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）：促凝途径与蛋白酶激活的受体（PARS）相关。通过体内PAR1或PAR2的炎症性TF依赖性信号传导知之甚少。拟议的研究是基于我们最近的发现，即TF细胞质结构域删除的小鼠表现出解析的PAR信号传导，但TF细胞质结构域在调节TF的Procagulant活性方面没有明显的功能。 TF细胞质结构域删除的小鼠在TF信号传导中鉴定出的不平衡为定义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细胞质结构域删除的小鼠的炎症反应增强，以评估肺和肾脏的炎症加剧，并确定增强的炎症反应是否导致实验性内毒素血症的致死性增加。 AIM 2是使用具有PAR1或PAR2缺乏动物的遗传杂交来定义PAR1和PAR2信号对这些小鼠增强炎症反应的贡献。该假设检验了TF细胞质结构域删除的小鼠的功能表型的增益被PAR1或PAR2的缺失消除。这些实验将定义炎症中TF依赖性信号传导下游的体内相关PAR。目的3是利用生成的遗传小鼠模型进行原代细胞分离来表征体外TF下游PAR1和PAR2的作用，然后在体内炎症模型中进行验证。通过将TF反应与凝血酶信号传导进行比较，这些实验将阐明血管细胞中TF和凝血酶信号的差异。一个长期目标是开发新型的诊断方法，以区分上游和下游凝血信号传导。此类标记物的识别将促进体内凝血信号传导的定量，从而提高临床努力，以设计凝结剂细胞信号通路的治疗干预。