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：
8040658
负责人：
JOHN H GRIFFIN
金额：
$ 47.38万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8040658
关键词：
1-Phosphatidylinositol 3-Kinase Abbreviations Adaptor Signaling Protein Adult Alkaline Phosphatase Animal Model Anti-Inflammatory Agents Anti-inflammatory Anticoagulants Apoptotic Basic Science Binding Blood Cells Blood Proteins Blood Vessels Blood coagulation Cells Clinic Clinical Data Defect Diagnostic Disabled Persons Dissection Elements Endothelial Cells Engineering F2R gene Factor Va Future Gene Expression Alteration Glycogen Synthase Kinases Goals Human In Vitro Injury Knowledge Left Ligation Low Density Lipoprotein Receptor Mediating Medicine Membrane Proteins Modeling Molecular Morbidity - disease rate Mus PAR-1 Receptor Pathway interactions Patients Physiological Plasma Proteins Protein C Protein S Protein S Deficiency Proteins Reaction Reagent Receptor Cell Recombinants Relative (related person)Reporter Research Role Sepsis Signal Pathway Signal Transduction Sphingosine-1-Phosphate Receptor Structural Protein Structure Surface Plasmon Resonance System Therapeutic Thrombosis Translating Translational Research VLDL receptor activated Protein C apolipoprotein E receptor 2 base citrate carrier cofactor human C4BPA protein in vitro activity in vivo inhibitor/antagonist insight men mortality mutant novel pre-clinical preclinical study prevent receptor receptor binding research study sphingosine 1-phosphate src-Family Kinases tool

项目摘要

DESCRIPTION (provided by applicant): Protein C and protein S deficiencies contribute to morbidity and mortality in men and mice. Recombinant activated 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, this project uses genetically modified mice, murine injury models, and novel recombinant murine proteins. APC can exert two major, 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 APC's cell signaling involves binding of APC by endothelial protein C receptor (EPCR) combined with protease activated receptor-1 (PAR1) 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, Dab1, and Src-family kinases with downstream activation of the PI3K-Akt survival pathway. Engineering of murine APC and apoER2 mutants will allow interrogation of the protein surfaces that mediate binding and signal initiation by APC:apoER2 interactions and will provide reagents for in vivo proof of principle studies for mechanisms of APC's action in murine sepsis. Studies of mice genetically modified in apoER2 and Dab1 will establish whether apoER2 and Dab1 mediate APC's mortality reduction activities in sepsis. Protein S deficient mice will be subjected to thrombotic provocation and treated with combinations of recombinant wild type and mutant murine protein S and APC or other agents to define the relative 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 diagnostic or therapeutic advances involving the protein C and protein S systems. PUBLIC HEALTH RELEVANCE: Previous basic research on plasma Protein C, a naturally occurring plasma protein, was translated into diagnostic and therapeutic tools now used in the clinic. The proposed basic research studies on activated protein C will provide proof of principle and definitive insights into molecular mechanisms by which activated protein C acts on blood cells to prevent damage that can be fatal. The findings may well be translatable into future clinical advances.

描述（由申请人提供）：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) 疗法可降低成年严重脓毒症患者的死亡率。迫切需要对 APC 和蛋白 S 的生理和药理学作用机制有新的见解。为了建立此类机制的体内原理证明，该项目使用转基因小鼠、小鼠损伤模型和新型重组小鼠蛋白。 APC 可以发挥两种主要的、不同的活性：(1) 抗凝活性和 (2) 对细胞的直接有益作用，包括多种细胞保护作用。后一种活性对于 APC 在小鼠脓毒症模型中降低死亡率至关重要。当前 APC 细胞信号转导的范例涉及 APC 通过内皮蛋白 C 受体 (EPCR) 与蛋白酶激活受体 1 (PAR1) 蛋白水解激活相结合。我们发现 APC 启动了另一条信号通路，涉及载脂蛋白 E 受体 2 (apoER2) 的连接，通过接头蛋白、Dab1 和 Src 家族激酶发出信号，并激活下游 PI3K-Akt 存活通路。鼠类 APC 和 apoER2 突变体的工程设计将允许对通过 APC:apoER2 相互作用介导结合和信号启动的蛋白质表面进行询问，并将为 APC 在鼠类脓毒症中作用机制的体内原理研究证明提供试剂。对 apoER2 和 Dab1 基因修饰小鼠的研究将确定 apoER2 和 Dab1 是否介导 APC 在脓毒症中降低死亡率的活性。蛋白S缺陷小鼠将受到血栓激发，并用重组野生型和突变型鼠蛋白S和APC或其他试剂的组合进行治疗，以确定依赖于APC或独立于APC的蛋白S抗血栓活性的相对功效。这些临床前动物模型研究建立的新原理可能最终转化为涉及蛋白 C 和蛋白 S 系统的诊断或治疗进展。公共健康相关性：之前对血浆蛋白 C（一种天然存在的血浆蛋白）的基础研究已转化为目前在临床中使用的诊断和治疗工具。拟议的关于活化蛋白 C 的基础研究将为活化蛋白 C 作用于血细胞以防止可能致命的损伤的分子机制提供原理证明和明确的见解。这些发现很可能转化为未来的临床进展。