Please use this identifier to cite or link to this item: http://hdl.handle.net/2381/31977
Title: Human lung myofibroblast TGFβ1-dependent Smad2/3 signalling is Ca(2+)-dependent and regulated by KCa3.1 K(+) channels.
Authors: Roach, Katy M.
Feghali-Bostwick, C.
Wulff, H.
Amrani, Yassine
Bradding, Peter
First Published: 26-Mar-2015
Publisher: BioMed Central
Citation: Fibrogenesis Tissue Repair, 2015, 8:5
Abstract: Background Idiopathic pulmonary fibrosis (IPF) is a common and invariably lethal interstitial lung disease with poorly effective therapy. Blockade of the K+ channel KCa3.1 reduces constitutive α-SMA and Smad2/3 nuclear translocation in IPF-derived human lung myofibroblasts (HLMFs), and inhibits several transforming growth factor beta 1 (TGFβ1)-dependent cell processes. We hypothesized that KCa3.1-dependent cell processes also regulate the TGFβ1-dependent Smad2/3 signalling pathway in HLMFs. HLMFs obtained from non-fibrotic controls (NFC) and IPF lungs were grown in vitro and examined for αSMA expression by immunofluorescence, RT-PCR, and flow cytometry. Two specific and distinct KCa3.1 blockers (TRAM-34 200 nM and ICA-17043 [Senicapoc] 100 nM) were used to determine their effects on TGFβ1-dependent signalling. Expression of phosphorylated and total Smad2/3 following TGFβ1 stimulation was determined by Western blot and Smad2/3 nuclear translocation by immunofluorescence. Results KCa3.1 block attenuated TGFβ1-dependent Smad2/3 phosphorylation and nuclear translocation, and this was mimicked by lowering the extracellular Ca2+ concentration. KCa3.1 block also inhibited Smad2/3-dependent gene transcription (αSMA, collagen type I), inhibited KCa3.1 mRNA expression, and attenuated TGFβ1-dependent αSMA protein expression. Conclusions KCa3.1 activity regulates TGFβ1-dependent effects in NFC- and IPF-derived primary HLMFs through the regulation of the TGFβ1/Smad signalling pathway, with promotion of downstream gene transcription and protein expression. KCa3.1 blockers may offer a novel approach to treating IPF. Keywords: Human lung myofibroblast; Idiopathic pulmonary fibrosis; Potassium channel KCa3.1
DOI Link: 10.1186/s13069-015-0022-0
ISSN: 1755-1536
eISSN: 1755-1536
Links: http://www.fibrogenesis.com/content/8/1/5
http://hdl.handle.net/2381/31977
Version: Publisher Version
Status: Peer-reviewed
Type: Journal Article
Rights: © 2015 Roach et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0) (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Description: PMCID: PMC4379608 PMID: 25829947 [PubMed]
Appears in Collections:Published Articles, Dept. of Infection, Immunity and Inflammation

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