Systematic Identification and Functional Validation of <scp>CASP10</scp> as a <scp>DNA</scp> ‐Damage‐Responsive Driver of Endothelial Pyroptosis in Atherosclerosis

Atherosclerosis (AS) is a chronic inflammatory disease driven by endothelial dysfunction and plaque instability. The DNA damage response (DDR) has been implicated in endothelial cell fate; its precise role in AS remains unclear. This study aims to identify DDR-related biomarkers associated with AS and elucidate their mechanisms in endothelial pyroptosis. Our analysis identified 66 DDR-related genes, which achieved over 80% accuracy in discriminating early from advanced lesions and hemorrhagic from non-hemorrhagic plaques in external datasets, and reached 100% accuracy in differentiating plaque stability. CASP10 emerged as a top diagnostic biomarker (AUC = 0.991) compared to other DDR genes. Single-cell analysis confirmed elevated CASP10 expression in endothelial cells of AS plaques. Functional experiments revealed that CASP10 is both necessary and sufficient for ox-LDL-induced DNA damage and pyroptosis in HUVECs. CASP10 overexpression exacerbated γH₂AX accumulation, NLRP3 expression, GSDMD-N cleavage and IL-1β release, while CASP10 knockdown attenuated these effects. Overall, CASP10 serves as a reliable biomarker for identifying unstable plaques and functions as a pivotal mediator linking DNA damage to endothelial pyroptosis. Targeting CASP10 may represent a novel therapeutic strategy to reduce endothelial cell death and stabilise atherosclerotic plaques.