A Pathogenic Smoke Associated Neutrophilic Exosomal Pathway.

项目摘要 慢性阻塞性肺疾病(COPD)是一种慢性炎症性疾病，据信由 由蛋白酶-抗蛋白酶失衡所致。导致这种不平衡的机制尚未完全形成。 明白了。最近的研究表明，外小体(由细胞释放的小纳米囊泡)来自 激活的中性粒细胞(PMN)包被来自脱颗粒的PMN的中性粒细胞弹性蛋白酶(NE)，这 外切体相关的NE使其免受其天然抗蛋白酶-α-1-抗胰蛋白酶(α1AT)的保护。 这种对α1AT的抵抗力使外切体相关NE在导致慢性阻塞性肺疾病方面的效力提高了几倍 小鼠模型中的疾病表型比溶液中游离NE的表型要好。这些PMN外切体可以结合到 I型胶原和降解结构性细胞外基质(ECM)蛋白。更重要的是，这些 中性粒细胞来源的NE+外切体可引起小鼠模型的肺泡破坏，这些NE+外切体 在COPD患者的BALF中可以发现，但不是健康的从不吸烟的对照组，这表明 外切体相关NE在COPD疾病进展中的重要作用。这笔赠款将确定 去甲肾上腺素与中性粒细胞外切体表面结合的机制及对分子作用的研究 破坏这种联系，使NE容易受到α1AT失活的影响。此外，这笔赠款将 建立NE+PMN外切体产生吸烟小鼠模型并将疾病转移给幼稚小鼠， 有效地创建了一种老鼠-老鼠传播疾病的模型。此外，这笔赠款将与 COPD患者BALF中中性粒细胞NE+外切体的存在与COPD的其他重要指标 严肃性。此外，PMN来源于其他侵入性较小的患者体液样本、血清中的NE+外切体 和痰，将被量化，它们将疾病传播到COPD小鼠模型的能力将是 与患者的BALF相比。此外，所研究的物质可以将NE从 外切体表面可以发展成为潜在的治疗靶点。

Project Summary Chronic Obstructive Pulmonary Disease (COPD) is a chronic inflammatory disease believed to be driven by protease-antiprotease imbalance. The mechanisms leading to this imbalance have yet to be fully understood. Recent work has suggested that exosomes (small nanovesicles released by cells) from activated neutrophils (PMNs) are coated in neutrophil elastase (NE) from degranulated PMNs and this exosome associated NE renders it protected from its native antiprotease, alpha-1-antitrypsin (α1AT). This resistance to α1AT makes exosome associated NE several log-fold more potent in causing a COPD disease-like phenotype in mouse models than free NE in solution. These PMN exosomes can bind to type I collagen and degrade structural extracellular matrix (ECM) proteins. Of bigger significance, these PMN derived NE+ exosomes can cause alveolar destruction in a mouse model and these NE+ exosomes can be found in the BALF of COPD patients, but not healthy never smoker controls, indicating an important role for exosome associated NE in COPD disease progression. This grant will identify the mechanism of NE association to the surface of PMN exosomes as well as focusing on molecules to disrupt this association, rendering the NE susceptible to α1AT inactivation. Furthermore, this grant will develop a smoking mouse model of NE+ PMN exosome production and disease transfer to naïve mice, effectively creating a mouse-mouse transfer model of disease. Additionally, this grant will correlate the presence of PMN NE+ exosomes in COPD patient BALF with other significant parameters of COPD severity. Moreover, PMN derived NE+ exosomes from other, less invasive patient fluid samples, serum and sputum, will be quantified and their ability to transfer disease to mouse model of COPD will be compared to those from patient BALF. Additionally, substances studied that can dissociate NE from the exosome surface can be developed into potential therapeutic targets.