DNA Methylation-Guided Prediction and Validation of TAAD Molecular Targets

Thoracic aortic aneurysms and dissection are pivotal cardiovascular conditions necessitating accurate diagnostics. DNA methylation, a crucial epigenetic mediator, is implicated in early disease biomarkers. Our analysis of GSE84274 and GSE202047 datasets pinpointed 498 DEGs with promoter DMPs and DMRs. Outliers were detected via DIvisive ANAlysis (DIANA) and Orthogonal Projection to Latent Structures-Discriminant Analysis (OPLS-DA) (P < 0.05). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses emphasized vascular and TNF signaling. Utilizing the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, Nuclear Receptor Subfamily 2 Group F Member 2 (NR2F2) and GATA-Binding Protein 2 (GATA2) were identified as core differentially expressed genes (DEGs). Transcriptomic validation confirmed that promoter methylation modulates transcriptional activity (P < 0.05). Enzyme-Linked Immunosorbent Assay (ELISA) indicated heightened 5-mC levels in β-aminopropionitrile (BAPN)-challenged mice (P < 0.05), and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) confirmed the modulation of DNA Methyltransferase 3 Alpha (DNMT3A), DNMT3B, NR2F2, and GATA2. Methylation-Specific PCR (MSP) and Bisulfite Sequencing PCR (BSP) substantiated the hypermethylation of NR2F2 and GATA2 promoters in TAAD (P < 0.05). Our study correlates heightened promoter methylation of NR2F2 and GATA2 with TAAD, proposing them as novel diagnostic biomarkers.