Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes

Abstract

BackgroundAirway fibrosis is one of the pathological features of chronic obstructive pulmonary disease (COPD), and recent studies revealed that acetylcholine plays an important role in the development of airway remodeling by stimulating proliferation and collagen synthesis of lung fibroblasts. This study was designed to examine the effects of a long-acting muscarinic receptor antagonist (LAMA) glycopyrronium and a long-acting β2 adrenergic receptor agonist (LABA) indacaterol on acetylcholine-mediated fibrotic responses in lung fibroblasts.MethodsAfter carbachol (CCh) or transforming growth factor-β1 (TGF-β1) exposure, the response to glycopyrronium and indacaterol was determined in vitro in fibroblasts isolated from mild-to-moderate COPD lung tissue. The ability of fibroblasts to mediate the contraction of collagen gels was assessed. The expression of α-smooth muscle actin (α-SMA) and the phosphorylation of extracellular-signal-regulated kinase 5 (ERK5) were determined by immunoblot. TGF-β1 was quantified by ELISA and acetylcholine was quantified by liquid chromatography tandem-mass spectrometry.ResultsCCh stimulated fibroblast-mediated collagen gel contraction and α-SMA expression and TGF-β1 release by fibroblasts. Blockade of autocrine TGF-β1 attenuated CCh-mediated fibrotic responses, while TGF-β1 did not stimulate acetylcholine release. Glycopyrronium plus indacaterol significantly attenuated CCh- and TGF-β1-mediated fibrotic responses through inhibition of ERK5 phosphorylation. Notably, the magnitudes of CCh- and TGF-β1-stimulated gel contraction, CCh-induced TGF-β1 release, and ERK5 phosphorylation were greater in fibroblasts isolated from COPD subjects than in those from non-smokers.ConclusionsCCh induced TGF-β1 self-sustaining signaling loops by potentiating ERK5 signaling and promoted myofibroblast activity. This autocrine signaling mechanism may be an attractive therapeutic target to block the fibrotic response, which was modulated by the combination of glycopyrronium and indacaterol.

Specific Features of Fibrotic Lung Fibroblasts Highly Sensitive to Fibrotic Processes Mediated via TGF-β–ERK5 Interaction

“…In conclusion, the results presented here provide evidence of the high sensitivity of fibrotic fibroblasts to the canonical TGF-β1/Smad3 signaling pathway, which mediates fibrotic processes that can be suppressed by ERK5 inhibitor treatment; this extends our previous findings that ERK5 inhibitor suppressed airway fibrosis in COPD lung fibroblasts [12]. The screened genes and TGF-β1-ERK5 interaction analysis provide detailed insights into the molecular mechanism underlying the progression of lung fibrosis mediated by fibrotic fibroblasts.…”

“…BIX02189, a pharmacological inhibitor of MAPK kinase 5/ERK5, blocked TGF-β signaling and subsequent profibrogenic responses in a bleomycin-induced lung fibrosis model [11]. We previously demonstrated that BIX02189 attenuates acetylcholine and TGF-β1-mediated ERK5 signaling, reducing the contractility of lung fibroblasts present in clinical samples from patients with chronic obstructive pulmonary disease (COPD), which suggests prevention of airway remodeling [12].…”

Pirfenidone attenuates lung fibrotic fibroblast responses to transforming growth factor-β1

“…The two wells of the Boyden blind well chamber were separated by an 8-μM pore filter (Nucleopore, Pleasanton, CA, USA). The chambers were incubated at 37 °C in a humid atmosphere containing 5% CO 2 for 8 h, after which the cells on top of the filter were removed by scraping [8, 11, 23]. The cells that had migrated through the filter were then fixed, stained with DiffQuick Sysmex (16920), and mounted on glass microscope slides.…”

The concept of chronic obstructive pulmonary disease clinical control as a decision - making tool in real clinical practice for optimizing of basic pharmacotherapy

“…Показано, что терапия с использованием фиксированной комбинации индакатерол (ИНД)/гликопирроний (ГЛИ) может предотвратить «клинически значимое ухудшение» ХОБЛ на 28% по сравнению с тиотропием и на 33% по сравнению с ИГКС/ДДБА («клинически значимое ухудшение ХОБЛ» определяется временем/риском развития одного из изменений: снижение объема форсированного выдоха за 1-ю секунду (ОФВ 1 ) ≥100 мл, ухудшение показателя SGRQ ≥4 баллов, CAT ≥2 баллов, TDI ≥1 балла, возникновение умеренных /тяжелых обострений) [13]. Подтверждена способность ИНД/ГЛИ in vitro предупреждать развитие ремоделирования бронхов и перибронхиального фиброза на ранних стадиях ХОБЛ [14]. Таким образом, ДДБА/ДДАХ -наиболее эффективная медикаментозная терапия для достижения и поддержания контроля как симптомов, так и риска обострений у большинства пациентов с ХОБЛ [при отсутствии сопутствующей БА и выраженной эозинофилии (ЭОЗ) в крови].…”