Tspan14 expression and function in cardiovascular disease

Tspan14在心血管疾病中的表达和功能

• 批准号: 10851296
• 负责人: Vivian Lee-Kim
• 金额: $ 5.91万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10851296
• 关键词: Adaptor Signaling Protein; Advisory Committees; Affect; Alleles; Animal Model; Blood Vessels; Cardiovascular Diseases; Cell membrane; Cell model; Cells; Cholesterol; Chromosomes; Coronary Arteriosclerosis; Coronary heart disease; DNA; Dedications; Development; Disease; Enhancers; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Enhancer Element; Genetic Predisposition to Disease; Genetic Variation; Hematological Disease; Human; Hypertension; Immune; Lead; Mentors; Minor; Modeling; Pathogenesis; Pathway interactions; Phase; Population Genetics; Regulation; Regulatory Element; Risk Factors; Role; Supervision; Training; Untranslated RNA; Variant; Vascular Endothelial Cell; activation-induced cytidine deaminase; cell type; differential expression; disorder risk; effective therapy; genetic approach; genome wide association study; monocyte; mortality; new therapeutic target; notch protein; protective effect; protein transport; risk variant; success; transcriptome sequencing

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.

项目总结/摘要 遗传易感性是冠状动脉疾病（CAD）的重要危险因素，CAD是导致冠心病的主要原因。 mortality.在全基因组关联研究（GWAS）中发现的大多数CAD风险变异是非编码的 DNA，这对识别疾病关键细胞类型中的靶基因构成了重大挑战。使用 活性接触模型预测细胞类型特异性基因增强子对，我们确定了顺式调节 在染色体10 q23基因座中的元件携带多种CAD风险变异。我们的初步研究显示 10 q23基因座中的这些增强子调节四跨膜蛋白14（TSPAN 14）基因表达，特别是在 血管内皮细胞（EC）和单核细胞。我们已经建立了强有力的群体遗传学证据， 较高的TSPAN 14表达降低了CAD风险，并且对次要等位基因具有一致的保护作用 在领先的变种。TSPAN 14是一种衔接蛋白，有助于运输蛋白质，如ADAM 10，一种Notch 受体激活剂，到质膜。由这种TSPAN 14 - 15所赋予的Notch途径活化的作用是： 尚未探索CAD发病机制中的ADAM 10相互作用。此外，我们进行了RNA-seq， 在TSPAN 14缺陷细胞中发现差异表达的基因，这将有助于发现Notch非依赖性 TSPAN 14功能。这些发现为核心假设提供了前提，即 TSPAN 14通过10 q23增强子序列的变异通过Notch- 血管内皮细胞和单核细胞的依赖和非依赖机制。在目标1中，李金博士将 确定10 q23增强子序列变异对TSPAN 14表达的细胞类型特异性影响 调控在目标2中，她将确定TSPAN 14表达在血管中的Notch依赖性作用， 免疫细胞在目标3中，她将描述Notch非依赖性TSPAN 14在血管和 免疫细胞。这些研究的结果将在功能上验证非编码变体的基因靶点 在疾病相关细胞类型中与CAD风险相关，并阐明TSPAN 14在CAD中的功能 发病机制这些研究将在导师Rajat Gupta博士和Dr. 斯蒂芬·布莱克洛和一个致力于李金博士成功的顾问委员会。也提供了其他协助 从MOSAIC UE 5获奖者赞助的专业发展机会，继续培训， K99阶段将为李金博士成功过渡到独立做好准备。