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

<b>Background</b>: 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. <b>Methods</b>: 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. <i>In vitro</i> experiments were conducted to further investigate the effects of ALKBH5, WTAP and METTL3 on atherosclerosis. <b>Results</b>: <i>ALKBH5</i> upregulated in endothelial cells induced cell proliferation and migration involved in sprouting angiogenesis. In smooth muscle cells, upregulation of <i>WTAP</i> enhanced proliferation, migration and phenotypic transformation. Upregulation of <i>METTL3</i> and <i>YTHDF2</i> 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. <b>Conclusion</b>: 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.