Peli1, regulated by m 6 A modification, suppresses atherosclerosis progression by inhibiting YB-1-mediated NLRP3 inflammasome activation

Abstract Background The activation of NLRP3 inflammasome in macrophages is a risking factor accelerating atherosclerosis (AS) progression. Here, the function of Peli1 in regulating NLRP3 inflammasome activation during AS progression were investigated. Methods ApoE −/− mice were subjected to high fat diet to construct AS model in vivo . HE, Oil red O and Sirius red staining were adopted to analyze histopathological changes and lipid accumulation. Raw264.7 cells stimulated by ox-LDL were used as AS cellular model. Pro-inflammatory cytokines secretion was assessed using ELISA. Total m6A level was examined by m 6 A dot blot assay, and Peli1 m 6 A level was assessed using MeRIP assay. The interactions between METTL3, YTHDC2, Peli1, YB-1 and NLRP3 were analyzed by ChIP, dual-luciferase reporter gene, CoIP, RIP and/or RNA pull down assays. Results YB-1 knockdown could inhibit AS progression in vivo , and YB-1 silencing repressed ox-LDL-mediated lipid accumulation and inflammation in macrophages by inactivating NLRP3 inflammasome. E3 ubiquitination ligase Peli1 mediated ubiquitination degradation of YB-1 during AS progression. Moreover, it was found that YTHDC2 recognized METTL3-mediated Peli1 m 6 A modification and mediated Peli1 mRNA degradation. Rescue studies revealed that YB-1 upregulation abrogated Peli1 upregulation’s repression on AS progression both in vitro and in vivo . Conclusion Peli1, regulated by m 6 A modification, inhibited YB-1-mediated NLRP3 inflammasome activation in macrophages by promoting YB-1 ubiquitination to suppress AS progression.