Pharmacological inhibition of β-catenin/CBP interaction with the small molecule ICG-001 inhibits proliferation and extracellular matrix production in airway smooth muscle

Rationale Airway hyperresponsiveness is a principle feature of asthma, explained in part by remodeling of airway smooth muscle (ASM), including muscle thickening and increased extracellular matrix (ECM) protein production by the ASM. Current therapies are largely successful in targeting the inflammatory component of asthma, but lack the necessary means to reverse or prevent the remodeling process. β-catenin is a potential therapeutic target in this regard, as we have shown previously its critical role in the regulation of ASM proliferation and ECM production. Accordingly we have tested several small-molecule signaling modulators that attenuate the nuclear functions of β-catenin. Methods Three small-molecule compounds were employed in this study that each selectively target different components of β-catenin signaling: ICG-001 (inhibits β-catenin/CBP interaction), IQ-1 (prevents β-catenin/p300 interaction) and XAV-939 (a WNT-specific β-catenin antagonist that inhibits Tankyrase 1 and 2 to stabilize Axin). Immortalized human bronchial smooth muscle cell lines were treated with Fetal Bovine Serum (FBS) or Transforming Growth Factor-β1 (TGF-β1) in combination with increasing concentrations of ICG-001, IQ-1 or XAV-939. We assessed cell proliferation using an Alamar Blue conversion assay and Western blotting. ECM production was determined via RT-qPCR and Western blotting. Results ICG-001 dose dependently (0,1-10 μM) inhibited FBS-induced airway smooth muscle proliferation (58% at 3 μM,

Caveolae and Caveolins in the respiratory system

10.2174/156652408786733720Current Molecular Medicine88741-753CMMU

Immunomodulatory innate defence regulator (IDR) peptide alleviates airway inflammation and hyper-responsiveness

Pathogenesis and treatment of chronic pulmonary disease

Combination of IL-17A/F and TNF-alpha uniquely alters the bronchial epithelial cell proteome to enhance proteins that augment neutrophil migration

Background: The heterodimer interleukin (IL)-17A/F is elevated in the lungs in chronic respiratory disease such as severe asthma, along with the pro-inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). Although IL-17A/F and TNF-alpha are known to functionally cooperate to exacerbate airway inflammation, proteins altered by their interaction in the lungs are not fully elucidated.Results: We used Slow Off-rate Modified Aptamer-based proteomic array to identify proteins that are uniquely and/or synergistically enhanced by concurrent stimulation with IL-17A/F and TNF-alpha in human bronchial epithelial cells (HBEC). The abundance of 38 proteins was significantly enhanced by the combination of IL-17A/F and TNF-alpha, compared to either cytokine alone. Four out of seven proteins that were increased > 2-fold were those that promote neutrophil migration; host defence peptides (HDP; Lipocalin-2 (LCN-2) and Elafin) and chemokines (IL-8, GRO alpha). We independently confirmed the synergistic increase of these four proteins by western blots and ELISA. We also functionally confirmed that factors secreted by HBEC stimulated with the combination of IL-17A/F and TNF-alpha uniquely enhances neutrophil migration. We further showed that PI3K and PKC pathways selectively control IL-17A/F + TNF-alpha-mediated synergistic production of HDPs LCN-2 and Elafin, but not chemokines IL-8 and GRO alpha. Using a murine model of airway inflammation, we demonstrated enhancement of IL-17A/F, TNF-alpha, LCN-2 and neutrophil chemokine KC in the lungs, thus corroborating our findings in-vivo.Conclusion: This study identifies proteins and signaling mediated by concurrent IL-17A/F and TNF-alpha exposure in the lungs, relevant to respiratory diseases characterized by chronic inflammation, especially neutrophilic airway inflammation such as severe asthma