Unraveling the molecular mechanisms underlying the coronary artery disease risk gene REST

Abstract Background/Introduction Genome-wide association studies (GWAS) have identified two novel coronary artery disease (CAD) risk variants, rs17087335 [1] and rs72627509 [2], located near the REST gene, which encodes the RE-1 silencing transcription factor. While REST is recognised as a transcriptional regulator of neuronal genes, its role in atherosclerosis and CAD is poorly understood. Purpose To investigate the molecular mechanisms by which the CAD risk variants rs17087335 and rs72627509 influence REST gene expression and contribute to atherosclerotic phenotypes. Methods In silico analyses were performed to identify variants in linkage disequilibrium with rs17087335 and rs72627509. Reporter gene assays were used to assess regulatory activity at these loci. Mass spectrometry was employed to examine transcription factor binding to variant alleles. Expression quantitative trait loci (eQTL) data were obtained from the GTEx database, and VSMC siRNA knockdown studies were conducted to evaluate cellular effects. Results Linkage disequilibrium (LD) analysis revealed that rs17087335 and rs72627509 are in LD with 14 other variants. To narrow the number of putative causal variants, we performed reporter gene assays that confirmed regulatory activity specifically at rs17087335 and rs72627509, as well as at rs3796529. Mass spectrometry analysis revealed differential transcription factor binding to the alleles of these variants, potentially influencing REST transcription. Indeed, eQTL data demonstrated that CAD patients carrying the risk alleles of rs17087335, rs72627509, and rs3796529, respectively, had significantly decreased REST mRNA levels in the aorta. Functional studies on vascular smooth muscle cells (VSMCs) showed that REST siRNA knockdown increased cell migration and lipid accumulation under inflammatory conditions. RNA sequencing revealed ATF6 and PLA2G4C as potential downstream targets of REST, potentially mediating the observed changes in VSMC migration and lipid accumulation. Conclusion(s) This study provides the first insights into the role of REST in atherosclerosis. The CAD risk variants rs17087335, rs72627509, and rs3796529 influence REST expression, and decreased REST levels promote atherosclerotic phenotypes in VSMCs. These effects are likely mediated through altered lipid metabolism and cell migration, driven by REST targets such as ATF6 and PLA2G4C. Our findings suggest that targeting REST could offer new therapeutic strategies for CAD prevention and treatment.