Section of Pulmonary, Critical Care and Sleep Medicine, Yale School of Medicine, New Haven, Conn.
Section of Pulmonary, Critical Care and Sleep Medicine, Yale School of Medicine, New Haven, Conn; Yale School of Nursing, Orange, Conn.
J Allergy Clin Immunol. 2020 Feb;145(2):550-562. doi: 10.1016/j.jaci.2019.10.031.
Airway eosinophilia is a prominent feature of asthma and chronic rhinosinusitis (CRS), and the endothelium plays a key role in eosinophil trafficking. To date, microRNA-1 (miR-1) is the only microRNA known to be regulated in the lung endothelium in asthma models.
We sought to determine the role of endothelial miR-1 in allergic airway inflammation.
We measured microRNA and mRNA expression using quantitative RT-PCR. We used ovalbumin and house dust mite models of asthma. Endothelium-specific overexpression of miR-1 was achieved through lentiviral vector delivery or induction of a transgene. Tissue eosinophilia was quantified by using Congo red and anti-eosinophil peroxidase staining. We measured eosinophil binding with a Sykes-Moore adhesion chamber. Target recruitment to RNA-induced silencing complex was assessed by using anti-Argonaute2 RNA immunoprecipitation. Surface P-selectin levels were measured by using flow cytometry.
Serum miR-1 levels had inverse correlations with sputum eosinophilia, airway obstruction, and number of hospitalizations in asthmatic patients and sinonasal tissue eosinophilia in patients with CRS. IL-13 stimulation decreased miR-1 levels in human lung endothelium. Endothelium-specific overexpression of miR-1 reduced airway eosinophilia and asthma phenotypes in murine models and inhibited IL-13-induced eosinophil binding to endothelial cells. miR-1 recruited P-selectin, thymic stromal lymphopoietin, eotaxin-3, and thrombopoietin receptor to the RNA-induced silencing complex; downregulated these genes in the lung endothelium; and reduced surface P-selectin levels in IL-13-stimulated endothelial cells. In our asthma and CRS cohorts, miR-1 levels correlated inversely with its target genes.
Endothelial miR-1 regulates eosinophil trafficking in the setting of allergic airway inflammation. miR-1 has therapeutic potential in asthmatic patients and patients with CRS.
气道嗜酸性粒细胞增多是哮喘和慢性鼻-鼻窦炎（CRS）的一个显著特征，内皮在嗜酸性粒细胞迁移中起关键作用。迄今为止，miR-1 是唯一已知在哮喘模型中肺内皮中受调控的 microRNA。
我们旨在确定内皮 miR-1 在过敏性气道炎症中的作用。
我们使用定量 RT-PCR 测量 microRNA 和 mRNA 的表达。我们使用卵清蛋白和屋尘螨模型来模拟哮喘。通过慢病毒载体转导或诱导转基因来实现内皮特异性 miR-1 过表达。使用刚果红和抗嗜酸性粒细胞过氧化物酶染色来定量组织嗜酸性粒细胞。我们使用 Sykes-Moore 黏附室来测量嗜酸性粒细胞的黏附。通过抗 Argonaute2 RNA 免疫沉淀来评估靶基因对 RNA 诱导沉默复合物的招募。通过流式细胞术测量表面 P 选择素水平。
哮喘患者的血清 miR-1 水平与痰嗜酸性粒细胞、气道阻塞和住院次数呈负相关，与 CRS 患者的鼻-鼻窦组织嗜酸性粒细胞也呈负相关。IL-13 刺激降低了人肺内皮中的 miR-1 水平。在小鼠模型中，内皮特异性过表达 miR-1 可减少气道嗜酸性粒细胞和哮喘表型，并抑制 IL-13 诱导的嗜酸性粒细胞与内皮细胞的黏附。miR-1 将 P 选择素、胸腺基质淋巴细胞生成素、嗜酸性粒细胞趋化因子-3 和血小板生成素受体招募到 RNA 诱导沉默复合物；下调肺内皮中的这些基因；并降低 IL-13 刺激的内皮细胞表面 P 选择素水平。在我们的哮喘和 CRS 队列中，miR-1 水平与其靶基因呈负相关。
内皮 miR-1 调节过敏性气道炎症中的嗜酸性粒细胞迁移。miR-1 在哮喘患者和 CRS 患者中具有治疗潜力。
