Analysis of the Correlation Between Cuproptosis and Instability of Atherosclerotic Plaques

<b>Background/Objectives</b>: Cuproptosis, a newly discovered form of programmed cell death, is dependent on the regulation of copper ions. The roles and mechanisms of cuproptosis-related genes (CRGs) in the instability of atherosclerotic plaques are still unclear. <b>Methods</b>: GEO microarray datasets were downloaded to analyze stable and unstable human carotid artery plaques. Differential expression analysis was performed to screen for CRGs from the Molecular Signatures Database (MSigDB). Machine learning was applied to identify key genes and cluster unstable plaque genes. The identified genes were verified by immunohistochemistry (IHC) of human carotid plaque samples, and the effect of ATOX1 on cuproptosis was detected in human umbilical vein endothelial cells (HUVEC). <b>Results</b>: This study identified 27 CRGs differentially expressed between stable and unstable plaques. Five characteristic genes (LC3A, ATP7B, ATOX1, CTR1, and NLRP3) were selected by machine learning. A diagnostic model for unstable plaques was developed based on these genes. The expression of ATOX1 and NLRP3 was increased, while LC3A and ATP7B were decreased in unstable plaques. However, there was no significant change in CTR1. The Cell Counting Kit-8 (CCK-8) assay indicated that inhibiting ATOX1 reduced CuSO₄-induced HUVEC death. <b>Conclusions</b>: CRGs appear to influence atherosclerotic plaque formation. Five key genes (LC3A, ATP7B, ATOX1, CTR1, NLRP3) were identified as being differentially expressed in unstable plaques. Cluster analysis uncovered two subtypes (C1, C2) linked to cuproptosis and immune infiltration in unstable plaques. These genes likely affect atherosclerosis progression by influencing immune cell infiltration, thus impacting plaque stability. Furthermore, the cuproptosis-related gene ATOX1 can regulate CuSO₄-induced HUVEC death. This study contributes to predicting plaque instability and offers potential diagnostic and therapeutic targets.