The Institute of Infection and Inflammation, Department of Microbiology and Immunology, Medical College, Key Laboratory of Ischemic Cardiovascular and Cerebrovascular Disease Translational Medicine, China Three Gorges University, Yichang, Hubei 443002, PR China; Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Center for Human Genome Research, Cardio-X Institute, Huazhong University of Science and Technology, Wuhan 430074, PR China.
BayerAG, Drug Discovery, 42096 Wuppertal, Germany.
Gene. 2020 Aug 30;753:144805. doi: 10.1016/j.gene.2020.144805. Epub 2020 May 20.
Genomic variants in both ADTRP and TFPI genes are associated with risk of coronary artery disease (CAD). ADTRP regulates TFPI expression and endothelial cell functions involved in the initiation of atherosclerotic CAD. ADTRP also specifies primitive myelopoiesis and definitive hematopoiesis by upregulating TFPI expression. However, the underlying molecular mechanism is unknown. Here we show that transcription factor POU1F1 is the key by which ADTRP regulates TFPI expression. Luciferase reporter assays, chromatin-immunoprecipitation (ChIP) and electrophoretic mobility shift assay (EMSA) in combination with analysis of large and small deletions of the TFPI promoter/regulatory region were used to identify the molecular mechanism by which ADTRP regulates TFPI expression. Genetic association was assessed using case-control association analysis and phenome-wide association analysis (PhenGWA). ADTRP regulates TFPI expression at the transcription level in a dose-dependent manner. The ADTRP-response element was localized to a 50 bp region between -806 bp and -756 bp upstream of TFPI transcription start site, which contains a binding site for POU1F1. Deletion of POU1F1-binding site or knockdown of POU1F1 expression abolished ADTRP-mediated transcription of TFPI. ChIP and EMSA demonstrated that POU1F1 binds to the ADTRP response element. Genetic analysis identified significant association between POU1F1 variants and risk of CAD. PhenGWA identified other phenotypic traits associated with the ADTRP-POU1F1-TFPI axis such as lymphocyte count (ADTRP), waist circumference (TFPI), and standing height (POU1F1). These data identify POU1F1 as a transcription factor that regulates TFPI transcription in response to ADTRP, and link POU1F1 variants to risk of CAD for the first time.
ADTRP 和 TFPI 基因中的基因组变异与冠心病 (CAD) 的风险相关。ADTRP 调节 TFPI 的表达和内皮细胞功能,这些功能涉及动脉粥样硬化性 CAD 的起始。ADTRP 还通过上调 TFPI 的表达来指定原始髓系造血和定型造血。然而,潜在的分子机制尚不清楚。在这里,我们表明转录因子 POu1F1 是 ADTRP 调节 TFPI 表达的关键。使用荧光素酶报告基因检测、染色质免疫沉淀 (ChIP) 和电泳迁移率变动分析 (EMSA) 以及 TFPI 启动子/调控区的大、小缺失分析,确定了 ADTRP 调节 TFPI 表达的分子机制。使用病例对照关联分析和表型全基因组关联分析 (PhenGWA) 评估遗传关联。ADTRP 以剂量依赖的方式在转录水平上调节 TFPI 的表达。ADTRP 反应元件定位于 TFPI 转录起始位点上游的-806bp 至-756bp 之间的 50bp 区域,该区域包含一个结合 POu1F1 的结合位点。POu1F1 结合位点缺失或 POu1F1 表达敲低均消除了 ADTRP 介导的 TFPI 转录。ChIP 和 EMSA 表明 POu1F1 结合到 ADTRP 反应元件上。遗传分析确定了 POu1F1 变异与 CAD 风险之间的显著关联。PhenGWA 确定了与 ADTRP-POu1F1-TFPI 轴相关的其他表型特征,如淋巴细胞计数 (ADTRP)、腰围 (TFPI) 和站立身高 (POu1F1)。这些数据表明 POu1F1 是一种转录因子,它可以响应 ADTRP 调节 TFPI 的转录,并首次将 POu1F1 变异与 CAD 风险联系起来。