Circulating extracellular vesicles as a marker of human Type 1 Diabetes pathogenesis

About This Grant

PROJECT SUMMARY/ABSTRACT (30 lines or less) The premise for the proposed research stems from precedence in other diseases, such as cancer, cardiovascular, neurodegenerative, and most relevant to the current proposal, autoimmune diseases, in which circulating extracellular vesicles (cEVs) play a role in pathophysiology and are important biomarkers for early detection. However, little is known concerning the role of cEVs in human type 1 diabetes (T1D). Our overall hypothesis is that cEVs have the potential to be used as biomarkers for early pre-disease detection of T1D, based on their distinct molecular and functional phenotype in T1D and pre-disease stages, compared to healthy or low risk individuals. Our specific aims are: 1) To identify the distinct protein and RNA cargo unique to cEVs at different stages of T1D disease development; and 2) To investigate the effect of cEVs from subjects at different stages of disease progression on immune and beta cell phenotypes and elucidate the functional relevance of their distinct molecular cargo. To address these aims, we have assembled a team of investigators with highly relevant expertise and techniques. We propose to use longitudinal samples from The Environmental Determinants of Diabetes in the Young (TEDDY) study of children with T1D associated genetic risk to identify critical timepoints and underlying mechanisms mediating, A) the earliest stages of pathogenesis preceding AAb appearance of the first islet autoantibody, and B) the period of seroconversion from single to multipe AAb+ or remaining single AAb+ and C) the period after multiple AAb appearance with a highly variable rate of progression to hyperglycemia. We have the expertise and technical ability to isolate cEVs from plasma, perform proteomic and RNAseq analysis on EVs, and perform immune and beta cell related functional assays. Our Research Plan is to generate cEVs from donors at different stages of T1D disease progression, to identify the uniquely packaged protein and RNA cargo from these cEVs, to evaluate the effects of the cEVs on immune cell functional phenotype and islet health and elucidate the functional relevance of the distinct molecular cargo targets. These studies will yield novel mechanistic insights into early disease pathogenesis and identify potential novel biomarkers for T1D initiation and progression.