Tspan14 expression and function in cardiovascular disease

Tspan14在心血管疾病中的表达和功能

基本信息

批准号：
10656419
负责人：
Vivian Lee-Kim
金额：
$ 9.94万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
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/10656419
关键词：
Adaptor Signaling Protein Address Advisory Committees Affect Alleles Animal Model Apoptosis Architecture Basement membrane Binding Binding Sites Bioinformatics Biological Biological Process Biology Blood Vessels CD36 gene Cardiovascular Diseases Cell membrane Cell model Cells Cholesterol Chromatin Chromosomes Committee Members Complement Complex Coronary Arteriosclerosis Coronary heart disease DNA Data Dedications Development Diabetes Mellitus Disease Endothelial Cells Endothelium Enhancers Extracellular Matrix Fostering Future Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Enhancer Element Genetic Predisposition to Disease Genetic Variation Grant Growth Hematological Disease Heritability Human Hyperlipidemia Hypertension Immune Introns Laboratories Lead Macrophage Maps Measures Mediating Membrane Membrane Microdomains Mentors Mesenchymal Minor Modeling Nucleic Acid Regulatory Sequences Pathogenesis Pathway interactions Peptide Hydrolases Phase Population Genetics Proteins Receptor Activation Regulation Regulator Genes Regulatory Element Research Research Personnel Risk Factors Role Scientist Signal Transduction Supervision Techniques Testing Tissues Training Untranslated RNA Variant Vascular Diseases Vascular Endothelial Cell Writing Zebrafish activation-induced cytidine deaminase causal variant cell type differential expression disorder risk effective therapy epigenetic regulation epigenomic profiling epigenomics genetic approach genome editing genome wide association study genomic locus in vivo insight model organism monocyte mortality new therapeutic target notch protein novel novel strategies prime editing programs protective effect protein function protein transport response risk variant skills success trafficking transcription factor transcriptome sequencing transcriptomics transdifferentiation

项目摘要

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.

项目概要/摘要遗传易感性是冠心病（CAD）的一个重要危险因素，而冠心病是导致冠心病的主要原因。全基因组关联研究 (GWAS) 中发现的大多数 CAD 风险变异都是非编码的。 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。通过接触活动模型来预测细胞类型特异性基因增强子对，我们确定了顺式调节我们的初步研究表明，染色体 10q23 位点中的元件含有多种 CAD 风险变异。 10q23 位点中的这些增强子调节四跨膜蛋白 14 (TSPAN14) 基因表达，特别是在我们已经建立了强有力的群体遗传学证据表明，血管内皮细胞（EC）和单核细胞。较高的 TSPAN14 表达可降低 CAD 风险，并且对次要等位基因具有一致的保护作用 TSPAN14 是一种接头蛋白，有助于运输 ADAM10（一种 Notch）等蛋白。受体激活剂，对质膜的 Notch 通路激活的作用。 ADAM10 在 CAD 发病机制中的相互作用尚未得到探索。此外，我们还进行了 RNA 测序。鉴定出 TSPAN14 缺陷细胞中的差异表达基因，这将有助于发现 Notch 独立性这些发现为基因调控的中心假设提供了前提。 TSPAN14 通过 10q23 增强子序列的变异通过 Notch-影响 CAD 发病机制在目标 1 中，Lee-Kim 博士将研究血管 EC 和单核细胞中的依赖和独立机制。确定 10q23 增强子序列变异对 TSPAN14 表达的细胞类型特异性影响在目标 2 中，她将确定 TSPAN14 表达在血管和神经系统中的 Notch 依赖性效应。在目标 3 中，她将描述血管和免疫细胞中不依赖于 Notch 的 TSPAN14 功能。这些研究的结果将在功能上验证非编码变异的基因靶点。与疾病相关细胞类型的 CAD 风险相关，并阐明 TSPAN14 在 CAD 中的功能这些研究将在导师 Rajat Gupta 博士和 Dr. Rajat 的监督下进行。斯蒂芬·布莱克洛 (Stephen Blacklow) 以及致力于李金博士成功的咨询委员会提供了额外支持。来自 MOSAIC UE5 获奖者赞助的专业发展机会，继续培训 K99 阶段将为 Lee-Kim 博士成功过渡到独立做好准备。