NAT10 Regulates the Migration, Invasion, and Angiogenesis of Human Umbilical Vein Endothelial Cells Through ac4C Modification of MEOX2 in Gestational Diabetes Mellitus

Gestational diabetes mellitus (GDM) is a metabolic disorder during pregnancy with significant implications for maternal and fetal health. N4-acetylcytidine (ac⁴C) RNA modification, catalyzed by N-acetyltransferase 10 (NAT10), plays a critical role in regulating mRNA stability and translation. This study aimed to investigate the role of NAT10-mediated ac⁴C modification in migration, invasion, and angiogenesis in GDM. Bioinformatics analysis of the GSE203346 dataset identified differentially expressed genes in GDM placental tissues. In vitro experiments using human umbilical vein endothelial cells (HUVECs) included NAT10 knockdown, high-glucose (HG) treatment, and functional assays (cell counting kit-8, wound healing, Transwell, and tube formation). Molecular mechanisms were explored using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot, methylated RNA immunoprecipitation (MeRIP)-qPCR, RNA immunoprecipitation (RIP), dual-luciferase reporter, and RNA stability assays. NAT10 was significantly upregulated in GDM placental tissues at both mRNA and protein levels (P < 0.01), accompanied by increased global ac⁴C modification levels. In HG-treated HUVECs, NAT10 knockdown enhanced cell viability, migration, invasion, and tube formation (P < 0.01). Mechanistically, NAT10 directly bound to mesenchyme homeobox 2 (MEOX2) mRNA, and NAT10 silencing reduced ac⁴C modification levels at a specific site (nucleotides 409-423) of MEOX2 (P < 0.01). This reduction in ac⁴C modification decreased MEOX2 mRNA stability and subsequent expression (P < 0.01). Rescue experiments showed that MEOX2 overexpression reversed the promoting effects of NAT10 inhibition on viability, migration, invasion, and tube formation in HG-treated HUVECs (P < 0.01). NAT10 regulated the migration, invasion, and angiogenesis of HUVECs via ac⁴C modification of MEOX2, emphasizing its potential as a therapeutic target for GDM-related placental complications.

Ac4C; Angiogenesis; Gestational diabetes mellitus; Invasion; MEOX2; Migration; NAT10.