The regulation of Pseudomonas aeruginosa surface colonization

描述（由申请人提供）：在囊性纤维化（CF）患者中，铜绿假单胞菌在肺部的慢性定植导致频繁住院和过早死亡。铜绿假单胞菌可以通过多种方式在表面定植：蜂群运动、滑动运动和生物膜的形成。细菌表面组分（如IV型菌毛、外多糖、鞭毛和鼠李糖脂）的表达对这些活性很重要，但目前对这些因子的相互作用和共同调节的了解尚不完整。默里博士有初步证据表明，铜绿假单胞菌产生的某些调节蛋白控制着对定植很重要的多个因素。他的研究目标是：1)确定由这些蛋白质共同调节的表面定植所需的成分（使用不同程度缺乏这些调节蛋白的铜绿假单胞菌菌株，并测量在蜂群、滑动和生物膜形成过程中的毛蛋白、鞭毛蛋白、外多糖和鼠李糖脂的水平）；2)定量这些菌株在CF感染组织培养模型中的表面定植（通过共聚焦显微镜观察细菌/上皮细胞相互作用并使用COMSTAT软件分析这些图像）；3)将体外表面定植缺陷与体内异常慢性定植（通过测量感染这些菌株的大鼠肺部细菌的恢复、组织学和免疫反应）联系起来。了解这些过程可能会带来新的治疗方法。Murray博士将通过学习新的技术来补充他在微生物学方面的经验：实时PCR、共聚焦显微镜、CF组织培养模型和慢性肺炎大鼠模型。Murray博士的主要导师在动物感染模型方面具有专业知识，他的顾问委员会包括微生物发病机理方面的专家和耶鲁大学CF中心主任，后者将教授他这些技能。他将学习转化研究、免疫生物学和动物护理方面的课程。通过这次额外的培训，Murray博士将获得成为假单胞菌慢性感染发病机制独立研究者的工具。相关性（见说明）：铜绿假单胞菌在囊性纤维化患者的肺部存活多年，导致频繁住院。目前的抗生素治疗不能成功地消除细菌，需要新的治疗方法。该项目的目标是更好地了解铜绿假单胞菌的特性，使其能够在肺部存活，从而开发新的治疗方法。

DESCRIPTION (provided by applicant): In patients with cystic fibrosis (CF), chronic colonization of the lungs with Pseudomonas aeruginosa results in frequent hospitalizations and in premature mortality. P. aeruginosa can colonize surfaces in multiple ways: swarming motility, sliding motility and the formation of biofilms. Expression of components on the surface of the bacteria (e.g. type IV pili, exopolysaccharide, flagella, and rhamnolipids) is important for these activities, but current understanding of the interactions and co-regulation of these factors is incomplete. Dr. Murray has preliminary evidence that certain regulatory proteins produced by P. aeruginosa each controls multiple factors important for colonization. His research goals are to: 1) Determine the components required for surface colonization co-regulated by these proteins (using strains of P. aeruginosa that variously lack these regulatory proteins and measuring levels of pilin, flagellin, exopolysaccharide, and rhamnolipid during swarming, sliding and formation of biofilms); 2) To quantitate the colonization of surfaces by these strains in a CF tissue culture model of infection (by visualizing bacterial/epithelial cell interactions with confocal microscopy and analyzing these images with COMSTAT software); and 3) To correlate in vitro surface colonization defects with abnormal chronic colonization in vivo (by measuring the recovery of bacteria, histology, and the immune response in the lungs of rats infected with these strains). Understanding these processes may lead to novel therapies. Dr. Murray will complement his experience in microbiology by learning new techniques: real time PCR, confocal microscopy, a CF tissue culture model, and a rat model of chronic pneumonia. Dr Murray's primary mentor has expertise in animal infection models and his advisory committee includes experts in microbial pathogenesis and the director of Yale's CF Center who will teach him these skills. He will take courses in translational research, immunobiology, and animal care. Through this additional training, Dr. Murray will gain the tools to become an independent investigator in the pathogenesis of chronic infection by Pseudomonas. RELEVANCE (See instructions): The bacterium Pseudomonas aeruginosa lives in the lungs of people with cystic fibrosis for many years, resulting in frequent hospitalizations. Current antibiotic therapies are not successful in eliminating the bacteria and new approaches to therapy are needed. The goal of this project is to better understand the properties of P. aeruginosa that allow it to survive in the lung so new therapies may be developed.