Neutrophil Exosomes: New Pathogenic Entities in COPD

中性粒细胞外泌体：慢性阻塞性肺病的新致病实体

• 批准号: 10467984
• 负责人: AMIT GAGGAR
• 金额: --
• 依托单位: BIRMINGHAM VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10467984
• 关键词: Active Sites; Address; Aging; Binding; Biological Markers; Bronchoalveolar Lavage; Bypass; Cause of Death; Cells; Characteristics; Charge; Chronic; Chronic Obstructive Airway Disease; Cleaved cell; Collagen; Data; Development; Diagnosis; Disease; Enzymes; Experimental Models; Extracellular Matrix; Extracellular Matrix Degradation; Generations; Glycine; Health; Healthcare Systems; Hospitalization; Impairment; In Vitro; Incidence; Individual; Inflammatory; Journals; Lead; Leukocyte Elastase; Lung; Lung diseases; Measurement; Measures; Medicine; Military Personnel; Modeling; Molecular Target; Morbidity - disease rate; Mucociliary Clearance; Mus; Natural History; Natural Immunity; Nature; Pathogenesis; Pathogenicity; Pathologic; Peptide Hydrolases; Play; Population; Prevalence; Process; Proline; Protease Inhibitor; Proteoglycan; Publishing; Pulmonary Emphysema; Recombinants; Resistance; Role; Sampling; Seminal; Smoke; Smoker; Smoking; Surface; Technology; Therapeutic; Time; Treatment Cost; United States; Veterans; airway epithelium; airway inflammation; airway obstruction; airway remodeling; alpha 1-Antitrypsin; cigarette smoke; cohort; disease phenotype; exosome; former smoker; improved; in vivo; military veteran; mortality; mouse model; neutrophil; neutrophil elastase inhibitor; never smoker; novel; novel therapeutic intervention; patient population; prolyl oligopeptidase; pulmonary function; small molecule; therapeutic target; ventilation

COPD is a chronic inflammatory pulmonary condition which the 3rd leading cause of death in the United States and has significant impact on the US Veteran and active military population. Despite its prevalence and increased attributable morbidity/mortality, there are no specific COPD therapeutics which alter the natural history of the disorder. Our group and others have provided extensive evidence of the importance of proteolytic damage as a critical component to the progression of COPD. However, our ability to understand how proteases bypass the robust antiprotease shield within the lung is poorly understood. Here, we examine a new pathogenic entity, the neutrophil-derived exosome, which expresses the protease neutrophil elastase (NE) on its surface. We provide preliminary data demonstrating these exosomes have active NE enzymatic activity, capable of degrading components of the lung extracellular matrix (ECM). Importantly, we highlight that exosome-associated NE is resistant to its naturally occurring antiprotease, alpha-1 antitrypsin (A1AT), and can lead to fulminant emphysema when intratracheally administered in vivo. For this proposal, we will build on these seminal observations, first by examining the impact of smoke to induce the release of these proteolytic exosomes from PMNs (Specific Aim 1a) and then examining the antiprotease resistance of exosomes isolated from COPD and non-COPD subjects, with a focus of inducing NE disassociation from these exosomes to enhance endogenous antiprotease sensitivity (Specific Aim 1b). Next, we will use bronchoalveolar lavage (BAL) samples from 6-month smoking mouse model, isolate PMN-derived exosomes, and intratracheally deliver these into naïve mice to induce emphysema, inhibiting these effects via NE disassociation (Specific Aim 2). Finally, we will examine a cohort of COPD subjects (current or former smokers) and non-COPD subjects (smokers and never smokers) to determine the expression of NE-associated exosomes in these cohorts and stability of these measurements over time (Specific Aim 3). The successful completion of these aims will lead to an increased understanding of the PMN exosome as a critical pathogenic entity in COPD, with improved understanding of the downstream effects of its unfettered protease activity. Importantly, these studies will likely result in the development of a new biomarker and potential new therapeutic approaches for the treatment of Veterans who are diagnosed with COPD.

COPD 是一种慢性炎症性肺部疾病，是美国第三大死亡原因 州并对美国退伍军人和现役军人产生重大影响。尽管其 患病率和归因发病率/死亡率增加，没有具体的慢性阻塞性肺病治疗方法 这改变了疾病的自然史。我们的团队和其他人提供了广泛的证据 蛋白水解损伤作为 COPD 进展的关键组成部分的重要性。然而， 我们对蛋白酶如何绕过肺内强大的抗蛋白酶屏障的理解能力很差 明白了。在这里，我们检查了一种新的致病实体，即中性粒细胞衍生的外泌体，它 在其表面表达蛋白酶中性粒细胞弹性蛋白酶（NE）。我们提供初步数据 证明这些外泌体具有活跃的 NE 酶活性，能够降解 肺细胞外基质（ECM）。重要的是，我们强调外泌体相关的 NE 具有耐药性 其天然存在的抗蛋白酶，α-1 抗胰蛋白酶 (A1AT)，并可导致暴发性 体内气管内给药时出现肺气肿。对于这项提案，我们将在这些开创性的基础上 观察，首先通过检查烟雾对诱导这些蛋白水解物释放的影响 从 PMN 中提取外泌体（具体目标 1a），然后检查外泌体的抗蛋白酶抗性 从 COPD 和非 COPD 受试者中分离出来，重点是诱导 NE 与这些受试者分离 外泌体增强内源性抗蛋白酶敏感性（具体目标 1b）。接下来，我们将使用 来自 6 个月吸烟小鼠模型的支气管肺泡灌洗 (BAL) 样本，分离出 PMN 来源 外泌体，并将其气管内递送至幼鼠体内以诱导肺气肿，从而抑制这些 通过 NE 解离产生影响（具体目标 2）。最后，我们将检查一组慢性阻塞性肺病受试者 （当前或以前吸烟者）和非慢性阻塞性肺病受试者（吸烟者和从不吸烟者）以确定 这些队列中 NE 相关外泌体的表达以及这些测量值随时间的稳定性 （具体目标 3）。这些目标的成功实现将加深人们对 PMN 外泌体作为 COPD 的关键致病实体，随着人们对 PMN 外泌体的了解的加深 其不受限制的蛋白酶活性的下游影响。重要的是，这些研究可能会导致 开发新的生物标志物和潜在的新治疗方法 被诊断患有慢性阻塞性肺病的退伍军人。