The present proposal concentrates the activities of six European Laboratories working in the field of Regeneration. The ultimate aim will be a full understanding of the factors being involved in regenerative events in the central nervous system (CNS) and thereby providing the basis for the development of therapeutic tools. A central issue addressed in the project is the development of tools which allow to screen for bioactive compounds on an industrial scale and to selectively interfere with individual steps relevant for Regeneration. These tools will be optimised to allow the study of all steps in successful regeneration by comparison with appropriate in-vivo-models. These aims are reflected in the selection of participating groups which concentrate expertise in the fields of biotechnology, as well as in experimental approaches to individual steps relevant for Regeneration. As an integrative approach and an understanding of the interdependence of these steps is, as yet, not established, the project does not include industrial partners. However, it is the principal aim of the project to set the stage for an industrial exploitation by developing standardised model systems which fulfil the necessary perequisites for large-scale use and the application on different issues in Regeneration-research.
A variety of mechanisms have to interact closely in order to achieve successful regeneration in the CNS. Therefore we will combine research from several different fields and apply the results at a combinatorial and highly interactive level. The present proposal will address the following points: 1) Analysis and experimental modification of the key-processes in successful regeneration, namely neuronal survival, activation of growth programmes, axon elongation, and appropriate synapse formation.
2) Development of model systems which allow to monitor and influence regeneration and which may ultimately be used for industrial applications, e.g. development and screening of therapeutic agents.
3) Development and application of biotechnological tools for the experimental induction or support of mechanisms crucial for successful regeneration. The key-mechanisms for successful regeneration addressed in the project are: i) rescue of neurons after insult by suppression of apoptosis, ii) activation of genetic programmes related to axonal growth, iii) appropriate modification of the axonal growth environment, iv) maintenance or reinduction of mechanisms underlying axonal pathfinding, v) induction of activity-dependent refinement of axonal projections.
Each individual question will be addressed by at least one group highly specialised in the given field. By extensive collaborations these results will then be combined to investigate the interdependence of the individual mechanisms. In order to optimize the collaborative nature of the project we will focus on two model systems, the visual pathway and the olivo-cerebellar system. For both systems we will develop organotypic culture models which will be compared to and optimised according to in-vivo preparations. The initial experimental strategies for the improvement of Regeneration will be the overexpression of genes controlling cell death, e.g. bc1-2 (Drs. Maffei/Müller), the modification of the axonal growth environment by the controlled application of blocking antibodies/peptides (Dr. Schwab in collaboration with all groups) or growth factors from genetically engineered cell-lines (Dr. Arenas in collaboration with all groups), and the overexpression of genes involved in axonal growth, e.g. GAP-43 (Drs. Strata/Sotelo). Furthermore, we will search for additional molecules implicated in the support or suppression of regenerative events by comparing regenerating and non-regenerating pathways (Drs. Sotelo/Strata) and attempt to develop appropriate biotechnological tools for the selective manipulation of such factors.
Funding SchemeCSC - Cost-sharing contracts
171 77 Stockholm
10126 Torino (Turin)