NSUN2 promoted tumor growth and metastatic via m5C-regulation of YAP through ALYREF/YBX1 axis in NSCLC

Abstract NSUN2-dependent RNA m 5 C methylation is essential for RNA stability, cellular metabolism, and intracellular transport. Aberrant YAP expression is closely associated with tumorigenesis and progression in human cancers. However, the molecular mechanism by which m⁵C regulates the growth and metastasis of NSCLC through modulating YAP expression remains incompletely understood. Our results demonstrate that YAP and NSUN2 play analogous roles in regulating NSCLC cell growth, migration, invasion, and EMT. NSUN2 increased m 5 C modification of YAP mRNA. ALYREF and YBX1 combined and then interacted with YAP mRNA in an m 5 C-dependent manner to increase YAP stability and translation. Importantly, NSUN2, ALYREF and YBX1 bind to each other and affected their interaction with YAP mRNA. Mechanistically, NSUN2 first initiates the m 5 C within YAP mRNA and then ALYREF recognizes m 5 C modification on YAP mRNA, YBX1 was more likely to bind to the transitive m 5 C from ALYREF and then promoted YAP mRNA stability through impeding the combination between AGO2 and YAP mRNA whereby increasing the expression of YAP with interaction with eIF3a and thus excessive cell growth and metastasis via regulation of YAP’s target genes of CTGF, Cyr61 MMP2, MMP9 in NSCLC. Moreover, NSUN2 is transcriptionally activated by the YAP-TEAD2 complex, forming a positive feedback loop that promotes tumor growth and metastasis, a process effectively suppressed by m 5 C inhibitors both in vivo and in vitro. Furthermore, our presented findings suggest that NSUN2 promotes tumor growth and metastasis by increasing ALYREF/YBX1-mediated YAP expression in NSCLC and effective inhibition of m 5 C modification might provide a potential treatment strategy for NSCLC.