title: IL-6 trans-signaling increases expression of airways disease genes in airway smooth muscle.
abstract: Genetic data suggest that IL-6 trans-signaling may have a pathogenic role in the lung; however, the effects of IL-6 trans-signaling on lung effector cells have not been investigated. In this study, human airway smooth muscle (HASM) cells were treated with IL-6 (classical) or IL-6+sIL6R (trans-signaling) for 24 h and gene expression was measured by RNAseq. Intracellular signaling and transcription factor activation were assessed by Western blotting and luciferase assay, respectively. The functional effect of IL-6 trans-signaling was determined by proliferation assay. IL-6 trans-signaling had no effect on phosphoinositide-3 kinase and Erk MAP kinase pathways in HASM cells. Both classical and IL-6 trans-signaling in HASM involves activation of Stat3. However, the kinetics of Stat3 phosphorylation by IL-6 trans-signaling was different than classical IL-6 signaling. This was further reflected in the differential gene expression profile by IL-6 trans-signaling in HASM cells. Under IL-6 trans-signaling conditions 36 genes were upregulated, including PLA2G2A, IL13RA1, MUC1, and SOD2. Four genes, including CCL11, were downregulated at least twofold. The expression of 112 genes was divergent between IL-6 classical and trans-signaling, including the genes HILPDA, NNMT, DAB2, MUC1, WWC1, and VEGFA. Pathway analysis revealed that IL-6 trans-signaling induced expression of genes involved in regulation of airway remodeling, immune response, hypoxia, and glucose metabolism. Treatment of HASM cells with IL-6+sIL6R induced proliferation in a dose-dependent fashion, suggesting a role for IL-6 trans-signaling in asthma pathogenesis. These novel findings demonstrate differential effect of IL-6 trans-signaling on airway cells and identify IL-6 trans-signaling as a potential modifier of airway inflammation and remodeling.
authors.affiliation: Wake Forest School of Medicine, Center for Genomics and Personalized Medicine Research, Winston-Salem, North Carolina; Wake Forest School of Medicine, Department of Neurobiology and Anatomy, Winston-Salem, North Carolina;
authors.forename: Mac B
authors.initials: MB
authors.lastname: Robinson
authors.affiliation: Center for Translational Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania; and.
authors.forename: Deepak A
authors.initials: DA
authors.lastname: Deshpande
authors.affiliation: Wake Forest School of Medicine, Center for Public Health Genomics, Winston-Salem, North Carolina.
authors.forename: Jeffery
authors.initials: J
authors.lastname: Chou
authors.affiliation: Wake Forest School of Medicine, Center for Genomics and Personalized Medicine Research, Winston-Salem, North Carolina;
authors.forename: Wei
authors.initials: W
authors.lastname: Cui
authors.affiliation: Wake Forest School of Medicine, Center for Genomics and Personalized Medicine Research, Winston-Salem, North Carolina;
authors.forename: Shelly
authors.initials: S
authors.lastname: Smith
authors.affiliation: Wake Forest School of Medicine, Center for Public Health Genomics, Winston-Salem, North Carolina.
authors.forename: Carl
authors.initials: C
authors.lastname: Langefeld
authors.affiliation: Wake Forest School of Medicine, Center for Genomics and Personalized Medicine Research, Winston-Salem, North Carolina;
authors.forename: Annette T
authors.initials: AT
authors.lastname: Hastie
authors.affiliation: Wake Forest School of Medicine, Center for Genomics and Personalized Medicine Research, Winston-Salem, North Carolina;
authors.forename: Eugene R
authors.initials: ER
authors.lastname: Bleecker
authors.affiliation: Wake Forest School of Medicine, Center for Genomics and Personalized Medicine Research, Winston-Salem, North Carolina; ghawkins@wakehealth.edu.
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Hawkins
mesh_terms: D001249:Asthma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D015415:Biomarkers / Q000378:metabolism*; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D015850:Interleukin-6 / Q000494:pharmacology*; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012137:Respiratory System / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D014132:Trachea / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D015850:Interleukin-6; D012333:RNA, Messenger
keywords: IL-6; IL-6 trans-signaling; airway disease; gene expression; human airway smooth muscle