PAI-1 and Vitronectin in Failure of Coronary Revascularization

PAI-1 和玻连蛋白在冠状动脉血运重建失败中的作用

基本信息

批准号：
8518443
负责人：
William P Fay
金额：
$ 35.11万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8518443
关键词：
3-Dimensional Address Adhesives Affect Alteplase American Angioplasty Animal Model Animals Arterial Injury Arteries Atherosclerosis Biochemistry Blood Vessels Cardiovascular system Cell Culture System Cell Surface Receptors Clinical Sciences Collagen Consensus Coronary Coronary Arteriosclerosis Coronary artery Data Development Dominant-Negative Mutation Endothelial Cells Experimental Models Extracellular Matrix Failure Family suidae Glycoproteins Human Hyperplasia In Vitro Injury Knockout Mice Mediating Medicine Migration Assay Modeling Molecular Molecular Weight Molecular and Cellular Biology Mus Mutation Pathogenesis Pharmacology Physiological Plasma Plasminogen Activator Plasminogen Activator Inhibitor 1 Play Prevention Procedures Process Proliferating Proteins Proteolysis Reagent Recombinants Recurrence Research Personnel Role Site Smooth Muscle Myocytes Stents Structure Testing Therapeutic Thrombus Tissues Vascular Diseases Vascular remodeling Vitronectin arterial remodeling clinically relevant coronary angioplasty human disease in vivo inhibitor/antagonist neointima formation percutaneous coronary intervention prevent public health relevance research study restenosis small molecule stoichiometry two-dimensional urinary vascular smooth muscle cell migration

项目摘要

DESCRIPTION (provided by applicant): Plasminogen activator inhibitor-1 (PAI-1) is the primary inhibitor of tissue- and urinary-type plasminogen activators. Vitronectin (VN) is an adhesive glycoprotein present in extracellular matrix (ECM) and plasma that stabilizes PAI-1 and whose function is regulated by PAI-1. PAI-1 and VN play major roles in regulating vascular smooth muscle cell (VSMC) migration and intimal hyperplasia, which cause restenosis after percutaneous coronary intervention (PCI) and are central processes in the development of atherosclerosis and other human vascular diseases. Available studies suggest that PAI-1 and VN can either promote or inhibit VSMC migration and intimal hyperplasia, depending on experimental conditions. However, previous in vitro studies have been limited by the fact that they largely involved 2-dimensional cell culture systems and purified reagents, which do not adequately model the ECM in which VSMC migrate in vivo or assess the functions of PAI-1 and VN produced by VSMC themselves. We have developed an experimental model to study the migration of VSMC with genetic alterations in PAI-1 and VN expression through 3-dimensional matrices composed of collagen, VN, and other ECM molecules. Our preliminary data suggest that the stoichiometric relationship of PAI-1 and VN plays a critical role in determining PAI-1's pro- and anti-migratory effects, and that PAI-1 regulates VN expression by VSMC. Available in vivo data regarding the roles of PAI-1 and VN in regulating arterial remodeling have been derived nearly exclusively from experiments with knockout mice. While these studies have yielded a wealth of information, they have been controversial. Consequently, a clear consensus regarding the net effect of PAI-1 on arterial remodeling is lacking, which has hindered development of pharmacological strategies to target PAI-1 to treat or prevent human vascular disease. We hypothesize that elucidation of the authentic function of PAI-1 in human vascular diseases will require experiments involving clinically relevant forms of vascular injury in animals whose vascular size, structure, and function more closely resembles those of humans. Consequently, we have developed a porcine model of PCI and a panel of specific, pharmacological PAI-1 inhibitors that disrupt PAI-1 function by several distinct mechanisms. Our preliminary data with this large animal model suggest that a dominant-negative form of PAI-1 inhibits intimal hyperplasia after PCI. In the proposed experiments we will probe the functions of PAI-1 and VN in vascular remodeling by employing 3-dimensional VSMC migration assays, studying vascular remodeling in mice with a spectrum of PAI-1 and VN expression levels, and determining the effects of a panel of recombinant PAI-1 proteins and small molecule PAI-1 inhibitors in a murine vascular injury model and a clinically relevant porcine model of PCI. To achieve our objectives we have assembled a diverse team of investigators with considerable expertise in PAI-1 and VN biochemistry, cellular and molecular biology, and pharmacology, as well as in veterinary biomedical sciences and clinical cardiovascular medicine. We anticipate that the experiments outlined in this proposal will help to elucidate the vascular functions of PAI-1 and VN under physiologically and clinically relevant conditions and will help to define the role of PAI-1 targeting compounds as potential agents to prevent and treat human vascular disease. PUBLIC HEALTH RELEVANCE: Each year millions of American undergo percutaneous coronary intervention ("stent") procedures for treatment of coronary artery disease. However, percutaneous coronary interventions can fail due to recurrent narrowing of the artery at the site of angioplasty or formation of a thrombus within the stented segment. This application will study the roles of plasminogen activator inhibitor-1 and vitronectin in the failure of percutaneous coronary interventions.

描述（由申请人提供）：纤溶酶原激活剂抑制剂-1（PAI-1）是组织型和尿型纤溶酶原激活剂的主要抑制剂。玻连蛋白 (VN) 是一种存在于细胞外基质 (ECM) 和血浆中的粘附糖蛋白，可稳定 PAI-1，其功能受 PAI-1 调节。 PAI-1和VN在调节血管平滑肌细胞（VSMC）迁移和内膜增生中发挥重要作用，导致经皮冠状动脉介入治疗（PCI）后再狭窄，是动脉粥样硬化和其他人类血管疾病发展的核心过程。现有研究表明，PAI-1 和 VN 可以促进或抑制 VSMC 迁移和内膜增生，具体取决于实验条件。然而，之前的体外研究主要涉及二维细胞培养系统和纯化试剂，无法充分模拟 VSMC 在体内迁移的 ECM 或评估 PAI-1 和 VN 产生的功能。由 VSMC 自己提供。我们开发了一个实验模型，通过由胶原蛋白、VN 和其他 ECM 分子组成的 3 维基质来研究 PAI-1 和 VN 表达基因改变的 VSMC 的迁移。我们的初步数据表明，PAI-1 和 VN 的化学计量关系在确定 PAI-1 的促迁移和抗迁移作用中起着关键作用，并且 PAI-1 通过 VSMC 调节 VN 表达。关于 PAI-1 和 VN 在调节动脉重塑中的作用的现有体内数据几乎完全来自基因敲除小鼠的实验。虽然这些研究提供了大量信息，但也存在争议。因此，关于 PAI-1 对动脉重塑的净效应缺乏明确的共识，这阻碍了针对 PAI-1 治疗或预防人类血管疾病的药理学策略的开发。我们假设，要阐明 PAI-1 在人类血管疾病中的真实功能，需要在血管大小、结构和功能更接近人类的动物中进行涉及临床相关形式的血管损伤的实验。因此，我们开发了 PCI 猪模型和一组特异性药理学 PAI-1 抑制剂，这些抑制剂通过几种不同的机制破坏 PAI-1 功能。我们对该大型动物模型的初步数据表明，显性失活形式的 PAI-1 可抑制 PCI 后的内膜增生。在拟议的实验中，我们将通过采用 3 维 VSMC 迁移测定来探讨 PAI-1 和 VN 在血管重塑中的功能，研究具有一系列 PAI-1 和 VN 表达水平的小鼠血管重塑，并确定鼠血管损伤模型和临床相关猪 PCI 模型中的一组重组 PAI-1 蛋白和小分子 PAI-1 抑制剂。为了实现我们的目标，我们组建了一支多元化的研究人员团队，他们在 PAI-1 和 VN 生物化学、细胞和分子生物学、药理学以及兽医生物医学科学和临床心血管医学方面拥有丰富的专业知识。我们预计本提案中概述的实验将有助于阐明 PAI-1 和 VN 在生理和临床相关条件下的血管功能，并将有助于确定 PAI-1 靶向化合物作为预防和治疗人类血管疾病的潜在药物的作用疾病。公共卫生相关性：每年有数百万美国人接受经皮冠状动脉介入治疗（“支架”）手术来治疗冠状动脉疾病。然而，由于血管成形术部位的动脉反复狭窄或支架段内血栓的形成，经皮冠状动脉介入治疗可能会失败。该应用将研究纤溶酶原激活剂抑制剂-1 和玻连蛋白在经皮冠状动脉介入治疗失败中的作用。