Epigenetic signatures of cardiometabolic risk in men: accelerated aging and differential methylation replicated across cohorts

Men exhibit greater susceptibility to cardiovascular diseases and metabolic disorders, with an earlier onset and more aggressive progression, potentially driven by epigenetic modifications, particularly DNA methylation. Our goal was to comprehensively characterize the epigenetic landscape of a broad cardiometabolic burden in a cohort composed exclusively of men. We generated novel DNA methylation profiles from whole blood samples of men with cardiometabolic disturbances (hypertension, ischemic heart disease, obesity, dyslipidemia) and age-matched healthy controls. Cases demonstrated significant epigenetic age acceleration, most pronounced for second-generation clocks (GrimAge, GrimAge2) and pace of aging measures (DunedinPACE), accompanied by shortened epigenetic telomere length (DNAmTL). Notably, none of the 19 evaluated first-generation epigenetic clocks exhibited sensitivity to the studied diseases. Epigenome-wide association analysis identified differentially methylated positions (DMPs), predominantly hypomethylated in cases compared to controls. Gene set enrichment analysis of genes annotated to these DMPs revealed nine distinct biological pathway clusters that reflect the multifactorial processes associated with cardiometabolic burden, including chronic inflammation, GPCR signaling dysregulation, metabolic disturbances, mitochondrial dysfunction, vascular remodeling, and renal electrolyte regulation. Key findings, including GrimAge acceleration, DunedinPACE elevation, DNAmTL shortening, and enrichment of inflammatory and GPCR pathways, were replicated in an independent cohort of men with atherosclerosis. Men with cardiometabolic disturbances exhibit accelerated epigenetic aging and distinct DNA methylation signatures associated with cardiometabolic burden. Analysis of a broad battery of epigenetic clock models revealed that only the second-generation (GrimAge) and third-generation (DunedinPACE) models demonstrated a pronounced sensitivity to the uncomplicated diseases evaluated. In contrast, the first-generation models, trained to predict chronological age, failed to detect significant differences between the groups, suggesting limited applicability in these pathologies. The concordance of results across original and independent replication cohorts underscores the fundamental nature of these epigenetic alterations. Our findings suggest candidate biomarkers measurable in minimally invasive blood samples that may assist in early risk stratification and monitoring of disease progression in men, warranting further prospective evaluation to facilitate clinical translation.