Promoter hypomethylation of

Introduction: Cerebral small vessel disease (SVD), manifesting as white matter hyperintensities (WMH), lacunar infarctions, and cerebral microbleeds on magnetic resonance imaging (MRI), has been linked to developmental epigenetic alterations. This study aimed to identify and validate gene-specific promoter methylation changes as epigenetic markers associated with SVD, using MRI-defined imaging features and blood inflammatory cells. Methods: Genome-wide promoter methylation was profiled using the Infinium MethylationEPIC v2.0 array in peripheral inflammatory cells from 16 patients without SVD and 16 patients with all three imaging features, including WMH, lacunes, and microbleeds on MRI. Candidate CpGs were defined as consensus DMPs detected by both minfi and SeSAMe (nominal P < 0.05 in both pipelines with concordant direction), filtered by absolute delta beta >0.10 and promoter proximity (TSS200/TSS1500). Validation was performed to determine whether these gene-specific promoter methylations could serve as independent variables predicting the presence of SVD imaging features when combined with established cardiovascular risk factors, using data from 766 patients with ischemic stroke (53 [6.9%] without SVD and 713 [93.1%] with ≥1 SVD imaging feature). Hierarchical logistic regression analysis and a deep learning model were applied. Subgroup analyses using multinomial logistic regression were performed to assess whether gene-specific promoter methylation could independently predict WMH or lacunes. Results: EPIC profiling identified 17 promoter regions with significant differences between groups, corresponding to Conclusion: