NAT10-mediated ac4C modifications regulate glioblastoma progression.

N4-acetylcytidine (ac4C) is a recently identified mRNA modification, with N-acetyltransferase 10 (NAT10) being the sole known enzyme responsible for its catalysis. However, the biological functions and regulatory mechanisms of NAT10-mediated ac4C modification in glioblastoma (GBM) remain largely unclear. In this study, we aimed to elucidate the regulatory pathways and functional implications of NAT10 and ac4C modification in GBM. We found that NAT10 is significantly upregulated in GBM, and its elevated expression is associated with disease progression and poor patient prognosis. Functionally, NAT10 promotes glioblastoma cell proliferation and migration in vitro and accelerates tumor growth in vivo. Mechanistically, we identified BOC mRNA, a member of the immunoglobulin superfamily of cell adhesion molecules, as a direct target of NAT10-catalyzed ac4C modification. This modification enhances both the stability and translational efficiency of BOC mRNA, thereby contributing to GBM progression. Furthermore, we demonstrate that HIF1α, a key transcription factor in the hypoxic response, directly activates NAT10 transcription by binding to hypoxia response elements HRE1 and HRE2, leading to increased ac4C modification of BOC mRNA under hypoxic conditions. Notably, pharmacological inhibition of NAT10 effectively suppresses its enzymatic activity, particularly under hypoxia, underscoring its potential as a therapeutic target in GBM. In summary, our findings reveal a critical role for NAT10-mediated mRNA ac4C modification in GBM oncogenesis and highlight NAT10 as a promising target for therapeutic intervention. NAT10 was upregulated in GBM, and NAT10 facilitated GBM progression in vitro and in vivo. Mechanistically, NAT10 catalyzed ac4C modification of BOC mRNA and maintained its stability and promoted translation. Besides, HIF1α influenced NAT10 and its ac4C writer function through transcriptional activation.