Systems Genetics of Vascular Smooth Muscle Phenotypes

血管平滑肌表型的系统遗传学

• 批准号: 10559249
• 负责人: Mete Civelek
• 金额: $ 50.74万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10559249
• 关键词: ARVCF gene; Affect; African ancestry; Allelic Imbalance; Antihypertensive Agents; Aorta; Arterial Fatty Streak; Arteries; Asian ancestry; Atherosclerosis; Bayesian Network; Biological; Biological Assay; Blood Vessels; Cause of Death; Cell Culture Techniques; Cell Separation; Cell physiology; Characteristics; Cholesterol; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Computational Biology; Coronary; Coronary Arteriosclerosis; Coronary artery; Data; Diagnosis; Disease; Disease susceptibility; Donor person; Early Diagnosis; Enhancers; Environmental Risk Factor; European ancestry; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genetic Variation; Genetic study; Genome; Genotype; Goals; Heart Transplantation; Human; Human Resources; Immunohistochemistry; In Vitro; Individual; Inflammatory; Latinx; Lentivirus; Linkage Disequilibrium; Lipids; Maps; Measures; Modeling; Molecular; Myocardial Infarction; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Process; Proliferating; Protein Analysis; Proteins; Quantitative Trait Loci; RNA; Regulation; Regulator Genes; Regulatory Element; Reporter; Research; Resources; Risk; Risk Factors; Role; Series; Serum; Single Nucleotide Polymorphism; Smooth Muscle Myocytes; Specimen; Stimulus; Sudden Death; System; Testing; Time; Transcript; Translating; Transposase; United States; Untranslated RNA; Update; Variant; Vascular Smooth Muscle; Western World; ascending aorta; calcification; causal variant; cell type; chromatin remodeling; clinical care; cytokine; disorder prevention; disorder risk; drug discovery; experimental study; gain of function; gene function; gene interaction; gene network; gene regulatory network; genetic approach; genetic variant; genome wide association study; genomic locus; loss of function; migration; multi-ethnic; particle; public health relevance; response; transcription factor; transcriptome sequencing; transdifferentiation

PROJECT SUMMARY Coronary artery disease (CAD) remains the leading cause of death in the Western world despite significant advances in early detection and extensive use of lipid-lowering and anti-hypertensive drugs. To date, no single drug has been developed to target the primary disease process in the vessel wall. A complete understanding of the disease susceptibility is urgently needed to develop additional therapies. Common forms of atherosclerosis involve environmental factors, hundreds of genetic variations, and their interactions, each of which exert a relatively small effect on disease susceptibility. The most recent genome-wide association study (GWAS) in nearly six hundred fifty thousand individuals identified 175 independent variants associated with increased risk for CAD. However, most of the underlying genes and the related mechanisms of how these variants contribute to the disease process remain unknown. This proposal outlines an integrative genetics study in a unique resource of human aortic smooth muscle cells (SMCs) isolated from 151 genotyped multi- ethnic heart transplant donors to discover the CAD-associated variants that perturb SMC gene expression and their downstream functional consequences. In recent studies, we measured gene expression in quiescent and proliferative culture conditions representing the transdifferentiation of SMCs from a healthy to an atherogenic phenotype. We identified 84 genes whose expression was associated with CAD variants in GWAS loci. However, the causal genetic variants in these loci remain to be elucidated. Therefore, as part of the proposed studies, we will first perform massively parallel reporter assays to identify the variants that modulate gene expression in SMCs. We will also take advantage of the natural variation in gene expression to construct co- expression and Bayesian networks to understand how the predicted candidate causal genes function in SMCs. We will refine these networks by mapping regulatory elements to nascent RNA transcripts in response to pro- inflammatory cytokines. We will then validate our predictions in gain and loss-of-function experiments in cultured SMCs. We will also validate our predictions in well-phenotyped coronary artery specimens from cases of unexpected sudden death by performing immunohistochemical analysis of proteins encoded by genes that are predicted to play a key role in atherosclerosis-relevant SMC phenotypes. The overall goal of the proposed studies is to integrate systems genetics and computational biology leading to mechanistic predictions of the gene networks that are perturbed by CAD. Besides understanding CAD loci, these integrative genetics studies will provide a useful window into the flow of biological information from genetic variants to SMC gene expression and atherosclerosis-relevant phenotypes.

项目总结 冠状动脉疾病(CAD)仍然是西方世界的主要死亡原因，尽管 降脂降血压药物的早期发现和广泛应用进展。到目前为止，还没有一张 已经开发出针对血管壁中的原发疾病过程的药物。完全了解 目前迫切需要开发新的治疗方法，以减少疾病的易感性。常见形式的 动脉粥样硬化涉及环境因素、数百种遗传变异及其相互作用，每一种 它们对疾病易感性的影响相对较小。最新的全基因组关联研究 在近65万名个体中发现了175个与 增加了冠心病的风险。然而，大多数潜在的基因和相关的机制如何这些 导致疾病过程的变异体仍不清楚。这项建议勾勒出了一种综合遗传学 从151例多基因分型人主动脉平滑肌细胞分离获得独特来源的研究 少数民族心脏移植捐赠者发现扰乱SMC基因表达和 它们的下游功能后果。在最近的研究中，我们测量了静止和 代表SMC从健康细胞向致动脉粥样硬化细胞转分化的增殖培养条件 表型。我们鉴定了84个基因，这些基因的表达与Gwas基因座上的CAD变异相关。 然而，这些基因座的致病遗传变异仍有待阐明。因此，作为提议的一部分， 在研究中，我们将首先进行大规模的平行报告分析，以确定调节基因的变异 在SMC中的表达。我们还将利用基因表达的自然变异来构建联合 表达和贝叶斯网络，以了解预测的候选因果基因如何在SMC中发挥作用。 我们将通过将调控元件映射到新生的RNA转录产物来完善这些网络，以响应亲- 炎性细胞因子。然后，我们将在功能的增益和损耗实验中验证我们的预测 培养的SMC。我们还将在来自病例的表型良好的冠状动脉样本中验证我们的预测。 通过对由以下基因编码的蛋白质进行免疫组织化学分析 被预测在动脉粥样硬化相关的SMC表型中起关键作用。建议的总体目标是 研究的目的是将系统遗传学和计算生物学相结合，从而实现对 被CAD扰乱的基因网络。除了了解CAD基因座外，这些综合遗传学研究 将为从遗传变异到SMC基因的生物信息流提供一个有用的窗口 表达与动脉粥样硬化相关表型。