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

Abstract

ObjectiveWe investigated the relationship between plasma miRNAs levels and inflammatory characteristics in asthmatic patients.MethodsEligible 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.ResultsThe 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.ConclusionPlasma 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.

Phenotypical microRNA screen reveals a noncanonical role of CDK2 in regulating neutrophil migration

“…Here we provide evidence that miR-199 is a suppressor of cell migration in leukocytes, expanding its role to inflammation. Our observation is in line with a recent report that miR-199a-5p is increased in the plasma in patients with neutrophilic asthma 48 and inflammatory bowel disease 49 , suggesting that miR-199 may play physiological functions in inflammation. In addition, miR-199b suppress myeloid differentiation 50 , which can also contribute to its role in suppressing the innate immunity.…”

Phenotypical microRNA screen reveals a noncanonical role of CDK2 in regulating neutrophil migration

“…Here, we provide evidence that miR-199 is a suppressor of cell migration in leukocytes, expanding its role beyond cancer biology. The MIR-199a-5p levels were increased in the plasma of patients with neutrophilic asthma (53) and inflammatory bowel disease (54), suggesting that MIR-199 possibly plays physiological functions in inflammation. In addition, MIR-199b suppresses myeloid differentiation (55), which can also contribute to its role in suppressing the innate immunity.…”

MicroRNAs: Potential Biomarkers and Targets of Therapy in Allergic Diseases?

“…MiR-629-3p is expressed in bronchial epithelium and miR-223-3p and miR-142-3p—in neutrophils, monocytes and macrophages. Level of miR-199a-5p was significantly associated with impaired pulmonary function (Huang et al 2018). MiR-629-3p induces upregulation of IL-8 in epithelium, thus providing a mechanistic link between increased miR-629-3p expression and airway neutrophilia in patients with severe asthma (Maes et al 2016).…”