MicroRNA Mediated Chemokine Responses in Human Airway Smooth Muscle Cells.

Mythili Dileepan,Savita P Rao,Anne E Sarver,Reynold A Panettieri,Mathur S Kannan,Subbaya Subramanian,Neeraj Vij

doi:10.1371/journal.pone.0150842

Mythili Dileepan, Savita P Rao + Show 5 more

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https://doi.org/10.1371/journal.pone.0150842

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Journal: PloS one	Publication Date: Mar 21, 2016
Citations: 32	License type: CC BY 4.0

Affiliation: University of Minnesota, University of Pennsylvania

Abstract

Airway smooth muscle (ASM) cells play a critical role in the pathophysiology of asthma due to their hypercontractility and their ability to proliferate and secrete inflammatory mediators. microRNAs (miRNAs) are gene regulators that control many signaling pathways and thus serve as potential therapeutic alternatives for many diseases. We have previously shown that miR-708 and miR-140-3p regulate the MAPK and PI3K signaling pathways in human ASM (HASM) cells following TNF-α exposure. In this study, we investigated the regulatory effect of these miRNAs on other asthma-related genes. Microarray analysis using the Illumina platform was performed with total RNA extracted from miR-708 (or control miR)-transfected HASM cells. Inhibition of candidate inflammation-associated gene expression was further validated by qPCR and ELISA. The most significant biologic functions for the differentially expressed gene set included decreased inflammatory response, cytokine expression and signaling. qPCR revealed inhibition of expression of CCL11, CXCL10, CCL2 and CXCL8, while the release of CCL11 was inhibited in miR-708-transfected cells. Transfection of cells with miR-140-3p resulted in inhibition of expression of CCL11, CXCL12, CXCL10, CCL5 and CXCL8 and of TNF-α-induced CXCL12 release. In addition, expression of RARRES2, CD44 and ADAM33, genes known to contribute to the pathophysiology of asthma, were found to be inhibited in miR-708-transfected cells. These results demonstrate that miR-708 and miR-140-3p exert distinct effects on inflammation-associated gene expression and biological function of ASM cells. Targeting these miRNA networks may provide a novel therapeutic mechanism to down-regulate airway inflammation and ASM proliferation in asthma.

Highlights

Several recent reports have provided evidence that airway smooth muscle (ASM) has strong pro-inflammatory and immunomodulatory functions [1,2,3,4]
Recent reports show that ASM is capable of releasing cytokines such as IL-5 [16, 17], IL-6 [18, 19], IL-33 [20], TSLP [21], GM-CSF [22] and VEGF [23]; chemokines such as RANTES[16], Fractalkine [24], CCL11 [25], CXCL10[26,27,28], CXCL8[29]; adhesion molecules such as ICAM-1[30, 31], VCAM-1 [30, 32], CD44 [33] and LFA-1[34]; and growth factors such as IGF-1 [35, 36] and stem cell factor [37]
We performed a detailed analysis of the pattern of gene expression in human ASM cells (HASM) cells stimulated with TNF-α following transfection with miR-708

Summary

Introduction

Several recent reports have provided evidence that airway smooth muscle (ASM) has strong pro-inflammatory and immunomodulatory functions [1,2,3,4]. The immunomodulatory role of ASM may be more significant in establishing structural changes within the airways than its contractile function In this context, recent reports show that ASM is capable of releasing cytokines such as IL-5 [16, 17], IL-6 [18, 19], IL-33 [20], TSLP [21], GM-CSF [22] and VEGF [23]; chemokines such as RANTES[16], Fractalkine [24], CCL11 [25], CXCL10[26,27,28], CXCL8[29]; adhesion molecules such as ICAM-1[30, 31], VCAM-1 [30, 32], CD44 [33] and LFA-1[34]; and growth factors such as IGF-1 [35, 36] and stem cell factor [37]. Recent studies provide evidence for specific microRNAs in the regulation of ASM proliferation [43, 44], ASM phenotype [45] and airway inflammation [46, 47]

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