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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要心血管疾病是世界上导致死亡的主要原因，发展非脂质至关重要。治疗心血管疾病的方法，因为成功降脂后仍然存在显着的风险。使用人类疾病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、途径和治疗策略。 Stitziel 实验室发现了细胞外基质基因 SVEP1 中的一个变体，该变体与为了测试 SVEP1 是否是风险基因座中的致病基因，实验室首先生成了冠状动脉疾病的基因。 Svep1 单倍体不足的易患动脉粥样硬化的小鼠被发现表现出较少的动脉粥样硬化。类似地，成熟血管平滑肌细胞中 Svep1 的条件性缺失。小鼠的血管平滑肌细胞 (VSMC) 显着减少了斑块负担。越来越多的证据表明，血管平滑肌细胞 (VSMC) 可以显着减少斑块负担。在动脉粥样硬化中发挥着核心作用，除了这些细胞之外，还包括目前与这些细胞相关的几个疾病位点。产生 SVEP1 的 VSMC 含有 Notch 和整合素受体，我们追踪到它们与 SVEP1 结合。发现在重组 SVEP1 上生长的 VSMC 增加了 Notch 和整合素信号传导，并且参与细胞增殖和分化的基因转录增加可诱导强健。原代 VSMC 的增殖，依赖于 Notch 和整合素 α9β1 信号传导。初步研究结果证实 SVEP1 可能通过影响 VSMC 来促进动脉粥样硬化尽管有这些有希望的线索，但其机制仍以细胞自主方式进行增殖和分化。 SVEP1 及其变体导致疾病的机制尚未完全确定。该项目将通过以下方式回答有关分子和细胞机制的关键、悬而未决的问题其中 SVEP1 会促进动脉粥样硬化形成，我将使用互补的分子技术、细胞培养模型和我首先旨在确定 SVEP1 是否直接与 Notch 和整合素结合。受体，如果是的话，蛋白质的哪些区域有助于结合亲和力。这个实验也将阐明。每个信号通路对 SVEP1 对 VSMC 的总体影响的贡献。将被纳入这些研究中，因为变体残基位于推定的整联蛋白结合域内然后我将使用小鼠疾病来探究 SVEP1 在动脉粥样硬化形成中的细胞机制。这将包括在有或没有的情况下进行谱系追踪和单细胞 RNA 测序。新内膜 VSMC 内源性产生 SVEP1，这种体内方法补充了所提出的方法。分子技术通过关注其病理生理学背景的机制成功完成。这些目标将揭示 SVEP1 的常见和风险等位基因促进动脉粥样硬化的机制同时深入了解世界上最致命疾病的发病机制，有可能揭示新的治疗候选者。