IDENTIFICATION OF A NOVEL GENE EXPRESSED DURING MOUSE PANCREAS DEVELOPMENT

Cell therapy offers good prospects for the treatment of pancreatic diseases, for this reason more attention has been given to the pancreatic progenitor cells. Several studies show the improvement of differentiation of embryonic stem cells towards the definitive endoderm and pancreatic progenitors, but the identification of new molecular markers could further facilitate differentiation. The aim of this study is to find new molecular markers involved in pancreatic specification. Therefore, our efforts will be directed towards in vitro and in vivo assessment of signature genes that would target the specification formation of pancreatic progenitor cells . Our approach to identify in vivo pancreas progenitor cell markers will be based on the analysis of global gene expression profile of dorsal pancreatic buds isolated from E10.5 mouse embryos. There is a signi?cant gap in our knowledge on the endodermal players that act in the time window between the early stages of endoderm formation and the regionalized expression of the posterior-foregut/pancreas marker gene Pdx1. Development of more precise lineage-tracing tools allow a deeper focus on the functional delineation between stages and mechanisms of speci?cation, commitment, lineage allocation, and differentiation. In particular, conditional gene knockout strategy will be used to characterize spatio-temporal function of the putative markers during pancreatic specification. The study will be followed by in vitro characterization of a candidate during differentiation of embryonic stem cells into the pancreatic lineage. We propose to develop a protocol based on the sequential activation of signaling pathways that recapitulate pancreatic development in vitro, through the definitive endoderm formation, the pancreatic specification. We aim to generate genetically modified ES cell line expressing the beta-galactosidase under the promoter of the gene candidate to enable the stain and the collection of the cell population that expresses the novel putative marker of pancreatic cell specification. In this way, we would like to characterize both the global molecular signature of those cells, and eventually evaluate their ability to repopulate a diseased and/or damaged pancreas after current transplantation procedures.