The regulation of Pseudomonas aeruginosa surface colonization

铜绿假单胞菌表面定植的调控

• 批准号: 7572165
• 负责人: THOMAS SCOT MURRAY
• 金额: $ 13.19万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-04 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7572165
• 关键词: Advisory Committees; Affect; Anabolism; Animals; Antibiotic Therapy; Bacteria; Bacterial Infections; Cell Communication; Cell Death; Chronic; Communities; Complement; Complex; Computer software; Confocal Microscopy; Cystic Fibrosis; Defect; Educational process of instructing; Epithelial Cells; Exotoxins; Fimbriae Proteins; Flagella; Flagellin; Genetic Transcription; Goals; Histology; Hospitalization; Image Analysis; Immune response; Immunobiology; In Vitro; Infection; Instruction; Lactate Dehydrogenase; Lead; Learning; Life; Lung; Lung Inflammation; Measures; Mentors; Microbial Biofilms; Microbiology; Modeling; Morbidity - disease rate; Mutation; Organelles; Organism; Pathogenesis; Patients; Pilum; Pneumonia; Premature Mortality; Principal Investigator; Process; Production; Property; Proteins; Pseudomonas Infections; Pseudomonas aeruginosa; Rattus; Recovery; Regulation; Research; Research Personnel; Slide; Surface; Techniques; Testing; Tight Junctions; Time; Training; Translational Research; Virulence Factors; animal care; cell motility; cystic fibrosis patients; cytokine; experience; genetic regulatory protein; in vitro Model; in vivo; in vivo Model; microbial; mortality; mutant; novel; novel strategies; novel therapeutic intervention; pathogen; research study; rhamnolipid; skills; surfactant; tissue culture; tool

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.

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。铜绿假单胞菌可以以多种方式在表面定殖：群集运动、滑动运动和生物膜的形成。细菌表面组分（如IV型皮利、胞外多糖、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调节的理解还不完整。Murray博士有初步证据表明，铜绿假单胞菌产生的某些调节蛋白各自控制着对定植至关重要的多种因素。他的研究目标是：1）确定由这些蛋白质共调节的表面定殖所需的组分（使用不同程度地缺乏这些调节蛋白的铜绿假单胞菌菌株，并在群集、滑动和生物膜形成期间测量菌毛蛋白、鞭毛蛋白、外泌多糖和鼠李糖脂的水平）; 2）在感染的CF组织培养模型中定量这些菌株在表面的定殖（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）;和3）将体外表面定植缺陷与体内异常慢性定植相关联（通过测量细菌的恢复、组织学和感染这些菌株的大鼠肺中的免疫应答）。了解这些过程可能会导致新的治疗方法。Murray博士将通过学习新技术来补充他在微生物学方面的经验：真实的时间PCR，共聚焦显微镜，CF组织培养模型和慢性肺炎大鼠模型。默里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中心的主任，他们将教他这些技能。他将参加转化研究，免疫生物学和动物护理课程。通过这次额外的培训，Murray博士将获得工具，成为假单胞菌慢性感染发病机制的独立研究者。相关性（参见说明）：铜绿假单胞菌在囊性纤维化患者的肺部生活多年，导致频繁住院。目前的抗生素疗法在消除细菌方面并不成功，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，以便开发新的治疗方法。