Creatine Transporter, Reduced in Colon Tissues From Patients With Inflammatory Bowel Diseases, Regulates Energy Balance in Intestinal Epithelial Cells, Epithelial Integrity, and Barrier Function.

Abstract Background & Aims Patients with inflammatory bowel diseases (IBD) have intestinal barrier dysfunction. Creatine regulates energy distribution within cells and reduces the severity of colitis in mice. We studied the functions of the creatine transporter solute carrier family 6 member 8 (SLC6A8, also called CRT) in intestinal epithelial cells (IECs) and mice, and measured levels in mucosal biopsies from patients with IBD. Methods Colon biopsies from patients with IBD (30 with Crohn’s disease and 27 with ulcerative colitis) and 30 patients without IBD (controls), and colon tissues from mice (with and without disruption of Crt), were analyzed by immunofluorescence, immunoblots, and/or quantitative reverse-transcription PCR (qPCR). CRT was knocked down or overexpressed in T84 cells, which were analyzed by immunofluorescence, immunoblots, high-performance liquid chromatography (to measure creatine levels), qPCR, transepithelial electrical resistance, barrier function, actin localization, wound healing, mitochondrial oxygen consumption, and glycolysis extracellular acidification rate assays. Organoids from colon cells of CRT-knockout mice and control mice were analyzed by qPCR, immunoblot and transepithelial electrical resistance. Results CRT localized around tight junctions (TJs) of T84 IECs. In analyses of IECs with CRT knockdown or overexpression, we found that CRT regulates intracellular creatine, barrier formation, and wound healing. CRT-knockout organoids also had diminished barrier formation. In the absence of adequate creatine, IECs transition toward a stressed, glycolysis-predominant form of metabolism; this resulted in leaky TJs and mislocalization of actin and TJ proteins. Colon tissues from patients with IBD had reduced levels of CRT mRNA compared with controls. Conclusions In an analysis of IEC cell lines and colonoids derived from CRT-knockout mice, we found that CRT regulates energy balance in IECs, and thereby epithelial integrity and barrier function. Mucosal biopsies from patients with ulcerative colitis and inactive Crohn’s disease have lower levels of CRT, which might contribute to the reduced barrier function observed in patients with IBD.