Clostridioides difficile Toxins Alter Host Metabolic Pathway and Bile Acid Homeostasis Gene Expression In Colonic Epithelium

Abstract

A major risk factor for acquiring Clostridioides difficile is antibiotic usage that disrupts a healthy microbial gut community facilitating establishment of infection. Once established, C. difficile secretes endotoxins (TcdA and TcdB) that are internalized into host colonic epithelial cells where they disrupt gut barrier function and induce hyper inflammation resulting in severe diarrhea and possibly leading to death. We employed three different platforms to explore gene expression of cells in the gut when exposed to C. difficile or its toxins, TcdA and TcdB. An antibiotic treated mouse model of C. difficile infection (CDI) was used to identify differential gene expression with a NanoString Technologies mouse inflammatory gene panel consisting of 770 genes including a subset of bile acid (BA) homeostasis and nuclear receptor genes. In the cecal tissue of mice with CDI, significant down expression was observed for genes involved in PPAR signaling, cholesterol and glucose metabolism, while a significant increase in expression was observed for IL-17 related inflammatory genes. Similarly, Caco-2 cell culture and primary human colonocytes (hCE) exposed to toxins for 24 hours showed altered expression in several PPAR regulated and cholesterol metabolic genes similar to those found in mice. These cell culture experiments also revealed significant alterations in gene expression of the FXR BA regulatory pathway. Together these data suggest that exposure to C. difficile and its toxins may alter host cholesterol metabolic processes, including BA transport and synthesis.