In Vivo System - Screening Anti-Inflammatory Compounds

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.