The proposed experiments aim to identify novel anti-bacterial drug targets for treatment of chronic wound infections and to carry out screens for small molecule inhibitors of several such targets. To accomplish this goal, we exploit three tools: 1) a newly developed diabetic mouse wound model in which healing is delayed due to a Pseudomonas aeruginosa biofilm infection, 2) a genome-scale genetic approach (Tn- seq) that can identify bacterial functions required for wound infection persistence, and 3) highly sensitive methodology employing mass spectrometry to identify proteins induced in wound infections that may include persistence functions missed by Tn-seq. Potential drug targets will be prioritized using a battery of tests thought to reflect infection. Screening strains designed to detect inhibitors of the three top targets will then be constructed and employed for screening a 200,000 chemical library.
The proposed research seeks to identify therapeutic targets for treatment of chronic Pseudomonas aeruginosa infections of diabetic wounds. The work will employ novel genome-scale technologies to identify and prioritize targets. Inhibitors of several high priority targets will be identified by screening specific reporter strains using a large libray of discrete small molecules and natural products.