Direct Cellular Effects of Blood Coagulation Proteases

凝血蛋白酶的直接细胞效应

基本信息

批准号：
7545924
负责人：
Laurent Olivier Mosnier
金额：
$ 24.9万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-12-01 至 2011-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7545924
关键词：
Achievement Active Sites Antibodies Anticoagulants Antithrombins Apoptosis Apoptotic Binding Biochemical Blood coagulation Cell membrane Cells Cellular biology Characteristics Charge Clinical Clinical Trials Coagulation Process Complex Data Defect Disease Dissociation Endocytosis Endothelial Cells Engineering Enzymes Factor IX Functional disorder Funding Future Generations Goals Heparin Inflammation Inflammatory K-Series Research Career Programs Knowledge Lead Mediating Membrane Methods Microscopy Molecular Mutagenesis PAR-1 Receptor Pathogenesis Pathway interactions Patients Peptide Hydrolases Phase III Clinical Trials Phospholipids Physiological Platelet Factor 4 Property Protease Domain Protein C Proteomics Prothrombin Publishing Reaction Regulation Research Personnel Risk Role Sepsis Solid Specificity Surface TFPI Techniques Testing Therapeutic Thrombin Thrombocytopenia Thrombomodulin Thromboplastin Thrombosis United States National Institutes of Health Variant Vitamin K activated Protein C base career development cell growth regulation citrate carrier clinically relevant cofactor experience improved in vivo meizothrombin mortality novel receptor receptor binding success unpublished works

项目摘要

The major scientific goal of this Pathway to Independence (K99/ROO) Career Development Award application is to understand the molecular pathophysiology of thrombotic and inflammatory disorders by studying novel mechanisms for cytoprotective actions of vitamin K-dependent coagulation proteases. This application focuses initially on the role of membrane receptors in the regulation of the cellular protein C pathway and later on the exploration of novel mechanisms for cytoprotective activities of coagulation proteases. The major career development goal of the applicant is to expand his technical and academic experience required for a successful transition into an independent investigator. These studies will provide the opportunity and solid basis to apply successfully for future independent NIH R01 funding focused on the molecular mechanistic studies centered on the crossroads of coagulation and inflammation. Novel hypotheses on the functional proteomics of cytoprotective actions by blood coagulation proteases will be tested using biochemical and cellular biology methods. The clinical and therapeutic implications of the proposed studies are clear from the large clinical trials, where activated protein C (ARC), but not other anticoagulants reduced mortality in severe sepsis patients and implied that the unique combination of APC's anticoagulant activity and direct activity on cells is the basis for APC's success. My published work and unpublished preliminary data lead directly to the proposed studies and provide strong support for my hypotheses. In testing these hypotheses, I propose: 1) To characterize the formation of endothelial cell membrane receptor complexes between thrombomodulin, endothelial protein C receptor and protease activated receptor-1 required for APC generation and APC's direct effects on cells; 2) To clarify the potential beneficial and detrimental functional properties of platelet factor 4 for APC generation and APC's direct effects on cells; 3) To identify novel themes and mechanisms for APC and fVlla cytoprotective actions on cells by exploration of the similarities and differences between APC and fVlla anti-apoptotic activities; and 4) To establish whether meizothrombin has anti-apoptotic activity, as predicted, and if this activity requires cofactor-dependent and PAR-dependent mechanisms. If the proposed studies are successful, they will increase our knowledge and may lead to improved treatment of a variety of disorders in which thrombosis, apoptosis and inflammation contribute to pathogenesis.

独立之路（K99/ROO）职业发展奖申请的主要科学目标是通过研究新的机制来了解血栓和炎症性疾病的分子病理生理学维生素 K 依赖性凝血蛋白酶的细胞保护作用机制。这个应用程序最初关注膜受体在细胞蛋白 C 途径调节中的作用，随后探索了凝血蛋白酶细胞保护活性的新机制。主要申请人的职业发展目标是扩大其所需的技术和学术经验成功转型为独立调查员。这些研究将提供机会和坚实的为成功申请未来专注于分子机制的独立 NIH R01 资金奠定基础研究集中在凝血和炎症的十字路口。关于泛函的新假设凝血蛋白酶的细胞保护作用的蛋白质组学将使用生化和细胞生物学方法。从以下数据中可以清楚地看出拟议研究的临床和治疗意义大型临床试验，其中活性蛋白 C (ARC) 而不是其他抗凝剂可降低重症患者的死亡率败血症患者并暗示 APC 的抗凝活性和直接活性的独特组合细胞是APC成功的基础。我已发表的作品和未发表的初步数据直接导致提出的研究并为我的假设提供了强有力的支持。在检验这些假设时，我建议：1）为了表征血栓调节蛋白之间内皮细胞膜受体复合物的形成， APC 生成和 APC 所需的内皮蛋白 C 受体和蛋白酶激活受体 1 对细胞的直接影响； 2) 阐明血小板潜在的有益和有害的功能特性 APC 生成的因子 4 以及 APC 对细胞的直接影响； 3）确定新的主题和机制通过探索 APC 和 fVIIa 对细胞的异同 APC 和 fVIIa 抗凋亡活性； 4) 确定酶促凝血酶是否具有抗细胞凋亡作用活性，如预测的那样，以及该活性是否需要辅因子依赖性和 PAR 依赖性机制。如果拟议的研究是成功的，它们将增加我们的知识，并可能导致改善治疗血栓形成、细胞凋亡和炎症参与发病机制的多种疾病。