MicroRNA-155-5p Differentially Regulates IL-13Rα1 and IL-13Rα2 Expression and Signaling Driving Abnormal Lung Epithelial Cell Phenotype in Severe Asthma.

IF 5.9 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY American Journal of Respiratory Cell and Molecular Biology Pub Date : 2024-11-01 DOI:10.1165/rcmb.2024-0089OC
Martin Klein, Pierre-Alexandre Gagnon, Mabrouka Salem, Mahmoud Rouabhia, Jamila Chakir
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Abstract

MicroRNA (miR)-155-5p increases in innate and adaptive immune cells in response to IL-13 and is associated with the severity of asthma. However, little is known about its role in airway structural cells. Bronchial epithelial cells (BECs) isolated from healthy donors and patients with severe asthma were stimulated with IL-13. miR-155-5p expression and release were measured by real-time (RT)-PCR in BECs and in their derived exosomes. Modulation of miR-155-5p in BECs was performed using transfection of miR-155-5p inhibitor and mimic. IL-13 receptor α1 (IL-13Rα1), IL-13Rα2, MUC5AC, IL-8, and eotaxin-1 expression was measured by RT-PCR and Western blot analysis. The BEC repair process was assessed by a wound-healing assay. IL-13Rα1 and IL-13Rα2 expression and downstream pathways were evaluated by Western blot analysis. A dual luciferase assay was used to identify miR-155-5p target genes associated with IL-13R signaling. BECs from patients with severe asthma showed increased expression and exosomal release of miR-155-5p at baseline with amplification by IL-13 stimulation. BECs from patients with asthma expressed more IL-13Rα1 and less IL-13Rα2 than those from healthy donors, and IL-13Rα1 but not IL-13Rα2 induced miR-155-5p expression under IL-13 stimulation. miR-155-5p overexpression favored MUC5AC, IL-8, and Eotaxin-1 through the IL-13Rα1/SOCS1/STAT6 pathway while delaying the repair process by downregulating IL-13Rα2/MAPK14/c-Jun/c-fos signaling. The dual luciferase assay confirmed that miR-155-5p modulates both IL-13R pathways by directly targeting SOCS1, c-fos, and MAPK14. miR-155-5p is overexpressed in BECs from patients with severe asthma and regulates IL-13Rα1 and IL-13Rα2 expression and signaling, favoring expression of mucin- and eosinophil-related genes to the detriment of airway repair. These results show that miR-155-5p may contribute to airway epithelial cell dysfunction in patients with severe asthma.

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MiR-155-5p 可差异化调控 IL-13Rα1 和 IL-13Rα2 的表达和信号传导,导致严重哮喘患者肺上皮细胞表型异常。
众所周知,MiR-155-5p 在先天性和适应性免疫细胞中对 IL-13 的反应会增加,并与哮喘的严重程度有关。然而,人们对其在气道结构细胞中的作用知之甚少。用 IL-13 刺激从健康供体和严重哮喘患者体内分离出的 BECs。通过 RT-PCR 测定了 BECs 及其衍生外泌体中 MiR-155-5p 的表达和释放。利用转染 miR-155-5p 抑制剂和模拟物对 BECs 中的 miR-155-5p 进行调节。RT-PCR和Western blot检测了IL-13Rα1、IL-13Rα2、MUC5AC、IL-8和Eotaxin-1的表达。伤口愈合试验评估了 BECs 的修复过程。IL-13Rα1 和 IL-13Rα2 的表达及下游通路通过 Western 印迹进行评估。双荧光素酶测定法用于确定与IL-13受体信号转导相关的miR-155-5p靶基因。重症哮喘患者的 BECs 在基线期显示出 miR-155-5p 的表达和外泌体释放增加,并在 IL-13 刺激下放大。与健康供体相比,哮喘患者的 BECs 表达了更多的 IL-13Rα1,而较少的 IL-13Rα2;在 IL-13 刺激下,IL-13Rα1 而不是 IL-13Rα2 会诱导 miR-155-5p 的表达。通过 IL-13Rα1/SOCS1/STAT6 通路,MiR-155-5p 的过量表达有利于 MUC5AC、IL-8 和 Eotaxin-1,而 IL-13Rα2/MAPK14/c-Jun/c-Fos 信号下调则不利于延迟修复过程。双荧光素酶测定证实,miR-155-5p 通过直接靶向 SOCS1、c-Fos 和 MAPK14 来调节 IL-13 受体的两条通路。MiR-155-5p 在重症哮喘 BECs 中过表达,并调控 IL-13Rα1 和 IL-13Rα2 的表达和信号传导,有利于粘蛋白和嗜酸性粒细胞相关基因的表达,不利于气道修复。这些结果表明,miR-155-5p 可能会导致严重哮喘患者的气道上皮细胞功能障碍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
11.20
自引率
3.10%
发文量
370
审稿时长
3-8 weeks
期刊介绍: The American Journal of Respiratory Cell and Molecular Biology publishes papers that report significant and original observations in the area of pulmonary biology. The focus of the Journal includes, but is not limited to, cellular, biochemical, molecular, developmental, genetic, and immunologic studies of lung cells and molecules.
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