Decoding the lactylation-immune link in calcific aortic valve disease: a comprehensive bioinformatics approach

Calcific aortic valve disease is a progressive disorder characterized by pathological leaflet thickening and calcification. Early disease features include immune cell infiltration and metabolic dysregulation, which persist throughout disease progression. Recent studies implicate aberrant glycolysis and lactate accumulation in CAVD pathogenesis, though mechanistic insights remain limited. We analyzed bulk RNA-seq data (GSE76717, GSE235995, GSE153555) and applied machine learning algorithms to identify lactylation-related hub genes in calcific aortic valve disease. Functional enrichment, correlation analysis, single-gene Gene Set Enrichment Analysis, and immune infiltration profiling were performed to elucidate the functional roles of lactylation-related hub genes. Furthermore, single-cell RNA sequencing data were integrated to evaluate lactylation-related gene expression across cell types, quantify lactylation scores, and correlate these scores with Kyoto Encyclopedia of Genes and Genomes pathways. Besides, single-cell samples were stratified by calcification severity (mild vs. severe) to assess the lactylation levels of distinct cell populations during disease progression. Our multi-dimensional analysis revealed eight lactylation-related hub genes (CRABP2, SOD1, PFN1, MBD2, ARGLU1, ALB, ALDH1A1, THRAP3) with significant differential expression in calcific aortic valve disease. These genes correlated with RUNX2 upregulation, immune cell infiltration, and metabolic pathways (glycolysis, oxidative phosphorylation). Furthermore, single-cell RNA sequencing analysis of 47,328 cells revealed elevated lactylation scores in valvular interstitial cells, dendritic cells and M1 macrophages during severe calcification, thereby establishing a correlation between lactylation and MYC-driven metabolic reprogramming. This study systematically elucidates the dynamic relationship between immune cell infiltration and lactylation levels during calcific aortic valve disease progression. These findings provide novel perspectives for understanding the molecular mechanisms of calcific aortic valve disease and highlight potential therapeutic targets for clinical intervention.