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（由申请方提供）：促凝血和抗凝途径通过蛋白酶激活受体（PAR）与细胞信号传导相关。体内通过PAR1或PAR2的炎性TF依赖性信号传导知之甚少。这些研究是基于我们最近发现TF胞质结构域缺失的小鼠表现出PAR信号失调，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胞质结构域缺失小鼠所表现出的TF信号传导的鉴定不平衡为确定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研究TF胞浆区缺失小鼠炎症反应增强的特点，评价肺、肾炎症反应的加重程度，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。目的2是使用与PAR1或PAR2缺陷动物的遗传杂交来确定PAR1和PAR2信号传导对这些小鼠中增强的炎症反应的贡献。验证了TF胞质结构域缺失小鼠的功能表型获得通过PAR1或PAR2缺失而消除的假设。这些实验将确定炎症中TF依赖性信号传导下游的体内相关PAR。目的3是利用所产生的遗传小鼠模型进行原代细胞分离，以在体外表征TF下游的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TF和凝血酶信号传导在血管细胞中的差异。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。这些标志物的鉴定将有助于体内凝血信号的定量，从而推进临床努力，以设计凝血细胞信号通路的治疗干预。