Murine Protein C and Protein S proof of Principle Research

鼠蛋白 C 和蛋白 S 原理研究证明

基本信息

批准号：
8380735
负责人：
JOHN H GRIFFIN
金额：
$ 41.94万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-12-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8380735
关键词：
1-Phosphatidylinositol 3-Kinase Abbreviations Adaptor Signaling Protein Adult Alkaline Phosphatase Animal Model Anti-Inflammatory Agents Anti-inflammatory Anticoagulants Apolipoproteins Apoptotic Basic Science Binding Blood Blood Clot Blood Vessels Blood coagulation Cells Cessation of life Clinical Data Defect Diagnostic Disabled Persons Dissection Elements Endothelial Cells Engineering F2R gene FDA approved Factor Va Gene Expression Alteration Glycogen Synthase Kinase 3 Goals Human In Vitro Injury Intervention Knowledge Left Ligation Low Density Lipoprotein Receptor Mediating Medicine Membrane Proteins Modeling Molecular Morbidity - disease rate Mus PAR-1 Receptor Pathway interactions Patients Peptide Hydrolases Pharmaceutical Preparations Physiological Principal Investigator Protein C Protein S Protein S Deficiency Proteins Reagent Receptor Cell Recombinants Relative (related person)Reporter Research Role Sepsis Signal Pathway Signal Transduction Structural Protein Structure Surface Plasmon Resonance System The Sun Therapeutic Thrombosis Thrombus Translating Translational Research VLDL receptor Work activated Protein C apolipoprotein E receptor 2 base citrate carrier cofactor human C4BPA protein in vivo inhibitor/antagonist insight men mortality mutant novel pre-clinical preclinical study receptor receptor binding sphingosine 1-phosphate src-Family Kinases

项目摘要

Protein C and protein S deficiencies contribute to morbidity and mortality in men and mice. Recombinant acfivated protein C (APC) therapy reduces mortality in adult severe sepsis patients. There is a major need for new insights into the physiologic and pharmacologic mechanisms of action of APC and protein S. To establish in vivo proof of principle for such mechanisms. Project 1 uses genetically modified mice, murine injury models, and novel recombinant murine proteins and involves extensive collaborafions with Drs. Ruggeri and Ruf APC exerts two distinct activities: (1) anticoagulant activity and (2) direct beneficial effects on cells comprising a variety of cytoprotective actions. This latter activity is critical for mortality reduction by APC in murine sepsis models. The current paradigm for APCs cell signaling involves binding of APC by endothelial protein C receptor (EPCR) combined with protease activated receptor-1 (PARI) proteolytic activation. We found that there is another signaling pathway initiated by APC that involves ligation of apolipoprotein E Receptor 2 (apoER2), signaling via the adaptor protein. Dabi, and Src-family kinases with downstream acfivation of the PI3K-Akt survival pathway. Engineering of murine APC and apoER2 mutants will allow interrogation ofthe protein surfaces that mediate binding and signal inifiation by APC:apoER2 interactions and will provide reagents for in vivo proof of principle studies for mechanisms of APCs acfion in murine sepsis. Studies of mice genefically modified in apoER2 and Dabi will establish whether apoER2 and Dabi mediate APCs mortality reducfion acfivities in sepsis. Protein S deficient mice will be subjected to thrombofic provocafion and treated with combinations of recombinant wild type and mutant murine protein S and APC or other agents to define the relafive efficacies for protein S antithrombotic activity that is either dependent on APC or independent of APC. Novel Principles that are established by these preclinical animal model studies may ultimately be translated into diagnosfic or therapeufic advances involving the protein C and protein S systems. RELEVANCE (See instmctions): Protein C or protein S deficiency contributes to excessive blood clotting. Recombinant acfivated protein C (APC) is FDA-approved for reducing death in adult severe sepsis, but we don't understand how this drug works. To gain new insights, we will use engineered APC and protein S plus mice that are genefically modified. Novel principles that are established by these preclinical animal model studies may ulfimately be translated into diagnosfic or therapeutic advances involving the protein C and protein S systems.

蛋白质 C 和蛋白质 S 缺乏会导致男性和小鼠的发病率和死亡率。重组活化蛋白 C (APC) 治疗可降低成年严重脓毒症患者的死亡率。有一个重大需求对 APC 和蛋白质 S 的生理和药理学作用机制有新的见解。为此类机制建立体内原理证明。项目1使用转基因小鼠、小鼠损伤模型和新型重组鼠蛋白，并涉及与博士的广泛合作。 Ruggeri 和 Ruf APC 具有两种不同的活性：(1) 抗凝活性和 (2) 直接有益作用对细胞具有多种细胞保护作用。后一项活动对于降低死亡率至关重要小鼠脓毒症模型中的 APC。目前 APC 细胞信号转导的范式涉及 APC 的结合内皮蛋白 C 受体 (EPCR) 联合蛋白酶激活受体 1 (PARI) 蛋白水解激活。我们发现还有一条由 APC 启动的信号通路，涉及连接载脂蛋白 E 受体 2 (apoER2)，通过衔接蛋白发出信号。 Dabi 和 Src 家族激酶 PI3K-Akt 生存途径的下游激活。小鼠 APC 和 apoER2 突变体的工程改造将允许通过 APC:apoER2 询问介导结合和信号启动的蛋白质表面相互作用，并将为 APC 作用机制的体内原理研究证明提供试剂鼠败血症。对 apoER2 和 Dabi 进行基因修饰的小鼠的研究将确定 apoER2 和 Dabi 是否 Dabi 介导败血症中 APC 的死亡率降低活性。蛋白质S缺乏的小鼠将受到血栓激发并用重组野生型和突变型鼠蛋白 S 组合治疗和 APC 或其他药物来定义蛋白 S 抗血栓活性的相对功效，即依赖于 APC 或独立于 APC。这些临床前动物建立的新原理模型研究最终可能转化为涉及蛋白 C 的诊断或治疗进展和蛋白质S系统。相关性（参见说明）：蛋白质 C 或蛋白质 S 缺乏会导致过度凝血。重组活化蛋白C (APC) 已获得 FDA 批准用于减少成人严重败血症的死亡，但我们不明白这种药物的作用作品。为了获得新的见解，我们将使用基因工程 APC 和 Protein S plus 小鼠修改的。这些临床前动物模型研究建立的新原理最终可能是转化为涉及蛋白 C 和蛋白 S 系统的诊断或治疗进展。