Integration of Multi-Omics and Machine Learning Identifies TGFB1 and SERPINE1 as Biomarkers of Vascular Smooth Muscle...

Intracranial aneurysms (IAs) are severe cerebrovascular disorders lacking effective gene therapy or pharmacological interventions. Vascular smooth muscle cell (VSMC) senescence has been associated with various cardiovascular and cerebrovascular diseases; however, its role in IA pathogenesis remains unexplored, and the key regulatory genes remain unidentified. By integrating multi-omics data, machine learning, and experimental validation, we identified two senescence-associated genes (SAGs) that may serve as VSMC senescence biomarkers in IAs. We integrated single-cell RNA sequencing data from a mouse elastase-induced IA model (GSE193533) with human bulk RNA sequencing and microarray datasets to investigate VSMC senescence in IA. Our analyses included cell clustering, cell-cell communication, differential expression, functional enrichment, pseudotime trajectory inference, and high-dimensional weighted gene co-expression network analysis (hdWGCNA). Machine learning identified candidate SAGs, which were validated using external datasets and immunohistochemical analysis of human IA tissues. Single-cell analysis of the mouse IA model revealed 26 clusters across 10 cell types, with IA samples showing VSMC depletion, immune cell enrichment, and fibroblast expansion. VSMCs were categorized into five subsets (VSMC1-5), with the extracellular matrix remodeling and synthetic-inflammatory subtype (VSMC1) significantly increased. Differential expression analysis of VSMCs intersecting with senescence-associated genes from the Aging Atlas database identified 159 SAGs enriched in inflammatory and apoptotic pathways. Integrating hdWGCNA and bulk transcriptomics identified 45 key SAGs. TGFB1 and SERPINE1 emerged as robust biomarkers of senescence-associated VSMCs, showing high diagnostic accuracy (AUC > 0.75) and upregulation in IA tissues. Our study provides new insights into the pathological mechanisms of intracranial aneurysms. VSMC senescence may contribute to IA progression. TGFB1 and SERPINE1 may serve as biomarkers and promising therapeutic targets for VSMC senescence in IA. However, their specific mechanisms require further investigation and validation.