NEW GENETIC MODELS FOR TISSUE FACTOR SIGNALING

组织因子信号转导的新遗传模型

• 批准号: 6908126
• 负责人: WOLFRAM RUF
• 金额: $ 42.23万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-16 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6908126
• 关键词: biological signal transduction; cell surface receptors; endotoxins; gene expression; gene expression profiling; genetic models; genetically modified animals; immune response; inflammation; kidney; laboratory mouse; lipopolysaccharides; lung; model design /development; phosphorylation; thrombin; thromboplastin

DESCRIPTION (provided by applicant): The pro- and anti-coagulant pathways are linked to cell signaling through protease-activated receptors (PARs). The inflammatory TF-dependent signaling through PAR1 or PAR2 in vivo is poorly understood. The proposed studies are based on our recent finding that TF cytoplasmic domain deleted mice exhibit deregulated PAR signaling, but the TF cytoplasmic domain has no apparent function in regulating TF's procoagulant activity. The identified imbalance in TF signaling exhibited by TF cytoplasmic domain deleted mice provides novel opportunities to define the role of TF-dependent signaling to inflammatory responses associated with activation of coagulation. Aim 1 is to characterize the enhanced inflammatory response of TF cytoplasmic domain deleted mice, to evaluate the inflammatory exacerbation in lung and kidney, and to determine whether the enhanced inflammatory response results in increased lethality in experimental endotoxemia. Aim 2 is to define the contributions of PAR1 and PAR2 signaling to the enhanced inflammatory response in these mice, using genetic crosses with PAR1 or PAR2 deficient animals. The hypothesis is tested that the gain of function phenotype of TF cytoplasmic domain deleted mice is ablated by deletion of PAR1 or PAR2. These experiments will define the in vivo relevant PAR(s) downstream of TF-dependent signaling in inflammation. Aim 3 is to employ the generated genetic mouse models for primary cell isolation to characterize the role of PAR1 and PAR2 downstream of TF in vitro, followed by validation in inflammatory models in vivo. By comparing the TF response with thrombin signaling, these experiments will elucidate the differences between TF and thrombin signaling in vascular cells. A long-term goal is to develop novel diagnostic approaches to distinguish between upstream and downstream coagulation signaling. Identification of such markers will facilitate the quantitation of coagulation signaling in vivo and thus advance clinical efforts to design therapeutic intervention in coagulant cell signaling pathways .

描述(由申请人提供)：促凝血和抗凝血途径通过蛋白酶激活受体(PARs)与细胞信号相联系。活体内通过PAR1或PAR2的炎性因子依赖信号转导机制尚不清楚。这些研究是基于我们最近的发现，即Tf胞浆结构域缺失的小鼠表现出非调控的PAR信号，但Tf胞质结构域在调节Tf的促凝血活性方面没有明显的功能。Tf胞浆结构域缺失的小鼠表现出的Tf信号的不平衡为确定依赖Tf的信号在与凝血激活相关的炎症反应中的作用提供了新的机会。目的1研究Tf胞浆结构域缺失小鼠炎症反应增强的特点，评价肺、肾组织炎症加重情况，探讨炎症反应增强是否导致实验性内毒素血症致死率升高。目的2是利用PAR1或PAR2缺陷动物的遗传杂交，确定PAR1和PAR2信号在这些小鼠增强炎症反应中的作用。假设Tf胞浆结构域缺失的小鼠的功能表型的获得通过PAR1或PAR2的缺失而被消融。这些实验将确定炎症中TF依赖信号的下游与体内相关的PAR(S)。目的3是利用建立的小鼠遗传模型进行原代细胞分离，在体外研究PAR1和PAR2在Tf下游的作用，并在体内炎症模型中进行验证。通过比较转铁蛋白反应和凝血酶信号，这些实验将阐明转铁蛋白和凝血酶信号在血管细胞中的差异。一个长期的目标是开发新的诊断方法来区分上游和下游的凝血信号。这些标志物的识别将有助于体内凝血信号的定量，从而推动临床努力设计凝血细胞信号通路的治疗干预。