CHARACTERIZATION OF PSEUDOMONAS PHOSPHOLIPASES INVOLVED IN VIRULENCE

参与毒力的假单胞菌磷脂酶的表征

• 批准号: 8168184
• 负责人: MATTHEW J WARGO
• 金额: $ 3.84万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168184
• 关键词: Computer Retrieval of Information on Scientific Projects Database; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Enzymes; Failure; Functional disorder; Funding; Grant; Infection; Institution; Knockout Mice; Lead; Lipase; Lung; Measures; Mechanics; Mediator of activation protein; Modeling; Morbidity - disease rate; Mus; Phospholipase; Phospholipase C; Pseudomonas; Pseudomonas aeruginosa; Pulmonary Surfactants; Research; Research Personnel; Resources; Respiratory physiology; Role; Source; System; Testing; United States National Institutes of Health; Virulence; Virulence Factors; clinically significant; cystic fibrosis patients; mortality; research study; surfactant; therapeutic target

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The most common cause of morbidity and mortality in patients with Cystic Fibrosis (CF) is pulmonary system failure caused by Pseudomonas aeruginosa infection. One potential mechanism of P. aeruginosa damage to the lung is destruction of lung surfactant. Surfactant dysfunction can lead to small airway closure and severely decreased lung function. While the clinical significance of surfactant dysfunction in the CF lung is not fully understood, surfactant dysfunction may result in reduced lung function during exacerbations. We propose that several virulence factors secreted by P. aeruginosa function to degrade surfactant during infection resulting in decreased lung function. Preliminary evidence from a mouse lung infection model shows that P. aeruginosa infected mice show extensive airway closure. Some of this closure is due to the secreted hemolytic phospholipase C, PlcH, but in a plcH deletion strain there remains significant surfactant dysfunction. It is our hypothesis that the secreted lipase encoded by PA4921 is one of the additional mediators of surfactant degradation during P. aeruginosa lung infection. In this study we will verify the predicted enzymatic activity of PA4921 and determine the transcriptional regulatory mechanism governing its induction in pulmonary surfactant. Following the initial characterization, we will examine the role of PA4921 in alteration of lung mechanics in both wild type and Cystic Fibrosis Transmembrane Regulator knock-out (CFTR KO) mice. Lung mechanics, surfactant composition, and surfactant function will be measured during P. aeruginosa infection. Pulmonary instillation of purified PA4921 enzyme will be tested for sufficiency to alter lung mechanics and catalyze surfactant degradation. These experiments will allow us to determine if PA4921 is a potential therapeutic target applicable to CF exacerbations

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 囊性纤维化（CF）患者发病和死亡的最常见原因是铜绿假单胞菌感染引起的肺系统衰竭。铜绿假单胞菌对肺损伤的一种潜在机制是破坏肺表面活性物质。表面活性剂功能障碍可导致小气道闭合和肺功能严重下降。虽然 CF 肺中表面活性剂功能障碍的临床意义尚未完全了解，但表面活性剂功能障碍可能会导致病情加重期间肺功能下降。我们提出，铜绿假单胞菌分泌的几种毒力因子在感染过程中起到降解表面活性剂的作用，导致肺功能下降。 小鼠肺部感染模型的初步证据表明，铜绿假单胞菌感染的小鼠表现出广泛的气道闭合。这种封闭的部分原因是由于分泌的溶血磷脂酶 C、PlcH，但在 plcH 缺失菌株中，仍然存在显着的表面活性剂功能障碍。我们假设 PA4921 编码的分泌脂肪酶是铜绿假单胞菌肺部感染期间表面活性剂降解的附加介质之一。在本研究中，我们将验证 PA4921 的预测酶活性，并确定控制其在肺表面活性剂中诱导的转录调控机制。在初步表征之后，我们将研究 PA4921 在野生型和囊性纤维化跨膜调节因子敲除 (CFTR KO) 小鼠肺力学改变中的作用。将在铜绿假单胞菌感染期间测量肺力学、表面活性剂成分和表面活性剂功能。将测试纯化的 PA4921 酶的肺部滴注是否足以改变肺力学和催化表面活性剂降解。这些实验将使我们能够确定 PA4921 是否是适用于 CF 恶化的潜在治疗靶点