P. aeruginosa virulence in cystic fibrosis lung disease

描述(由申请人提供)： 这份提案描述了一项为期5年的培训计划，旨在发展肺部医学的转化性研究。拟议的计划将囊性纤维化(CF)的临床研究与功能基因组学结合起来。作为导师，铜绿假单胞菌生物学专家斯蒂芬·洛里博士将监督铜绿假单胞菌基因分析的各个方面。作为共同导师，CF临床研究专家Mary Ellen Wohl博士将监督涉及临床研究设计的各个方面。作为共同导师，杰拉尔德·皮尔博士，一位病原体-宿主相互作用的专家，将监督涉及数据分析和宿主反应应用的各个方面。 铜绿假单胞菌感染与CF患者的肺功能下降有关。铜绿假单胞菌微阵列提供了对编码在铜绿假单胞菌基因组中的所有潜在毒力因子的表达进行全面分析的手段。这项拟议的研究的目标是使用功能基因组学方法来确定与CF肺部疾病进展相关的毒力因素。我们的初步数据表明，我们可以通过高通量转录图谱来表征临床分离的铜绿假单胞菌的基因表达，并且老年CF受试者定植于经历了几个关键毒力因子缺失的减毒菌株。 我们推测，慢性肺病的严重程度与铜绿假单胞菌毒力因子的差异表达有关。我们的假设有以下几个目的：1)通过微阵列分析确定铜绿假单胞菌在肺部疾病进展的不同阶段表达的毒力因子；2)确认已识别的铜绿假单胞菌基因的表达与肺部疾病进展之间的关联。首先，我们将使用微阵列分析来获得不同疾病严重程度的受试者中铜绿假单胞菌的全球转录图谱。我们将使用RT-PCR对从铜绿假单胞菌分离株和直接从大量受试者的痰样本中获得的RNA进行验证微阵列实验的结果。这些实验将为铜绿假单胞菌毒力因子在体外和体内对CF的影响提供洞察力，这些毒力因子可能成为降低CF疾病特异性发病率和死亡率的新疗法的重要靶点。

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.