In Vivo System - Screening Anti-Inflammatory Compounds

描述(由申请人提供)：急性炎症是必要的 宿主防御，但也可造成器官损伤和功能障碍。核因子-kB 转录因子复合体对急性炎症的调节至关重要 通过控制一些关键细胞因子、黏附分子和 酵素。我们已经培育出了表达石楠荧光素酶的转基因小鼠 在依赖于核因子-kB的启动子的控制下。在这项提案中，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 非侵入性体内测量核因子-kB依赖炎症的工具 这些动物可以被用作高通量的工具 抗炎化合物的筛选。我们提出了两个具体目标：1) 使用INTERNAL优化核因子-kB活性的重复性、非侵入性测量 转基因荧光素酶报告鼠，以及2)验证这一方法的实用性 使用细菌小鼠模型筛选核因子-kB抑制剂的系统 脂多糖诱导的肺部炎症。当前可用的方法 筛选抗炎化合物需要进行初步的体外研究 随后是涉及大量动物的研究。我们的方法论 通过将每个鼠标用作自己的鼠标，表示此过程中的显著改进 控制，并允许多个测量超时。这些改进应该 允许在小鼠身上进行初步测试，从而简化和加快了 抗炎药物开发。 建议的商业应用： 发现具有抗炎治疗潜力的新药物需要 在动物模型中进行筛选，但当前的模型是劳动密集型的，并且效率低下，如 筛查工具。由于核因子-kB的激活在宿主的发病机制中是常见的 对于以中性粒细胞炎症为特征的疾病，一种体内快速检测方法 核因子-kB的检测将是一项有价值的筛查工具。我们建议完善一项 无创、连续、定量、器官特异性测量的小鼠模型 可以激活核因子-kB。该模型具有很高的药学应用价值 公司等开发的消炎治疗剂作为高通量 用于识别潜在有效药物和定义抗炎的筛选工具 药代动力学。

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.