N⁶-Methyladenosine (m⁶A) Methylation Is Associated with the Immune Microenvironments in Acute Intracerebral...

Researchers have recently found that N⁶-methyladenosine (m⁶A) is a type of internal posttranscriptional modification that is essential in mammalian mRNA. However, the features of m⁶A RNA methylation in acute intracerebral hemorrhage (ICH) remain unknown. To explore differential methylations and to discover their functions in acute ICH patients, we recruited three acute ICH patients, three healthy controls, and an additional three patients and healthy controls for validation. The m⁶A methylation levels in blood samples from the two groups were determined by ultrahigh-performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS). Methylated RNA immunoprecipitation sequencing (MeRIP-seq) was employed to identify differences in m⁶A modification, and the differentially expressed m⁶A-modified genes were confirmed by MeRIP-qPCR. We found no significant differences in the total m⁶A levels between the two groups but observed differential methylation peaks. Compared with the control group, the coding genes showing increased methylation following acute ICH were mostly involved in processes connected with osteoclast differentiation, the neurotrophin signaling pathway, and the spliceosome, whereas genes with reduced m⁶A modification levels after acute ICH were found to be involved in the B-cell and T-cell receptor signaling pathways. These results reveal that differentially m⁶A-modified genes may influence the immune microenvironments in acute ICH.