Small molecule focal adhesion kinase activator alters macrophage chemotaxis and polarization 3055

Abstract Description A novel drug-like, water-soluble molecule, M64HCl, specifically activates focal adhesion kinase (FAK). M64HCl increases FAK Y397 phosphorylation in intestinal epithelial cells and leads to epithelial sheet migration and intestinal wound healing. Macrophages also aid in intestinal wound healing. M1 macrophages phagocytize microbes and trigger the inflammatory response, M2 macrophages release anti-inflammatory cytokines and promote tissue repair and angiogenesis. In macrophages, FAK regulates cell adhesion, migration, and proliferation under basal or inflammatory conditions. We treated mouse RAW 264.7 and human PMA-stimulated THP-1 macrophage cell lines with M64HCl and confirmed an increase in FAK Y397 phosphorylation. When RAW 264.7 or THP-1 cells were treated with M64HCl, less macrophages polarized to M1 while there was an increase in M2 macrophage polarization. Next, M64HCl treatment led to an increase in RAW 264.7 or THP-1 cell transwell chemotaxis towards injured Caco2 cells. This increase in macrophage chemotaxis could be due to the correlated increase of VEGF and TGFβ protein expressed by M64HCl-treated Caco2 cells. Confocal microscopy of intestinal tissues injured by indomethacin in CX3CR1GFP/+ CCR2RFP/+ mice +/- M64HCl, showed that CX3CR1+ CCR2 + (transition) and CX3CR1+ CCR2- (M2) cells were in greater number and diffused in with the epithelial cells in the control mice whereas M64HCl had fewer cells that were more confined to the lamina propria. Funding Sources Supported by UND internal funds Topic Categories Mucosal and Regional Immunology (MUC)