Direct Cellular Effects of Blood Coagulation Proteases

凝血蛋白酶的直接细胞效应

• 批准号: 7224029
• 负责人: Laurent Olivier Mosnier
• 金额: $ 8.59万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7224029
• 关键词: Abbreviations; Achievement; Active Sites; Antibodies; Anticoagulants; Antithrombins; Apoptosis; Apoptotic; Binding; Binding Sites; Biochemical; Biology; Blood coagulation; Cell membrane; Cells; Cellular biology; Characteristics; Charge; Clinical; Clinical Trials; Coagulation Process; Complex; Data; Defect; Development; Disease; Dissociation; Endocytosis; Endopeptidases; Endothelial Cells; Engineering; Enzymes; Factor IX; Factor VIIa; Functional disorder; Funding; Future; GAG Gene; Generations; Glutamic Acid; Glycosaminoglycans; 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; Principal Investigator; 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; abstracting; activated Protein C; base; career; cell growth regulation; chymotrypsin; citrate carrier; clinically relevant; cofactor; experience; improved; in vivo; meizothrombin; mortality; novel; programs; receptor; receptor binding; success; unpublished works

DESCRIPTION (provided by applicant): The major scientific goal of this Pathway to Independence (K99/R00) 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. (End of Abstract)

描述（由申请人提供）： 这个独立之路（K99/R 00）职业发展奖申请的主要科学目标是通过研究维生素K依赖性凝血蛋白酶的细胞保护作用的新机制来了解血栓形成和炎症性疾病的分子病理生理学。该应用程序最初侧重于膜受体在细胞蛋白C途径的调节中的作用，后来探索凝血蛋白酶的细胞保护活性的新机制。申请人的主要职业发展目标是扩大成功过渡到独立调查员所需的技术和学术经验。这些研究将为成功申请未来独立的NIH R 01基金提供机会和坚实的基础，该基金专注于分子机制。 研究集中在凝血和炎症的交叉点上。将使用生物化学和细胞生物学方法测试凝血蛋白酶的细胞保护作用的功能蛋白质组学的新假设。所提出的研究的临床和治疗意义从大型临床试验中是清楚的，其中活化蛋白C（ARC）而不是其他抗凝剂降低了严重脓毒症患者的死亡率，并暗示APC的抗凝活性和对细胞的直接活性的独特组合是APC成功的基础。我发表的工作和未发表的初步数据直接导致了拟议的研究，并为我的假设提供了强有力的支持。为了验证这些假设，我提出：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， （2）阐明血小板第4因子对APC生成的潜在有利和不利的功能特性以及APC对细胞的直接作用;第三章通过探索APC和fVIIa之间的相似性和差异，确定APC和fVIIa对细胞的细胞保护作用的新主题和机制。fVIIa抗细胞凋亡活性;和4）确定甲唑凝血酶是否如预测的那样具有抗细胞凋亡活性，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，他们将增加我们的知识，并可能导致改善治疗的各种疾病，其中血栓形成，细胞凋亡和炎症有助于发病机制。 (End摘要）