Thursday, April 25, 2024
4/25/2024
PROJECT SUMMARY: Type 2 diabetes is associated with loss of activity and expression of pancreas-enriched transcription factors (TFs) that control gene expression programs necessary for mature ß-cell function. Among these is Pdx1, which plays key roles during early pancreas development, islet endocrine cell differentiation, and ß-cell development and function. Pdx1, like most TFs, requires additional regulation from recruited coregulators to modulate specific regulatory programs. Numerous coregulators have been identified as Pdx1-interacting partners including the ATP-dependent Swi/Snf chromatin remodeling complex. In the early developing pancreas, Swi/Snf plays a critical role in progenitor cell expansion, where early pancreatic progenitor cell deletion of one essential Swi/Snf ATPase subunit, Brg1, results in a 50% reduction in final pancreas mass. In the mature ß-cell, deletion of both Swi/Snf ATPase subunits, Brg1 and Brm, impairs whole-body glucose tolerance through severe loss of insulin production, which is largely driven by a loss of Pdx1 occupancy on the insulin gene promoter. While Swi/Snf plays an essential role in the early developing pancreas and the mature ß-cell, the contribution of Swi/Snf chromatin remodeling activity to islet endocrine progenitor development has not yet been explored. Herein, I will test the hypothesis that the Pdx1-recruited Swi/Snf chromatin remodeling complex dynamically controls the chromatin landscape and expression of genes essential for endocrine progenitor cell development and postnatal islet function. Mutant mice with endocrine-specific deletions of either Brg1, Brm, or both subunits were generated to determine the mechanistic actions of Swi/Snf at this stage of islet development and evaluate the postnatal consequences of losing Swi/Snf during islet endocrine cell development. Preliminary results demonstrate that loss of the Brg1 subunit, but not Brm, from endocrine progenitors leads to severe glucose dyshomeostasis beginning at 4 weeks of age with a reduction in plasma insulin levels, suggesting that Brg1 is essential for proper islet development and function. Remarkably, no mice deficient for both subunits have been recovered at weaning, indicating that total loss of Swi/Snf activity results in postnatal lethality. Aim 1 will explore the anatomical and physiological attributes driving the postnatal phenotype observed in the Swi/Snf mutants through quantitation of islet cell mass and evaluation of islet function through perifusion analysis on isolated islets. Aim 2 will investigate the mechanistic actions of Swi/Snf in controlling chromatin accessibility, TF recruitment, and gene expression programs in endocrine progenitor cells with RNA-sequencing, ATAC-sequencing, and ChIP-qPCR approaches. With this F31 Predoctoral Fellowship, I will be able to commit my time to completing the research described in the Aims outlined in this application, while also focusing on career development and enhancing my scientific skillset through attending seminars and workshops. Indiana University School of Medicine is equipped with experienced faculty and state-of-the-art core facilities to assist in carrying out this research proposal and provide guidance in my training.
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