medicine/plesnila/index.html
7. Prof. Hartmut Wekerle (Manx-Plank Institute, Martinsried, DE; http://www.neuro.mpg.de/wekerle)
8. Prof. Melitta Schachner (University of Hamburg, DE; http://www.zmnh.uni-hamburg.de/)
9. Prof. Piotr Bregestovski (INSERM, Marseille, FR; http://www.inserm.fr/ezantidot/search)
10. Prof. Frank Kirchhoff (University of Saarland in Homburg, DE; http://www.kirchhoff-lab.de/)
11. Prof. Frauke Zipp (University Medical Centre Mainz, Germany http://www.uni-mainz.de/universitaet/681_DEU_HTML.php)
12. Prof. Thomas Brocker (Ludwig Maximilians Universitaet Muenchen, Germany; http://www.sfb914.med.uni-muenchen.de/principal_investigators/principal_investigators/brocker_thomas/index.html)
In practise, there were some deviations regarding the originally proposed time period of secondments and the number of secondments. These alterations were made because: a) the necessary gain of know-how was obtained by shorter than originally proposed secondments of members of our groups to our collaborators, and b) several of the incoming secondments from our collaborators to our groups did not finally take place or were supported by other sources. More specifically, the outgoing secondments to our collaborators were performed as originally proposed but these in most cases lasted the half time period originally estimated, since no more was actually needed. Regarding the incoming secondments, the most important indeed took place. These dealt with secondments of experts from our collaborators’ groups to HPI so as to help us establish the new Electrophysiology Unit and set up the acquired two-photon instrumentation (major acquired equipment). Several of the remaining incoming secondments did not take place (e.g. secondments from Corringer, Charneaue, Shorte and Mrsic-Flogel groups), since our collaborators continued guiding us via e-mail and skype communication. We should mention the fact that this alteration did not affect the expected results from the secondments originally stated since the results have been obtained successfully. The remaining budget of this WP was reallocated to the “Personnel costs” for researchers and technicians after approval from the project officer, as will be discussed under the Management paragraph.
In total, during the Neurosign project, 12 outgoing secondments of ten of Neurosign researchers from HPI to external collaborators took place (see list below). The seconded HPI researchers gained expertise in a) intravital imaging and two-photon microscopy, b) electrophysiology, c) recombinant vector technology and dynamic bioimaging, d) modelling and analyzing spinal cord injury in mice and e) neural-glial signalling.
Outgoing secondments:
HPI researcher: Dr Maria Evangelidou; Host lab: Dr Hartmut Wekerle, Max Planck Institute of Neurobiology, Germany; Duration: 16/03/2011-18/04/2011; Expertise gained: Cell transfer and tracking techniques in mice for the study of neuron-immune cell interactions
HPI researcher: Dr Era Taufik; Host lab: Dr Nikolaus Plesnila, Royal College of Surgeons, Ireland; Duration: 20/07/2011-08/08/2011; Expertise gained: Modelling and analyzing cerebral stroke and spinal cord injury in mice.
HPI researcher: Dr Georgia Kouroupi; Host lab: Dr Spencer Shorte, Institut Pasteur, France; Duration: 10/10/2011-10/01/2012; Expertise gained: Human induced Pluripotent Stem (iPS) cell generation and training in two-photon microscopy
HPI researcher: Dr Dimitra Thomaidou; Host lab: Dr Frank Kirchhoff and Dr Nikolaus Plesnila, Univerity of Munich, Germany; Duration: 26/02/2012-08/03/2012; Expertise gained: Intravital imaging and training in two-photon microscopy.
HPI researcher: Dr Petros Giastas; Host lab: Dr Wolfram Welte, University of Konstanz, Germany; Duration: 01/04/2012-30/04/2012; Expertise gained: Membrane protein purification for structural studies
HPI researcher: Rebecca Matsas, Ph.D.; Host lab: Dr. Pierre Charneau and Dr. Spencer Shorte, Molecular Virology and Dynamic Imaging, Institut Pasteur, Paris, France; Duration: 22/10/2011-30/10/2011; Expertise gained: Induced pluripotent stem cells and imaging
HPI researcher: Florentia Papastefanaki, Ph.D.; Host lab: Prof. Melitta Schachner, Center for Molecular Neurobiology, Hamburg, Germany; Duration: 12/09/2012-12/12/2012; Expertise gained: Spinal cord injury
HPI researcher: Dafni Chroni, MSc; Host lab: Prof. Piotr Bregestovski, Brain Dynamics Institute, Aix-Marseille University, France; Duration: 1-21/11/2012; Expertise gained: Electrophysiology
HPI researcher: Paris Koutsoudaki, Ph.D.; Host lab: Prof. Frank Kirchhoff, University of Saarland, Germany; Duration: 14/03/2013-14/05/2013; Expertise gained: Intravital imaging
HPI researcher: Rebecca Matsas, Ph.D.; Host lab: Dr Mrsic-Flogel in UCL, London, UK; Duration: 19/6/2013-25/6/2013; Expertise gained: Induced pluripotent stem cells and imaging
HPI researcher: Lesley Probert, Ph.D.; Host lab: Prof. David Attwell, UCL Neuroscience, UK; Duration: 15/09/2013-29/09/2013; 03/02/2014-21/02/2014; Expertise gained: 2-photon imaging
The secondments of our researchers to the collaborators’ groups were very beneficial for the seconded researchers themselves and for the other members of Neurosign group, since the knowledge acquired was transferred to the latter through detailed presentations in our monthly meetings (http://www.neurosign.gr/index.php?page=meetings). Moreover, the gain of knowledge accomplished through these secondments was very catalytic for the successful progress of Neurosign members’ research projects and for the most effective exploitation of the new state-of-the-art instrumentation purchased through Neurosign (i.e. two-photon microscope and electrophysiology equipment).
Regarding the incoming secondments from collaborators to HPI, these were mainly scheduled for gaining expertise in multiphoton microscopy, electrophysiology and spinal cord injury. In total, during NeuroSign, 6 incoming secondments of five of our collaborators took place (see Table below) which helped us setting up experimental approaches in a) electrophysiology in cell lines, neurons and Xenopus oocytes, b) intravital imaging and two-photon microscopy and c) cerebral stroke and spinal cord injury models. The duration of these incoming secondments and the expertise transferred are shown below.
Incoming secondments:
Seconded Scientist: Dr Piotr Bregestovski; Host Lab: Dr Socrates Tzartos; Period: 17/03/12-24/03/2012; Expertise transferred: Electrophysiology in cell lines
Seconded Scientist: Dr Igor Jakovcevski; Host Lab: Dr Rebecca Matsas; Period: 30/03/2012-09/04/2012; Expertise transferred: Spinal cord injury
Seconded Scientist: Dr Frank Kirchhoff; Host Lab: Dr Dimitra Thomaidou; Period: 21/11/2012-24/11/2012; 01//10/2013-05/10/2013; Expertise transferred: Intravital imaging and two-photon microscopy
Seconded Scientist: Dr Nikolaus Plesnila; Host Lab: Dr Lesley Probert; Period: 04/12/2012-08/12/2012; Expertise transferred: Cerebral stroke and spinal cord injury
Seconded Scientist: Dr Naoto Kawakami; Host Lab: Dr Lesley Probert; Period: 04/12/2012-07/12/2012; 01/10/2013-04/10/2013; Expertise transferred: Intravital imaging
Seconded Scientist: Dr Piotr Bregestovski; Host Lab: Dr Rebecca Matsas; Period: 01/10/2013-10/10/2013; Expertise transferred: Electrophysiology in neurons
The gain of knowledge accomplished through these secondments was very catalytic for the successful progress of Neurosign members’ research projects and for the most effective exploitation of the new state-of-the-art instrumentation purchased through Neurosign (i.e. two-photon microscope and Electrophysiology setups).
Conclusively, the objectives of this WP were met with full success regarding a) collaboration with top quality research groups of related research interests and b) transfer of know-how mainly from collaborators to our group and subsequent adaptation of the state-of-the art methodological approaches in our groups.
The established collaborations are maintained after completion of Neurosign project and many complementary studies with our Neurosign collaborators are now continuing.
WP2 yielded four deliverables (D2.1: First report on the secondments of HPI staff members to collaborating EU labs and the transfer of know-how; D2.2: First report on the secondments of collaborators to our groups and the transfer of know-how; D2.3: Second report on the secondments of HPI staff members to collaborating EU labs and the transfer of know-how; D2.4: Second report on the secondments of collaborators to our groups and the transfer of know-how) and reached two milestones (MS3: Establishment of strategic partnerships; MS4: Selection of researchers for secondments) within the originally proposed time period.
Work package 3: Recruitment of experienced researchers (WP leader: Dr Lesley Probert)
The purpose of WP3 was to recruit researchers with proven experience in the field of:
• Advanced imaging and functioning of multi-photon confocal microscope
• Advanced imaging and neurophysiology
• Live mouse imaging
• Ca2+ imaging
• Electrophysiology
• Molecular Biology and various protein expression systems
• Expression of soluble and membrane proteins
After defining the qualifications and specialization of the researchers to be recruited in each position, we proceeded to advertise the available positions on the Institute website, in the international scientific press (i.e. Nature jobs), collaborating Institutes (sent e-mails to Research and Academic Institutes) and e-press (FENS web site, Hellenic Pasteur Institute web site, etc). A uniform procedure was followed for the recruitment of candidates which included initial evaluation of their CVs and references and invitation of shortlisted candidates for interview. We applied an equal opportunities policy concerning gender and nationality. As defined in the NeuroSign Annex I, the expertise required and the corresponding researchers recruited are as follows:
• Calcium imaging and signalling in neurons for evaluating the efficacy of neuroprotective strategies. Dr Maria Evangelidou was recruited in Probert’s lab for full time position for 36 person-months. Her expertise is in cell signalling analyses in vitro and in vivo using transgenic mice and experimentally-inducible models for multiple sclerosis.
• Advanced imaging and stem cells. Dr Georgia Kouroupi was recruited in Matsas’ lab for 36 person-months full time position. Her expertise is in neural stem cell culture and differentiation as well as in advanced imaging and stem cells.
• Molecular Biology and various protein expression systems. Dr Paraskevi Zisimopoulou was recruited in Tzartos’ lab for 36 person-months full time position. Her expertise is in molecular biology several protein expression systems: bacteria, yeast, mammalian cells, baculo-, adeno- and retro- virus systems.
• Expression of soluble and membrane proteins. Dr Marios Zouridakis was recruited in Tzartos’ lab for 29 person-months full time position. His expertise is in the expression of both soluble and membrane proteins expressed in prokaryotic and eukaryotic host cells followed by a variety of protein purification methods and biochemical and biophysical approaches for protein characterization. He also spent 7 person-months in acting as the Scientific Manager of the project.
• Soluble and membrane protein crystallization and crystallography. Dr Petros Giastas was recruited in Tzartos’ lab for 36 person-months full time position. His expertise is in crystallization of water-soluble proteins, data collection, and structure solution. His expertise further extends to other scientific areas including protein expression in E. coli and in yeast P. pastoris, protein purification, many chromatographic techniques and protein characterization through biochemical methods.
• Advanced imaging and neurophysiology. Dr Paraskevi Koutsoudaki was recruited in Matsas’ lab for 36 person-months full time position. Her expertise is in the neurophysiology of animal models of neurodegeneration and imaging.
• Live mouse imaging. Dr Vivian Tseveleki was recruited in Probert’s lab for 36 person-months full time position. Her expertise is in T cell biology and the analysis of neuroimmune interactions in the CNS, as well in vivo imaging.
• Expression and purification of recombinant membrane proteins. Dr Nikos Kouvatsos was recruited in Tzartos’ lab for 19 person-months full time position. His expertise is in the expression of soluble and membrane proteins expressed in prokaryotic and eukaryotic host cells and in protein purification methods.
• Molecular Neurobiology and Immunology. Dr Konstantinos Lazaridis was recruited in Tzartos’ lab for 19 person-months full time position. His expertise is in the molecular design of constructs and their subsequent use for protein expression in a variety of systems, ranging from yeast and insect cells to mammalian expression systems.
• In vivo monitoring of stem cells following neurotrauma at the whole animal level. Dr Kanella Prodromidou was recruited in Matsas’ lab for 28 person-months full time position as a researcher with expertise in the study of prion disease in mouse models as well as the function of prion protein in neural stem cells.
• Electrophysiology. We faced major difficulties in attracting an experienced electrophysiologist to establish our Electrophysiology Unit. Despite the extensive advertisement of this post-doc position (Nature jobs, Hellenic Pasteur Institute web site, FENS web site, Scientifica web site, etc), only a few candidates came forward but none proved to be suitable for taking over related responsibility. In order to achieve the establishment and successful use of the Electrophysiology Unit, which is one of the main goals of NeuroSign-REGPOT, we decided after official approval from our project officer to recruit Mrs Dafni Chroni as an Electrophysiology technician in Tzartos’ lab for 24 person-months full time position. Mrs Chroni is a physicist with Masters’ degree in Biophysics and expertise in cell cultures, who together with Dr Marios Zouridakis (Neurosign recruited researcher) and strong collaboration with our collaborator Dr Piotr Bregestovski, established a functional Electrophysiology Unit.
We also recruited three other technicians during Neurosign project:
• Maria Karamita, BSc. Miss Karamita assisted the other personnel in organizational activities, particularly those concerning the installment and up-keep of new equipment. She also assisted in a research project investigating inflammation and demyelination in mice conditionally deficient in a major neuroprotection signaling pathway (Probert lab). Full-time position for 19 person-months.
• Vasiliki Kyrargyri, BSc. Miss Kyrargyri assisted the other personnel in organizational activities, particularly those concerning the introduction of basic imaging protocols. She also assisted in a research project involving the functional imaging of neurons by confocal microscopy and live cell imaging techniques (Probert lab). Full-time position for 19 person-months.
• Maria Belimezi, Ph.D. She was recruited to complement the personnel for dissemination activities. Dr Belimezi prepared and uploaded updated information for the NeuroSign web site (www.neurosign.gr) and relevant scientific info material and assisted in the NeuroSign dissemination and promotional activities (e.g. organization of lectures, open day events, etc) and corresponding reports. Full-time position for 15 person-months.
In total, 10 post-doctoral researchers (6: female, 4: male) and 4 technicians (female) were recruited during Neurosign. For six of these recruited scientists (four post-doctoral fellows, namely Drs M. Evangelidou, P. Giastas, K. Lazaridis and M. Zouridakis and two technicians, namely, Ms D. Chroni and Ms M. Karamita) we extended their recruitment for 6 months (up to month 42), after approval from the project officer (in compliance with the 6-month extension of the Neurosign project).
The recruitment of these highly experienced researchers in our group proved to be very beneficial for the upgrade of our research potential. The recruited staff boosted the research conducted in our groups by introducing novel and cutting-edge methodological approaches to our group. More precisely, the recruited researchers with expertise in advanced imaging, live mouse imaging, Ca2+ imaging, advanced imaging and stem cells, together with the recently purchased and installed multi-photon microscope and their secondments to EU groups of collaborators initiated research projects involving high-tech imaging methodologies. This significantly contributed to the achievement of our main research goal, which dealt with development of a) novel diagnostics for neuromuscular and neurodegenerative diseases using imaging techniques and b) treatment approaches related to these diseases as well as spinal cord injury. The recruited researchers with expertise in molecular biology, soluble and membrane protein expression, purification and crystallization significantly contributed to the crystallization of proteins with a major pharmacological importance (i.e. two subtypes of neuronal acetylcholine receptor). Elucidation of their 3D X-ray structure opens the way for the rational design of drugs towards treatment of neurological diseases or disorders related to them. Also, the establishment of Electrophysiology Unit within our group and its current full functionality complements our structural studies by confirming the functionality of the crystallized proteins/membrane receptors and serves as a platform for screening potent drugs to the expressed proteins.
Apparently, the objectives of WP3 met with full success regarding a) the recruitment of researchers with highly competitive skills to contribute to the establishment of our group as a Centre of Excellence in Neuroimaging, b) the enhancement of the research skills of our seconded researchers to our top quality collaborators’ groups, c) the adaptation of the new methodological research approaches of the recruited researchers to our group and d) the best exploitation of the introduced research skills of the Neurosign recruited researchers.
All of the recruited Neurosign researchers and technicians continued their research after completion of the Neurosign project, since they are now funded by other research grants (National and European grants) acquired by the three Neurosign group leaders (Drs S. Tzartos, R. Matsas and L. Probert). Therefore, sustainability of the Neurosign recruited researchers is ensured for at least the two next years.
WP3 yielded five deliverables (D3.1: Report on the recruitment of researchers; D3.2: First year progress reports from the recruited researchers; D3.3: Second year progress reports from the recruited researchers; D3.4: Third year progress reports from the recruited researchers; D3.5: Interim report on recruitment) and reached its milestone (MS2: Recruitment of researchers) with a short delay due to the unforeseen problem dealing with recruitment of expert electrophysiologist.
Work package 4: Acquisition of equipment (WP leader: Dr Dimitra Thomaidou)
In WP4 the aim was to purchase the following instrumentation:
1. Multiphoton confocal microscope for advanced live cell and tissue imaging
2. Patch-clamp equipment for establishment of the electrophysiology unit
3. Fast protein liquid chromatography for high throughput protein purification, accompanied by a nano-spectrophotometer
4. Upgrade of current crystallization robot
All of the originally proposed instrumentation was already purchased during the first period of Neurosign. Furthermore, we also purchased some extra instrumentation and renovated the Imaging Unit (in order to meet with the special requirements of the multi-photon microscope) after approval from the project officer, always within the budget. The upgrade of our crystallization robot finally did not take place, because we already acquired scientific results with the current set up. The budget for this upgrade was used to contribute to the rest of equipment.
In brief, the acquired instrumentation is as follows:
Multiphoton Confocal Microscope System
Multiphoton confocal microscope for advanced live cell and tissue imaging (Leica, Germany). The system is comprised of a Leica-SP5 II modular laser scanning inverted microscope equipped with multiple visual (VIS) laserlines (450-640nm) and near UV lasers (400-440nm) for single-photon confocal microscopyand a Mai-Tai (Spectra-Physics) infrared (IR) oscillating Ti/Saphire femtosecond laser, allowing Multiphoton Confocal microscopy.
Two complete cell patch-clamp set ups
Electrophysiology equipment: The equipment includes the following devices: Vibration isolation table with faraday cage (Thorlabs, France), Micromanipulators (Narishige, Japan), Microelectrode amplifier, Oscilloscope, Stimulator, Electrode puller (HEKA, Germany), Microelectrode holder and two accompanying fluorescent microscopes (Olympus CXK41 for mammalian cells and IX51 for oocytes, Japan).
Preparative protein liquid chromatography (FPLC) and NanoDrop
ÄKTApurifier 10 FPLC system (GE Healthcare, UK), that allows purification of microgram to milligram quantities of protein, operating at flow rates of 0.001 to 10 ml/min at pressures of 0 to 25 MPa. A Nanodrop 2000 spectrophotometer (Thermo Scientific, USA) for the precise and fast determination of the concentration of the purified proteins effectively complements the FPLC instrument.
Multi-crystal gamma counter
Multi Crystal LB 2111 gamma counter (Berthold Technologies, UK) is a compact instrument for most applications where emitting isotopes are used.
Automated agarose gel electrophoresis multiparametric system
Sebia’s (Sebia, USA) second generation semi-automated agarose gel electrophoresis system with scanner for the diagnosis of MS.
All the purchased equipment are labelled with badges declaring “Equipment purchased through FP7-REGPOT-NEUROSIGN No264083” accompanied by the EU logo.
All the originally proposed equipment, except the upgrade of the crystallization robot, were successfully purchased and installed in HPI by month 24 of the Neurosign project. The upgrade of the crystallization robot was judged to be not essential after the impressive accumulation of protein crystals from our research team using the current set up. Therefore, the saved amount of money was reallocated for the purchase of other equipment.
Apart from the successful acquisition of the state-of-the-art equipment (mostly referring to the multi-photon confocal microscope and the electrophysiology equipment), there had been considerable training of Neurosign researchers and members on the acquired instrumentation, mainly through outgoing and incoming secondments. More specifically, for the multi-photon microscope, two collaborators of ours (Drs F. Kirchhoff and N. Kawakami) were seconded to HPI for short time periods and for the electrophysiology set-up one collaborator of ours (Dr P. Bregestovski) visited us twice. Also, five HPI researchers were seconded to collaborators in order to gain additional expertise in intravital imaging, namely Drs P. Koutsoudaki, D. Thomaidou, R. Matsas, L. Probert and M. Evangelidou and one recruited technician was seconded to one of our collaborators to gain expertise in electrophysiology. The seconded researchers presented the acquired knowledge regarding use of this instrumentation to all other members of Neurosign and HPI through lectures and hands-on demonstrations.
The above purchased instrumentation is now fully functional and has entered effectively in our daily experimental protocols. The state-of-the-art multiphoton confocal microscope is fully exploited by Neurosign researchers yielding scientific results published in high-impact journals. The multiphoton microscope is now used also as a service provider to other Academic/Research Institutions and we plan to use the Electrophysiology equipment for providing service to such Institutions and the Pharmaceutical Industry.
WP4 yielded two deliverables (D4.1: Report on the purchase and installation of the equipment and D4.2: Report on the training of recruited and experienced researchers in the acquired equipment) and reached its milestone (MS8: Purchase of equipment) with a short delay compared to the originally proposed time period, mainly due to the time-consuming actions regarding the purchase of the multi-photon microscope (eg: extensive market search and the international open tender for its purchase).
Work package 5: Organization of workshops and conference (WP leader: Dr Alexandros Lavdas)
In WP5, we had proposed three workshops and one final conference to take place in HPI for disseminating the knowledge acquired through Neurosign to the regional and European scientific community, as well to the public of general interest. In the original proposal we had proposed one Workshop to take place each year of Neurosign and a final conference at the end of the project. The order and titles of these workshops and conference were:
1. 1st Workshop: “Animal models of neurodegeneration and behavioral tests for assessment of motor and cognitive function”
2. 2nd Workshop: “Advanced Imaging and Electrophysiology in the mammalian CNS”
3. 3rd Workshop: “Functional analysis of CNS-infiltrating immune cells during the development of CNS autoimmunity, injury and repair”
4. Final Conference on “Advances in the treatment of neurodegenerative and neuromuscular disorders”
A deviation regarding the originally proposed schedule of Workshops and Conference took place. We changed the order between the 2nd and 3rd Workshop. We had to take this action for a couple of reasons: a) there was been a significant delay in the installation of the Multiphoton Microscope in our Institute. The reasons were two-fold: Firstly, the time-consuming procedures for extensive market search and the international open tender for its purchase, as dictated by the relatively high cost (over 300K €) of this major equipment. Secondly, the renovation of the Imaging Unit space to meet the installation requirements of the new Multiphoton microscope could only be performed after we obtained permission to use part of the NeuroSign budget towards this goal; b) we faced major difficulties in attracting an experienced electrophysiologist to establish our Electrophysiology Unit. This Unit became functional only in month 19 of Neurosign project after the secondment of the electrophysiology expert Professor Piotr Bregestovski (Neurosign external collaborator) to our lab and the recruitment of Mrs Dafni Chroni, as an electrophysiology technician.
The first of the three Neurosign workshops took place at the early start of the second year of the project (11th -14th January 2012) and was entitled “Animal models of neurodegeneration and behavioural tests for assessment of motor and cognitive function”. This was a 4-day workshop, whose aim was to discuss the most appropriate animal models for selected neurodegenerative disorders and neurotrauma, as well as to illustrate simple and more sophisticated tests for assessing motor and cognitive function after cell transplantation or other treatment. Lectures were held in the morning and practical sessions in the afternoon. During the practicals, the students had the chance to observe stereotactic operations on mice, permanent middle cerebral artery occlusion (a model of thrombotic stroke), the rotarod test for balance, behavioral tests involving flies and the Morris water maze cognitive test for mice (video recorded). Twenty students participated in the practicals, mostly undergraduate and early pre-doctoral students (2 from Barcelona and 18 from various Institutions in Greece) while many more students and scientists attended the lectures at the Hellenic Pasteur Institute lecture theatre. In total, this Workshop was attended by 100 participants. Twelve speakers (4 from abroad and 8 National) who have proven expertise in the field of Neurosciences gave the lectures and nine instructors, most of them highly experienced researchers taught in the practicals. Twenty synoptic handouts were printed, and the students were also given USB sticks including full details and ppt presentations. List of lecturers: Drs Pico Caroni, Spiros Efthimiopoulos, Spiros Georgopoulos, Rebecca Matsas, Ada Mitsacos, Nektarios Tavernarakis, Nikolaus Plesnila, Makis Skoulakis, Fotini Stylianopoulou, Rebecca Trueman, Vivian Tseveleki and Luca Turin.
The 2nd Neurosign workshop took place at the early start of the second period of the project (4th-7th December, 2012) and was entitled “Functional analysis of CNS-infiltrating immune cells during the development of CNS autoimmunity, injury and repair”. This was a 4-day workshop involving theoretical and practical parts and took place at the Hellenic Pasteur Institute (HPI). The program of this workshop was prepared as an overview of the state-of-the-art knowledge in the development of neuroimmune pathologies of the central nervous system together with hands-on practicals in current investigative techniques. The aim of this workshop was to promote and encourage awareness in the field and cooperation amongst aspiring and established neuroimmunologists. During the practicals, the students had the chance to participate in and observe experimental processes in the following sessions: EAE: induction and evaluation, neuropathological analysis, isolation andanalysis of spinal cord infiltrates and ex vivo T-cell imaging. The participants were mostly undergraduate and early pre-doctoralstudents from the universities and research institutes across Greece (Crete, Ioannina, Athens, Alexandroupoli, Thessaloniki). The number of students participating in both the theoretical and practical parts of the workshop was 28. The number of participants registered only for the theoretical part was 16. In total, ~100 people attended the theoretical part of the Workshop (since this session was open to the local scientific community). Twelve speakers (8 from abroad and 4 National) who have proven expertise in the field of CNS autoimmunity gave the lectures and six instructors, most of them highly experienced researchers, taught in the practicals. List of lecturers: Dr Christopher Linington, Hans Lassmann, David Baker, Roland Liblau, Edgar Meinl, George Kollias, HartmutWekerle, Maria Evangelidou, Roberto Furlan, Krysztof Selmaj, Antonio Uccelli, Nikolaos Grigoriadis and Vivian Tseveleki.
The 3rd workshop was entitled «Live Cell Imaging and Electrophysiology”. This was a 4-day workshop (1st- 4th October 2013) involving theoretical and practical parts and took place at the Hellenic Pasteur Institute (HPI). The aim was to discuss the cutting edge live-cell imaging technology and electrophysiological approaches to study protein receptors. Lectures were held in the morning and practical sessions in the afternoon. During the practicals, the participants had the chance to observe video recorded experiments and most importantly to perform hands-on experiments including surgical procedures for creating brain and spinal cord windows for intravital Multiphoton imaging and whole-cell and single-channel recordings, using the patch-clamp technique applied on mammalian cells. Twenty students participated in the practicals, mostly undergraduate and early predoctoral students (1 from Russia, 1 from France, 1 from UK, 1 from Cyprus, 1 from Germany and 15 from various Institutions in Greece), while many more students and scientists attended the lectures at the Hellenic Pasteur Institute lecture theatre. They attended the practicals in rotation of 4 groups of 5 people each. In total, ~100 people attended the theoretical part of the Workshop (since this session was open to the local scientific community). Ten speakers (5 from abroad and 5 National) who have proven expertise in the field of live imaging and electrophysiology gave the lectures and six instructors, most of them highly experienced researchers, taught in the practicals. List of speakers: Drs Piotr Bregestovski, Dimitrios Davalos, Naoto Kawakami, Frank Kirchhoff, Rebecca Matsas, Cornelia Poulopoulou, Davide Ragozzino, Dimitra Thomaidou, Irini Skaliora and Pavlos Rigas.
Regarding the Final Conference, we decided to join it with the 26th Meeting of the Hellenic Neuroscience Society in order to have increased visibility. It was entitled “Understanding Brain Function to Treat Dysfunction” and was held on the 29/11-1/12/2013 at the Eugenides Foundation in Athens (http://www.eugenfound.edu.gr). The most recent developments in stem cell research, cellular and gene therapy approaches for the treatment of neurodegenerative diseases and traumas and in structure and function of several ligand-gated ion channels were presented and discussed by 21 invited lecturers, 14 of which from abroad. Three poster sessions were also included, where 154 posters were presented with full attendance and 11 posters were selected for oral presentations. The thematics were diverse and covered a large area of the Neuroscience field (Nervous System Development, Synaptic transmission /Signaling /ion and molecule channels, Stem Cells and Nervous System Regeneration, Neuroendocrinology and Neuroimmunology, Behavior and Cognition, Neurodegenerative Diseases, Nervous System Disorders and Neuropsychopharmacology, Systems and computational Neuroscience , Other). The participation at the Final Conference was very high (total 509 participants) and was secured by the extensive advertisement from i) the website that was created specifically for the Meeting (2013.hsfn.gr) but was linked to the NeuroSign, Pasteur Institute and Hellenic Society for Neuroscience websites, that also had a banner at their front page advertising the event ii) e-mails to all academic bodies and institutions in Greece and all relevant Greek researchers and iii) posters and programs that were distributed to other relevant meetings and academic and research institutions in Greece. A very important feature that also secured this unexpectedly high participation was that students and post-doctoral researchers attended the meeting without any registration fees and this was particularly important at this time period of economic crisis in Greece.
After successful implementation of these Workshops and the Final Conference, our group has gained a good expertise in organizing such scientific events. As a Center of Excellence, we will organize additional Workshops and Conferences in the future, so as to educate people in the newly entered technologies in HPI through Neurosign (i.e.: multiphoton microscopy, Electrophysiology, induced pluripotent stem cells, models for spinal cord injury and stroke).
WP5 yielded three deliverables (D5.1: Report on the 1st and 2nd Workshops; D5.2: Report on the 3rd Workshop; D5.3: Report on the Final Conference) and reached one milestone (MS10: Decision about the organization of the Final Conference) within its originally proposed time schedule.
Work package 6: Dissemination activities (WP leader: Dr Era Taufik)
During Neurosign, we a) organized monthly meetings in HPI with open access where the progress of the recruited researchers and the knowledge gained by secondments were presented, b) participated in 23 International and 14 National conferences or workshops with oral and poster presentations (in total: 37), c) maintained and regularly updated our website (www.neurosign.gr) d) published our research results in sixteen (Neurosign acknowledged) peer-reviewed scientific journals, e) organized an open-day event and f) provided two press releases to the social media. Further details on these dissemination actions are provided in the following section.
In the future, we will continue disseminating the knowledge acquired in our groups by publishing our research results (which have been boosted by our significant R/T upgrade through Neurosign) in top quality journals, by maintaining and updating our website, by participating in Conferences and Workshops, by organizing open-day events and workshops and by providing press releases.
Potential Impact:
Neurosign project had an important impact on the regional socio-economic development.
1. During a severe period of financial crisis in Greece, Neurosign supported the recruitment of 10 post-doctoral fellows (experienced researchers) and 4 technicians. Equally important was the accomplishment of the sustainability of these recruited staff after the completion of the Neurosign project. Due to our significant R&T upgrade through implementation of the project, the three group leaders (Drs S. Tzartos, R. Matsas and L. Probert) secured additional funding for their research projects by National and International and EC grants. In total, the three group leaders were granted 2,790,000 euro from other financial sources { ~900,000 (MDA2013-16; AFM20012-14; Aristeia, Thalis); 210,000 (Aristeia II + IKYDA Greek-German collaboration, both directly linked to CNS Intravital Imaging); ~1,100,000 (NOISEPLUS COOPERATION, ARISTEIA I, BNP PARIBAS, PTR, BODOSSAKI FOUNDATION, EMPEIRIKION FOUNDATION, Thalis, KRHPIS); 580,000 (Synergasia)}. Some of these grants were awarded during the mid-term implementation of the Neurosign project and some others at the end of the project, assuring the continuation of the contracts of the Neurosign recruited staff for at least two years after the end of Neurosign project (and in some cases up to 3 years after the end of the Neurosign project). From the recruited staff during Neurosign (in total 14), 4 are men and 10 are women. Also, 10 PhD students (6 women and 4 men) were indirectly benefited, since they were supported scientifically by the supervision of the experienced staff.
2. The acquisition of state-of-the-art equipment of a total cost of ~650,000 euro contributed to the sustainability of scientific equipment suppliers mainly in Europe. The maintenance of this equipment contributes to the sustainability of the regional representatives of the suppliers.
3. As a Center of Excellence in Neurosignalling and neurological diseases and disorders we put much of our efforts in supporting several related patients’ associations. We have been instrumental in the establishment of the Hellenic MG Association (HMGA; web site: www.myasthenia.gr) whose full-members are exclusively myasthenic patients. We provide: i) space for office and meetings, ii) we bridge them with the relevant clinicians for expert’s advice, and iii) provide them with reliable information on the research progress towards the most effective treatment approaches of MG. In addition, we are collaborating with the MDA-Hellas. Given the expected expansion of our translational research, we intend to contribute to the establishment and/or to the strengthening of other similar societies of patients suffering from chronic neurological disorders relevant to our research interests and translational aims. One such society is ELEPAP-GR (www.elepap.gr) which aims at supporting children with movement disorders resulting from neurological disease and neurotrauma. Apparently, given that most of our research regards the best diagnosis and treatment procedures of autoimmune diseases such as myasthenia gravis, neuromyelitis optica, multiple sclerosis, amyotrophic lateral sclerosis and stem cells therapies for spinal cord injuries and stroke, there is a huge social impact of the Neurosign project.
4. During implementation of the Neurosign project, we increased our collaborations with National hospitals (Alexandra Hospital, Laiko Hospital, Eginitio Hospital, Red Cross Hospital) for the study of several human autoimmune diseases and for developing the best strategies to treat them.
5. Our significant R&T upgrade through Neurosign has led to attracting an increased number of PhD applicants compared to the past. There has been ~70% increase in such applications and we have already accepted many young PhD students to get enrolled with our projects. This also contributes to the regional economical development, since we support the PhD students by the additional grants awarded to us after Neurosign.
6. Finally, we envisage attracting additional funding by Pharmaceutical Industries and SMEs (either local or European), with which we are collaborating in order to commercialize our developed diagnostic kits for several autoimmune diseases and peptide vaccines to treat multiple sclerosis. This will ensure the sustainability of the recruited Neurosign staff for many years, since it may lead to their permanent employment.
Dissemination activities during Neurosign:
As an established Center of Excellence, we are enrolled in several activities ensuring the widespread dissemination of knowledge acquired in our groups. We have taken the following actions through implementation of Neurosign project, and are continuing most of them:
1. We set up the Neurosign website (www.neurosign.gr) which is a well informative and easy-to-use website, whose sessions were described in full detail in the First Periodic Report. The website was updated regularly in order to a) disseminate the acquired knowledge through Neurosign to both the scientific community and to the general public, b) promote important events associated with the program (i.e. forthcoming meetings, workshops, conferences, publications, open day events, etc) and c) advertise services offered by Neurosign group and emerged job openings. The Neurosign website is still active after the end of the project and we intend to continuously update it. There are also hyperlinks to the websites of the Hellenic Pasteur Institute (HPI) and its Neurosign participant laboratories (Tzartos’ lab, Matsas’ lab and Probert’s lab), whose research aims and projects are discussed in detail, as well links to our external collaborators’ research groups. Photos of the state-of-the-art equipment acquired through Neurosign (mainly referring to the multiphoton confocal microscope and the Electrophysiology apparatus) are uploaded to the site with a brief description of its applications. It is declared that this apparatus is available for use for any Academic/Research Institute that might find useful. The most representative publications of Neurosign scientists are also uploaded on the site; we included reviews in this part, so that to be informative as much as possible to scientists of another research field. Our website also contains information on the monthly meetings, annual meetings and workshops of the Neurosign project. In addition, there is a special section for disseminating the knowledge acquired through Neurosign to the National (in Greek language) and International (in English) general public. In this section, the various neurological and neuromuscular disorders under study by the three Neurosign participants are discussed in non-technical terms for the lay audience (pathology, symptoms, diagnosis, progress towards therapy, etc, are discussed). Finally, in all sections of Neurosign website, EC FP7 is acknowledged through the FP7 logo linking to the CORDIS website.
2. NeuroSign Meetings: For disseminating reasons, we have arranged monthly meetings participated by Neurosign members and all other lab members inside the HPI. In each of these meetings, a different Neurosign scientist presents his/her research interests and results, as well as detailed research protocols and techniques used. In these meetings, the seconded Neurosign researchers to the groups of our external collaborators present the knowledge acquired and transfer the hain of know-how to all other Neurosign members. The abstracts of these presentations are uploaded in the corresponding section of the web site.
3. Publications of research papers: During Neurosign, sixteen publications (in which Neurosign was acknowledged) in peer-reviewed scientific journals took place. Six from Tzartos’ group (1. Lazaridis K, Zisimopoulou P, Giastas P, Bitzopoulou K, Evangelakou P, Sideri A, Tzartos SJ. Expression of human AChR extracellular domain mutants with improved characteristics Int J Biol Macromol. 2014 Feb;63:210-7; 2. Lazaridis K, Zisimopoulou P, Lagoumintzis G, Skriapa L, Trakas N, Evangelakou P, Kanelopoulos I, Grapsa E, Poulas K, Tzartos S. Antigen-specific apheresis of autoantibodies in myasthenia gravis Ann N Y Acad Sci. 2012 Dec;1275:7-12; 3. Zisimopoulou P, Brenner T, Trakas N, Tzartos SJ. Serological diagnostics in myasthenia gravis based on novel assays and recently identified antigens Autoimmun Rev. 2013 Jul;12(9):924-30; 4. Tzartos J, Zisimopoulou P, Rentzos M, Karandreas N, Zouvelou V, Evangelakou P, Tsonis A, Thomaidis T, Lauria G, Andreetta F, Mantegazza R, Tzartos S. LRP4 antibodies in serum and CSF from amyotrophic lateral sclerosis patients Annals of Clinical and Translational Neurology, in press; 5. Niarchos A, Zouridakis M, Douris V, Georgostathi A, Kalamida D, Sotiriadis A, Poulas K, Iatrou K, Tzartos SJ Expression of a highly antigenic and native-like folded extracellular domain of the human α1 subunit of muscle nicotinic acetylcholine receptor, suitable for use in antigen specific therapies for myasthenia gravis. PLoS One. 2013 Dec 20;8(12):e84791. doi: 10.1371/journal.pone.0084791; 6. Zouridakis M, Giastas P, Zarkadas E, Tzartos SJ Crystal structures of the free and antagonist-bound states of the extracellular domain of human α9 nAChR Nature Structural and Molecular Biology (under revision), three from Matsas’ group (1. Tsioras K, Papastefanaki F, Politis PK, Matsas R, Gaitanou M. Functional Interactions between BM88/Cend1, Ran-binding protein M and Dyrk1B kinase affect cyclin D1 levels and cell cycle progression/exit in mouse neuroblastoma cells. PLoSOne. 2013 Nov 28;8(11):e82172. doi: 10.1371/journal.pone.0082172; 2. Prodromidou K, Papastefanaki F, Sklaviadis T, Matsas R Functional cross-talk between the cellular prion protein and the neural cell adhesion molecule NCAM is critical for neuronal differentiation of neural stem/precursor cells. Stem Cells. 2014 Feb 4. doi: 10.1002/stem.1663; 3. Miltiadous P, Kouroupi G, Stamatakis A, Koutsoudaki PN, Matsas R, Stylianopoulou F. Subventricular zone-derived neural stem cell grafts protect against hippocampal degeneration and restore cognitive function in the mouse following intrahippocampal kainic acid administration. Stem Cells Transl Med. 2013 Mar;2(3):185-98) and two from Proberts’ group (1. Voulgari-Kokota A, Fairless R, Karamita M, Kyrargyri V, Tseveleki V, Evangelidou M, Delorme B, Charbord P, Diem R, Probert L Mesenchymal stem cells protect CNS neurons against glutamate excitotoxicity by inhibiting glutamate receptor expression and function. Exp Neurol. 2012 Jul; 236(1):161-70. doi: 10.1016 / j.expneurol.2012.04.011; 2. Evangelidou M, Karamita M, Vamvakas SS, Szymkowski DE, Probert L. (2014) Altered expression of oligodendrocyte and neuronal marker genes predicts the clinical onset of autoimmune encephalomyelitis and indicates the effectiveness of multiple sclerosis-directed therapeutics. J Immunol. May 1;192(9):4122-33. doi: 10.4049/jimmunol.1300633)
4. Participation in Conferences/Workshops. During the second period of Neurosign project, Neurosign researchers participated in 23 International Conferences and 14 National Conferences (in total: 37. They presented their research results either in oral or printed presentations (posters) and communicated them with International and National Neuroscientists.
5. Open day events. We organized one open day for school pupils, students, researchers and medical doctors, which apart from disseminating the gathered knowledge on cutting-edge technology experimental approaches through our secondments (e.g. stem cells, advanced live animal imaging, electrophysiology), it also promoted the services available in HPI, after purchase of this major equipment (multiphoton and electrophysiology). During this open day, regional researchers/Academics/students/school pupils had the chance to take a tour to this Neurosign-acquired equipment and be informed about the applications of the instrumentation.
6. Interviews in newspapers/social media. The Coordinator, Prof. Socrates Tzartos, was interviewed by a couple of local media regarding the Neurosign project and its results. The interviews were published at newspapers (http://www.kathimerini.gr/755815/article/epikairothta/episthmh/neo-e8niko-kentro-aristeias-gia-tis-neyroepisthmes) and web newspapers (http://www.goodnews.gr/Articles/I-symmetochi-tis-Elladas-stin-pagkosmia-ereyna_2059.html). The interviews were addressed both to the scientific and general interests audience.
Exploitation of results:
During Neurosign, we yielded important scientific results, mainly due to our significant upgrade through recruitment of very experienced researchers, collaborations with top-quality EC research groups and acquisition of state-of-the-art equipment. Exploitation of these results has already started and is expected to be fully accomplished in the forthcoming couple of years and beyond. More specifically:
1. We patented three of our scientific results (Patent Application numbers: EP11724450; WO2013057599 and WO2009093143) and intend to commercialize them in the near future. More specifically, we are collaborating with a local SME (Tzartos Neurodiagnostics) for the commercialization of novel diagnostic assays for a more sensitive and reliable diagnosis method for the myasthenia gravis, neuromyelitis optica and amyotrophic lateral sclerosis diseases and with two Pharmaceutical Industries for the commercialization of peptide vaccines for the treatment of multiple sclerosis. Especially for the latter, pre-clinical studies completed (submitted for publication), toxicology tests underway (results expected summer 2014); development of patient screening and selection assay is underway.
2. We plan to commercialize immunoadsorption columns for the specific treatment of myasthenia gravis patients. We had patented this specific treatment procedure before the Neurosign project (EP 1509605 B1), but significant progress took place during implementation of the Neurosign project. We are collaborating with a local SME (Tzartos Neurodiagnostics) for this project and plan to collaborate with an emerging spin-off SME within HPI organized by Prof. Socrates Tzartos. Clinical trials are pending, which will take place in the near future in collaboration with a relevant company with experience in the field.
3. We have initiated service provision to other Academic/Research Institutions based on some of the equipment purchased through Neurosign. Therefore, our newly acquired multi-photon microscope is now used by many external users. We are planning to provide service to Academic/Research Institutions and Pharmaceutical Industry using our newly established Electrophysiology setups (applicable to membrane proteins expressed in cell lines and Xenopus oocytes) for the evaluation of the potency and efficacy of drugs under development. Both instrumentation (multiphoton microscope for live animal imaging and Electrophysiology setups) is unique in the Athens area.
4. We plan to collaborate with Pharmaceutical Industries in order to design specific drugs for the treatment of chronic pain and epilepsy, based on the elucidation of the X-ray 3D structure of two subtypes of the human neuronal acetylcholine receptor, which was accomplished during Neurosign.
5. In the frame of the above, we have significantly increased (~300% increase compared to before Neurosign project) our collaborations with several enterprises to get involved in further collaborative actions. The SMEs and Pharmaceutical Industries with which we are now collaborating are Alomone; Tzartos Neurodiagnostics; Syneuro, Moscow; Myomedix, Germany; Euroimmune AG, Germany; VIANEX; Eldrug; Redoxis; Serono; Xencor; FPRT Bio and AbbVie.
6. Improvement of the potential of our participation in FP7, beyond-FP7 and other funding projects. Our improved RTD capacity and capability, the significant upgrade of our Imaging Unit, the establishment of a new Unit of Electrophysiology, the established professional management, the development of collaborations with leading European research groups and the increase of our publications in top quality journals have now led to our increased visibility in the European research area. All above have significantly increased our potential to receive further research grants (e.g. HORIZON 2020) either as the leading partner or member of a large consortium.
List of Websites:
Website of Neurosign project: www.neurosign.gr
Contact details of NeuroSign partners:
1. Prof. Socrates Tzartos, Coordinator. Head of the Lab of Molecular Neurobiology and Immunology, Hellenic Pasteur Institute
2. Dr Rebecca Matsas. Head of the Lab of Cellular and Molecular Neurobiology, Hellenic Pasteur Institute
3. Dr Lesley Probert. Head of the Lab of Molecular Genetics, Hellenic Pasteur Institute
4. Dr Marios Zouridakis, Scientific Project Manager, Hellenic Pasteur Institute
The goal of NeuroSign FP7-REGPOT 2010-1 project was to create a National Centre of Excellence for the study of Neurosignalling during nervous system function and dysfunction, through capacity development of the scientific potential of the Hellenic Pasteur Institute (HPI). HPI hosts three large Laboratories (Molecular Neurobiology and Immunology; Cellular and Molecular Neurobiology; Molecular Genetics) dealing with research in the study of neuromuscular function and neuroimmune interactions, stem cell biology and differentiation as well as the processes of neurodegeneration. Our studies focus primarily on multiple sclerosis, myasthenia gravis, stroke and neurotrauma and extend to a multitude of neurological disorders ranging through Alzheimer’s and Parkinson’s diseases, amyotrophic lateral sclerosis, neuromyelitis optica, schizophrenia, depression, epilepsy, as well as addiction to smoking. Given the broad range and the complementarity of our research topics, a new Department of Neurobiology was established in HPI. NeuroSign aimed at developing this Department into a national Centre of Excellence for the study of neuronal signalling and the consequences of its interruption during nervous system function and dysfunction. The results of Neurosign met successfully with its objectives for achieving excellence in Neurosignalling and are the following:
1. Recruitment of 10 highly-qualified researchers and 3 technicians with expertise in neuroimaging in whole animal level, calcium imaging, electrophysiology, membrane protein expression and crystallization, inflammation, spinal cord injury and stroke models and stem cell biology and transplantation.
2. Establishment of strong collaborations with 12 front-line European research groups (7 in Germany, 1 in UK and 4 in France) of complementary research interests and accomplishment of two-way secondments of research staff for the mutual transfer of knowledge and expertise. The main gain of know how was on in vivo imaging, electrophysiology, membrane protein purification and crystallization, gene transfer, disease modelling and cell transplantation in mice.
3. Acquisition of state-of-the-art scientific equipment. The main instrumentation purchased was a cutting edge technology multiphoton confocal microscope and two Electrophysiology setups. This instrumentation significantly upgraded our research capacities and synchronized us with major European Centers in allowing for advanced light microscopy studies in vivo whole animal imaging and for electrophysiology recordings of human membrane proteins of pharmacological interest expressed in cell lines or Xenopus oocytes. The multiphoton confocal microscope and the specific electrophysiology set ups are unique in the greater Athens area and invaluable to many scientists in the whole region. Neurosign also supported the renovation of the Imaging Unit in order to meet with the special requirements for the best use of the 2-photon microscope. We also purchased a modern FPLC system for protein purification, a nanophotometer, a multi-crystal gamma counter and an automated agarose gel electrophoresis multiparametric system.
4. Dissemination of knowledge. As a Center of Excellence, we put much effort on disseminating the acquired knowledge to both the scientific and the public of general interests. We organized three workshops dealing with experimental models for neurological diseases, injury and repair, live cell imaging and electrophysiology, and an International Conference dealing with the advances in the treatment of neurodegenerative and neuromuscular disorders, which were attended by 1,000 participants. We published 16 research articles (acknowledging Neurosign) in peer-reviewed journals, and participated in 23 International and 14 National conferences or workshops with oral and poster presentations of our scientific results. We organized 20 lectures in HPI with open access in which Neurosign researchers presented their scientific progress and the knowledge gained by their secondments to collaborators and organized an open-day event for school pupils, students and the public of general interest Dissemination of knowledge was also accomplished by the Neurosign website and press releases.
5. Establishment of professional management. We recruited a professional Manager from abroad, who trained a Scientific and Administrative Manager from HPI so as to further strengthen our management skills and our potential to participate in further funding programs. With help from the Managers we improved our research quality, management strategy, dissemination and promotional activities and infrastructure exploitation.
Project Context and Objectives:
The goal of NeuroSign project was to create a Center of Excellence for the study of Neurosignalling during nervous system function and dysfunction, through capacity development of the neuroscience potential of the Hellenic Pasteur Institute (HPI). Our Neuroscience group consists of three labs (Tzartos, Matsas and Probert labs) and conducts research in topical issues ranging from the study of neuromuscular and neuroimmune interactions to neurodevelopment, stem cell biology and the processes of neurodegeneration. Neuroimaging is a key component underlying these studies. Research projects of our group aim at a) gaining basic understanding of the mechanisms involved in neuronal degeneration, protection and regeneration during the development of a number of neurodegenerative and neuromuscular disorders, and b) using this knowledge to identify new targets and strategies for the diagnosis and treatment of such disorders. NeuroSign aimed at further developing our group to foster its integration in the European area of excellence in neuroscience research.
The objectives of the Neurosign Action Plan were:
a) To acquire cutting-edge instrumentation which will allow advanced light microscopy in the whole animal level, electrophysiology for the functional studies of heterologously expressed membrane proteins, fast performance protein purification, accurate measurement of protein concentration, ligand-binding studies and rapid diagnosis system
b) To recruit highly-qualified researchers and technicians with the relevant experience to implement the new technologies
c) To establish collaborations with pioneer European research groups and to promote two-way secondments of research staff for the exchange of know-how in neuroimaging at the whole animal level, electrophysiology, membrane protein purification and crystallization, stem cell biology and transplantation in human disease models and analysis of neurological functional recovery
d) To disseminate the acquired knowledge through organization of Workshops, Conference, open-day events, website, publications, participation in Conferences, and press releases
e) To establish a professional management mechanism to meet the EC requirements.
The Neurosign Action Plan consisted of six work packages (WP 1-6). Listed below are the objectives of Neurosign for each WP.
WP1: Management. Objectives:
1. To set up an effective management for NeuroSign with help from a recruited research Manager, who will ensure the progress of the project towards its planned objectives.
2. To act as the interface between the NeuroSign researchers and the European Commission.
3. To ensure that all actions are performed correctly and within the rules and regulations established by the EC, including financial management, and to ensure that the received funds are correctly distributed and accounted for.
4. To ensure that the work plan and tasks are performed on time, within budget and to the highest quality and create an early warning system.
5. To keep each NeuroSign scientist, the External Advisory Board (EAB) and the Commission, fully informed about the project status, scientific issues, the work planning (adjustments) and all other issues which are important and relevant to the members, in order to obtain maximum transparency; to ensure that all researchers are informed on all important and impacting information that can influence the outcome of the project.
6. To follow up the recommendations of the EAB for further improvement and future development.
WP2: Strategic partnerships. Objectives:
1. To collaborate with 12 top quality research groups of related research interests.
2. To support the transfer of know-how between the collaborating research groups through two-way secondments of research staff.
3. To adopt the state-of-the art methodological approaches of the collaborators in our groups.
WP3: Recruitment of experienced researchers. Objectives:
1. To motivate the recruitment of researchers with highly competitive skills. The recruited researchers should have a proven expertise in the fields of advanced imaging, electrophysiology, membrane protein expression, purification and crystallization, immunology, stem cells and inflammation.
2. To select the candidate researchers on the basis of their research background and relevant experience in the new methods and techniques we wish to adopt correlated to the acquired equipment.
3. To foster the full development of the research skills of incoming researchers in the new working environment.
WP4: Acquisition of equipment. Objectives:
1. To acquire and successfully set up the new state-of-the art equipment. More precisely, the main instrumentation to be purchased is a cutting edge technology multiphoton confocal microscope and two Electrophysiology setups. This instrumentation would significantly upgrade our research capacities and synchronize us with major European Centers in allowing for advanced light microscopy studies in vivo whole animal imaging and for electrophysiology recordings of human membrane proteins of pharmacological interest expressed in cell lines or Xenopus oocytes. The multiphoton confocal microscope and the specific electrophysiology set ups are unique in the greater Athens area and invaluable to many scientists in the whole region. Other purchased instrumentation is a fast performance liquid chamatography system for protein purification, a nanophotometer, a multi-crystal gamma counter and an automated agarose gel electrophoresis multiparametric system.
2. To educate users on the capacities of the new equipment and promote the updating of their knowledge regarding the new equipment. Training on the acquired instrumentation will be based mainly upon mutual secondments of research staff between HPI and 12 European collaborators with relevant expertise.
3. To adopt effectively the new technology in the experimental strategies of our research projects.
WP5: Organization of Workshops and final Conference. Objectives:
1. To communicate our acquired technological expertise and research results with other research groups and spread knowledge among neuroscientists through the organization of 3 workshops and 1 final conference. More specifically, the titles of the 3 Workshops and Final Conference are: 1st Workshop: “Animal models of neurodegeneration and behavioral tests for assessment of motor and cognitive function”; 2nd Workshop: “Functional analysis of CNS-infiltrating immune cells during the development of CNS autoimmunity, injury and repair”; 3rd Workshop: “Advanced Imaging and Electrophysiology in the mammalian CNS”; Final Conference: “Advances in the treatment of neurodegenerative and neuromuscular disorders”.
2. To educate and train interested people in the new technologies introduced to our groups during the practical sections of the workshops.
3. To attract and enforce the participation of private sector in the organized workshops and Final Conference.
WP6: Dissemination. Objectives:
1. To establish a website for Neurosign project, in which the project will be described in full detail. The website will advertise all dissemination activities of our group (Forthcoming Neurosign meetings, Workshops, Conferences, open-day events) and will inform about the acquired infrastructure (new equipment, methodologies, etc). It will also make the publications of our group available to all visitors.
2. To secure dissemination of knowledge acquired by our seconded staff, upon their return, to all other members of our Institute. Monthly meetings within HPI are organized for this reason.
3. To communicate our research results to the scientific community by publications of our research results in peer-reviewed journals and participation of our research staff in International Conferences/Workshops.
4. To communicate our research results to social groups of a wide range of interests by press releases and open-day events.
5. To promote important findings of our research projects in a more effective way and gain a wider recognition as a Centre of Excellence.
6. To advertise our service provision (eg in vivo imaging, electrophysiology, diagnosis, protein expression and crystallization, etc), so as to broaden our customer list.
7. To attract funding from the public and private sector.
Project Results:
The main goal of Neurosign project was to enhance the research potential of the Neurosciences Department of the Hellenic Pasteur Institute (HPI) and establish a Centre of Excellence in Neurosignalling. This was accomplished by a) recruitment of highly experienced researchers, b) establishment of an effective Management, c) acquisition of state-of-the-art equipment, d) two-way secondments between our staff members and EU collaborating labs of top quality, e) dissemination of the acquired knowledge by publications in peer-reviewed journals, website, open-day events, participation in international conferences and workshops, press releases, lectures and the organization of three Workshops and a Final Conference.
Neurosign project was a 3.5 year-funded project (September 2010-February 2014), since it received a 6-month extension. It yielded 27 deliverables and reached 10 milestones. The main results for each corresponding work package follow:
Work package 1: Management (WP leader: Dr Socrates Tzartos)
A. Establishment of an effective Management for Neurosign.
The Management was conducted by the Coordinator (Prof. Socrates Tzartos) who with the help of a recruited very experienced Manager from abroad (Dr Nicole Kerlero De Rosbo) and a recruited Scientific (Dr Marios Zouridakis) and Administrative Manager (Ms Vassiliki Staikou) within HPI organized the actions of the six WP leaders in order to ensure the progress of the project towards its planned objectives. The Scientific Managers and the Administrative Manager were recruited from the start month of the Neurosign project and were in close collaboration with each other. Thus, this management scheme ensured that all actions were performed correctly and within the rules and regulations established by the European Commission, including financial management and that the received funds were correctly distributed to the participating three research groups and between the workpackages. In addition, the Neurosign work plans and tasks were successfully coordinated and performed on time, within budget and to the highest quality. Also, the Neurosign Management has kept the External Advisory Board, the Commission and the three participating group leaders (Drs Tzartos, Matsas and Probert) fully informed about the project status (periodic reporting), scientific issues, adjustments to the work plans, remaining budget and all other important issues relevant to the research group and WP leaders.
B. Establishment of the Steering Committee and External Advisory Board.
One of the first tasks of the Management was to establish the Steering Committee of Neurosign, which consisted of the Coordinator (chair), the 6 WP leaders and by the Managers. Its task was to supervise the progress of NeuroSign, to coordinate the overall work within the project, to discuss major problems and delays and find appropriate solutions, and to assist the Coordinator in ensuring that deliverables and milestones were reached. The Steering Committee met every 6 months and minutes of the meetings were kept as records. Also, an External Advisory Board (EAB) was established consisting of four eminent scientists, namely Prof Jean-Pierre Changeux (College de France and Institut Pasteur, Paris), Prof Dame Louise N. Johnson (R.I.P.) (University of Oxford and Diamond Synchrotron, UK), Prof Melitta Schachner (University of Hamburg, Germany and Rutgers University, New Jersey, USA), and Prof Angela Vincent (Honorary Consultant in Immunology, University of Oxford). Additionally, a Representative of National Authority (GSRT), Dr Sissy Kolyva was included in the EAB. All members of the EAB were informed about the progress of the project through reports sent to them before the Annual Meetings of Neurosign.
C. Organization of the kick-off meeting, 1st, 2nd and 3rd Annual Meetings and six 6-monthly meetings of the Steering Committee.
Before the Annual Meetings, extensive reporting to the members of the EAB was sent together with invitation to these meetings. Dr Greg Ambroziewicz, Coordinator of REGPOT programs attended the final meeting, representing the European Commission. A pre-final meeting also took place the day before the Third Annual Meeting (Monday, February 24th, 2014) between Neurosign Coordinator (Dr S. Tzartos), WP leaders (Drs R. Matsas, L. Probert, E. Taoufik, D. Thomaidou), Managers (Drs N. Kerlero De Rosbo and M. Zouridakis) and REGPOT Coordinator (Dr G. Ambroziewicz). This pre-final meeting took place at the Coordinator’s office at HPI. During this meeting, Neurosign Coordinator, WP leaders and Scientific Manager discussed with Dr Ambroziewicz the results of the Neurosign project in full detail. The success indicators were presented to Dr Ambroziewicz and sustainability of the project was discussed extensively. Afterwards he visited the three Neurosign labs (Tzartos, Matsas and Probert) and was guided to the purchased NeuroSign equipment.
D. Sustainability of recruited experienced researchers and exploitation of the state-of-the-art scientific equipment.
Management of Neurosign successfully coordinated the actions of the correspondent WP leader for recruiting 10 experienced researchers and 4 technicians. Most of the recruited Neurosign persons continued their research after Month 36 of the Neurosign project, since they were funded from other research grants (National and European grants) acquired by the three Neurosign group leaders (Drs S. Tzartos, R. Matsas and L. Probert). However, for 6 of the recruited persons (4 post-docs and 2 technicians), a 6-month extension of their contracts with Neurosign was catalytic (approved by the project officer together with a 6-month extension of the project) in order to get enrolled in other contracts (supported by other grants) beginning in March 2014 (exactly after Month 42 of the Neurosign project). Regarding the newly acquired equipment, this is now adopted in each day research activities of the associated research groups and is available to other researchers from other Academic/Research Institutions. Furthermore, the multi-photon microscope, the crystallization robot and the Electrophysiology equipment will be used for offering service to clients from Academic/Research Institutions or from Pharmaceutical Industry in the near future.
The establishment of a Professional Management within our group and HPI will help us implementing more effectively and successfully future EC grants.
WP1 yielded six deliverables (D1.1: Establishment of the Steering Committee and External Advisory Board; D1.2: Neurosign Brochure; D1.3: Report on the kick-off meeting; D1.4: Report on the First Annual Review; D1.5: Report on the Second Annual Review; D1.6: Report on the Third Annual Review) and reached its milestone (MS1: kick-off meeting) within the scheduled time.
Work package 2: Strategic partnerships (WP leader: Dr Rebecca Matsas)
We established collaborations with 12 top-quality EU laboratories and arranged two-way secondments to gain the know-how in a) advanced imaging (calcium and in vivo imaging), b) stem cells biology, c) electrophysiology, d) membrane protein purification and crystallography, e) spinal cord injury and stroke. In the original proposal we had proposed the establishment of collaborations with seven EU labs but during the first period of the project we added other five EU collaborators (in total: 12 collaborators) of top research quality. The list of the Neurosign collaborators and contact details follows:
NeuroSign list of external collaborators
1. Prof. Wolfram Welte (University of Konstanz, DE; http://strucbio.biologie.uni-konstanz.de/strucbio/index.php?article_id=2&clang=0)
2. Dr Pierre-Jean Corringer (Institut Pasteur, Paris, FR; http://www.eni-net.org/organization/members/dr-pierre-jean-corringer/)
3. Dr Pierre Charneau (Institut Pasteur, Paris, FR; http://www.pasteur.fr/ip/easysite/pasteur/fr)
4. Dr Spencer Shorte (Institut Pasteur, Paris, FR; http://www.openmicroscopy.org/site/about/development-teams/spencer)
5. Dr Tom Mrsic-Flogel (UCL, London, UK; https://iris.ucl.ac.uk/iris/browse/profile?upi=TMRSI56)
6. Prof. Nikolaus Plesnila (LMU Munich Medical School, Munich, DE; http://www.amgenscholars.mcn.uni-muenchen.de/faculty/molecular_medicine/plesnila/index.html
7. Prof. Hartmut Wekerle (Manx-Plank Institute, Martinsried, DE; http://www.neuro.mpg.de/wekerle)
8. Prof. Melitta Schachner (University of Hamburg, DE; http://www.zmnh.uni-hamburg.de/)
9. Prof. Piotr Bregestovski (INSERM, Marseille, FR; http://www.inserm.fr/ezantidot/search)
10. Prof. Frank Kirchhoff (University of Saarland in Homburg, DE; http://www.kirchhoff-lab.de/)
11. Prof. Frauke Zipp (University Medical Centre Mainz, Germany http://www.uni-mainz.de/universitaet/681_DEU_HTML.php)
12. Prof. Thomas Brocker (Ludwig Maximilians Universitaet Muenchen, Germany; http://www.sfb914.med.uni-muenchen.de/principal_investigators/principal_investigators/brocker_thomas/index.html)
In practise, there were some deviations regarding the originally proposed time period of secondments and the number of secondments. These alterations were made because: a) the necessary gain of know-how was obtained by shorter than originally proposed secondments of members of our groups to our collaborators, and b) several of the incoming secondments from our collaborators to our groups did not finally take place or were supported by other sources. More specifically, the outgoing secondments to our collaborators were performed as originally proposed but these in most cases lasted the half time period originally estimated, since no more was actually needed. Regarding the incoming secondments, the most important indeed took place. These dealt with secondments of experts from our collaborators’ groups to HPI so as to help us establish the new Electrophysiology Unit and set up the acquired two-photon instrumentation (major acquired equipment). Several of the remaining incoming secondments did not take place (e.g. secondments from Corringer, Charneaue, Shorte and Mrsic-Flogel groups), since our collaborators continued guiding us via e-mail and skype communication. We should mention the fact that this alteration did not affect the expected results from the secondments originally stated since the results have been obtained successfully. The remaining budget of this WP was reallocated to the “Personnel costs” for researchers and technicians after approval from the project officer, as will be discussed under the Management paragraph.
In total, during the Neurosign project, 12 outgoing secondments of ten of Neurosign researchers from HPI to external collaborators took place (see list below). The seconded HPI researchers gained expertise in a) intravital imaging and two-photon microscopy, b) electrophysiology, c) recombinant vector technology and dynamic bioimaging, d) modelling and analyzing spinal cord injury in mice and e) neural-glial signalling.
Outgoing secondments:
HPI researcher: Dr Maria Evangelidou; Host lab: Dr Hartmut Wekerle, Max Planck Institute of Neurobiology, Germany; Duration: 16/03/2011-18/04/2011; Expertise gained: Cell transfer and tracking techniques in mice for the study of neuron-immune cell interactions
HPI researcher: Dr Era Taufik; Host lab: Dr Nikolaus Plesnila, Royal College of Surgeons, Ireland; Duration: 20/07/2011-08/08/2011; Expertise gained: Modelling and analyzing cerebral stroke and spinal cord injury in mice.
HPI researcher: Dr Georgia Kouroupi; Host lab: Dr Spencer Shorte, Institut Pasteur, France; Duration: 10/10/2011-10/01/2012; Expertise gained: Human induced Pluripotent Stem (iPS) cell generation and training in two-photon microscopy
HPI researcher: Dr Dimitra Thomaidou; Host lab: Dr Frank Kirchhoff and Dr Nikolaus Plesnila, Univerity of Munich, Germany; Duration: 26/02/2012-08/03/2012; Expertise gained: Intravital imaging and training in two-photon microscopy.
HPI researcher: Dr Petros Giastas; Host lab: Dr Wolfram Welte, University of Konstanz, Germany; Duration: 01/04/2012-30/04/2012; Expertise gained: Membrane protein purification for structural studies
HPI researcher: Rebecca Matsas, Ph.D.; Host lab: Dr. Pierre Charneau and Dr. Spencer Shorte, Molecular Virology and Dynamic Imaging, Institut Pasteur, Paris, France; Duration: 22/10/2011-30/10/2011; Expertise gained: Induced pluripotent stem cells and imaging
HPI researcher: Florentia Papastefanaki, Ph.D.; Host lab: Prof. Melitta Schachner, Center for Molecular Neurobiology, Hamburg, Germany; Duration: 12/09/2012-12/12/2012; Expertise gained: Spinal cord injury
HPI researcher: Dafni Chroni, MSc; Host lab: Prof. Piotr Bregestovski, Brain Dynamics Institute, Aix-Marseille University, France; Duration: 1-21/11/2012; Expertise gained: Electrophysiology
HPI researcher: Paris Koutsoudaki, Ph.D.; Host lab: Prof. Frank Kirchhoff, University of Saarland, Germany; Duration: 14/03/2013-14/05/2013; Expertise gained: Intravital imaging
HPI researcher: Rebecca Matsas, Ph.D.; Host lab: Dr Mrsic-Flogel in UCL, London, UK; Duration: 19/6/2013-25/6/2013; Expertise gained: Induced pluripotent stem cells and imaging
HPI researcher: Lesley Probert, Ph.D.; Host lab: Prof. David Attwell, UCL Neuroscience, UK; Duration: 15/09/2013-29/09/2013; 03/02/2014-21/02/2014; Expertise gained: 2-photon imaging
The secondments of our researchers to the collaborators’ groups were very beneficial for the seconded researchers themselves and for the other members of Neurosign group, since the knowledge acquired was transferred to the latter through detailed presentations in our monthly meetings (http://www.neurosign.gr/index.php?page=meetings). Moreover, the gain of knowledge accomplished through these secondments was very catalytic for the successful progress of Neurosign members’ research projects and for the most effective exploitation of the new state-of-the-art instrumentation purchased through Neurosign (i.e. two-photon microscope and electrophysiology equipment).
Regarding the incoming secondments from collaborators to HPI, these were mainly scheduled for gaining expertise in multiphoton microscopy, electrophysiology and spinal cord injury. In total, during NeuroSign, 6 incoming secondments of five of our collaborators took place (see Table below) which helped us setting up experimental approaches in a) electrophysiology in cell lines, neurons and Xenopus oocytes, b) intravital imaging and two-photon microscopy and c) cerebral stroke and spinal cord injury models. The duration of these incoming secondments and the expertise transferred are shown below.
Incoming secondments:
Seconded Scientist: Dr Piotr Bregestovski; Host Lab: Dr Socrates Tzartos; Period: 17/03/12-24/03/2012; Expertise transferred: Electrophysiology in cell lines
Seconded Scientist: Dr Igor Jakovcevski; Host Lab: Dr Rebecca Matsas; Period: 30/03/2012-09/04/2012; Expertise transferred: Spinal cord injury
Seconded Scientist: Dr Frank Kirchhoff; Host Lab: Dr Dimitra Thomaidou; Period: 21/11/2012-24/11/2012; 01//10/2013-05/10/2013; Expertise transferred: Intravital imaging and two-photon microscopy
Seconded Scientist: Dr Nikolaus Plesnila; Host Lab: Dr Lesley Probert; Period: 04/12/2012-08/12/2012; Expertise transferred: Cerebral stroke and spinal cord injury
Seconded Scientist: Dr Naoto Kawakami; Host Lab: Dr Lesley Probert; Period: 04/12/2012-07/12/2012; 01/10/2013-04/10/2013; Expertise transferred: Intravital imaging
Seconded Scientist: Dr Piotr Bregestovski; Host Lab: Dr Rebecca Matsas; Period: 01/10/2013-10/10/2013; Expertise transferred: Electrophysiology in neurons
The gain of knowledge accomplished through these secondments was very catalytic for the successful progress of Neurosign members’ research projects and for the most effective exploitation of the new state-of-the-art instrumentation purchased through Neurosign (i.e. two-photon microscope and Electrophysiology setups).
Conclusively, the objectives of this WP were met with full success regarding a) collaboration with top quality research groups of related research interests and b) transfer of know-how mainly from collaborators to our group and subsequent adaptation of the state-of-the art methodological approaches in our groups.
The established collaborations are maintained after completion of Neurosign project and many complementary studies with our Neurosign collaborators are now continuing.
WP2 yielded four deliverables (D2.1: First report on the secondments of HPI staff members to collaborating EU labs and the transfer of know-how; D2.2: First report on the secondments of collaborators to our groups and the transfer of know-how; D2.3: Second report on the secondments of HPI staff members to collaborating EU labs and the transfer of know-how; D2.4: Second report on the secondments of collaborators to our groups and the transfer of know-how) and reached two milestones (MS3: Establishment of strategic partnerships; MS4: Selection of researchers for secondments) within the originally proposed time period.
Work package 3: Recruitment of experienced researchers (WP leader: Dr Lesley Probert)
The purpose of WP3 was to recruit researchers with proven experience in the field of:
• Advanced imaging and functioning of multi-photon confocal microscope
• Advanced imaging and neurophysiology
• Live mouse imaging
• Ca2+ imaging
• Electrophysiology
• Molecular Biology and various protein expression systems
• Expression of soluble and membrane proteins
After defining the qualifications and specialization of the researchers to be recruited in each position, we proceeded to advertise the available positions on the Institute website, in the international scientific press (i.e. Nature jobs), collaborating Institutes (sent e-mails to Research and Academic Institutes) and e-press (FENS web site, Hellenic Pasteur Institute web site, etc). A uniform procedure was followed for the recruitment of candidates which included initial evaluation of their CVs and references and invitation of shortlisted candidates for interview. We applied an equal opportunities policy concerning gender and nationality. As defined in the NeuroSign Annex I, the expertise required and the corresponding researchers recruited are as follows:
• Calcium imaging and signalling in neurons for evaluating the efficacy of neuroprotective strategies. Dr Maria Evangelidou was recruited in Probert’s lab for full time position for 36 person-months. Her expertise is in cell signalling analyses in vitro and in vivo using transgenic mice and experimentally-inducible models for multiple sclerosis.
• Advanced imaging and stem cells. Dr Georgia Kouroupi was recruited in Matsas’ lab for 36 person-months full time position. Her expertise is in neural stem cell culture and differentiation as well as in advanced imaging and stem cells.
• Molecular Biology and various protein expression systems. Dr Paraskevi Zisimopoulou was recruited in Tzartos’ lab for 36 person-months full time position. Her expertise is in molecular biology several protein expression systems: bacteria, yeast, mammalian cells, baculo-, adeno- and retro- virus systems.
• Expression of soluble and membrane proteins. Dr Marios Zouridakis was recruited in Tzartos’ lab for 29 person-months full time position. His expertise is in the expression of both soluble and membrane proteins expressed in prokaryotic and eukaryotic host cells followed by a variety of protein purification methods and biochemical and biophysical approaches for protein characterization. He also spent 7 person-months in acting as the Scientific Manager of the project.
• Soluble and membrane protein crystallization and crystallography. Dr Petros Giastas was recruited in Tzartos’ lab for 36 person-months full time position. His expertise is in crystallization of water-soluble proteins, data collection, and structure solution. His expertise further extends to other scientific areas including protein expression in E. coli and in yeast P. pastoris, protein purification, many chromatographic techniques and protein characterization through biochemical methods.
• Advanced imaging and neurophysiology. Dr Paraskevi Koutsoudaki was recruited in Matsas’ lab for 36 person-months full time position. Her expertise is in the neurophysiology of animal models of neurodegeneration and imaging.
• Live mouse imaging. Dr Vivian Tseveleki was recruited in Probert’s lab for 36 person-months full time position. Her expertise is in T cell biology and the analysis of neuroimmune interactions in the CNS, as well in vivo imaging.
• Expression and purification of recombinant membrane proteins. Dr Nikos Kouvatsos was recruited in Tzartos’ lab for 19 person-months full time position. His expertise is in the expression of soluble and membrane proteins expressed in prokaryotic and eukaryotic host cells and in protein purification methods.
• Molecular Neurobiology and Immunology. Dr Konstantinos Lazaridis was recruited in Tzartos’ lab for 19 person-months full time position. His expertise is in the molecular design of constructs and their subsequent use for protein expression in a variety of systems, ranging from yeast and insect cells to mammalian expression systems.
• In vivo monitoring of stem cells following neurotrauma at the whole animal level. Dr Kanella Prodromidou was recruited in Matsas’ lab for 28 person-months full time position as a researcher with expertise in the study of prion disease in mouse models as well as the function of prion protein in neural stem cells.
• Electrophysiology. We faced major difficulties in attracting an experienced electrophysiologist to establish our Electrophysiology Unit. Despite the extensive advertisement of this post-doc position (Nature jobs, Hellenic Pasteur Institute web site, FENS web site, Scientifica web site, etc), only a few candidates came forward but none proved to be suitable for taking over related responsibility. In order to achieve the establishment and successful use of the Electrophysiology Unit, which is one of the main goals of NeuroSign-REGPOT, we decided after official approval from our project officer to recruit Mrs Dafni Chroni as an Electrophysiology technician in Tzartos’ lab for 24 person-months full time position. Mrs Chroni is a physicist with Masters’ degree in Biophysics and expertise in cell cultures, who together with Dr Marios Zouridakis (Neurosign recruited researcher) and strong collaboration with our collaborator Dr Piotr Bregestovski, established a functional Electrophysiology Unit.
We also recruited three other technicians during Neurosign project:
• Maria Karamita, BSc. Miss Karamita assisted the other personnel in organizational activities, particularly those concerning the installment and up-keep of new equipment. She also assisted in a research project investigating inflammation and demyelination in mice conditionally deficient in a major neuroprotection signaling pathway (Probert lab). Full-time position for 19 person-months.
• Vasiliki Kyrargyri, BSc. Miss Kyrargyri assisted the other personnel in organizational activities, particularly those concerning the introduction of basic imaging protocols. She also assisted in a research project involving the functional imaging of neurons by confocal microscopy and live cell imaging techniques (Probert lab). Full-time position for 19 person-months.
• Maria Belimezi, Ph.D. She was recruited to complement the personnel for dissemination activities. Dr Belimezi prepared and uploaded updated information for the NeuroSign web site (www.neurosign.gr) and relevant scientific info material and assisted in the NeuroSign dissemination and promotional activities (e.g. organization of lectures, open day events, etc) and corresponding reports. Full-time position for 15 person-months.
In total, 10 post-doctoral researchers (6: female, 4: male) and 4 technicians (female) were recruited during Neurosign. For six of these recruited scientists (four post-doctoral fellows, namely Drs M. Evangelidou, P. Giastas, K. Lazaridis and M. Zouridakis and two technicians, namely, Ms D. Chroni and Ms M. Karamita) we extended their recruitment for 6 months (up to month 42), after approval from the project officer (in compliance with the 6-month extension of the Neurosign project).
The recruitment of these highly experienced researchers in our group proved to be very beneficial for the upgrade of our research potential. The recruited staff boosted the research conducted in our groups by introducing novel and cutting-edge methodological approaches to our group. More precisely, the recruited researchers with expertise in advanced imaging, live mouse imaging, Ca2+ imaging, advanced imaging and stem cells, together with the recently purchased and installed multi-photon microscope and their secondments to EU groups of collaborators initiated research projects involving high-tech imaging methodologies. This significantly contributed to the achievement of our main research goal, which dealt with development of a) novel diagnostics for neuromuscular and neurodegenerative diseases using imaging techniques and b) treatment approaches related to these diseases as well as spinal cord injury. The recruited researchers with expertise in molecular biology, soluble and membrane protein expression, purification and crystallization significantly contributed to the crystallization of proteins with a major pharmacological importance (i.e. two subtypes of neuronal acetylcholine receptor). Elucidation of their 3D X-ray structure opens the way for the rational design of drugs towards treatment of neurological diseases or disorders related to them. Also, the establishment of Electrophysiology Unit within our group and its current full functionality complements our structural studies by confirming the functionality of the crystallized proteins/membrane receptors and serves as a platform for screening potent drugs to the expressed proteins.
Apparently, the objectives of WP3 met with full success regarding a) the recruitment of researchers with highly competitive skills to contribute to the establishment of our group as a Centre of Excellence in Neuroimaging, b) the enhancement of the research skills of our seconded researchers to our top quality collaborators’ groups, c) the adaptation of the new methodological research approaches of the recruited researchers to our group and d) the best exploitation of the introduced research skills of the Neurosign recruited researchers.
All of the recruited Neurosign researchers and technicians continued their research after completion of the Neurosign project, since they are now funded by other research grants (National and European grants) acquired by the three Neurosign group leaders (Drs S. Tzartos, R. Matsas and L. Probert). Therefore, sustainability of the Neurosign recruited researchers is ensured for at least the two next years.
WP3 yielded five deliverables (D3.1: Report on the recruitment of researchers; D3.2: First year progress reports from the recruited researchers; D3.3: Second year progress reports from the recruited researchers; D3.4: Third year progress reports from the recruited researchers; D3.5: Interim report on recruitment) and reached its milestone (MS2: Recruitment of researchers) with a short delay due to the unforeseen problem dealing with recruitment of expert electrophysiologist.
Work package 4: Acquisition of equipment (WP leader: Dr Dimitra Thomaidou)
In WP4 the aim was to purchase the following instrumentation:
1. Multiphoton confocal microscope for advanced live cell and tissue imaging
2. Patch-clamp equipment for establishment of the electrophysiology unit
3. Fast protein liquid chromatography for high throughput protein purification, accompanied by a nano-spectrophotometer
4. Upgrade of current crystallization robot
All of the originally proposed instrumentation was already purchased during the first period of Neurosign. Furthermore, we also purchased some extra instrumentation and renovated the Imaging Unit (in order to meet with the special requirements of the multi-photon microscope) after approval from the project officer, always within the budget. The upgrade of our crystallization robot finally did not take place, because we already acquired scientific results with the current set up. The budget for this upgrade was used to contribute to the rest of equipment.
In brief, the acquired instrumentation is as follows:
Multiphoton Confocal Microscope System
Multiphoton confocal microscope for advanced live cell and tissue imaging (Leica, Germany). The system is comprised of a Leica-SP5 II modular laser scanning inverted microscope equipped with multiple visual (VIS) laserlines (450-640nm) and near UV lasers (400-440nm) for single-photon confocal microscopyand a Mai-Tai (Spectra-Physics) infrared (IR) oscillating Ti/Saphire femtosecond laser, allowing Multiphoton Confocal microscopy.
Two complete cell patch-clamp set ups
Electrophysiology equipment: The equipment includes the following devices: Vibration isolation table with faraday cage (Thorlabs, France), Micromanipulators (Narishige, Japan), Microelectrode amplifier, Oscilloscope, Stimulator, Electrode puller (HEKA, Germany), Microelectrode holder and two accompanying fluorescent microscopes (Olympus CXK41 for mammalian cells and IX51 for oocytes, Japan).
Preparative protein liquid chromatography (FPLC) and NanoDrop
ÄKTApurifier 10 FPLC system (GE Healthcare, UK), that allows purification of microgram to milligram quantities of protein, operating at flow rates of 0.001 to 10 ml/min at pressures of 0 to 25 MPa. A Nanodrop 2000 spectrophotometer (Thermo Scientific, USA) for the precise and fast determination of the concentration of the purified proteins effectively complements the FPLC instrument.
Multi-crystal gamma counter
Multi Crystal LB 2111 gamma counter (Berthold Technologies, UK) is a compact instrument for most applications where emitting isotopes are used.
Automated agarose gel electrophoresis multiparametric system
Sebia’s (Sebia, USA) second generation semi-automated agarose gel electrophoresis system with scanner for the diagnosis of MS.
All the purchased equipment are labelled with badges declaring “Equipment purchased through FP7-REGPOT-NEUROSIGN No264083” accompanied by the EU logo.
All the originally proposed equipment, except the upgrade of the crystallization robot, were successfully purchased and installed in HPI by month 24 of the Neurosign project. The upgrade of the crystallization robot was judged to be not essential after the impressive accumulation of protein crystals from our research team using the current set up. Therefore, the saved amount of money was reallocated for the purchase of other equipment.
Apart from the successful acquisition of the state-of-the-art equipment (mostly referring to the multi-photon confocal microscope and the electrophysiology equipment), there had been considerable training of Neurosign researchers and members on the acquired instrumentation, mainly through outgoing and incoming secondments. More specifically, for the multi-photon microscope, two collaborators of ours (Drs F. Kirchhoff and N. Kawakami) were seconded to HPI for short time periods and for the electrophysiology set-up one collaborator of ours (Dr P. Bregestovski) visited us twice. Also, five HPI researchers were seconded to collaborators in order to gain additional expertise in intravital imaging, namely Drs P. Koutsoudaki, D. Thomaidou, R. Matsas, L. Probert and M. Evangelidou and one recruited technician was seconded to one of our collaborators to gain expertise in electrophysiology. The seconded researchers presented the acquired knowledge regarding use of this instrumentation to all other members of Neurosign and HPI through lectures and hands-on demonstrations.
The above purchased instrumentation is now fully functional and has entered effectively in our daily experimental protocols. The state-of-the-art multiphoton confocal microscope is fully exploited by Neurosign researchers yielding scientific results published in high-impact journals. The multiphoton microscope is now used also as a service provider to other Academic/Research Institutions and we plan to use the Electrophysiology equipment for providing service to such Institutions and the Pharmaceutical Industry.
WP4 yielded two deliverables (D4.1: Report on the purchase and installation of the equipment and D4.2: Report on the training of recruited and experienced researchers in the acquired equipment) and reached its milestone (MS8: Purchase of equipment) with a short delay compared to the originally proposed time period, mainly due to the time-consuming actions regarding the purchase of the multi-photon microscope (eg: extensive market search and the international open tender for its purchase).
Work package 5: Organization of workshops and conference (WP leader: Dr Alexandros Lavdas)
In WP5, we had proposed three workshops and one final conference to take place in HPI for disseminating the knowledge acquired through Neurosign to the regional and European scientific community, as well to the public of general interest. In the original proposal we had proposed one Workshop to take place each year of Neurosign and a final conference at the end of the project. The order and titles of these workshops and conference were:
1. 1st Workshop: “Animal models of neurodegeneration and behavioral tests for assessment of motor and cognitive function”
2. 2nd Workshop: “Advanced Imaging and Electrophysiology in the mammalian CNS”
3. 3rd Workshop: “Functional analysis of CNS-infiltrating immune cells during the development of CNS autoimmunity, injury and repair”
4. Final Conference on “Advances in the treatment of neurodegenerative and neuromuscular disorders”
A deviation regarding the originally proposed schedule of Workshops and Conference took place. We changed the order between the 2nd and 3rd Workshop. We had to take this action for a couple of reasons: a) there was been a significant delay in the installation of the Multiphoton Microscope in our Institute. The reasons were two-fold: Firstly, the time-consuming procedures for extensive market search and the international open tender for its purchase, as dictated by the relatively high cost (over 300K €) of this major equipment. Secondly, the renovation of the Imaging Unit space to meet the installation requirements of the new Multiphoton microscope could only be performed after we obtained permission to use part of the NeuroSign budget towards this goal; b) we faced major difficulties in attracting an experienced electrophysiologist to establish our Electrophysiology Unit. This Unit became functional only in month 19 of Neurosign project after the secondment of the electrophysiology expert Professor Piotr Bregestovski (Neurosign external collaborator) to our lab and the recruitment of Mrs Dafni Chroni, as an electrophysiology technician.
The first of the three Neurosign workshops took place at the early start of the second year of the project (11th -14th January 2012) and was entitled “Animal models of neurodegeneration and behavioural tests for assessment of motor and cognitive function”. This was a 4-day workshop, whose aim was to discuss the most appropriate animal models for selected neurodegenerative disorders and neurotrauma, as well as to illustrate simple and more sophisticated tests for assessing motor and cognitive function after cell transplantation or other treatment. Lectures were held in the morning and practical sessions in the afternoon. During the practicals, the students had the chance to observe stereotactic operations on mice, permanent middle cerebral artery occlusion (a model of thrombotic stroke), the rotarod test for balance, behavioral tests involving flies and the Morris water maze cognitive test for mice (video recorded). Twenty students participated in the practicals, mostly undergraduate and early pre-doctoral students (2 from Barcelona and 18 from various Institutions in Greece) while many more students and scientists attended the lectures at the Hellenic Pasteur Institute lecture theatre. In total, this Workshop was attended by 100 participants. Twelve speakers (4 from abroad and 8 National) who have proven expertise in the field of Neurosciences gave the lectures and nine instructors, most of them highly experienced researchers taught in the practicals. Twenty synoptic handouts were printed, and the students were also given USB sticks including full details and ppt presentations. List of lecturers: Drs Pico Caroni, Spiros Efthimiopoulos, Spiros Georgopoulos, Rebecca Matsas, Ada Mitsacos, Nektarios Tavernarakis, Nikolaus Plesnila, Makis Skoulakis, Fotini Stylianopoulou, Rebecca Trueman, Vivian Tseveleki and Luca Turin.
The 2nd Neurosign workshop took place at the early start of the second period of the project (4th-7th December, 2012) and was entitled “Functional analysis of CNS-infiltrating immune cells during the development of CNS autoimmunity, injury and repair”. This was a 4-day workshop involving theoretical and practical parts and took place at the Hellenic Pasteur Institute (HPI). The program of this workshop was prepared as an overview of the state-of-the-art knowledge in the development of neuroimmune pathologies of the central nervous system together with hands-on practicals in current investigative techniques. The aim of this workshop was to promote and encourage awareness in the field and cooperation amongst aspiring and established neuroimmunologists. During the practicals, the students had the chance to participate in and observe experimental processes in the following sessions: EAE: induction and evaluation, neuropathological analysis, isolation andanalysis of spinal cord infiltrates and ex vivo T-cell imaging. The participants were mostly undergraduate and early pre-doctoralstudents from the universities and research institutes across Greece (Crete, Ioannina, Athens, Alexandroupoli, Thessaloniki). The number of students participating in both the theoretical and practical parts of the workshop was 28. The number of participants registered only for the theoretical part was 16. In total, ~100 people attended the theoretical part of the Workshop (since this session was open to the local scientific community). Twelve speakers (8 from abroad and 4 National) who have proven expertise in the field of CNS autoimmunity gave the lectures and six instructors, most of them highly experienced researchers, taught in the practicals. List of lecturers: Dr Christopher Linington, Hans Lassmann, David Baker, Roland Liblau, Edgar Meinl, George Kollias, HartmutWekerle, Maria Evangelidou, Roberto Furlan, Krysztof Selmaj, Antonio Uccelli, Nikolaos Grigoriadis and Vivian Tseveleki.
The 3rd workshop was entitled «Live Cell Imaging and Electrophysiology”. This was a 4-day workshop (1st- 4th October 2013) involving theoretical and practical parts and took place at the Hellenic Pasteur Institute (HPI). The aim was to discuss the cutting edge live-cell imaging technology and electrophysiological approaches to study protein receptors. Lectures were held in the morning and practical sessions in the afternoon. During the practicals, the participants had the chance to observe video recorded experiments and most importantly to perform hands-on experiments including surgical procedures for creating brain and spinal cord windows for intravital Multiphoton imaging and whole-cell and single-channel recordings, using the patch-clamp technique applied on mammalian cells. Twenty students participated in the practicals, mostly undergraduate and early predoctoral students (1 from Russia, 1 from France, 1 from UK, 1 from Cyprus, 1 from Germany and 15 from various Institutions in Greece), while many more students and scientists attended the lectures at the Hellenic Pasteur Institute lecture theatre. They attended the practicals in rotation of 4 groups of 5 people each. In total, ~100 people attended the theoretical part of the Workshop (since this session was open to the local scientific community). Ten speakers (5 from abroad and 5 National) who have proven expertise in the field of live imaging and electrophysiology gave the lectures and six instructors, most of them highly experienced researchers, taught in the practicals. List of speakers: Drs Piotr Bregestovski, Dimitrios Davalos, Naoto Kawakami, Frank Kirchhoff, Rebecca Matsas, Cornelia Poulopoulou, Davide Ragozzino, Dimitra Thomaidou, Irini Skaliora and Pavlos Rigas.
Regarding the Final Conference, we decided to join it with the 26th Meeting of the Hellenic Neuroscience Society in order to have increased visibility. It was entitled “Understanding Brain Function to Treat Dysfunction” and was held on the 29/11-1/12/2013 at the Eugenides Foundation in Athens (http://www.eugenfound.edu.gr). The most recent developments in stem cell research, cellular and gene therapy approaches for the treatment of neurodegenerative diseases and traumas and in structure and function of several ligand-gated ion channels were presented and discussed by 21 invited lecturers, 14 of which from abroad. Three poster sessions were also included, where 154 posters were presented with full attendance and 11 posters were selected for oral presentations. The thematics were diverse and covered a large area of the Neuroscience field (Nervous System Development, Synaptic transmission /Signaling /ion and molecule channels, Stem Cells and Nervous System Regeneration, Neuroendocrinology and Neuroimmunology, Behavior and Cognition, Neurodegenerative Diseases, Nervous System Disorders and Neuropsychopharmacology, Systems and computational Neuroscience , Other). The participation at the Final Conference was very high (total 509 participants) and was secured by the extensive advertisement from i) the website that was created specifically for the Meeting (2013.hsfn.gr) but was linked to the NeuroSign, Pasteur Institute and Hellenic Society for Neuroscience websites, that also had a banner at their front page advertising the event ii) e-mails to all academic bodies and institutions in Greece and all relevant Greek researchers and iii) posters and programs that were distributed to other relevant meetings and academic and research institutions in Greece. A very important feature that also secured this unexpectedly high participation was that students and post-doctoral researchers attended the meeting without any registration fees and this was particularly important at this time period of economic crisis in Greece.
After successful implementation of these Workshops and the Final Conference, our group has gained a good expertise in organizing such scientific events. As a Center of Excellence, we will organize additional Workshops and Conferences in the future, so as to educate people in the newly entered technologies in HPI through Neurosign (i.e.: multiphoton microscopy, Electrophysiology, induced pluripotent stem cells, models for spinal cord injury and stroke).
WP5 yielded three deliverables (D5.1: Report on the 1st and 2nd Workshops; D5.2: Report on the 3rd Workshop; D5.3: Report on the Final Conference) and reached one milestone (MS10: Decision about the organization of the Final Conference) within its originally proposed time schedule.
Work package 6: Dissemination activities (WP leader: Dr Era Taufik)
During Neurosign, we a) organized monthly meetings in HPI with open access where the progress of the recruited researchers and the knowledge gained by secondments were presented, b) participated in 23 International and 14 National conferences or workshops with oral and poster presentations (in total: 37), c) maintained and regularly updated our website (www.neurosign.gr) d) published our research results in sixteen (Neurosign acknowledged) peer-reviewed scientific journals, e) organized an open-day event and f) provided two press releases to the social media. Further details on these dissemination actions are provided in the following section.
In the future, we will continue disseminating the knowledge acquired in our groups by publishing our research results (which have been boosted by our significant R/T upgrade through Neurosign) in top quality journals, by maintaining and updating our website, by participating in Conferences and Workshops, by organizing open-day events and workshops and by providing press releases.
Potential Impact:
Neurosign project had an important impact on the regional socio-economic development.
1. During a severe period of financial crisis in Greece, Neurosign supported the recruitment of 10 post-doctoral fellows (experienced researchers) and 4 technicians. Equally important was the accomplishment of the sustainability of these recruited staff after the completion of the Neurosign project. Due to our significant R&T upgrade through implementation of the project, the three group leaders (Drs S. Tzartos, R. Matsas and L. Probert) secured additional funding for their research projects by National and International and EC grants. In total, the three group leaders were granted 2,790,000 euro from other financial sources { ~900,000 (MDA2013-16; AFM20012-14; Aristeia, Thalis); 210,000 (Aristeia II + IKYDA Greek-German collaboration, both directly linked to CNS Intravital Imaging); ~1,100,000 (NOISEPLUS COOPERATION, ARISTEIA I, BNP PARIBAS, PTR, BODOSSAKI FOUNDATION, EMPEIRIKION FOUNDATION, Thalis, KRHPIS); 580,000 (Synergasia)}. Some of these grants were awarded during the mid-term implementation of the Neurosign project and some others at the end of the project, assuring the continuation of the contracts of the Neurosign recruited staff for at least two years after the end of Neurosign project (and in some cases up to 3 years after the end of the Neurosign project). From the recruited staff during Neurosign (in total 14), 4 are men and 10 are women. Also, 10 PhD students (6 women and 4 men) were indirectly benefited, since they were supported scientifically by the supervision of the experienced staff.
2. The acquisition of state-of-the-art equipment of a total cost of ~650,000 euro contributed to the sustainability of scientific equipment suppliers mainly in Europe. The maintenance of this equipment contributes to the sustainability of the regional representatives of the suppliers.
3. As a Center of Excellence in Neurosignalling and neurological diseases and disorders we put much of our efforts in supporting several related patients’ associations. We have been instrumental in the establishment of the Hellenic MG Association (HMGA; web site: www.myasthenia.gr) whose full-members are exclusively myasthenic patients. We provide: i) space for office and meetings, ii) we bridge them with the relevant clinicians for expert’s advice, and iii) provide them with reliable information on the research progress towards the most effective treatment approaches of MG. In addition, we are collaborating with the MDA-Hellas. Given the expected expansion of our translational research, we intend to contribute to the establishment and/or to the strengthening of other similar societies of patients suffering from chronic neurological disorders relevant to our research interests and translational aims. One such society is ELEPAP-GR (www.elepap.gr) which aims at supporting children with movement disorders resulting from neurological disease and neurotrauma. Apparently, given that most of our research regards the best diagnosis and treatment procedures of autoimmune diseases such as myasthenia gravis, neuromyelitis optica, multiple sclerosis, amyotrophic lateral sclerosis and stem cells therapies for spinal cord injuries and stroke, there is a huge social impact of the Neurosign project.
4. During implementation of the Neurosign project, we increased our collaborations with National hospitals (Alexandra Hospital, Laiko Hospital, Eginitio Hospital, Red Cross Hospital) for the study of several human autoimmune diseases and for developing the best strategies to treat them.
5. Our significant R&T upgrade through Neurosign has led to attracting an increased number of PhD applicants compared to the past. There has been ~70% increase in such applications and we have already accepted many young PhD students to get enrolled with our projects. This also contributes to the regional economical development, since we support the PhD students by the additional grants awarded to us after Neurosign.
6. Finally, we envisage attracting additional funding by Pharmaceutical Industries and SMEs (either local or European), with which we are collaborating in order to commercialize our developed diagnostic kits for several autoimmune diseases and peptide vaccines to treat multiple sclerosis. This will ensure the sustainability of the recruited Neurosign staff for many years, since it may lead to their permanent employment.
Dissemination activities during Neurosign:
As an established Center of Excellence, we are enrolled in several activities ensuring the widespread dissemination of knowledge acquired in our groups. We have taken the following actions through implementation of Neurosign project, and are continuing most of them:
1. We set up the Neurosign website (www.neurosign.gr) which is a well informative and easy-to-use website, whose sessions were described in full detail in the First Periodic Report. The website was updated regularly in order to a) disseminate the acquired knowledge through Neurosign to both the scientific community and to the general public, b) promote important events associated with the program (i.e. forthcoming meetings, workshops, conferences, publications, open day events, etc) and c) advertise services offered by Neurosign group and emerged job openings. The Neurosign website is still active after the end of the project and we intend to continuously update it. There are also hyperlinks to the websites of the Hellenic Pasteur Institute (HPI) and its Neurosign participant laboratories (Tzartos’ lab, Matsas’ lab and Probert’s lab), whose research aims and projects are discussed in detail, as well links to our external collaborators’ research groups. Photos of the state-of-the-art equipment acquired through Neurosign (mainly referring to the multiphoton confocal microscope and the Electrophysiology apparatus) are uploaded to the site with a brief description of its applications. It is declared that this apparatus is available for use for any Academic/Research Institute that might find useful. The most representative publications of Neurosign scientists are also uploaded on the site; we included reviews in this part, so that to be informative as much as possible to scientists of another research field. Our website also contains information on the monthly meetings, annual meetings and workshops of the Neurosign project. In addition, there is a special section for disseminating the knowledge acquired through Neurosign to the National (in Greek language) and International (in English) general public. In this section, the various neurological and neuromuscular disorders under study by the three Neurosign participants are discussed in non-technical terms for the lay audience (pathology, symptoms, diagnosis, progress towards therapy, etc, are discussed). Finally, in all sections of Neurosign website, EC FP7 is acknowledged through the FP7 logo linking to the CORDIS website.
2. NeuroSign Meetings: For disseminating reasons, we have arranged monthly meetings participated by Neurosign members and all other lab members inside the HPI. In each of these meetings, a different Neurosign scientist presents his/her research interests and results, as well as detailed research protocols and techniques used. In these meetings, the seconded Neurosign researchers to the groups of our external collaborators present the knowledge acquired and transfer the hain of know-how to all other Neurosign members. The abstracts of these presentations are uploaded in the corresponding section of the web site.
3. Publications of research papers: During Neurosign, sixteen publications (in which Neurosign was acknowledged) in peer-reviewed scientific journals took place. Six from Tzartos’ group (1. Lazaridis K, Zisimopoulou P, Giastas P, Bitzopoulou K, Evangelakou P, Sideri A, Tzartos SJ. Expression of human AChR extracellular domain mutants with improved characteristics Int J Biol Macromol. 2014 Feb;63:210-7; 2. Lazaridis K, Zisimopoulou P, Lagoumintzis G, Skriapa L, Trakas N, Evangelakou P, Kanelopoulos I, Grapsa E, Poulas K, Tzartos S. Antigen-specific apheresis of autoantibodies in myasthenia gravis Ann N Y Acad Sci. 2012 Dec;1275:7-12; 3. Zisimopoulou P, Brenner T, Trakas N, Tzartos SJ. Serological diagnostics in myasthenia gravis based on novel assays and recently identified antigens Autoimmun Rev. 2013 Jul;12(9):924-30; 4. Tzartos J, Zisimopoulou P, Rentzos M, Karandreas N, Zouvelou V, Evangelakou P, Tsonis A, Thomaidis T, Lauria G, Andreetta F, Mantegazza R, Tzartos S. LRP4 antibodies in serum and CSF from amyotrophic lateral sclerosis patients Annals of Clinical and Translational Neurology, in press; 5. Niarchos A, Zouridakis M, Douris V, Georgostathi A, Kalamida D, Sotiriadis A, Poulas K, Iatrou K, Tzartos SJ Expression of a highly antigenic and native-like folded extracellular domain of the human α1 subunit of muscle nicotinic acetylcholine receptor, suitable for use in antigen specific therapies for myasthenia gravis. PLoS One. 2013 Dec 20;8(12):e84791. doi: 10.1371/journal.pone.0084791; 6. Zouridakis M, Giastas P, Zarkadas E, Tzartos SJ Crystal structures of the free and antagonist-bound states of the extracellular domain of human α9 nAChR Nature Structural and Molecular Biology (under revision), three from Matsas’ group (1. Tsioras K, Papastefanaki F, Politis PK, Matsas R, Gaitanou M. Functional Interactions between BM88/Cend1, Ran-binding protein M and Dyrk1B kinase affect cyclin D1 levels and cell cycle progression/exit in mouse neuroblastoma cells. PLoSOne. 2013 Nov 28;8(11):e82172. doi: 10.1371/journal.pone.0082172; 2. Prodromidou K, Papastefanaki F, Sklaviadis T, Matsas R Functional cross-talk between the cellular prion protein and the neural cell adhesion molecule NCAM is critical for neuronal differentiation of neural stem/precursor cells. Stem Cells. 2014 Feb 4. doi: 10.1002/stem.1663; 3. Miltiadous P, Kouroupi G, Stamatakis A, Koutsoudaki PN, Matsas R, Stylianopoulou F. Subventricular zone-derived neural stem cell grafts protect against hippocampal degeneration and restore cognitive function in the mouse following intrahippocampal kainic acid administration. Stem Cells Transl Med. 2013 Mar;2(3):185-98) and two from Proberts’ group (1. Voulgari-Kokota A, Fairless R, Karamita M, Kyrargyri V, Tseveleki V, Evangelidou M, Delorme B, Charbord P, Diem R, Probert L Mesenchymal stem cells protect CNS neurons against glutamate excitotoxicity by inhibiting glutamate receptor expression and function. Exp Neurol. 2012 Jul; 236(1):161-70. doi: 10.1016 / j.expneurol.2012.04.011; 2. Evangelidou M, Karamita M, Vamvakas SS, Szymkowski DE, Probert L. (2014) Altered expression of oligodendrocyte and neuronal marker genes predicts the clinical onset of autoimmune encephalomyelitis and indicates the effectiveness of multiple sclerosis-directed therapeutics. J Immunol. May 1;192(9):4122-33. doi: 10.4049/jimmunol.1300633)
4. Participation in Conferences/Workshops. During the second period of Neurosign project, Neurosign researchers participated in 23 International Conferences and 14 National Conferences (in total: 37. They presented their research results either in oral or printed presentations (posters) and communicated them with International and National Neuroscientists.
5. Open day events. We organized one open day for school pupils, students, researchers and medical doctors, which apart from disseminating the gathered knowledge on cutting-edge technology experimental approaches through our secondments (e.g. stem cells, advanced live animal imaging, electrophysiology), it also promoted the services available in HPI, after purchase of this major equipment (multiphoton and electrophysiology). During this open day, regional researchers/Academics/students/school pupils had the chance to take a tour to this Neurosign-acquired equipment and be informed about the applications of the instrumentation.
6. Interviews in newspapers/social media. The Coordinator, Prof. Socrates Tzartos, was interviewed by a couple of local media regarding the Neurosign project and its results. The interviews were published at newspapers (http://www.kathimerini.gr/755815/article/epikairothta/episthmh/neo-e8niko-kentro-aristeias-gia-tis-neyroepisthmes) and web newspapers (http://www.goodnews.gr/Articles/I-symmetochi-tis-Elladas-stin-pagkosmia-ereyna_2059.html). The interviews were addressed both to the scientific and general interests audience.
Exploitation of results:
During Neurosign, we yielded important scientific results, mainly due to our significant upgrade through recruitment of very experienced researchers, collaborations with top-quality EC research groups and acquisition of state-of-the-art equipment. Exploitation of these results has already started and is expected to be fully accomplished in the forthcoming couple of years and beyond. More specifically:
1. We patented three of our scientific results (Patent Application numbers: EP11724450; WO2013057599 and WO2009093143) and intend to commercialize them in the near future. More specifically, we are collaborating with a local SME (Tzartos Neurodiagnostics) for the commercialization of novel diagnostic assays for a more sensitive and reliable diagnosis method for the myasthenia gravis, neuromyelitis optica and amyotrophic lateral sclerosis diseases and with two Pharmaceutical Industries for the commercialization of peptide vaccines for the treatment of multiple sclerosis. Especially for the latter, pre-clinical studies completed (submitted for publication), toxicology tests underway (results expected summer 2014); development of patient screening and selection assay is underway.
2. We plan to commercialize immunoadsorption columns for the specific treatment of myasthenia gravis patients. We had patented this specific treatment procedure before the Neurosign project (EP 1509605 B1), but significant progress took place during implementation of the Neurosign project. We are collaborating with a local SME (Tzartos Neurodiagnostics) for this project and plan to collaborate with an emerging spin-off SME within HPI organized by Prof. Socrates Tzartos. Clinical trials are pending, which will take place in the near future in collaboration with a relevant company with experience in the field.
3. We have initiated service provision to other Academic/Research Institutions based on some of the equipment purchased through Neurosign. Therefore, our newly acquired multi-photon microscope is now used by many external users. We are planning to provide service to Academic/Research Institutions and Pharmaceutical Industry using our newly established Electrophysiology setups (applicable to membrane proteins expressed in cell lines and Xenopus oocytes) for the evaluation of the potency and efficacy of drugs under development. Both instrumentation (multiphoton microscope for live animal imaging and Electrophysiology setups) is unique in the Athens area.
4. We plan to collaborate with Pharmaceutical Industries in order to design specific drugs for the treatment of chronic pain and epilepsy, based on the elucidation of the X-ray 3D structure of two subtypes of the human neuronal acetylcholine receptor, which was accomplished during Neurosign.
5. In the frame of the above, we have significantly increased (~300% increase compared to before Neurosign project) our collaborations with several enterprises to get involved in further collaborative actions. The SMEs and Pharmaceutical Industries with which we are now collaborating are Alomone; Tzartos Neurodiagnostics; Syneuro, Moscow; Myomedix, Germany; Euroimmune AG, Germany; VIANEX; Eldrug; Redoxis; Serono; Xencor; FPRT Bio and AbbVie.
6. Improvement of the potential of our participation in FP7, beyond-FP7 and other funding projects. Our improved RTD capacity and capability, the significant upgrade of our Imaging Unit, the establishment of a new Unit of Electrophysiology, the established professional management, the development of collaborations with leading European research groups and the increase of our publications in top quality journals have now led to our increased visibility in the European research area. All above have significantly increased our potential to receive further research grants (e.g. HORIZON 2020) either as the leading partner or member of a large consortium.
List of Websites:
Website of Neurosign project: www.neurosign.gr
Contact details of NeuroSign partners:
1. Prof. Socrates Tzartos, Coordinator. Head of the Lab of Molecular Neurobiology and Immunology, Hellenic Pasteur Institute
2. Dr Rebecca Matsas. Head of the Lab of Cellular and Molecular Neurobiology, Hellenic Pasteur Institute
3. Dr Lesley Probert. Head of the Lab of Molecular Genetics, Hellenic Pasteur Institute
4. Dr Marios Zouridakis, Scientific Project Manager, Hellenic Pasteur Institute
