Projet ANR-07-BLAN-0078 – CRUMBS : Role of a polarity complex in epithelial morphogenesis
Programme : Blanc (BLANC) 2007
Année de financement : 2007
Coordinateur du projet : André LE BIVIC (CNRS DELEGATION REGIONALE PROVENCE ET CORSE)
Partenaires : Centre national de la recherche scientifique
Montant : 250000.0 euros
Date de début : 2007
Durée : 36 mois
Résumé : The goal of our project is to try to understand the integrated role of a polarity complex in epithelial cells as a model of a morphogenetic protein complex in Cell Biology. We have decided to focus on the role of the Crumbs 3 (CRB3) complex in mammalian epithelial cells since we have developed expertise and tools both in the study of this complex and in epithelial cell biology. Our group is the only one in France to have started the study of this essential apical complex some years ago and we have been able to put our mark in this field with original findings. Now that the identification of the molecular components of this complex is almost completed we want to replace this complex in a cellular network of connections with the goal to understand its role in epithelial morphogenesis and homeostasis since the role of the CRB3 goes far beyond its direct effect on apical junctions but extend to the regulation of apical membrane traffic, primary cilium formation and cell survival and apoptosis. Finally, we will start a search for new regulators of mammalian epithelial morphogenesis with the long-term goal to reconstruct the molecular networks that are necessary and sufficient to build a fully functional epithelial cell. In the project described here we will use a combination of classical immuno-confocal and electron microscopy, live imaging of fluorescent tagged-proteins, RNA interference with siRNA or inducible shRNA and protein-protein interaction techniques. A bio-informatic approach will be used to try to build models of Crb (and other polarity complexes) function in epithelial cells. A further step will be to test in vivo the prediction using mouse retina and kidney as models together with targeted loss of function with the Cre-Lox technology but this will be done on a longer frame of time than the proposal submitted for this application. To develop this project we have the scientific and technical environment at the IBDML in particular we will make use of our Imaging platform PICSL and of the Bio-informatics platform. We hope that we will be able to decipher the mechanisms used by one polarity complex to establish and maintain a proper epithelial organization and by comparing our findings to those obtained in Drosophila to have a view of how evolution has made use of these mechanisms.To this aim we will hire a post-doc fellow for the first two years (Carole Baron-Gaillard) and we will need to use the 3 year of salary in 2008 to hire two short-term post-doc (2x 5 months) to set-up some biochemical and cellular experiments described in the project. Finally the system biology analysis will be parformed by a post-doc hired in 2009 on an other fincial support (EC grant 2008-2010).                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                              ...