Matrix Metalloprotease-PAR1 Regulation of Atherosclerosis

基质金属蛋白酶-PAR1对动脉粥样硬化的调节

• 批准号: 10064145
• 负责人: ATHAN KULIOPULOS
• 金额: $ 64.61万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-15 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064145
• 关键词: Adhesions; Adoptive Transfer; Agonist; American; Apolipoprotein E; Arterial Fatty Streak; Atherosclerosis; Behavior; Blood Platelets; Blood Vessels; Bone Marrow; CCL2 gene; Cause of Death; Cells; Chronic; Cleaved cell; Clinical Data; Coronary Arteriosclerosis; Coronary artery; Data; Development; Disease; Endothelial Cells; Endothelium; Event; Exhibits; Future; G-Protein Signaling Pathway; Genetic; Genetic Polymorphism; Goals; Health Care Costs; Heart; High Fat Diet; In Vitro; Incidence; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-12; Lead; Lesion; Leukocytes; Life; Ligands; Link; Mediator of activation protein; Metalloproteases; Modeling; Mus; New Agents; Outcome; PAR-1 Receptor; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Phenotype; Regulation; Reporting; Role; Rupture; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Stenosis; Stimulation of Cell Proliferation; System; Testing; Thrombin; Thrombin Receptor; Work; acute coronary syndrome; atherogenesis; atherosclerosis risk; base; cell type; collagenase; coronary lesion; experimental study; interest; macrophage; monocyte; monolayer; mortality; mouse model; novel; percutaneous coronary intervention; promoter; receptor; recruit; vascular inflammation

Project Summary Emerging evidence suggests that inappropriate matrix metalloprotease (MMP) activity may underlie the pathogenesis of atherosclerosis and vascular inflammation. Despite data that MMPs contribute to atherosclerotic lesion remodeling and poor outcomes, essentially nothing is known regarding the role of MMPs as active signaling molecules in controlling the behavior of vascular cells in the context of atherosclerotic disease. We recently made the unanticipated discovery that MMP-1 cleaves and activates protease-activated receptor-1 (PAR1) signaling in blood vessels. This is of major import as it was the first report of a direct signaling function of the principal collagenase in blood vessels. Notably, we found that MMP-1 activates PAR1 by cleaving the receptor at a distinct site from the canonical thrombin cleavage site which generates a longer tethered ligand that is biased towards a different spectrum of G protein signaling pathways. The studies in this proposal will focus on the completely unexplored role of MMP1-PAR1 in the development of atherosclerotic plaques. Furthermore, the involvement of PAR1 in atherosclerosis (regardless of the protease agonist) as a chronic evolving inflammatory disease, is essentially unknown. The central hypothesis to be tested in aim 1 is that endothelial MMP-1 first acts as an active signaling molecule via PAR1 to trigger endothelial inflammation and monocyte entry into early plaques. In advanced lesions, MMP1 from macrophages autostimulates PAR1 to perpetuate a chronic inflammatory and mitogenic state by secretion of MCP-1 and other mediators. We will use both cell-based experiments, and hyperlipidemic mouse models with genetic deficiency of Mmp1a or Par1. We will determine the requirement of MMP1a and PAR1 in mouse endothelial cells for monocyte adhesion and transmigration under shear flow conditions in vitro using mouse heart endothelial cells isolated from of Mmp1a-/- and Par1-/- mice, and monocytes from ApoE-/- mice after high fat diet. Aim 2 will examine the role of the MMP1-PAR1 system on circulating monocytes in subjects with coronary artery disease and acute coronary syndromes undergoing percutaneous coronary interventions (PCI). Monocytes from patients at baseline prior to PCI and from those being treated with a novel PAR1 pepducin, PZ-128, will be used in Parallel Plate (arterial-shear) flow chambers to determine transmigration through endothelial monolayers. We will determine whether subjects with a super-active Mmp1 promoter polymorphism exhibit higher expression of MMP1 that contributes to a `MMP1-PAR1' phenotype vs `TF-PAR1' phenotype on their monocytes. Aim 3 will use adoptive transfer of bone marrow-derived cells from Par1-/-ApoE-/- and Mmp1a-/-ApoE-/- mice to help define the cell- type specific pathobiology (e.g. endothelium vs leukocytes) of the MMP1-PAR1 system in atherogenesis. Our understanding of the pathophysiologic relevance of MMP1-PAR1 signaling on endothelium and monocytes/ macrophages, and the mechanism linking these events to atherosclerotic plaque development will provide a framework to advance future therapies that could halt or reverse the progression of atherosclerosis.

项目摘要 新兴证据表明，不适当的基质金属蛋白酶（MMP）活动可能是 动脉粥样硬化和血管炎症的发病机理。尽管数据表明MMP有助于 动脉粥样硬化病变的重塑和不良的结果，基本上尚无MMP的作用 作为在动脉粥样硬化中控制血管细胞行为时的主动信号分子 疾病。我们最近做出了一个意外的发现，即MMP-1裂解并激活蛋白酶激活 血管中的受体1（PAR1）信号传导。这是重要的重要性，因为它是直接的第一个报告 血管中主要胶原酶的信号传导功能。值得注意的是，我们发现MMP-1激活PAR1 通过将受体在不同的位点与规范凝血酶裂解位点切割，该位点产生更长的 束缚的配体偏向不同的G蛋白信号通路。关于这一点的研究 提案将集中于MMP1-PAR1在动脉粥样硬化发展中的完全未开发的作用 斑块。此外，PAR1参与动脉粥样硬化（无论蛋白酶激动剂如何） 慢性发展的炎症性疾病本质上是未知的。 AIM 1中要检验的中心假设是 该内皮MMP-1首先通过PAR1充当活动信号分子，以触发内皮炎症 和单核细胞进入早期斑块。在晚期病变中，来自巨噬细胞的MMP1自动刺激PAR1至 通过MCP-1和其他介体的分泌使慢性炎症和有丝分裂状态永久存在。我们将使用 两个基于细胞的实验和具有MMP1A或PAR1遗传缺陷的高脂小鼠模型。我们 将确定小鼠内皮细胞中MMP1A和PAR1对单核细胞粘附和 使用小鼠心脏内皮细胞在体外剪切流条件下进行迁移 高脂肪饮食后，MMP1A - / - 和PAR1-/ - 小鼠以及ApoE-/ - 小鼠的单核细胞。 AIM 2将检查 冠状动脉疾病和急性冠状动脉受试者循环单核细胞的MMP1-PAR1系统 接受经皮冠状动脉干预（PCI）的综合征。基线先验的患者的单核细胞 对PCI以及接受新型PAR1遍布Pepducin PZ-128的人，将用于并行板 （动脉剪切）流室，以确定通过内皮单层移民。我们将确定 具有超活动MMP1启动子多态性的受试者是否表现出更高的MMP1表达 有助于单核细胞上的“ MMP1-PAR1”表型与`tf-par1'表型。 AIM 3将使用收养 骨髓衍生的细胞从PAR1 - / - APOE-/ - 和MMP1A - / - APOE-/ - 小鼠转移，以帮助定义细胞 - 在动脉粥样硬化中，MMP1-PAR1系统的类型特定病理生物学（例如内皮与白细胞）。我们的 了解MMP1-PAR1信号在内皮和单核细胞/ 巨噬细胞以及将这些事件与动脉粥样硬化斑块发育联系起来的机制将提供 框架可以推进可能停止或扭转动脉粥样硬化进展的未来疗法。

项目成果

Deficiency of MMP1a (Matrix Metalloprotease 1a) Collagenase Suppresses Development of Atherosclerosis in Mice: Translational Implications for Human Coronary Artery Disease.

• DOI: 10.1161/atvbaha.120.315837
• 发表时间: 2021-05-05
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Fletcher EK;Wang Y;Flynn LK;Turner SE;Rade JJ;Kimmelstiel CD;Gurbel PA;Bliden KP;Covic L;Kuliopulos A
• 通讯作者: Kuliopulos A

PAR1 (Protease-Activated Receptor 1) Pepducin Therapy Targeting Myocardial Necrosis in Coronary Artery Disease and Acute Coronary Syndrome Patients Undergoing Cardiac Catheterization: A Randomized, Placebo-Controlled, Phase 2 Study.

• DOI: 10.1161/atvbaha.120.315168
• 发表时间: 2020-12
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Kuliopulos A;Gurbel PA;Rade JJ;Kimmelstiel CD;Turner SE;Bliden KP;Fletcher EK;Cox DH;Covic L;TRIP-PCI Investigators
• 通讯作者: TRIP-PCI Investigators

Enhanced potency of prasugrel on protease-activated receptors following bivalirudin treatment for PCI as compared to clopidogrel.

与氯吡格雷相比，比伐卢定治疗 PCI 后普拉格雷对蛋白酶激活受体的效力增强。

• DOI: 10.1016/j.thromres.2019.01.017
• 发表时间: 2019
• 期刊: Thrombosis research
• 影响因子: 7.5
• 作者: Kimmelstiel,Carey;Stevenson,Ryan;Nguyen,Nga;VanDoren,Layla;Zhang,Ping;Perkins,James;Kapur,NavinK;Weintraub,Andrew;Castaneda,Vilma;Kuliopulos,Athan;Covic,Lidija
• 通讯作者: Covic,Lidija

Lipid Receptor GPR31 (G-Protein-Coupled Receptor 31) Regulates Platelet Reactivity and Thrombosis Without Affecting Hemostasis.

• DOI: 10.1161/atvbaha.120.315154
• 发表时间: 2021-01
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Van Doren L;Nguyen N;Garzia C;Fletcher EK;Stevenson R;Jaramillo D;Kuliopulos A;Covic L
• 通讯作者: Covic L

Noncanonical Matrix Metalloprotease 1-Protease-Activated Receptor 1 Signaling Drives Progression of Atherosclerosis.

• DOI: 10.1161/atvbaha.118.310967
• 发表时间: 2018-06
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Rana R;Huang T;Koukos G;Fletcher EK;Turner SE;Shearer A;Gurbel PA;Rade JJ;Kimmelstiel CD;Bliden KP;Covic L;Kuliopulos A
• 通讯作者: Kuliopulos A