Atherogenic mechanisms of SVEP1, a Novel Human Coronary Artery Disease Locus

人类冠状动脉疾病基因座 SVEP1 的致动脉粥样硬化机制

• 批准号: 10449595
• 负责人: Jared Scott Elenbaas
• 金额: $ 0.25万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10449595
• 关键词: Address; Adhesions; Affect; Affinity; Animal Model; Arterial Fatty Streak; Atherosclerosis; Binding; Biological Assay; Cardiovascular Diseases; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cells; Clinical; Clonal Expansion; Complement; Coronary Arteriosclerosis; Data; Development; Differentiation and Growth; Disease; Disease model; Embryo; Exhibits; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Foundations; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Genomics; Goals; Histologic; Human; Immobilization; Immunofluorescence Microscopy; Integrin Binding; Integrins; Investigation; Knock-out; Knowledge; Label; Link; Lipids; Measures; Methods; Modeling; Molecular; Mus; Nucleosides; Pathogenesis; Pathway interactions; Phenotype; Physicians; Plasma; Play; Preparation; Process; Production; Protein Region; Proteins; Recombinants; Resources; Risk; Role; SARS-CoV-2 B.1.1.7; Scientist; Serum; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Surface Plasmon Resonance; System; Techniques; Testing; Therapeutic; Variant; Vascular Smooth Muscle; atherogenesis; behavior influence; biobank; career; causal variant; cell behavior; cell growth; clinical application; disorder risk; drug development; experimental study; genome-wide; human disease; improved; in vivo; insight; integrin alpha9 beta1; interest; mortality; notch protein; novel; novel strategies; novel therapeutic intervention; novel therapeutics; overexpression; prevent; receptor; risk variant; single-cell RNA sequencing; small molecule; small molecule inhibitor; therapeutic candidate; vascular smooth muscle cell proliferation

PROJECT SUMMARY/ABSTRACT Cardiovascular disease is the leading cause of mortality in the world. It is critical to develop non-lipid therapies to address cardiovascular disease since significant risk remains after successful lipid reduction. By using human disease findings as a starting point for experimental investigation, we can focus our resources on the mechanisms, pathways and therapeutic strategies that are the most applicable to human disease. The Stitziel Lab discovered a variant in the extracellular matrix gene, SVEP1, that positively associates with coronary artery disease. To test if SVEP1 is the causal gene in the risk locus, the lab first generated athero-prone mice that were haploinsufficient for Svep1. These mice were found to exhibit less atherosclerotic plaque burden than controls. Similarly, conditional deletion of Svep1 in mature vascular smooth muscle cells (VSMCs) of mice resulted in dramatically less plaque burden. There is a growing body of evidence that VSMCs play a central role in atherosclerosis, including several disease loci now linked to these cells. In addition to producing SVEP1, VSMCs contain Notch and integrin receptors that we hypothesize bind to SVEP1. I discovered that VSMCs grown on recombinant SVEP1 have increased Notch and integrin signaling, as well as increased transcription of genes involved in cell proliferation and differentiation. SVEP1 induces robust proliferation of primary VSMCs, which is dependent on both Notch and integrin α9β1 signaling. These preliminary findings confirm the contribution of SVEP1 to atherosclerosis, potentially by influencing VSMC proliferation and differentiation in a cell-autonomous manner. Despite these promising leads, the mechanisms by which SVEP1 and its variants contribute to disease have yet to be fully characterized. This project will answer critical, outstanding questions about the molecular and cellular mechanisms by which SVEP1 promotes atherogenesis. I will use complementary molecular techniques, cell culture models and animal models to address these questions. I first aim to determine if SVEP1 binds directly to Notch and integrin receptors and, if so, which regions of the protein contribute to binding affinity. This experiment will also clarify the contribution of each signaling pathway to the overall effects of SVEP1 on VSMCs. The leading risk variant will be included in these studies, since the variant residue is within the putative integrin binding domain of SVEP1. I will then interrogate the cellular mechanisms of SVEP1 in atherogenesis using a murine disease model. This will include performing lineage tracing and single cell RNA sequencing with and without the endogenous production of SVEP1 by neointimal VSMCs. This in vivo approach complements the proposed molecular techniques by focusing on mechanisms in their pathophysiologic context. Successful completion of these aims will reveal the mechanisms by which the common and risk allele of SVEP1 promote atherosclerosis while providing insight into the pathogenesis of the world’s deadliest disease with potential to reveal new therapeutic candidates.

项目摘要/摘要 心血管疾病是世界上死亡率的主要原因。发展非脂质至关重要 解决心血管疾病的疗法，因为成功脂质降低后仍然存在重大风险。经过 使用人类疾病的发现作为实验研究的起点，我们可以将资源集中在 最适用于人类疾病的机制，途径和理论策略。 Stitziel Lab在细胞外基质基因SVEP1中发现了一个变体，该变体积极地关联 冠状动脉疾病。为了测试SVEP1是否是风险基因座的因果基因，实验室首先生成 对于SVEP1的动脉繁殖小鼠，易于使用。发现这些小鼠表现出较少的动脉粥样硬化 斑块伯恩比对照组。同样，成熟的血管平滑肌细胞中SVEP1的条件缺失 （VSMC）的小鼠导致斑块负担较小。越来越多的证据表明VSMC 在动脉粥样硬化中起着核心作用，其中包括与这些细胞相关的几个疾病基因座。此外 产生SVEP1，VSMC包含我们假设与SVEP1结合的Notch和整联蛋白受体。我 发现在重组SVEP1上生长的VSMC具有增加的Notch和整联蛋白信号传导，以及 参与细胞增殖和分化的基因转录增加。 SVEP1诱导健壮 原代VSMC的增殖，这取决于Notch和整合蛋白α9β1信号传导。这些 初步发现证实了SVEP1对动脉粥样硬化的贡献，可能会受到影响VSMC 以细胞自治方式扩散和分化。尽管有这些有希望的线索，但机制还是 SVEP1及其变体对疾病的贡献尚未充分表征。 该项目将通过 SVEP1促进了动脉粥样硬化。我将使用完整的分子技术，细胞培养模型和 动物模型以解决这些问题。我首先要确定SVEP1是否直接与Notch和Intemin结合 受体等，如果是这样，蛋白质的哪个区域有助于结合亲和力。该实验也将澄清 每个信号通路对SVEP1对VSMC的总体影响的贡献。领先的风险变体 这些研究将包括在这些研究中，因为变体居住在推定的整联蛋白结合域内 SVEP1。然后，我将使用鼠类疾病在动脉粥样硬化中询问SVEP1的细胞机制 模型。这将包括执行有或没有和不带有和没有 通过新内膜VSMC对SVEP1的内源性产生。这种体内方法使提出的 分子技术通过在病理生理环境中的机制重点关注。成功完成 这些目标将揭示SVEP1常见和风险等位基因促进动脉粥样硬化的机制 同时洞悉世界上最致命疾病的发病机理，有可能揭示新的 治疗候选人。