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，被认为是由 蛋白酶-抗蛋白酶失衡导致这种不平衡的机制尚未得到充分的研究。 明白最近的研究表明，外来体（由细胞释放的小纳米囊泡）， 活化的中性粒细胞（PMN）被来自脱颗粒的PMN的中性粒细胞弹性蛋白酶（NE）包被， 外泌体相关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的抗性使得外泌体相关的NE在引起COPD方面的效力增加几个对数倍 疾病样表型的小鼠模型中比溶液中的游离NE。这些PMN外泌体可以结合到 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 PMN衍生的NE+外泌体可以在小鼠模型中引起肺泡破坏，并且这些NE+外泌体可以在小鼠模型中引起肺泡破坏。 可以在COPD患者的BALF中发现，但在健康的从不吸烟者对照中没有发现，这表明 外泌体相关NE在COPD疾病进展中重要作用这笔赠款将确定 NE与PMN外泌体表面结合的机制以及关注 破坏这种联系，使NE易受α1AT失活的影响。此外，这笔赠款将 开发NE+ PMN外泌体产生和疾病转移至幼稚小鼠的吸烟小鼠模型， 有效地建立了一个疾病的小鼠-小鼠传播模型。此外，这笔赠款将与 COPD患者BALF中PMN NE+外来体的存在以及COPD的其他重要参数 严重性。此外，PMN来源于来自其他侵入性较小的患者体液样品、血清和外周血的NE+外泌体。 和痰，将被量化，并且将它们将疾病转移到COPD小鼠模型的能力被量化。 与患者BALF中的相比。此外，所研究的可以将NE从 外泌体表面可被开发为潜在的治疗靶点。

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.