miRNA expression profiling of intestinal epithelial cell-derived extracellular vesicles from mice following Paneth cell ablation by dithizone

Paneth cells, critical sentinels of the innate immune system within the intestinal epithelium, play a vital role in maintaining intestinal homeostasis and defending against pathogens. Their dysfunction is implicated in the pathogenesis of sepsis. However, the specific mechanisms by which Paneth cells communicate with and regulate the surrounding intestinal microenvironment, particularly via extracellular vesicles (EVs), remain poorly understood. This study aims to characterize the global miRNA expression profile of intestinal epithelial cell-derived EVs (IEC-EVs) in a mouse model of acute Paneth cell loss, to identify key intercellular signaling molecules that may mediate intestinal barrier response to injury. We employed a well-established model of Paneth cell ablation by a single intraperitoneal injection of dithizone in C57BL/6 mice. Control mice received a vehicle solution. Intestinal epithelial cells were isolated six hours post-injection, and IEC-EVs were subsequently purified from these cells. Small RNA from the purified IEC-EVs was extracted and subjected to next-generation sequencing to comprehensively profile the miRNA cargo. Our comparative analysis reveals a distinct set of miRNAs that are differentially expressed in IEC-EVs following Paneth cell ablation. These candidate miRNAs are predicted to target signaling pathways crucial for epithelial cell proliferation, apoptosis, and inflammatory responses. We hypothesize that the alteration of the miRNA landscape in IEC-EVs represents a novel mechanism of intercellular communication that orchestrates the intestinal epithelial response to damage. This dataset provides the comprehensive resource of miRNA expression in IEC-EVs under conditions of Paneth cell deficiency. The findings are expected to elucidate novel pathways in intestinal pathophysiology and may identify potential miRNA biomarkers or therapeutic targets for conditions involving intestinal barrier dysfunction, such as sepsis. Overall design: miRNA sequencing of intestinal epithelial cell-derived extracellular vesicles from C57BL/6 mice (n=3 per group) following Paneth cell ablation with dithizone versus vehicle control.