Protease-activated receptors in vascular integrity and dysfunction

血管完整性和功能障碍中的蛋白酶激活受体

• 批准号: 7883323
• 负责人: ATHAN KULIOPULOS
• 金额: $ 40.25万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7883323
• 关键词: Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Arterial Injury; Atherosclerosis; Biochemical; Biochemical Genetics; Biology; Blood Platelets; Blood Vessels; Cell Proliferation; Cell physiology; Cells; Cleaved cell; Communication; Coronary Arteriosclerosis; Data; Disease; Endothelial Cells; Endothelium; Functional disorder; Heart Diseases; Hyperplasia; Inflammation; Inflammatory; Kinetics; Lead; Leukocytes; Life; Ligands; Malignant Epithelial Cell; Mediating; Metalloproteases; Modeling; Molecular; Mouse Strains; Myocardial Infarction; Myocardial dysfunction; Output; PAR-1 Receptor; PAWR gene; Paracrine Communication; Pathogenesis; Pathway interactions; Patients; Physiological; Play; Process; Proteinase-Activated Receptors; Public Health; RNA Interference; Role; Sepsis; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Staging; Stroke; Techniques; Transactivation; Vascular Diseases; Vascular Endothelium; Vasomotor; Wild Type Mouse; Wound Healing; autocrine; base; cell type; chemokine; chemokine receptor; in vivo; injury and repair; migration; novel; novel therapeutics; prevent; protective effect; restenosis; rho

DESCRIPTION (provided by applicant): Emerging evidence suggests that matrix metalloproteases (MMPs) play a pivotal role in vascular integrity and wound healing, however, inappropriate MMP activity may underlie the pathogenesis of several diseases including restenosis, atherosclerosis, and myocardial dysfunction. Selectively controlling defined aspects of MMP activity in restenosis and end-stage coronary artery disease may be a valid target for therapy. We recently made the unanticipated discovery that the matrix metalloprotease MMP-1, cleaves and activates protease-activated receptor 1 (PAR1) in a variety of cells including endothelium, vascular SMCs, platelets and carcinoma cells. Both MMP-1 and PAR1 are highly upregulated by several cell types in the blood vessel wall in the context of intimal hyperplasia and inflammatory states. However, it is completely unknown what role(s), MMP1-PAR1 signaling plays in either normal blood vessel biology or in vascular disease. Moreover, activation of PAR1 can lead to decreased barrier function, dysregulated vasomotor tone and control of SMC functions through inter-cellular communication pathways. In aim 1, we will investigate the molecular mechanism and (patho)physiologic relevance of MMP1-PAR1 signaling in SMC proliferation, migration and invasion, survival and de-differentiation in the arterial wall. Modulation of PAR1 signaling with cell- penetrating pepducins will be used to evaluate the function(s) of MMP1-PAR1 signaling in in vivo arterial injury and restenosis models using wild-type mice and strains deficient in PAR1, PAR2 and PAR4. Furthermore, prior studies suggested that PAR1 promotes the conversion of the endothelium to a pro- thrombotic and pro-inflammatory state. We made the recent observation that PAR1 can also confer protective effects in endothelial barrier function and survival in animal models of systemic inflammation by transactivation of the structurally-related PAR2. Our preliminary data suggest that the signaling outputs of PAR1 and PAR2 are entwined and need to be examined as a potential functional unit. Aim 2 will use a combination of biochemical, pharmacological and RNA interference techniques to determine how PAR1 and PAR2 regulate Rac versus Rho signaling. We will delineate the components and kinetics of downstream pro- inflammatory versus anti-inflammatory signaling pathways emanating from PAR1 and PAR2 in the context of barrier disruption and leukocyte transmigration. Critical downstream chemokine pathways known to mediate both autocrine and paracrine signaling during these processes will be interrupted with our newly validated chemokine receptor pepducins. Lastly, we will investigate the molecular basis of PAR1-PAR2 interactions and liganding using biochemical and genetic approaches. Relevance to public health: The studies outlined here may lead to the eventual discovery of novel therapeutics that may prevent heart attacks and stroke in patients with heart disease, and could be used in other conditions such as life-threatening sepsis.

描述（由申请人提供）：新出现的证据表明，基质金属蛋白酶（MMP）在血管完整性和伤口愈合中起关键作用，但是，不适当的MMP活性可能是多种疾病的发病机理，包括再危害，动脉粥样硬化，动脉粥样硬化和心肌功能障碍。选择性控制MMP活性在再狭窄和末期冠状动脉疾病中的定义方面可能是治疗的有效靶标。我们最近做出了一个意想不到的发现，即基质金属蛋白酶MMP-1，裂解并激活蛋白酶激活的受体1（PAR1）在包括内皮，血管SMC，血小板和癌细胞在内的多种细胞中。在内膜增生和炎症态的背景下，血管壁中的几种细胞类型都高度上调了MMP-1和PAR1。但是，完全未知在正常血管生物学或血管疾病中有什么作用，MMP1-PAR1信号传导起着什么作用。此外，PAR1的激活可以导致屏障功能降低，血管舒缩张力失调以及通过细胞间通信对SMC功能的控制。在AIM 1中，我们将研究MMP1-PAR1信号在SMC增殖，迁移和侵袭，生存和脱离动脉壁中MMP1-PAR1信号的分子机制和（病原）生理相关性。使用野生型小鼠和pAR1，PAR2和PAR4缺乏的菌株，使用细胞穿透性腹膜素对PAR1信号传导的调节将用于评估体内动脉损伤中MMP1-PAR1信号传导的功能。此外，先前的研究表明，PAR1促进了内皮的转化为血栓形成和促炎状态。我们最近的观察结果是，PAR1还可以通过与结构相关的PAR2的反式激活进行全身炎症的动物模型中赋予内皮屏障功能的保护作用和生存。我们的初步数据表明，PAR1和PAR2的信号输出被缠绕，需要检查为潜在功能单元。 AIM 2将使用生化，药理学和RNA干扰技术的组合来确定PAR1和PAR2如何调节RAC与RHO信号传导。我们将在屏障破坏和白细胞传播的背景下描述下游促炎性与抗炎信号通路的组件和动力学。在这些过程中，已知可以介导自分泌和旁分泌信号传导已知的关键下游趋化因子途径将被我们新验证的趋化因子受体遍布蛋白质中断。最后，我们将使用生化和遗传方法研究PAR1-PAR2相互作用和配体的分子基础。与公共卫生的相关性：这里概述的研究可能导致最终发现新型治疗剂，这些疗法可能会阻止心脏病患者的心脏病发作和中风，并且可以在其他危及生命的败血症等疾病中使用。

项目成果

Mouse matrix metalloprotease-1a (Mmp1a) gives new insight into MMP function.

• DOI: 10.1002/jcp.24650
• 发表时间: 2014-12
• 期刊: JOURNAL OF CELLULAR PHYSIOLOGY
• 影响因子: 5.6
• 作者: Foley, Caitlin J.;Kuliopulos, Athan
• 通讯作者: Kuliopulos, Athan

Role of the PAR1 receptor 8th helix in signaling: the 7-8-1 receptor activation mechanism.

PAR1 受体第八螺旋在信号传导中的作用：7-8-1 受体激活机制。

• DOI: 10.1074/jbc.m509525200
• 发表时间: 2006
• 期刊: The Journal of biological chemistry
• 作者: Swift,Steven;Leger,AndrewJ;Talavera,Joyce;Zhang,Lei;Bohm,Andrew;Kuliopulos,Athan
• 通讯作者: Kuliopulos,Athan

Pharmacology, biodistribution, and efficacy of GPCR-based pepducins in disease models.

基于 GPCR 的 pepducins 在疾病模型中的药理学、生物分布和功效。

• DOI: 10.1007/978-1-60761-919-2_19
• 发表时间: 2011
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Tressel,SarahL;Koukos,Georgios;Tchernychev,Boris;Jacques,SuzanneL;Covic,Lidija;Kuliopulos,Athan
• 通讯作者: Kuliopulos,Athan

Platelet matrix metalloprotease-1 mediates thrombogenesis by activating PAR1 at a cryptic ligand site.

• DOI: 10.1016/j.cell.2009.02.018
• 发表时间: 2009-04-17
• 期刊: Cell
• 影响因子: 64.5
• 作者: Trivedi V;Boire A;Tchernychev B;Kaneider NC;Leger AJ;O'Callaghan K;Covic L;Kuliopulos A
• 通讯作者: Kuliopulos A

Protease-activated receptor-2 modulates protease-activated receptor-1-driven neointimal hyperplasia.

• DOI: 10.1161/atvbaha.111.238261
• 发表时间: 2011-12
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Sevigny LM;Austin KM;Zhang P;Kasuda S;Koukos G;Sharifi S;Covic L;Kuliopulos A
• 通讯作者: Kuliopulos A