TY - JOUR T1 - RNA-seq analysis of transforming growth factor-β-induced glucocorticoid resistance in human bronchial epithelial cells JF - European Respiratory Journal JO - Eur Respir J VL - 42 IS - Suppl 57 SP - P680 AU - Christine Keenan AU - Guillermo Lopez-Campos AU - Saad Salem AU - Trudi Harris AU - Michael Schuliga AU - Cameron Johnstone AU - Alastair Stewart Y1 - 2013/09/01 UR - http://erj.ersjournals.com/content/42/Suppl_57/P680.abstract N2 - Introduction: Glucocorticoid (GC) resistance limits the successful treatment of chronic inflammatory diseases. We have identified Transforming Growth Factor-β (TGF-β) as a novel inducer of GC insensitivity in the epithelial cell lines A549 and BEAS-2B. This resistance is not prevented by inhibiting known non-canonical TGF-β signalling pathways, but may be partially due to decreased GRα nuclear localisation in A549 cells, but not in BEAS-2B cells.Aim: To use RNA-seq to facilitate efforts to reveal the mechanism of TGF-β-induced GC resistance.Methods: BEAS-2B cells pre-treated for 24h with 40pM TGF-β were treated with 30nM dexamethasone (Dex) for 4h then total RNA was extracted. RNA-seq was performed using an Illumina HiSeq™ 2000 sequencer. Changes from control of more than 2.5 fold were analysed as significant changes and a subset of the observed expression changes were confirmed by RT-qPCR.Results: RNA-seq analysis detected 108 genes with expression up-regulated by Dex. Six of these that were up-regulated by TGF-β alone were removed to prevent confounding analyses. Sixty-six genes were only up-regulated by Dex in the absence of TGF-β, and 36 genes were still up-regulated by Dex in the presence of TGF-β.Conclusions: TGF-β impairs a subset of GC gene regulatory effects in BEAS-2B cells. RNA-seq analysis identified 2 sets of genes up-regulated by GCs, one of which remains inducible and the other which is rendered insensitive to GC activation in the presence of TGF-β. Understanding the differences in regulation of these two groups of GC-sensitive genes may lead to therapeutic strategies to reactivate their expression in chronic inflammatory and fibrotic disease. ER -