Protecting the beta cell in type 1 diabetes mellitus through better understanding of the intracellular signaling cascades: interventions in pre-clinical models.

Our main goal in the current project is to bridge the in vitro knowledge on beta-cell death and dysfunction to pre-clinical in vivo models of type 1 diabetes. The use of genetically-manipulated animal models as well as targeted interventions (using gene-therapy and anti-sense methodologies) in these and spontaneous models of diabetes mellitus will allow studying the role of key molecules in the effector pathways of beta-cell death and dysfunction in vivo. Genes and their products involved in allogeneic islet graft rejection and autoimmune destruction will be studied using molecular (quantitative RT-PCR) and imaging (multicolour flow cytometry and confocal microscopy) techniques. The introduction of proteomics (two-dimensional difference gel electrophoresis (2D DIGE)) will further provide information on the regulation of key proteins, their biochemical functions and their potential interaction partners in beta-cell damage. Moreover, using histological and flow cytometry techniques, we will be able to evaluate the influence of changes induced in the transplanted beta cells on the nature of the infiltrating immune cells. This information should lead to the development of novel therapeutic strategies that render beta cells more resistant against immune-mediated beta-cell damage improving early and long-term beta-cell survival. Specific aims of the project: 1. Improving early and late beta-cell survival by blocking distinct transcription factors involved in inflammation-mediated beta-cell destruction 2. Understanding the function of ER stress-regulated proteins, CHOP and BiP, in beta-cell death and protection in vitro and in vivo by use of full knock out models and by construction of new transgenic mice 3. Use of proteomics to confirm/compare changes in mRNA expression observed during immune-mediated beta-cell death and dysfunction

Keywords:viral gene therapy, proteomics, intracellular pathways, beta-cell death

Disciplines:Immunology, Endocrinology and metabolic diseases