Tspan14 expression and function in cardiovascular disease

项目摘要/摘要 遗传易感性是冠状动脉疾病(CAD)的重要危险因素，冠心病是 死亡率。在全基因组关联研究中发现的大多数冠心病风险变异都是非编码的 DNA，这对确定疾病关键细胞类型中的目标基因构成了重大挑战。使用 通过接触活性模型预测细胞类型特异性基因-增强子对，我们确定了顺式调控 染色体10q23基因座上含有多种易患冠心病的变异体。我们的初步研究表明 10q23基因座上的这些增强子调节TSPAN14(TSPAN14)基因的表达，特别是在 血管内皮细胞(ECs)和单核细胞。我们已经建立了强有力的群体遗传学证据 TSPAN14的高表达降低了冠心病的风险，对次要等位基因有一致的保护作用 处于领先地位的变种。TSPAN14是一种接头蛋白，有助于运输像ADAM10这样的Notch蛋白 受体激活剂，到质膜。TSPAN14--对Notch通路激活的影响 ADAM10在冠心病发病机制中的相互作用尚未被探讨。此外，我们还进行了RNA-seq和 在TSPAN14缺陷细胞中发现差异表达基因将有助于发现Notch非依赖性 TSPAN14函数。这些发现为中心假说提供了一个前提，即 TSPAN14通过10q23增强子序列的变异影响冠心病的发病。 血管内皮细胞和单核细胞的依赖和独立机制。在目标1中，Lee-Kim博士将 确定10q23增强子序列变异对TSPAN14表达的细胞类型特异性影响 监管。在目标2中，她将确定TSPAN14在血管和血管中表达的Notch依赖效应。 免疫细胞。在目标3中，她将描述血管和血管中非依赖Notch的TSPAN14功能 免疫细胞。这些研究的结果将从功能上验证非编码变体的基因靶点 与疾病相关细胞类型中的冠心病风险相关，并阐明TSPAN14在冠心病中的作用 发病机制。这些研究将在导师拉贾特·古普塔博士和拉贾特·古普塔博士的监督下进行。 斯蒂芬·布莱克洛和一个专门为李金博士的成功而成立的咨询委员会。提供更多支持 从马赛克UE5获奖者赞助的专业发展机会中，继续培训 K99阶段将为Lee-Kim博士成功过渡到独立做好准备。

PROJECT SUMMARY/ABSTRACT Genetic predisposition is a significant risk factor for coronary artery disease (CAD), the leading cause of mortality. Most CAD risk variants identified in genome-wide association studies (GWAS) are in noncoding DNA, which poses a major challenge in identifying the target genes in the disease-critical cell types. Using the Activity-by-Contact Model to predict cell type-specific gene-enhancer pairs, we identified cis-regulatory elements in the chromosome 10q23 locus harboring multiple CAD-risk variants. Our preliminary studies show that these enhancers in the 10q23 locus regulate tetraspanin 14 (TSPAN14) gene expression, specifically in vascular endothelial cells (ECs) and monocytes. We have established strong population genetics evidence that higher TSPAN14 expression reduces CAD risk and there is a consistent protective effect for the minor alleles at the lead variants. TSPAN14 is an adaptor protein that aids in trafficking proteins like ADAM10, a Notch receptor activator, to the plasma membrane. The role of Notch pathway activation imparted by this TSPAN14- ADAM10 interaction in CAD pathogenesis has not been explored. In addition, we performed RNA-seq and identified differentially expressed genes in TSPAN14-deficient cells that will help discover Notch-independent TSPAN14 functions. These findings provide a premise for the central hypothesis that the genetic regulation of TSPAN14 by variations in the 10q23 enhancer sequences affects CAD pathogenesis through Notch- dependent and -independent mechanisms in vascular ECs and monocytes. In Aim 1, Dr. Lee-Kim will determine the cell type-specific effect of 10q23 enhancer sequence variations on TSPAN14 expression regulation. In Aim 2, she will determine the Notch-dependent effects of TSPAN14 expression in vascular and immune cells. In Aim 3, she will characterize the Notch-independent TSPAN14 functions in vascular and immune cells. The results from these studies will functionally validate the gene target for noncoding variants associated with CAD-risk in the disease-relevant cell types and elucidate how TSPAN14 functions in CAD pathogenesis. These studies will be conducted under the supervision of mentors, Dr. Rajat Gupta and Dr. Stephen Blacklow, and an advisory committee dedicated to Dr. Lee-Kim’s success. With additional support from the MOSAIC UE5 awardee sponsored professional development opportunities, continued training in the K99 phase will prepare Dr. Lee-Kim for successful transition to independence.