NEW GENETIC MODELS FOR TISSUE FACTOR SIGNALING

组织因子信号转导的新遗传模型

• 批准号: 6815174
• 负责人: WOLFRAM RUF
• 金额: $ 46.93万
• 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/6815174
• 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（PAR）与细胞信号传导相关。人们对体内通过 PAR1 或 PAR2 的炎症 TF 依赖性信号传导知之甚少。拟议的研究基于我们最近的发现，即 TF 胞质结构域缺失的小鼠表现出 PAR 信号传导失调，但 TF 胞质结构域在调节 TF 促凝血活性方面没有明显的功能。 TF 胞质结构域缺失小鼠所表现出的 TF 信号传导失衡为定义 TF 依赖性信号传导对与凝血激活相关的炎症反应的作用提供了新的机会。目的 1 是表征 TF 胞质结构域缺失小鼠的炎症反应增强，评估肺和肾脏的炎症恶化，并确定炎症反应增强是否导致实验性内毒素血症致死率增加。目标 2 是通过与 PAR1 或 PAR2 缺陷动物进行遗传杂交，确定 PAR1 和 PAR2 信号传导对这些小鼠炎症反应增强的贡献。测试假设：TF 胞质结构域缺失小鼠的功能表型增益因 PAR1 或 PAR2 的缺失而被消除。这些实验将定义炎症中 TF 依赖性信号传导的体内相关 PAR 下游。目标 3 是利用生成的遗传小鼠模型进行原代细胞分离，以在体外表征 TF 下游 PAR1 和 PAR2 的作用，然后在体内炎症模型中进行验证。通过比较 TF 反应与凝血酶信号传导，这些实验将阐明血管细胞中 TF 和凝血酶信号传导之间的差异。长期目标是开发新的诊断方法来区分上游和下游凝血信号传导。这些标记物的鉴定将有助于体内凝血信号传导的定量，从而推进凝血细胞信号传导途径设计治疗干预的临床工作。