Tay Kai Yi, Wee Hannah Su-Ann, Nguyen Nhi, Autio Matias Ilmari, Wazny Vanessa Kristina, Lee Khang Leng, Tay Darwin, Wang Yuting, Gao Xu, Heng Chew Kiat, Yee Chan Mark Yan, Yin Foo Roger Sik, Lee Jimmy, Loh Marie, Cheung Christine
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
bioRxiv. 2025 May 15:2025.05.11.653298. doi: 10.1101/2025.05.11.653298.
Ischemic heart disease, particularly coronary artery disease (CAD), remain leading causes of mortality worldwide. The single nucleotide polymorphism rs6903956 on chromosome 6p24.1 has been identified as a susceptibility locus for CAD in East Asian populations through genome-wide association studies. However, its functional role has not been fully elucidated. This study investigates the mechanistic basis of rs6903956 and its contribution to CAD pathogenesis, focusing on endothelial cell dysfunction. We first conducted cohort studies, revealing an association between the rs6903956 'A' risk allele and blood pressure phenotypes, along with impaired endothelial responsiveness indicated by reduced flow-mediated dilation. Single-base editing of induced pluripotent stem cell-derived endothelial cells obtained from patients with CAD and expression quantitative trait loci analysis highlighted a cisacting impact of the 'A' allele on and expression, suggesting allele-specific regulatory effects. Using modeling by AlphaFold 3 platform, the 'A' allele exhibited enhanced binding affinity for HOXA4 and MEIS1 transcription factors, forming a stable ternary complex that promoted transcriptional activation of . Functional assays demonstrated the enhancer role of rs6903956 'A' in promoter activity, supporting its locus-specific regulatory function in endothelial cells. Under pathological flow conditions, endothelial cells harboring the 'A' allele display elevated ICAM-1 expression and increased monocyte adhesion compared to the 'G' allele, indicating allele-specific endothelial inflammatory activation. These findings propose a model in which rs6903956 influences expression via HOX-MEIS cooperative binding, thereby modulating endothelial function and contributing to CAD susceptibility. This study provides mechanistic insights into the role of rs6903956 in endothelial dysfunction and CAD, informing potential therapeutic targets arising from genetic determinants in cardiovascular pathogenesis.
缺血性心脏病，尤其是冠状动脉疾病（CAD），仍然是全球主要的死亡原因。通过全基因组关联研究，已确定位于6号染色体p24.1上的单核苷酸多态性rs6903956是东亚人群CAD的一个易感位点。然而，其功能作用尚未完全阐明。本研究调查了rs6903956的作用机制及其对CAD发病机制的影响，重点关注内皮细胞功能障碍。我们首先进行了队列研究，揭示了rs6903956“A”风险等位基因与血压表型之间的关联，以及血流介导的血管舒张减少所表明的内皮反应性受损。对从CAD患者获得的诱导多能干细胞衍生的内皮细胞进行单碱基编辑和表达数量性状位点分析，突出了“A”等位基因对[具体基因1]和[具体基因2]表达的顺式作用影响，表明存在等位基因特异性调控效应。使用AlphaFold 3平台进行的[具体分子]建模显示，“A”等位基因对HOXA4和MEIS1转录因子表现出增强的结合亲和力，形成稳定的三元复合物，促进[具体基因3]的转录激活。功能试验证明了rs6903956“A”在[具体基因3]启动子活性中的增强子作用，支持其在内皮细胞中的位点特异性调控功能。在病理血流条件下，与“G”等位基因相比，携带“A”等位基因的内皮细胞显示ICAM - 1表达升高和单核细胞黏附增加，表明存在等位基因特异性内皮炎症激活。这些发现提出了一个模型，其中rs6903956通过HOX - MEIS协同结合影响[具体基因3]表达，从而调节内皮功能并导致CAD易感性。本研究为rs6903956在内皮功能障碍和CAD中的作用提供了机制性见解，为心血管发病机制中遗传决定因素产生的潜在治疗靶点提供了信息。
