In Vivo System - Screening Anti-Inflammatory Compounds

描述（由申请人提供）：急性炎症是必要的， 但也可引起器官损伤和功能障碍。的NF-κ B 转录因子复合物对急性炎症的调节至关重要 通过控制许多关键细胞因子、粘附分子和 内切酶我们已经产生了转基因小鼠表达Photinus荧光素酶 在NF-kB依赖性启动子的控制下。在本提案中，我们 假设我们的转基因荧光素酶报告小鼠代表了一种强大的 用于非侵入性体内测量NF-kB依赖性炎症的工具 反应，这些动物可以用作高通量的工具 抗炎化合物的筛选。我们提出两个具体目标：1） 优化使用完整的NF-kB激活的重复性非侵入性测量， 转基因荧光素酶报告基因小鼠，和2）为了验证这种方法的实用性， 使用细菌感染小鼠模型筛选NF-κ B抑制剂的系统 脂多糖引起的肺部炎症现有方法 用于筛选抗炎化合物的方法包括最初的体外研究 随后是涉及大量动物的研究。我们的方法 通过将每只鼠标作为自己的鼠标使用， 控制和允许多个测量超时。这些改进应当 允许在小鼠中进行初步测试，从而简化和加速 消炎药的开发。 拟定商业应用： 鉴定具有抗炎治疗潜力的新药物需要 在动物模型中进行筛选，但目前的模型是劳动密集型的， 筛选工具 由于NF-kB的激活是宿主发病机制的共同点， 对于以嗜酸性炎症为特征的疾病， NF-kB的测定将是有价值的筛选工具。 我们建议完善一个 小鼠模型，其中非侵入性、连续、定量、器官特异性测量 NF-kB的激活。 该模型对制药行业具有重要的应用价值 公司和其他开发抗炎治疗剂作为高通量 用于鉴定潜在有效药物和定义抗炎药的筛选工具 药物动力学

DESCRIPTION (provided by the applicant): Acute inflammation is necessary for host defense but can also cause organ injury and dysfunction. The NF-kB transcription factor complex is critical for regulation of acute inflammation by controlling expression of a number of key cytokines, adhesion molecules and enzymes. We have generated transgenic mice that express Photinus luciferase under the control of an NF-kB dependent promoter. In this proposal, we hypothesize that our transgenic luciferase reporter mice represent a powerful tool for non-invasive in vivo measurements of NF-kB dependent inflammatory responses and that these animals can be used as a tool for high-throughput screening of anti-inflammatory compounds. We propose 2 specific aims: 1) to optimize repetitive, non-invasive measurement of NF-kB activation using intact transgenic luciferase reporter mice, and 2) to validate the utility of this system for screening NF-kB inhibitors using a mouse model of bacterial lipopolysaccharide-induced lung inflammation. Currently available methodologies for screening anti-inflammatory compounds involve initial in vitro studies followed by studies involving large numbers of animals. Our methodology represents a marked improvement in this process by using each mouse as its own control and allowing multiple measurements overtime. These improvements should allow initial testing in mice, thus simplifying and accelerating the process of anti-inflammatory drug development. PROPOSED COMMERCIAL APPLICATION: Identification of new agents with potential as anti-inflammatory therapeutics requires screening in animal models, but current models are labor intensive and inefficiect as screening tools. Since activation of NF-kB is common to pathogenesis of a host of diseases characterized by neutrophilic inflammation, an in vivo method for rapid determination of NF-kB would be a valuable screening tool. We propose to refine a mouse model in which non-invasive, serial, quantitative, organ specific measurements of NF-kB activation can be made. The model would be highly valuable to pharmaceutical companies and other developing anti-inflammatory therapeutic agents as a high throughput screening tool for identifying potentially efficacious agents and defining anti-inflammatory pharmacockinetics.