Abstract
<jats:p><p dir="ltr">Objective Cardiovascular complications are the leading cause of death in type 2 diabetes mellitus (T2DM). We investigated whether blood-based DNA methylation (DNAm) biomarkers could improve risk stratification for major adverse cardiovascular events (MACE) in individuals with T2DM. </p><p dir="ltr">Research design and methods Epigenome-wide DNAm was profiled in peripheral blood mononuclear cells obtained at baseline from 305 T2DM participants of the TOSCA.IT study, including 81 who experienced MACE within five years (T2DMACE) and 224 who did not (T2D). The population was randomly divided into training and internal test sets for model development and evaluation. Differential DNAm analyses were performed under five frameworks differing by covariate adjustment. Binomial LASSO logistic regression with stability selection was used to generate DNAm-based models to stratify individuals at higher risk of incident MACE over 5 years. </p><p dir="ltr">Results Differential DNAm analysis identified 23,839, 21,529, 21,500, 12,114 and 7,101 differentially methylated CpG sites across the five analytical frameworks. Using LASSO logistic regression, five DNAm-based models were developed. Stability selection retained 22-26 CpG predictors per model. When evaluated in the internal test cohort, the five models showed high discriminative performance, with ROC-AUC values ranging from 0.915 to 0.937 and AUPRC values from 0.866 to 0.940. Several CpGs were independently associated with either increased or decreased risk of MACE, including 12 CpGs identified across all of the models.</p><p dir="ltr">Conclusions The DNAm-based models we have generated may improve risk stratification for short-term cardiovascular events in individuals with T2DM, providing a promising approach to refine identification of those at increased risk of MACE.</p><p><br></p></jats:p>