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和蛋白质的缺陷有助于男性和小鼠的发病率和死亡率。重组激活的蛋白C（APC）疗法降低了成年严重败血症患者的死亡率。对APC和蛋白质的作用的生理和药理机理有主要需求，以建立这种机制的原理证明，该项目使用转基因的小鼠，鼠损伤模型和新型的重组鼠蛋白。 APC可以发挥两种主要的不同活性：（1）抗凝活性和（2）对包含多种细胞保护作用的细胞的直接有益作用。后一种活动对于鼠败血症模型中APC的死亡率降低至关重要。 APC细胞信号传导的当前范例涉及由内皮蛋白C受体（EPCR）结合蛋白酶活化受体-1（PAR1）蛋白水解活化的APC。我们发现，APC启动了另一种信号通路，涉及载脂蛋白E受体2（ApoER2）的连接，通过衔接蛋白，DAB1和SRC家庭激酶信号传导，并在PI3K-AKT生存途径下游激活。鼠APC和APOER2突变体的工程将允许询问通过APC：Apoer2相互作用介导结合和信号启动的蛋白质表面，并将为APC在鼠败血症中作用机制的原理研究提供试剂。对ApoER2和DAB1中基因修饰的小鼠的研究将确定ApoER2和DAB1是否介导APC的败血症死亡率降低活性。蛋白质的缺乏小鼠将受到血栓性挑衅的影响，并通过重组野生型和突变鼠蛋白S和APC或其他药物的组合处理，以定义依赖于APC或独立于APC的蛋白质抗强化活性的相对效率。这些临床前动物模型研究确定的新原理最终可以转化为涉及蛋白质C和蛋白质系统的诊断或治疗进展。公共卫生相关性：先前关于血浆蛋白C（一种天然存在的血浆蛋白）的基础研究，已转化为诊所中使用的诊断和治疗工具。提出的关于激活蛋白C的基础研究将提供原理和确定性见解，对分子机制，激活蛋白C对血细胞作用以防止可能致命的损害。这些发现很可能可以转化为未来的临床进展。