Disruption of CerS6-mediated sphingolipid metabolism by FTO deficiency aggravates ulcerative colitis

Background and aims Deregulation of RNA N6-methyladenosine (m 6 A) modification in intestinal epithelial cells (IECs) influences intestinal immune cells and leads to intestinal inflammation. We studied the function of fat mass-and obesity-associated protein (FTO), one of the m 6 A demethylases, in patients with ulcerative colitis (UC). Methods We analysed colon tissues of Fto flox/flox ; Villin-cre mice and their Fto flox/flox littermates with dextran sulfate sodium (DSS) using real-time PCR and 16s rRNA sequencing. RNA and methylated RNA immunoprecipitation sequencing were used to analyse immunocytes and IECs. Macrophages were treated with conditioned medium of FTO-knockdown MODE-K cells or sphingosine-1-phosphate (S1P) and analysed for gene expression. Liquid chromatograph mass spectrometry identified C 16 -ceramide. Results FTO downregulation was identified in our in-house cohort and external cohorts of UC patients. Dysbiosis of gut microbiota, increased infiltration of proinflammatory macrophages, and enhanced differentiation of Th17 cells were observed in Fto flox/flox ;Villin-cre mice under DSS treatment. FTO deficiency resulted in an increase in m 6 A modification and a decrease in mRNA stability of CerS6, the gene encoding ceramide synthetase, leading to the downregulation of CerS6 and the accumulation of S1P in IECs. Subsequentially, the secretion of S1P by IECs triggered proinflammatory macrophages to secrete serum amyloid A protein 1/3, ultimately inducing Th17 cell differentiation. In addition, through bioinformatic analysis and experimental validation, we identified UC patients with lower FTO expression might respond better to vedolizumab treatment. Conclusions FTO downregulation promoted UC by decreasing CerS6 expression, leading to increased S1P accumulation in IECs and aggravating colitis via m 6 A-dependent mechanisms. Lower FTO expression in UC patients may enhance their response to vedolizumab treatment.