Mechanism of RNAm6A methylation modification regulating NLRP3 inflammasome activation for hand, foot and mouth disease progression
During the life cycle of Enterovirus 71 (EV71), N6-methyladenosine (m6A) modification has an important impact on viral replication and the cellular response to viral infection, but the effect of m6A modification on cellular RNAs and pathways during infection is still unknown. The purpose of this research was to investigate the role of m6A in regulating host cell inflammatory response and disease progression during EV71 infection. Methylation RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) results show the m6A methylation modification map of control and EV71-infected groups of RD cells. And multilevel validation showed that decreased expression of demethylase FTO (fat mass and obesity-associated protein) was responsible for the elevated total m6A levels in EV71-infected RD cells and that TXNIP may be a target gene for demethylase FTO action. Further functional experiments showed that demethylase FTO silencing promoted TXNIP expression, activation of NLRP3 inflammasome and promoted the release of pro-inflammatory factors in vitro, and demethylase FTO overexpression showed the opposite result. Further tested in an animal model of severe EV71 infection, with results consistent with in vitro. In conclusion, our findings suggested that depletion of demethylase FTO in the presence of EV71 infection increased the m6A level of TXNIP mRNA 3' UTR, increased mRNA stability, and promoted TXNIP expression, which activated the NLRP3 inflammasome and led to the release of pro-inflammatory factors, ultimately promoted HFMD progression. This could help with the development and prevention of m6A modification inhibitor-based drugs for viral-related inflammation and disease. Overall design: The type of samples: RD cell. The experimental conditions and variables: Enterovirus 71 (EV71). Contains EV71-infected RD cell samples, three replicates; control RD cell samples, three replicates.