P. aeruginosa virulence in cystic fibrosis lung disease

囊性纤维化肺病中铜绿假单胞菌的毒力

• 批准号: 6816543
• 负责人: CLAUDIA ORDONEZ
• 金额: $ 12.64万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6816543
• 关键词: Pseudomonas aeruginosa; bacterial RNA; bacterial disease; bacterial genetics; clinical research; cystic fibrosis; functional /structural genomics; gene expression; gene expression profiling; host organism interaction; human subject; lung disorder; medical records; microarray technology; microorganism culture; pathologic process; patient oriented research; polymerase chain reaction; respiratory function; spirometry; sputum; virulence

DESCRIPTION (provided by applicant): This proposal describes a 5 year training program for development of a career in translational research in pulmonary medicine. The proposed program integrates clinical research in cystic fibrosis (CF) with functional genomics. As mentor, Dr. Stephen Lory, an expert in P. aeruginosa biology, will supervise aspects of P. aeruginosa genetic analysis. As co-mentor, Dr. Mary Ellen Wohl, an expert in CF clinical research, will supervise aspects involving clinical research design. As co-mentor, Dr. Gerald Pier, an expert in host-pathogen interactions in CF, will supervise aspects involving data analysis and applications to the host response. Infection with P. aeruginosa has been associated with decline in pulmonary function in CF. The P. aeruginosa microarray provides the means to undertake a comprehensive analysis of expression of all potential virulence factors encoded in the P. aeruginosa genome. The goal of the proposed research is to use a functional genomics approach to identify virulence factors associated with progression of CF lung disease. Our preliminary data show that we can characterize gene expression in clinical P. aeruginosa CF isolates by high throughput transcriptional profiling, and that older CF subjects are colonized with attenuated strains that have undergone deletions of several key virulence factors. We hypothesize that severity of CF lung disease is related to differential expression of specific P. aeruginosa virulence factors. We will address our hypothesis with the following aims: 1) to identify P. aeruginosa virulence factors expressed during different stages of lung disease progression by microarray analysis, and 2) to confirm the association between the expression of identified P. aeruginosa genes and progression of lung disease. Initially, we will employ microarray analysis to obtain global transcriptional profiles of P. aeruginosa in subjects with different disease severity. We will confirm the findings of the microarray experiments using RT-PCR on RNA obtained from P. aeruginosa isolates and directly from sputum samples from a larger number of subjects. These experiments will provide insights into the in vitro and in vivo effects of P. aeruginosa virulence factors in CF that may be important targets for new therapies with the potential to decrease disease-specific morbidity and mortality in CF.

描述（由申请人提供）： 该提案描述了一个为期5年的培训计划，用于发展肺部医学转化研究的职业生涯。该计划将囊性纤维化（CF）的临床研究与功能基因组学相结合。作为导师，铜绿假单胞菌生物学专家Stephen Lory博士将监督铜绿假单胞菌遗传分析的各个方面。作为共同导师，玛丽埃伦沃尔博士，在CF临床研究的专家，将监督涉及临床研究设计方面。作为共同导师，杰拉尔德码头，在CF宿主-病原体相互作用的专家，将监督涉及数据分析和应用到主机响应方面。 铜绿假单胞菌感染与CF的肺功能下降相关。铜绿假单胞菌微阵列提供了对铜绿假单胞菌基因组中编码的所有潜在毒力因子的表达进行全面分析的手段。这项研究的目的是使用功能基因组学方法来鉴定与CF肺病进展相关的毒力因子。我们的初步数据表明，我们可以通过高通量转录谱来表征临床铜绿假单胞菌CF分离株的基因表达，并且老年CF受试者被几个关键毒力因子缺失的减毒菌株定殖。 我们推测CF肺病的严重程度与特异性铜绿假单胞菌毒力因子的差异表达有关。我们将通过以下目的来解决我们的假设：1）通过微阵列分析来鉴定在肺部疾病进展的不同阶段表达的铜绿假单胞菌毒力因子，以及2）确认鉴定的铜绿假单胞菌基因的表达与肺部疾病进展之间的关联。最初，我们将采用微阵列分析来获得不同疾病严重程度受试者中铜绿假单胞菌的全局转录谱。我们将使用从铜绿假单胞菌分离株和直接从大量受试者的痰液样本中获得的RNA进行RT-PCR，证实微阵列实验的结果。这些实验将提供对铜绿假单胞菌毒力因子在CF中的体外和体内作用的见解，这些毒力因子可能是新疗法的重要靶点，具有降低CF中疾病特异性发病率和死亡率的潜力。