Tspan14 expression and function in cardiovascular disease

Tspan14在心血管疾病中的表达和功能

• 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
• 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

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.

项目摘要/摘要 遗传易感性是冠状动脉疾病（CAD）的重要危险因素，这是 死亡。在全基因组关联研究（GWAS）中确定的大多数CAD风险变体都在非编码 DNA在确定关键疾病细胞类型中的靶基因方面构成了重大挑战。使用 通过接触模型预测细胞类型特异性基因增强剂对，我们确定了顺式调控 染色体10q23基因座中具有多个CAD风险变体的元素。我们的初步研究表明 这些增强子在10q23基因座中调节四叠氮蛋白14（TSPAN14）基因表达，特别是在 血管内皮细胞（EC）和单核细胞。我们已经建立了强大的人口遗传学证据表明 较高的TSPAN14表达降低了CAD的风险，并且对小等位基因具有一致的保护作用 在铅变体中。 TSPAN14是一种适配蛋白，有助于ADAM10等运输蛋白 受体激活剂，到质膜。该TSPAN14-赋予Notch途径激活的作用 CAD发病机理中的ADAM10相互作用尚未探索。此外，我们进行了RNA-seq，并 在TSPAN14缺陷型细胞中鉴定出不同表达的基因，这将有助于发现与Notch无关的细胞 TSPAN14功能。这些发现为中心假设提供了前提 TSPAN14通过10q23增强子序列的变化会通过Notch-影响CAD发病 血管EC和单核细胞中的依赖性和非依赖性机制。在AIM 1中，Lee-Kim博士将 确定10q23增强子序列变化对TSPAN14表达的细胞类型特异性效应 规定。在AIM 2中，她将确定TSPAN14表达在血管和血管和 免疫细胞。在AIM 3中，她将表征与缺口无关的TSPAN14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标 与疾病相关的细胞类型中的CAD风险相关，并阐明TSPAN14在CAD中的功能 发病。这些研究将在导师Rajat Gupta博士和博士的指导下进行。 斯蒂芬·布莱克洛（Stephen Blacklow）和咨询委员会致力于李·金（Lee-Kim）的成功。提供额外的支持 从马赛克UE5获奖者赞助了专业发展机会，继续培训 K99阶段将使Lee-Kim博士成功过渡到独立。