title: Cytokine-induced loss of glucocorticoid function: effect of kinase inhibitors, long-acting β(2)-adrenoceptor [corrected] agonist and glucocorticoid receptor ligands.
abstract: Acting on the glucocorticoid receptor (NR3C1), glucocorticoids are widely used to treat inflammatory diseases. However, glucocorticoid resistance often leads to suboptimal asthma control. Since glucocorticoid-induced gene expression contributes to glucocorticoid activity, the aim of this study was to use a 2 × glucocorticoid response element (GRE) reporter and glucocorticoid-induced gene expression to investigate approaches to combat cytokine-induced glucocorticoid resistance. Pre-treatment with tumor necrosis factor-α (TNF) or interleukin-1β inhibited dexamethasone-induced mRNA expression of the putative anti-inflammatory genes RGS2 and TSC22D3, or just TSC22D3, in primary human airway epithelial and smooth muscle cells, respectively. Dexamethasone-induced DUSP1 mRNA was unaffected. In human bronchial epithelial BEAS-2B cells, dexamethasone-induced TSC22D3 and CDKN1C expression (at 6 h) was reduced by TNF pre-treatment, whereas DUSP1 and RGS2 mRNAs were unaffected. TNF pre-treatment also reduced dexamethasone-dependent 2×GRE reporter activation. This was partially reversed by PS-1145 and c-jun N-terminal kinase (JNK) inhibitor VIII, inhibitors of IKK2 and JNK, respectively. However, neither inhibitor affected TNF-dependent loss of dexamethasone-induced CDKN1C or TSC22D3 mRNA. Similarly, inhibitors of the extracellular signal-regulated kinase, p38, phosphoinositide 3-kinase or protein kinase C pathways failed to attenuate TNF-dependent repression of the 2×GRE reporter. Fluticasone furoate, fluticasone propionate and budesonide were full agonists relative to dexamethasone, while GSK9027, RU24858, des-ciclesonide and GW870086X were partial agonists on the 2×GRE reporter. TNF reduced reporter activity in proportion with agonist efficacy. Full and partial agonists showed various degrees of agonism on RGS2 and TSC22D3 expression, but were equally effective at inducing CDKN1C and DUSP1, and did not affect the repression of CDKN1C or TSC22D3 expression by TNF. Finally, formoterol-enhanced 2×GRE reporter activity was also proportional to agonist efficacy and functionally reversed repression by TNF. As similar effects were apparent on glucocorticoid-induced gene expression, the most effective strategy to overcome glucocorticoid resistance in this model was addition of formoterol to high efficacy NR3C1 agonists.
authors.affiliation: Airways Inflammation Research Group, Snyder Institute of Chronic Diseases, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Christopher F
authors.initials: CF
authors.lastname: Rider
authors.affiliation: Airways Inflammation Research Group, Snyder Institute of Chronic Diseases, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Suharsh
authors.initials: S
authors.lastname: Shah
authors.affiliation: AstraZeneca R&D Mölndal, Mölndal, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Miller-Larsson
authors.affiliation: Airways Inflammation Research Group, Snyder Institute of Chronic Diseases, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Giembycz
authors.affiliation: Airways Inflammation Research Group, Snyder Institute of Chronic Diseases, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Robert
authors.initials: R
authors.lastname: Newton
mesh_terms: D058666:Adrenergic beta-2 Receptor Agonists / Q000494:pharmacology*; D001249:Asthma / Q000188:drug therapy; D001980:Bronchi / Q000166:cytology; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology*; D004351:Drug Resistance; D004847:Epithelial Cells / Q000378:metabolism; D000068759:Formoterol Fumarate / Q000494:pharmacology*; D015870:Gene Expression; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D053583:Interleukin-1beta / Q000502:physiology*; D008024:Ligands; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*
chemical_list: D058666:Adrenergic beta-2 Receptor Agonists; D003907:Dexamethasone; D000068759:Formoterol Fumarate; D005938:Glucocorticoids; D051550:I-kappa B Kinase; D053583:Interleukin-1beta; D008024:Ligands; D019869:Phosphatidylinositol 3-Kinases; D047428:Protein Kinase Inhibitors; D011965:Receptors, Glucocorticoid; D014409:Tumor Necrosis Factor-alpha; C439524:TNF protein, human; C496560:IKBKB protein, human