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 新出现的证据表明，不适当的基质金属蛋白酶（MMP）的活性可能是基础， 动脉粥样硬化和血管炎症的发病机制。尽管有数据表明MMP有助于 动脉粥样硬化病变重塑和不良结局，基本上对MMPs的作用一无所知 在动脉粥样硬化的背景下作为控制血管细胞行为的活性信号分子 疾病我们最近意外地发现MMP-1可以切割并激活蛋白酶激活的 受体-1（PAR 1）信号在血管中的作用。这是重要的，因为它是第一个直接报告， 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PAR 1， 通过在不同于典型凝血酶切割位点的位点切割受体， 拴系配体偏向于不同的G蛋白信号传导途径谱。这方面的研究 一项提案将集中在MMP 1-PAR 1在动脉粥样硬化发展中完全未探索的作用上， 斑块此外，PAR 1参与动脉粥样硬化（不考虑蛋白酶激动剂）作为一种免疫调节剂， 慢性进展性炎症性疾病，基本上是未知的。目标1中要检验的中心假设是 内皮MMP-1首先通过PAR 1作为活性信号分子触发内皮炎症， 和单核细胞进入早期斑块。在晚期病变中，来自巨噬细胞的MMP 1自动刺激PAR 1， 通过分泌MCP-1和其它介质使慢性炎症和促有丝分裂状态永久化。我们将使用 基于细胞的实验和Mmp 1a或Par 1遗传缺陷的高脂血症小鼠模型。我们 将确定小鼠内皮细胞中MMP 1a和PAR 1对单核细胞粘附的需求， 使用分离自的小鼠心脏内皮细胞在体外剪切流条件下的迁移 Mmp 1a-/-和Par 1-/-小鼠以及来自高脂饮食后的ApoE-/-小鼠的单核细胞。目标2将审查 冠心病和急性冠脉综合征患者外周血单核细胞MMP 1-PAR 1系统的研究 经皮冠状动脉介入治疗（PCI）综合征。基线时患者的单核细胞， PCI和接受新型PAR 1 pepducin（PZ-128）治疗的患者将在平行板中使用 （动脉剪切）流动室，以确定通过内皮单层的迁移。我们将确定 具有超活性Mmp 1启动子多态性的受试者是否表现出更高的MMP 1表达， 导致其单核细胞上的“MMP 1-PAR 1”表型相对于“TF-PAR 1”表型。Aim 3将使用Adoptive 从Par 1-/-ApoE-/-和Mmp 1a-/-ApoE-/-小鼠转移骨髓来源的细胞，以帮助确定细胞 MMP 1-PAR 1系统在动脉粥样硬化形成中的类型特异性病理生物学（例如内皮细胞与白细胞）。我们 了解MMP 1-PAR 1信号在内皮细胞和单核细胞上的病理生理学相关性/ 巨噬细胞，以及将这些事件与动脉粥样硬化斑块发展联系起来的机制将提供一个 该框架旨在推进未来可能阻止或逆转动脉粥样硬化进展的疗法。

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

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

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