Targeting chronic Pseudomonas aeruginosa wound infections

描述(申请人提供)：拟议的实验旨在确定用于治疗慢性伤口感染的新的抗菌药物靶点，并对几个此类靶点的小分子抑制剂进行筛选。为了实现这一目标，我们利用了三种工具： 1)新开发的糖尿病小鼠伤口模型，在该模型中，由于铜绿假单胞菌生物被膜感染而延迟愈合；2)基因组规模的遗传方法(TN-SEQ)，可以确定伤口感染持续所需的细菌功能；以及3)高灵敏的方法，使用质谱仪来识别在伤口感染中诱导的可能包括TN-SEQ缺失的持续功能的蛋白质。潜在的药物靶点将使用一系列被认为反映感染的测试来确定优先顺序。然后，将构建用于检测这三个顶级目标的抑制剂的筛选菌株，并将其用于筛选20万个化学库。

DESCRIPTION (provided by applicant): 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.