METTL3‐mediated m6A modification of circPRKAR1B promotes Crohn's colitis by inducing pyroptosis via autophagy inhibition

Abstract

AbstractBackgroundThe roles of circRNA and N6‐methyladenosine (m6A) methylation in Crohn's disease (CD) have drawn much attention. Therefore, this investigation aimed to discover how the m6A modification of circRNAs contributes to CD progression.MethodsThe study performed circRNA sequencing on colon samples from four CD patients and four normal controls (NCs) to screen for dysregulated circRNAs. Quantitative real‐time polymerase chain reaction (qRT‐PCR) was performed to validate the candidate circRNA expression and determine its correlation to CD‐associated inflammatory indicators. In vivo and in vitro investigations were conducted to examine the functions and pathways of circPRKAR1B in CD, besides investigating the m6A modification role in circRNA expression modulation.ResultsThe RNA‐seq revealed that hsa_circ_0008039 (circPRKAR1B) was the most significant upregulated circRNA and was identified as the candidate circRNA for further examinations. Relative circPRKAR1B expression was significantly upregulated in CD colon tissues and closely related to CD‐associated inflammatory indices. The circPRKAR1B expression and function were regulated by methyltransferase‐like 3 (METTL3)‐mediated m6A methylation. In vitro studies indicated that circPRKAR1B promoted pyroptosis mediated by NLRP3 inflammasome (NLRP3; nucleotide‐binding oligomerization domain, leucine‐rich repeat and pyrin domain‐containing 3) and impaired autophagy by interacting with the RNA‐binding protein (RBP) SPTBN1, (SPTBN1; spectrin beta, non‐erythrocytic 1). The in vivo investigations revealed the treatment effects of si‐circPRKAR1B and si‐METTL3 in colitis models of IL‐10‐deficient mice.ConclusionOur study reveals that METTL3‐mediated m6A modification of circPRKAR1B promotes Crohn's colitis by aggravating NLRP3 inflammasome‐mediated pyroptosis via autophagy impairment in colonic epithelial cells.