Epithelial Muscarinic Receptor 3 Ablation Induces Sex-Specific Modulation of Colonic Intestinal Progenitor Cells and Response to Intestinal Injury [586CW]

In this study, we investigated regional and sex differences in the cholinergic modulation of intestinal epithelial cells in homeostasis and regeneration. Genetic intestinal epithelial ablation of the M3R employing Vil-Cre x M3R fl/fl mice interestingly resulted in the prominent reduction in Lgr5-expressing progenitor cells in male tissues, contrasting an expansion of these cells in the female intestinal epithelium. This prominent difference showed abrogated in young female mice with reduced circulating sex hormone levels. M3R ablation further induced the expansion of pEGFR+ tuft cells and induced the activation of the PI3K/Akt pathway. Importantly, sex-specific differences in Lgr5-expressing progenitor cells translated into the development of severe inflammation in male Vil-Cre x M3R fl/fl mice subjected to an acute colitis model, which did almost not affect female Vil-Cre x M3R fl/fl mice. In addition, sex-specific effects of modulations of cholinergic signaling on epithelial cells could be corroborated in murine and human colonoids. Collectively, our data reveal regional and sex differences in the cholinergic, muscarinic modulation of intestinal epithelial cells. Bulk gene expression data are presented for epithelial cells following treatment with Bethanechol (n=8) or no treatment (n=8) grown in female (n=4) and male (n=4) human colonic organoids.