Microbiome in Chronic Obstructive Pulmonary Disease

描述（由申请人提供）： 肺气道细菌感染是COPD的一些主要并发症的基础，其对疾病进展和结局有显著影响。急性加重（AE）是COPD患者发病、临床恶化和发病的主要促成因素之一。最重要的是，AE和COPD的严重程度相互关联，因为AE在更严重的COPD患者中更常见，相反，导致气流受限程度和疾病严重程度进一步恶化。下呼吸道细菌感染或定植可能是这些免疫和结构异常的触发机制。旨在扩增和测序编码细菌小亚基核糖体RNA（SSU-rRNA）的基因的新技术允许对肺部疾病中存在的细菌物种进行分子询问，例如囊性纤维化。该技术提供了显着的潜在优势，传统的方法，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患者的气道将包含定义的微生物群，这与患有轻微/轻度疾病的吸烟者不同。此外，我们提出COPD肺部的微生物群在整个支气管树中发生变化，并且与炎症标志物相关。具体目标1将使用基于编码SSU-rRNA的基因的高通量扩增和测序的激光捕获显微切割的LTRC样品，确定COPD患者、具有轻微或轻度疾病的吸烟者以及正常肺中沿气道和实质中的肺微生物群沿着。具体目标2将使用与通过测面积和/或体视学的定量相关联的免疫荧光将特定微生物群与COPD患者、具有轻微或轻度疾病的吸烟者和正常肺的肺中的炎症和细胞凋亡的组织标志物相关联。这一提议可能会导致高度相关的数据，对肺免疫的正常气道微环境的基本方面以及细菌在COPD发病机制中的作用具有潜在影响，并可能应用于非COPD肺部疾病。 公共卫生相关性：肺气道的细菌感染是COPD的一些主要并发症的基础，COPD是由香烟烟雾引起的疾病，其特征是进行性呼吸困难。严重COPD患者有恶化和获得病毒和细菌感染的风险。这些被称为加重，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，呼吸短促的临床症状显著恶化。这项研究是一种新的方法，可以发现COPD患者的小气道内是否存在细菌。我们建议使用LTRC收集的肺组织，并转化为一种新的最先进的方法，对可能存在于正常和患病肺部的细菌进行大规模筛选。 (End摘要）

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)