The Role and Diagnostic Efficacy of the METTL14/GADD45B m6A Methylation/BDNF Regulatory Axis in Acute Ischemic Stroke

Abstract Despite advances in acute ischemic stroke (AIS) research, identifying reliable biomarkers and regulatory mechanisms remains challenging. We first identified AIS-related genes via extensive literature review, retrieved dataset GSE16561 from the Gene Expression Omnibus (GEO, https://ncbi.nlm.nih.gov/geo/ ), and performed differential/enrichment analyses. Bioinformatics verified N6-methyladenosine (m 6 A) sites in target genes, focusing on the methyltransferase-like protein 14 (METTL14)/growth arrest and DNA damage-inducible β (GADD45B) m 6 A methylation/brain-derived neurotrophic factor (BDNF) axis. Peripheral blood samples, biochemical indicators, and demographic data were collected from Hongqi Hospital Affiliated to Mudanjiang Medical University. Human umbilical vein endothelial cells (HUVECs) underwent transfection and oxygen-glucose deprivation (OGD). METTL14, GADD45B, and BDNF were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR); BDNF protein by enzyme-linked immunosorbent assay (ELISA); global RNA m 6 A by Dot Blot; and GADD45B m 6 A by methylated RNA immunoprecipitation-quantitative polymerase chain reaction (MeRIP-RT-qPCR). Differential, diagnostic efficacy, logistic regression analyses, and nomogram validation were conducted. AIS patients showed increased METTL14, decreased GADD45B/BDNF, and increased levels of global m 6 A RNA and GADD45B m 6 A RNA ( P < 0.05). Receiver operating characteristic (ROC) analysis confirmed the three genes’ good diagnostic efficacy. The nomogram integrating these genes, globulin (GLOB), diabetes, high-density lipoprotein cholesterol (HDL-C), and hypertension performed excellently. This study highlights METTL14, GADD45B, and BDNF as key AIS biomarkers; METTL14 may indirectly regulate BDNF via GADD45B m 6 A methylation, providing potential therapeutic targets and novel mechanistic insights. Graphical Abstract