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）是组织和尿型纤溶酶原激活剂的主要抑制剂。玻染蛋白（VN）是存在于细胞外基质（ECM）和血浆中的粘合剂糖蛋白，可稳定PAI-1并且其功能受PAI-1调节。 PAI-1和VN在调节血管平滑肌细胞（VSMC）迁移和内膜增生方面起着重要作用，这些迁移和内膜增生在经皮冠状动脉干预（PCI）后引起再狭窄，并且是动脉粥样硬化和其他人血管疾病的发展中的中心过程。可用的研究表明，根据实验条件，PAI-1和VN可以促进或抑制VSMC迁移和内膜增生。然而，以前的体外研究受到了以下事实的限制：它们在很大程度上涉及二维细胞培养系统和纯化的试剂，而纯化的试剂并不能充分地对VSMC在体内迁移或评估VSMC本身产生的PAI-1和VN的功能的ECM进行建模。我们已经开发了一个实验模型，研究了通过由胶原蛋白，VN和其他ECM分子组成的3维矩阵在PAI-1和VN表达中具有遗传改变的VSMC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的促和抗迁移效应方面起着至关重要的作用，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体内数据，几乎完全是从敲除小鼠的实验中得出的。尽管这些研究产生了大量信息，但它们一直有争议。因此，缺乏关于PAI-1对动脉重塑的净影响的明确共识，这阻碍了针对PAI-1的药理策略的发展，以治疗或预防人类血管疾病。我们假设阐明PAI-1在人血管疾病中的真实功能将需要进行涉及临床相关形式的血管损伤形式的实验，而该动物的血管大小，结构和功能更像人类的血管大小，结构和功能。因此，我们开发了PCI的猪模型和由多种不同机制破坏PAI-1功能的特定药理PAI-1抑制剂。我们使用这种大动物模型的初步数据表明，PAI-1的显性阴性形式抑制PCI后内膜增生。 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 PCI的相关猪模型。为了实现我们的目标，我们组建了一支多样化的研究人员，在PAI-1和VN生物化学，细胞和分子生物学以及药理学以及兽医生物医学科学和临床心血管医学方面具有相当大的专业知识。我们预计，该提案中概述的实验将有助于在生理和临床相关条件下阐明PAI-1和VN的血管功能，并有助于将PAI-1靶向化合物作为预防和治疗人血管疾病的潜在药物的作用。公共卫生相关性：每年数以百万计的美国接受经皮冠状动脉干预（“支架”）治疗冠状动脉疾病的程序。但是，由于血管成形术部位的动脉复发或支架段内的血栓形成，经皮冠状动脉干预可能会失败。该应用将研究纤溶酶原激活剂-1和玻璃体蛋白在经皮冠状动脉干预措施中的作用。