BackgroundExtracellular vesicles (EVs) have been implicated in the pathogenesis of asthma, however, how EVs contribute to immune dysfunction and type 2 airway inflammation remains incompletely understood. We aimed to elucidate roles of airway EVs and their miRNA cargo in the pathogenesis of NSAID‐exacerbated respiratory disease (N‐ERD), a severe type 2 inflammatory condition.MethodsEVs were isolated from induced sputum or supernatants of cultured nasal polyp or turbinate tissues of N‐ERD patients or healthy controls by size‐exclusion chromatography and characterized by particle tracking, electron microscopy and miRNA sequencing. Functional effects of EV miRNAs on gene expression and mediator release by human macrophages or normal human bronchial epithelial cells (NHBEs) were studied by RNA sequencing, LC–MS/MS and multiplex cytokine assays.ResultsEVs were highly abundant in secretions from the upper and lower airways of N‐ERD patients. N‐ERD airway EVs displayed profoundly altered immunostimulatory capacities and miRNA profiles compared to airway EVs of healthy individuals. Airway EVs of N‐ERD patients, but not of healthy individuals induced inflammatory cytokine (GM‐CSF and IL‐8) production by NHBEs. In macrophages, N‐ERD airway EVs exhibited an impaired potential to induce cytokine and prostanoid production, while enhancing M2 macrophage activation. Let‐7 family miRNAs were highly enriched in sputum EVs from N‐ERD patients and mimicked suppressive effects of N‐ERD EVs on macrophage activation.ConclusionAberrant airway EV miRNA profiles may contribute to immune dysfunction and chronic type 2 inflammation in N‐ERD. Let‐7 family miRNAs represent targets for correcting aberrant macrophage activation and mediator responses in N‐ERD.

中文翻译：

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细胞外囊泡 miRNA 在 NSAID 加剧的呼吸系统疾病中驱动巨噬细胞异常反应

背景细胞外囊泡 (EV) 与哮喘的发病机制有关，然而，细胞外囊泡如何导致免疫功能障碍和 2 型气道炎症仍不完全清楚。我们的目的是阐明气道 EV 及其 miRNA 负载在 NSAID 加剧的呼吸系统疾病 (N-ER​​D)（一种严重的 2 型炎症性疾病）发病机制中的作用。方法从培养的鼻息肉或鼻甲组织的诱导痰或上清液中分离出 EV。 N-ERD 患者或健康对照通过尺寸排阻色谱进行分析，并通过粒子追踪、电子显微镜和 miRNA 测序进行表征。通过 RNA 测序、LC-MS/MS 和多重细胞因子测定研究 EV miRNA 对人巨噬细胞或正常人支气管上皮细胞 (NHBE) 基因表达和介质释放的功能影响。结果 EV 在上、下分泌物中高度丰富N-ERD 患者的气道。与健康个体的气道 EV 相比，N-ERD 气道 EV 表现出显着改变的免疫刺激能力和 miRNA 谱。 N-ERD 患者（而非健康个体）的气道 EV 诱导 NHBE 产生炎症细胞因子（GM-CSF 和 IL-8）。在巨噬细胞中，N-ERD 气道 EV 诱导细胞因子和前列腺素生成的潜力受损，同时增强了 M2 巨噬细胞的激活。 Let-7家族miRNA在N-ERD患者的痰EV中高度富集，并且模拟了N-ERD EV对巨噬细胞活化的抑制作用。结论异常的气道EV miRNA谱可能导致N-ERD中的免疫功能障碍和慢性2型炎症。 Let-7 家族 miRNA 代表纠正 N-ERD 中异常巨噬细胞激活和介质反应的靶标。