title: Repression of inflammatory gene expression in human pulmonary epithelial cells by small-molecule IkappaB kinase inhibitors.
abstract: The airway epithelium is critical in the pathogenesis of chronic inflammatory diseases, such as asthma and chronic obstructive pulmonary disease, and, by expressing numerous inflammatory genes, plays a prominent role in disease exacerbations. Since inflammatory gene expression often involves the transcription factor nuclear factor (NF)-kappaB, this signaling pathway represents a site for anti-inflammatory intervention. As the airway epithelium is targeted by inhaled therapeutic agents, for example corticosteroids, human A549 pulmonary cells and primary human bronchial epithelial (HBE) cells were selected to evaluate inhibitor of kappaB kinase (IKK) inhibitors. In A549 cells, interleukin (IL)-1beta and tumor necrosis factor (TNF) alpha increased phosphorylation of IkappaBalpha, and this was followed by loss of IkappaBalpha, induction of NF-kappaB DNA binding, and the induction of NF-kappaB-dependent transcription. These events were repressed by the IKK-selective inhibitors, PS-1145 [N-(6-chloro-9H-beta-carbolin-8-ly) nicotinamide] and ML120B [N-(6-chloro-7-methoxy-9H-beta-carbolin-8-yl)-2-methyl-nicotinamide]. Inhibition of NF-kappaB-dependent transcription was concentration-dependent and correlated with loss of intercellular adhesion molecule (ICAM)-1 expression. Similarly, IL-1beta- and TNFalpha-induced expression of IL-6, IL-8, granulocyte macrophage-colony-stimulating factor (GM-CSF), regulated and activation normal T cell expressed and secreted (RANTES), growth-related oncogene alpha, and monocyte chemotactic protein-1 (MCP-1) was also significantly repressed. Likewise, PS-1145 and ML120B profoundly reduced NF-kappaB-dependent transcription induced by IL-1beta and TNFalpha in primary HBE cells. Parallel effects on ICAM-1 expression and a significant repression of IL-8 release were observed. In contrast, the corticosteroid, dexamethasone, was without effect on NF-kappaB-dependent transcription or the expression of ICAM-1. The above data provide strong support for an anti-inflammatory effect of IKK2 inhibitors acting on the pulmonary epithelium and suggest that such compounds may prove beneficial in situations where traditional corticosteroid therapies prove inadequate.
authors.affiliation: Department of Cell Biology and Anatomy, Respiratory Research Group, Faculty of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, Alberta T2N 4N1, Canada. rnewton@ucalgary.ca
authors.forename: Robert
authors.initials: R
authors.lastname: Newton
authors.forename: Neil S
authors.initials: NS
authors.lastname: Holden
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Catley
authors.forename: Wale
authors.initials: W
authors.lastname: Oyelusi
authors.forename: Richard
authors.initials: R
authors.lastname: Leigh
authors.forename: David
authors.initials: D
authors.lastname: Proud
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Barnes
mesh_terms: D002243:Carbolines / Q000494:pharmacology; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D006801:Humans; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors* / Q000502:physiology; D053583:Interleukin-1beta / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D016209:Interleukin-8 / Q000235:genetics; D008168:Lung / Q000378:metabolism*; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009536:Niacinamide / Q000031:analogs & derivatives / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D029424:Pulmonary Disease, Chronic Obstructive / Q000188:drug therapy; D011725:Pyridines / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D002243:Carbolines; D016207:Cytokines; D006575:Heterocyclic Compounds, 3-Ring; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; C514957:N-(6-chloro-7-methoxy-9H-beta-carbolin-8-yl)-2-methylnicotinamide; D016328:NF-kappa B; C456319:PS1145; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D014409:Tumor Necrosis Factor-alpha; D009536:Niacinamide; C496559:CHUK protein, human; D051550:I-kappa B Kinase