Plasma miR-199a-5p is increased in neutrophilic phenotype asthma patients and negatively correlated with pulmonary function

• Published in: PloS one Vol. 13; no. 3; p. e0193502
• Main Authors: Huang, Yali, Zhang, Shengding, Fang, Xiaoyu, Qin, Lu, Fan, Yu, Ding, Dandan, Liu, Xiansheng, Xie, Min
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 05.03.2018; Public Library of Science (PLoS)
• Subjects: Adults; Allergies; Asthma; Biology and life sciences; Biomarkers; c-Myc protein; Correlation; Correlation analysis; Critical care; Cytokines; Development and progression; Epithelial cells; Fluids; Gene expression; Health aspects; Hospitals; Hypertrophy; Immunology; Inflammation; Interleukin 4; Leukocytes (eosinophilic); Leukocytes (neutrophilic); Lipopolysaccharides; Lymphocytes; Macrophages; Major histocompatibility complex; Medicine; Medicine and Health Sciences; MicroRNA; MicroRNAs; miRNA; Muscles; Myc protein; Ontology; Pathogenesis; Patients; Peripheral blood; Phenotypes; Plasma; Polymerase chain reaction; Pulmonary functions; Pulmonary ventilation; Regression analysis; Respiratory function; Respiratory tract; Signal processing; Signal transduction; Signaling; Smooth muscle; Sputum; Stimulation; Studies; Transfection; Wnt protein; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0193502

We investigated the relationship between plasma miRNAs levels and inflammatory characteristics in asthmatic patients. Eligible adults with untreated asthma (n = 35) underwent a clinical assessment, sputum induction, and assessment of pulmonary function test and Asthma Control Test (ACT) scores. Asthma phenotypes were defined using the sputum cell count. miR-199a-5p expression was measured using quantitative real-time polymerase chain reaction (qPCR). Lipopolysaccharide (LPS) stimulation was used to detect miR-199a-5p secretion from peripheral blood-derived neutrophil, lymphocyte, macrophage and BEAS-2B cells. The correlation of miR-199a-5p expression with clinical parameters was analyzed using multiple linear regression analysis. In silico analysis predicted the target genes and signaling pathway of miR-199a-5p. Transfection of miR-199a-5p mimics in human airway smooth muscle cells (HASMCs) was performed in vitro. The miRNA-199a-5p levels in plasma and sputum increased significantly in patients with neutrophilic asthma compared to healthy subjects (ps = 0.014 and 0.006, respectively). Expression of miR-199a-5p in the plasma of asthmatic patients positively correlated with sputum miR-199a-5p expression (r = 0.511, p = 0.021). The miR-199a-5p level was only elevated with LPS stimulation in neutrophils but not macrophages, lymphocytes, or epithelial cells from healthy controls (p < 0.01). miR-199a-5p expression increased in response to LPS (p = 0.005) and LPS combined with IL-4 (p = 0.003), but not IL-4 alone. However, peripheral neutrophils from eosinophilic asthma patients did not respond to LPS with increased miR-199a-5p expression (n = 5, p > 0.05) in contrast to the significant response from neutrophilic patients (n = 4, p < 0.0001). miR-199a-5p negatively correlated with FEV1, FVC and PEF (r = -0.377, p = 0.026; r = -0.419, p = 0.012; and r = -0.392, p = 0.024, respectively). Multivariate correlation analysis confirmed that the plasma miR-199a-5p levels negatively correlated with FEV1 in patients with asthma (Adjusted R2 = 0.164, p = 0.015). In silico analysis suggested that the WNT signaling pathway participates in miR-199a-5p mediation of smooth muscle cell hypertrophy. In vitro experiment, miR-199a-5p mimics inhibited the protein expressions of WNT2 and WNT4, decreased the c-myc expression and dramatically increased the Sm-MHC expression in HASMCs. Plasma miR-199a-5p was increased in neutrophilic asthma and negatively correlated with pulmonary function, which suggests that miR-199a-5p actively contributes to disease pathogenesis by modulating the inflammatory process and transferring the signal from inflammatory cells to structure cells.