title: MicroRNA-570-3p regulates HuR and cytokine expression in airway epithelial cells.
abstract: Asthma is a chronic lung disease that affects people of all ages and is characterized by high morbidity. The mechanisms of asthma pathogenesis are unclear, and there is a need for development of diagnostic biomarkers and greater understanding of regulation of inflammatory responses in the lung. Post-transcriptional regulation of cytokines, chemokines, and growth factors by the action of microRNAs and RNA-binding proteins on stability or translation of mature transcripts is emerging as a central means of regulating the inflammatory response. In this study, we demonstrate that miR-570-3p expression is increased with TNFα stimuli in normal human bronchial epithelial cells (2.6 ± 0.6, p = 0.01) and the human airway epithelial cell line A549 (4.6 ± 1.4, p = 0.0068), and evaluate the functional effects of its overexpression on predicted mRNA target genes in transfected A549 cells. MiR-570-3p upregulated numerous cytokines and chemokines (CCL4, CCL5, TNFα, and IL-6) and also enhanced their induction by TNFα. For other cytokines (CCL2 and IL-8), the microRNA exhibited an inhibitory effect to repress their upregulation by TNFα. These effects were mediated by a complex pattern of both direct and indirect regulation of downstream targets by miR-570-3p. We also show that the RNA-binding protein HuR is a direct target of miR-570-3p, which has implications for expression of numerous other inflammatory mediators that HuR is known regulate post-transcriptionally. Finally, expression of endogenous miR-570-3p was examined in both serum and exhaled breath condensate (EBC) from asthmatic and healthy patients, and was found to be significantly lower in EBC of asthmatics and inversely correlated to their lung function. These studies implicate miR-570-3p as a potential regulator of asthmatic inflammation with potential as both a diagnostic and therapeutic target in asthma.
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, Section of Allergy and Immunology, The Pennsylvania State University Milton S Hershey Medical Center 500 University Dr, Hershey, PA 17033, USA.
authors.forename: Alanna N
authors.initials: AN
authors.lastname: Roff
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, Section of Allergy and Immunology, The Pennsylvania State University Milton S Hershey Medical Center 500 University Dr, Hershey, PA 17033, USA.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Craig
authors.affiliation: Department of Microbiology and Immunology, Cornell University Ithaca, NY 14853.
authors.forename: Avery
authors.initials: A
authors.lastname: August
authors.affiliation: University of Salerno Medicine Campus Via Salvador Allende, Baronissi (SA), 84081 Italy.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Stellato
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, Section of Allergy and Immunology, The Pennsylvania State University Milton S Hershey Medical Center 500 University Dr, Hershey, PA 17033, USA ; Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine 500 University Dr, Hershey, PA 17033, USA.
authors.forename: Faoud T
authors.initials: FT
authors.lastname: Ishmael
keywords: MicroRNA; airway; asthma; biomarker; cytokines; inflammation; post-transcriptional regulation