TY - JOUR T1 - Investigation of the role of a macromolecular complex of CFTR-NHERF2-LPA2 in the fluid hemostasis and inflammatory responses in intestinal epithelial cells JF - bioRxiv DO - 10.1101/186023 SP - 186023 AU - Shanshan Kong AU - Weiqiang Zhang Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/09/07/186023.abstract N2 - CFTR is a cAMP-regulated chloride channel located in the apical surface of intestinal epithelial cells; where it forms a macromolecular complex with NHERF2 and LPA2. CFTR has been shown to play a role in the pathogenies of several types of secretory diarrheas. Inflammatory bowel disease (IBD) is a chronic condition of intestine characterized by severe inflammation and mucosal destruction, genetic analysis has shown that LPA contribute to IBD and patients of cystic fibrosis also display the phenotype of diarrhea. The purpose of this study is to investigate if this complex plays a role in the pathogenesis of IBD, especially in the intestinal fluid homeostasis and inflammatory responses.To investigate the role(s) of CFTR-NHERF2-LPA2 complex in the pathogenesis of IBD, we first identified the existence of this complex in intestinal epithelial cells; our results showed that (1) CFTR, NHERF2, LPA2 are expressed in these cells evidenced by western blotting and Q-PCR; (2) NHERF2 and LPA2 can be co-immunoprecipated with CFTR; (3) NHERF2 and LPA2 co-localize with CFTR at the plasma membrane of these cells; (4) NHERF2 and LPA2 interact with CFTR evidenced by proximity ligation assay. We then explored the role of this complex in maintaining the integrity of tight junction and inflammatory responses in these cells. Our preliminary data showed that inhibition of CFTR disrupted the tight junction and elicited the secretion of IL-8, while intriguing LPA2 increased the expression of IL-8. Our data also show that RNA knockdown LPA2 can decrease the expression of IL-8. These data suggest that CFTR-NHERF2-LPA2 might play a role in the fluid hemostasis and inflammatory responses of intestinal epithelium, thus could play a role in the pathogenesis of IBD. ER -