Extracellular Vesicles from Enterocytes are Mediators of Inflammation in IBD - Inflammatory bowel disease is a chronic and debilitating condition with no cure. It has been challenging to understand the underlying molecular bases of intestinal inflammation. Investigation into proinflammatory signaling requires a more thorough understanding of cell-to-cell communication between enterocytes. Extracellular vesicles (EVs) and submicron sized particles that carry genetic material as a means of messenger delivery between cells. This project seeks to understand how EVs from enterocytes are involved in the pathogenesis of inflammation. The long-term goal is to understand how inflammation prevents induction and maintenance of remission in inflammatory bowel disease. The immediate goal of this project is to understand how EVs from enterocytes mediate cell-to-cell communication to function as effectors of inflammation in the intestinal epithelial barrier. In Specific Aim 1, we will identify the transcriptomic miRNA signature of EVs produced by enterocytes in inflammation which will be induced by dextran sodium sulfate or lipopolysaccharide. In Specific Aim 2, we will determine how EVs from the stimulated enterocytes alter the inflammatory signaling, barrier function, and migration of recipient enterocytes. This study will be conducted in an in vitro model of HT-29 and C2BBe1 cells which can be induced by pro-inflammatory stimuli to study to hallmarks of inflammatory bowel disease. Cells will be grown in transwells and EVs will be isolated from apical and basolateral media of differentiated monolayers for analysis of miRNA profile. To assess how inflammation alters the composition of EVs, the miRNA content of EVs grown in typical conditions will be compared to EVs produced from enterocytes treated with an acute inflammatory stimulus. Finally, we will assess how EVs from stimulated enterocytes alter the expression of proinflammatory mediators, permeability of the enterocyte monolayer, and wound healing in recipient HT-29 and C2BBe1 cells. This research is aimed to foster the development of the scientific workforce by recruiting six undergraduate biochemistry students for an immersive research experience alongside the PI. The findings from this research study will provide a greater understanding about the role of EVs in inflammatory signaling, and epithelial barrier & enterocyte functions. With novel knowledge of the mechanism of inflammation, researchers can better understand the pathogenesis of IBD, develop new targeted therapies, and provide enhanced care for patients. This research is important because it has the potential to improve outcomes for patients with intestinal inflammation including IBD to achieve sustained remission.
