m5C: Novel Diagnostic and Drug Repurposing Targets for Nonalcoholic Steatohepatitis

Background: Epigenetic medication, such as RNA 5-methylcytosine (m5C), is well-recognized as a key regulator in hepatic metabolism and immune responses. However, m5C regulatory mechanisms in NASH pathogenesis have not yet been clearly elucidated. Methods: By utilizing three bulk profiles of NASH patients acquired from GEO and integrative bioinformatic pipelines, such as Limma framework, consensus clustering, and machine learning, we first identified m5C-related molecular subgroups and hub genes for NASH patients. Besides, diagnostic performance and biological characteristics of m5C-related hub gene were estimated at bulk level. Indeed, the heterogeneity of m5C-related hub gene for NASH patients was deciphered in single-cell transcriptomic profiles at temporal and spatial manners, especially in artificial intelligence (AI)-driven virtual cells. Furthermore, potential therapeutic agents targeting m5C-associated hub genes for the treatment of NASH were enriched by AI-driven drug enrichment framework (DrugReflector) based on NASH bulk profile and then validated by molecular docking. Finally, in vitro studies quantified the expression of m5C-associated hub genes compared to normal control. Results: m5C can divide NASH patients into two various consensus groups with different molecular and immune patterns. Furthermore, ERCC2 and FOXC2 can be considered two upregulated m5C-associated hub genes involved in NASH pathogenesis, which were mainly distributed at cholangiocyte. BRD-K93672499 can be considered a multitarget therapeutic strategy targeting ERCC2 and FOXC2 for the treatment of NASH. Conclusion: Our study first deciphered the m5C in predictive and therapeutic potential for NASH patients, which gains more insight into their personalized and precision medicine.