Microbiome in Chronic Obstructive Pulmonary Disease

描述(由申请人提供)： 肺部呼吸道细菌感染是慢性阻塞性肺疾病一些主要并发症的基础，这些并发症对疾病的进展和预后有显著影响。急性加重(AE)是慢性阻塞性肺疾病(COPD)患者发病率、临床恶化和发病率的主要因素之一。最重要的是，AE与COPD的严重程度是相互关联的，因为更严重的COPD患者更容易发生AE，反过来，会导致气流受限程度和疾病严重程度的进一步恶化。下呼吸道细菌感染或定植可能是这些免疫学和结构异常的触发机制。针对细菌小亚单位核糖体RNA(SSU-rRNA)编码基因的扩增和测序的新技术允许对存在于肺部疾病中的细菌种类进行分子询问，例如囊性纤维化。与传统方法相比，这项技术具有显著的潜在优势，包括痰细菌培养、诱导痰或支气管肺泡灌洗采样。在这项提案中，我们假设COPD患者的呼吸道将包含明确的微生物区系，这与患有轻度/轻度疾病的吸烟者不同。此外，我们认为COPD肺中的微生物区系在整个支气管树中发生变化，并与标志物炎症相关。具体目标1将利用高通量扩增和SSU-rRNA编码基因测序的激光捕获显微解剖LTRC样本，确定COPD患者、患有轻微或轻度疾病的吸烟者以及正常肺中沿呼吸道和实质内的肺微生物区系。特定目标2将利用免疫荧光结合平面测量法和/或体视学定量，将特定微生物群与COPD患者、吸烟者轻度或轻度疾病患者和正常肺中炎症和凋亡的组织标记物相关联。这一建议可能会导致高度相关的数据，这些数据可能会对正常呼吸道微环境对肺免疫的基本方面产生影响，以及细菌在COPD发病机制中的作用，并有可能应用于非COPD肺部疾病。与公共卫生相关：肺部呼吸道细菌感染是慢性阻塞性肺疾病(COPD)一些主要并发症的基础，COPD是一种由香烟烟雾引起的疾病，其特征是呼吸逐渐困难。患有严重COPD的患者有恶化和感染病毒和细菌的风险。这些被称为病情恶化，迫使患者住院并接受抗生素和抑制免疫系统的药物。然后，这些患者的死亡风险显著增加，呼吸短促的临床情况明显恶化。这项拟议的研究是一种新的方法，可以发现慢性阻塞性肺疾病患者的小气道中是否存在细菌。我们建议使用LTRC收集的肺组织并翻译一种新的和最先进的方法来大规模筛选可能存在于正常和病变肺中的细菌。(摘要结束)

DESCRIPTION (provided by applicant): Bacterial infection of lung airways underlies some of the main complications of COPD, which impact significantly in disease progression and outcome. Acute exacerbations (AE) represent one of the major contributing factors of morbidity, clinical deterioration, and morbidity among patients with COPD. Most importantly, AE and severity of COPD are mutually interrelated as AE are more frequent with patients with more severe COPD and, conversely, cause further deterioration of the degree of airflow limitation and disease severity. Lower airway bacterial infection or colonization may be the triggering mechanism of these immunological and structural abnormalities. Novel techniques aimed at amplification and sequencing of the gene encoding the bacterial small subunit ribosomal RNA (SSU-rRNA) allow for a molecular interrogation of species of bacteria present in lung diseases, such as demonstrated in cystic fibrosis. This technique offers significant potential advantages over traditional approaches including bacterial culture of sputum, induced sputum, or bronchoalveolar lavaged sampling. In this proposal, we hypothesize that the airways of COPD patients will contain defined microbiota, which is different from that of smokers with minimal/mild disease. Furthermore, we propose that the microbiota in COPD lungs changes throughout the bronchial tree and it correlates with markers inflammation. Specific Aim 1 will determine the lung microbiota along the airways and in the parenchyma in patients with COPD, smokers with minimal or mild disease, and in normal lungs using laser-capture microdissected LTRC samples based on high throughput amplification and sequencing of the gene encoding the SSU-rRNA. Specific Aim 2 will correlate specific microbiota with tissue markers of inflammation and apoptosis in lungs of patients COPD, smokers with minimal or mild disease and normal lungs using immunofluorescence allied to quantification by planimetry and/or stereology. This proposal may lead to highly relevant data with potential impact on fundamental aspects of the normal airway microenvironment on lung immunity and the role of bacterial in the pathogenesis of COPD, with potential applications to non-COPD lung diseases. PUBLIC HEALTH RELEVANCE: Bacterial infection of lung airways underlies some of the main complications of COPD, which is a disease caused by cigarette smoke and characterized by progressive difficulty in breathing. Patients with severe COPD are at risk of getting worse and acquiring viral and bacterial infections. These are called exacerbations, which oblige patients to be hospitalized and received antibiotics and drugs that suppress the immune system. These patients are then at significantly increased risk of death and marked clinical worsening of the shortness of breath. The proposed study is a novel way to find out whether there are bacteria present in small airway tubes in patients with COPD. We propose to use the lung tissues collected by the LTRC and translate a novel and state of the art method to perform a large scale screen of the bacteria that may present in normal and diseased lungs. (End of Abstract)