WTAP-Driven m6A Modification of TIMP1 mRNA Promotes Sepsis-Induced Cardiomyocytes Injury.

Sepsis-induced myocardial injury, characterized by severe myocardial dysfunction, is one of the most common and lethal complications of sepsis in clinical settings. However, the underlying pathogenesis is not fully understood. In this study, we conducted a combined analysis of mRNA-seq and MeRIP-seq datasets obtained from GEO, identifying 110 differentially expressed genes at both the transcriptional and m6A modification levels. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that these genes were primarily enriched in inflammation and immune response-related signaling pathways. Additionally, we identified 20 key genes involved in regulating myocardial injury during sepsis using Protein-Protein Interaction (PPI) analysis combined with the K-means clustering algorithm and MCC algorithm. Subsequently, the expression patterns of these 20 hub genes and their relationship with cardiac function were verified using transcriptome datasets from different species. The results showed that TIMP1 was the only gene significantly up-regulated in myocardial injury across different species and was closely associated with reduced cardiac function. Furthermore, through bioinformatics analysis, MeRIP sequencing, and dual luciferase reporter assays, we demonstrated that the abnormal up-regulation of TIMP1 in septicemia-induced myocardial injury is mediated by WTAP-driven m6A modifications, which enhance the stability of TIMP1 mRNA. Finally, we used lipopolysaccharide (LPS)-induced human cardiomyocytes AC16 to simulate a sepsis-induced myocardial injury model and conducted gene loss-function or rescue experiments. The results showed that targeted inhibition of the WTAP-m6A-TIMP1 axis can significantly improve LPS-induced myocardial cell damage. Collectively, these findings highlight the role of WTAP-mediated m6A modification in driving TIMP1 expression in sepsis-induced cardiomyocyte injury, potentially providing a new therapeutic target for sepsis-induced myocardial injury.