Deciphering single-cell landscape unravels cell-type-specific functional roles of RNA m⁶A modification in...

Background: Atherosclerosis is a chronic inflammatory disease that is the major cause of mortality worldwide. Although several studies have assessed the function of m⁶A (N⁶-methyladenosine) modification in atherosclerosis, its regulatory mechanism at the single-cell level remains unclear. This study provides a comprehensive single-cell atlas of m⁶A modification regulating cell-type-specific functions in atherosclerosis. Methods: We analyzed single-cell sequencing data derived from atherosclerosis patients to elucidate the influence of m⁶A modification on diverse cell types. We demonstrated the potential regulatory functions of m⁶A regulators across various cell types and key transcription factors involved. Furthermore, we discovered m⁶A regulators mediated intercellular communication in important biological processes. In vitro experiments were conducted to further investigate the effects of ALKBH5, WTAP and METTL3 on atherosclerosis. Results: ALKBH5 upregulated in endothelial cells induced cell proliferation and migration involved in sprouting angiogenesis. In smooth muscle cells, upregulation of WTAP enhanced proliferation, migration and phenotypic transformation. Upregulation of METTL3 and YTHDF2 promoted macrophage activation and differentiation. Furthermore, we identified abnormally activated transcription factors could regulate m⁶A regulators in a cell-type-specific manner. Moreover, we revealed that m⁶A regulators were implicated in dysregulated intercellular communication in atherosclerosis. And a series of experimental validations supported the conclusion that m⁶A regulators exert cell-type-specific regulatory functions. Conclusion: Our study provided evidence for the roles of ALKBH5, WTAP and METTL3 in orchestrating atherosclerotic cell-type-specific functions, representing promising targets for precision medicine.