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: 10664846
• 负责人: Jared Scott Elenbaas
• 金额: $ 5.52万
• 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/10664846
• 关键词: Address; Adhesions; Affect; Affinity; Alleles; 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; Coronary Arteriosclerosis; Data; Development; Disease; Disease model; Embryo; Exhibits; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Foundations; Genes; Genetic; Genetic Complementation Test; 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; Proliferating; Protein Region; Proteins; Recombinants; Resources; Risk; Role; SARS-CoV-2 B.1.1.7; Scientist; Serum; Signal Induction; 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; comparison control; 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.

项目总结/摘要 心血管疾病是世界上主要的死亡原因。开发非脂 治疗心血管疾病，因为成功降脂后仍存在重大风险。通过 利用人类疾病的发现作为实验研究的起点，我们可以把资源集中在 最适用于人类疾病的机制、途径和治疗策略。 缝合实验室在细胞外基质基因SVEP 1中发现了一种变体， 冠状动脉疾病为了测试SVEP 1是否是风险基因座中的致病基因，实验室首先生成了 易患动脉粥样硬化的小鼠，Svep 1单倍不足。这些小鼠被发现表现出较少的动脉粥样硬化 牙菌斑负荷高于对照组。类似地，成熟血管平滑肌细胞中Svep 1的条件性缺失 小鼠的血管平滑肌细胞（VSMCs）导致斑块负荷显著减少。越来越多的证据表明， 在动脉粥样硬化中起核心作用，包括现在与这些细胞相关的几个疾病位点。除了 在产生SVEP 1的过程中，VSMC含有Notch和整合素受体，我们假设它们与SVEP 1结合。我 发现在重组SVEP 1上生长的VSMC增加了Notch和整合素信号传导， 参与细胞增殖和分化的基因转录增加。SVEP 1诱导稳健 原代VSMCs的增殖依赖于Notch和整合素α9β1信号传导。这些 初步研究结果证实SVEP 1可能通过影响VSMC而对动脉粥样硬化起作用 以细胞自主的方式增殖和分化。尽管有这些有希望的线索， SVEP 1及其变异体对疾病的作用机制尚未完全确定。 这个项目将回答关键的，悬而未决的问题，分子和细胞机制， SVEP 1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， 动物模型来解决这些问题。我的第一个目标是确定SVEP 1是否直接结合Notch和整合素 受体，如果是，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SVEP 1对VSMC的总体作用的贡献。主要风险变量 将包括在这些研究中，因为变体残基位于 SVEP 1.然后，我将使用小鼠疾病研究SVEP 1在动脉粥样硬化形成中的细胞机制。 模型这将包括进行谱系追踪和单细胞RNA测序， 新生内膜VSMC内源性产生SVEP 1。这种体内方法补充了所提出的 分子技术通过关注其病理生理学背景下的机制。成功完成 这些目标将揭示SVEP 1的常见和危险等位基因促进动脉粥样硬化的机制 同时提供了对世界上最致命疾病的发病机制的深入了解， 治疗候选人。

项目成果

Vascular smooth muscle- and myeloid cell-derived integrin α9β1 does not directly mediate the development of atherosclerosis in mice.

• DOI: 10.1016/j.atherosclerosis.2022.09.015
• 发表时间: 2022-11
• 期刊: ATHEROSCLEROSIS
• 影响因子: 5.3
• 作者: Jung, In-Hyuk;Elenbaas, Jared S.;Burks, Kendall H.;Amrute, Junedh M.;Zhang, Xiangyu;Alisio, Arturo;Stitziel, Nathan O.
• 通讯作者: Stitziel, Nathan O.