Call Identifier: EOI.FP6.2002

Study the genetic origin and molecular pathogenesis of corneal dystrophies

NEED & RELEVANCE: How the proposed research activities contribute to realising the objectives of a priority thematic area and why it requires a European mobilisation of activities and resources through the means of a Network of Excellence?

1.1Aims and background

The aim of the CORNEAGENE proposal is to establish Europe as the international scientific leader in the study of corneal dystrophies. Partners plan to bring together a collaborative group of investigators to study the molecular genetic basis, immunohistological and biochemical characteristics of corneal dystrophies, with special emphasis on the most frequently occurring BIGH3 (kerato-epithelin) related dystrophies, macular corneal dystrophy and keratoconus.

The proposed research activities of CORNEAGENE project fit into the priority research area 1.1.1.ii.a: Application-oriented genomic approaches to medical knowledge and technologies - Combating cardiovascular disease, diabetes and RARE DISEASES. The other Sub-Thematic Priority is 1.1.2.i. Applied IST research addressing major societal and economic challenges. Partners will build intelligent system to support medical research and health professionals and will integrate web based medical record and eForms for research protocols with BioChips (DNA-Microarrays) andBioInformatics (database searches, data mining) for the design and optimization of microarrays.

The corneal dystrophies to be studied are rare diseases with frequencies of occurrence of 1:300 000 to 1:1 000. At present, corneal transplantation is the only way of preventing severe visual deteration for patients suffering from corneal dystrophies. Altogether, this disease group represents the third most frequent indication of corneal transplantation worldwide. Although corneal transplantation has a fairly good prognosis in this disease group, recurrence of the disease and some other complications may occur in 10-30% of the cases. Additionally, even in an uncomplicated event, the complete socio-economical rehabilitation of the patient may be difficult, since a corneal-transplanted eye needs special care.

In the past few years, different research groups have gathered considerable information on the genetics of corneal dystrophies. Mutations in the BIGH3 gene responsible for 6 different types of corneal dystrophies, and in the CHST6 gene responsible for macular corneal dystrophy have been discovered. In autosomal dominant keratoconus, chromosomal localization to chromosome 21 has been found by genetic mapping studies and the VISX gene has been implicated in the pathogenesis of the disease.

1.2Expected outcomes

The CORNEAGENE proposal focuses on two major fields:

1. Widening the genetic knowledge on these diseases by integrating research activities of partners from several countries of Europe and outside Europe.

2. Post-genomic research for understanding the mechanisms leading to these diseases, developing suitable model systems for such studies.

Ad 1. The mutations occurring in several parts of Europe have not been studied yet. To collect the most possible information, joint effort by several excellent centers is needed. The goal is to collect blood and corneal samples form dystrophic patients in clinical centers and transfer DNA samples to the research centers having expertise in the genetic analysis of certain diseases. Analysis of DNA samples by PCR amplification, followed by direct automated DNA sequencing is planned. After finding the specific mutations, simpler tests, such as DHPLC (the new standard mutation screening method) will be used. Mutations in the BIGH3 gene, CHST6 gene and genes implicated in autosomal dominant keratoconus are to be studied by this procedure.

Ad 2. Post-genomic research involve three main fields:

- One problem to overcome is that the corneal buttons excised during corneal transplantation is too small for extensive biochemical studies and does not allow in vivo investigations. To overcome this problem, establishment of immortalized corneal cell lines from dystrophic and normal corneas is planned. Immortalized cell lines seem to be suitable model systems for the investigation of altered protein function and meabolism in these dystrophies.

-Investigation of the changes in protein metabolism and function caused by the mutations in BIGH3 related and macular corneal dystrophies.Advanced proteomics studies involving elaborated two dimensional gel electrophoretic studies and mass spectrometry will be helpful in determining the proteolytic steps involved in the metabolism of BIGH3 and identifying the site and the enzymes of proteolysis. On the long term, these studies may help the development of drugs being able to inhibit the accumulation of abnormal protein end-products.

-Genetic microarray systems for the study gene expression in keratoconus in order to identify genes and proteins implicated in the pathogenesis of this disease

1.3The added value of Europe

Establishment of a Network of Excellence involving the leading European research and clinical centers in the field will provide the following benefits:

-Involvement of the greatest number of rare disease (dystrophic) patients in each research field may be reached by unifying several expert partners in a Network of Excellence.

-For extensive and worldwide competitive study of these diseases, especially the post-translational part, addition of expertise available in different centers will be needed in order to reduce costs and increase the effectiveness of research.

EXCELLENCE:The feasibility of putting together the critical mass of resources and expertise needed to achieve the network's objectives and to be a world force on the research activities proposed

By putting together the expertise of leading research centers all over Europe, the CORNEAGENE project unifies sufficient resources to make this group the leading force in the research of corneal dystrophies.

2.1The role of the participants within the consortium and the specific skills of each partners

2.1.1Core Group:

Organization / Country / Chief scientist / Area of excellence / Role in the project
1 / Varimed Ltd. / Hungary / Dr. Sandor G. Vari / Management and coordination of EU projects, web based electronic record, eForms. / Creation of common platform for cornea research via WEB based virtual and interactive working methods Project NetBoard, web based electronic patient records, eForms for genomic and proteomic protocols. WEB based interactive education tools for young scientist and for the public.
Infonet Kranj Slovenia / Slovenia / Bostjan Bercic / Web based shared medical record, associated contractor
Vitamib Sarl. / France / Xavier Fabre / eForms, electronic database
2 / University of Debrecen, Dept. of Ophthalmology / Hungary / Prof. Dr. Andras Berta / Corneal research and transplantation center / Scientific co-ordination,
Blood and corneal sample collection, DNA analysis for BIGH3 mutations, corneal cell culture , immunohistology and biochemistry of corneal dystrophies
Dept. of Biophysics / G. Vereb / Basic research on cell biology / Cell culture and transformation
Dept. of Clinical Biochemistry and Molecular Path. / G. Losonczy / Molecular genetics / DNA extraction and mutation analysis
3 / Cedars-Sinai Medical Center, Ophthalmology Research Department of Surgery / California, US / Prof. Dr. Yaron Rabinowitz / Corneal research and transplantation center / Mutation analysis of BIGH3 and VSX genes, high output DNA sequencing, create and store lymphoblastoid cell lines, proteomics studies of corneal dystrophies. Accesses to the library of corneal genes at the National Eye Institutes of Health for studying the corneal dystrophy and high output sequencer from NIH
4 / Dept. Of Ophthalmology, Friedrich-Alexander-Universitat, Erlangen-Nürnberg / Germany / Prof. G.O.H. Naumann / Corneal transplantation and research center / Collection of blood and corneal samples
5 / Community Hospital, Arhus, Dept. of Ophthalmology / Denmark / Prof. Niels Ehlers / Corneal transplantation and research center / Collection of blood and corneal samples, proteomics study of BIGH3, (EU center for proteomics studies – Head of RA6)
Keratoconus genomics database, DNA microarrays
6 / Univ. of Helsinki, Central Hospital / Finland / Prof. Timo Tervo / Corneal transplantation and research center / Keratoconus research, collection of blood and corneal samples, genetic analysis, corneal confocal microscopy
7 / Hadassah University Hospital, Dept. of Ophthalmology / Israel / Prof. Joseph Frucht-Pery / Corneal transplantation and research center / Collection of blood and corneal samples
8 / Ophthalmic Dept., Hopital Jules Gonin
and Division of Medical Genetics / Schwitzerland / Dr. Francis L. Munier
Daniel F. Schorderet / Corneal transplantation and Oculogentics research center / Mutation analysis of BIGH3 gene through DHPLC screening and high output DNA sequencing, proteomics studies of corneal dystrophies. Models of BIGH3 (overexpression of mutants and knock-out). Analysis of cellular pathways involved in the pathogenesis of corneal dystrophies
9 / Lublin Univ. School of Medicine, Ophthalmology / Poland / Prof. Zbigniew Zagorski / Corneal transplantation and research center / Collection of blood and corneal samples, DNA extraction and mutation analysis, and keratoconus research
10 / Charles University, Dept. of Ophthalmology / Czech Republic / Prof. Martin Filipec / Corneal transplantation and research center / DNA microarrays and mutation analysis, Keratoconus research, corneal cell culture
11 / Slovak Postgradual Academy of Medicine, Ophthalmic Department / Slovakia / Prof. Andrej Cernak / Corneal transplantation and research center / Collection of blood and corneal samples
12 / Lambda GmbH / Austria / genomics and proteomics research / genetic analysis, standardisation of technologies
13 / The Filatov Institute of Eye Diseases and Tissue Therapy / Ukraine / Galina I. Drozhyna / Corneal transplantation and research center, / Keratoconus research, collection of blood and corneal samples
Department of Human Genomics Institute of Molecular Biology and Genetics National Ac. of Sci of Ukraine / Prof. Ludmila A. Livshits / Molecular genetics / Keratoconus research, collection of blood and corneal samples, genetic analysis, corneal confocal microscopy

2.1.2Assistants to core group

Organization / Country / Chief scientist / Area of excellence / Role in the project
1 / Charles University, inst. of Metabolic Diseases / Czech Republic / Dr. Stanislav Kmoch / Molecular genetics / DNA microarrays, mutation analysis

INTEGRATION AND STRUCTURING EFFECT: The means by which the Network of Excellence will have a structuring and integrating effect on European research and assist in spreading European scientific excellence

3.1Joint research activities:

-The basis of the joint research activities will be the collection of blood and corneal samples from dystrophic patients in all the clinical centers. Trans-European collecting and circulating the samples to the centers having the most expertise in the research of certain diseases will enhance effectiveness in itself, by providing more research material. Co-operation between relevant centers in each research area will allow more detailed approach to special problems. Blood samples will be taken from all patients and available relatives suffering from corneal dystrophies. Corneal buttons, excised during corneal transplantation will be halved- one half will be frozen for biochemical and histological studies, the other half will be stored fresh and used for establishment of immortalized cell lines

- the research activities will be subdivided into four research activities (RA) and into several workpackages tasks:

  • RA1 - Common platform for cornea research: adaptation of the partners, research activities in order to strengthen their complementary, development and use of joint research infrastructure, analysis of the scientific/society issues related in the cornea research network
  • RA2-collection of blood and corneal samples: from dystrophic patients in all the clinical centers
  • RA3 - genetic analysis: search for new mutations in corneal dystrophies – correlation of genetic data with histological and biochemical characteristics of the corneal dystrophies
  • RA4 - keratoconus study: gene expression study by DNA microarray systems, investigation of the role of the VSX genes in the pathogenesis of keratoconus
  • RA5 - cell culture and transformation: establishment of immortalized cell lines from dystrophic and normal corneas, to provide suitable model systems for in vivo studies
  • RA6 - proteomics: investigation of the effect of genetic mutations on protein function and protein-protein interaction
  • RA7 – WEB based virtual and interactive working methods: communication of the achievements of the network and the dissemination of knowledge, interactive web based training of researchers, support of technological innovation in SME

Each partner will participate in one or several WP-s according to his or her expertise in the following way:

3.2Integration activities:

-Partners will communicate with each other on a daily basis via the web site established for the participants of the project. The web site will be updated regularly and contain relevant information on the actual state of each WP, as well as the availability of samples in each centers

-Subgroups participating in the different WP-s will launch joint projects to address individual problems

-Personnel mobility will help in sharing expertise and establish long lasting interaction between centers

3.3The operation of the network

The workpackages (WPs) are the hubs for the step wise project implementation, and each WP will be completed by a group of participants. In order to do so the Working Groups must organise themselves as an organisation, combining or integrating work and learning. To provide structural frame to achieve the goals of the CORNEAGENE project the participants will form:

The governing council (GC) made up of senior representatives of the core group organizations; VARIMED Ltd. University of Debrecen, Dept. of Ophthalmology, Cedars-Sinai Medical Center, Ophthalmology Research Department of Surgery, Dept. of Ophthalmology, Friedrich-Alexander-Universitat, Erlangen-Nürnberg, Community Hospital, Arhus, Dept. of Ophthalmology, Charles University, Dept. of Ophthalmology, The Filatov Institute of Eye Diseases and Tissue Therapy, Lambda GmbH. The GC will co-ordinate and supervise the co-operation between different research centers and makes decision on future work. Members of the GC will use the project NetBoard for daily communication and have regular meetings.

The WP councils (WPC) made up of representatives taking part in the work packages. The WPC-s will co-ordinate the work of the participants of each WP-s. They will use the project NetBoard for communication and organise meetings when needed.

Management Structure and decision process

The GOVERNING COUNCIL assists the COORDINATOR for decisions concerning achievements of the project and overall quality in order to: monitor the work toward internal and external deliverables. They seek for consistency in the progress of the project phases; ensure adherence of events to the project plan; adapt the planning to unforeseen distortions of the work-plan; implement the continuous review and assessment of project achievements. They select the external end-users who will be sub-contracted to participate to demonstration tasks; handle contractual matter; ensure overall quality of the activities respective outputs; control the overall project development plan having incidence on the schedule of the deliverables. They promote the complementary of medical, scientific, and technological competencies; foster the synergy between medical actors and technology suppliers.

Co-operation between the partners is promoted through regular meetings of the GOVERNING COUNCIL every 3 months (more frequently during the first 6 months of the project). Every six months a mandatory self-evaluation will take place, where the partners and WP COUNCILS will review the deadlines and the progress of the project. The meetings are organised by the board members, one after the other, and will provide the opportunity to visit the places where significant work is done. Meeting cost is shared among attendants and incurred on their respective project budgets. Meeting dates and agenda are fixed in advance and, at the latest, from one meeting to the next.

3.4The plan for the dissemination

The medical participants in CORNEAGENE are participating in international medical societies, in which they often play a leading role, and are contributing to international works such as International Council of Ophthalmology, Societas Ophthalmologica Europaea, and European Eye Bank Association. They are in a key position to communicate medical results from the project and ensure that these results will be transferred to other organisations in EUROPE. To strengthen this communication, members of those scientific and health care organisations will be regularly invited to participate CORNEAGENE meetings to discuss achievements. The Web Database will be the core of the system and its information should be defined with the view to maximise the opportunity for co-operation, data sharing, optimisation of the resources, dissemination of the information, standardisation and build-up of a European dimension in this sector. Considering the high scientific and social value of the project an accurate communication and dissemination activity is particularly required in order to ensure the widest visibility of the project results to the public and to promote the contribution to the healthcare policy.

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EoI CORNEAGENE Final09/14/18