Tiotropium attenuates IL-13-induced goblet cell metaplasia of human airway epithelial cells

It has been shown that acetylcholine is both a neurotransmitter and acts as a local mediator, produced by airway cells including epithelial cells. In vivo studies have demonstrated an indirect role for acetylcholine in epithelial cell differentiation. Here, we aimed to investigate direct effects of...

Full description

Saved in:

Bibliographic Details
Published in:	Thorax Vol. 70; no. 7; p. 668
Main Authors:	Kistemaker, Loes E M, Hiemstra, Pieter S, Bos, I Sophie T, Bouwman, Susanne, van den Berge, Maarten, Hylkema, Machteld N, Meurs, Herman, Kerstjens, Huib A M, Gosens, Reinoud
Format:	Journal Article
Language:	English
Published:	England 01-07-2015
Subjects:	Acetylcholine - physiology Cell Differentiation - drug effects Cell Differentiation - physiology Cells, Cultured Cholinergic Antagonists - pharmacology Dose-Response Relationship, Drug Epithelial Cells - drug effects Epithelial Cells - pathology Gene Expression Regulation - drug effects Goblet Cells - drug effects Goblet Cells - pathology Humans Interleukin-13 - administration & dosage Interleukin-13 - antagonists & inhibitors Interleukin-13 - pharmacology Metaplasia - chemically induced Metaplasia - genetics Metaplasia - pathology Mucin 5AC - biosynthesis Mucin 5AC - genetics Respiratory Mucosa - drug effects Respiratory Mucosa - pathology Scopolamine Derivatives - pharmacology Tiotropium Bromide Transcription Factors - metabolism Asthma Pharmacology Airway Epithelium
Online Access:	Get more information
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	It has been shown that acetylcholine is both a neurotransmitter and acts as a local mediator, produced by airway cells including epithelial cells. In vivo studies have demonstrated an indirect role for acetylcholine in epithelial cell differentiation. Here, we aimed to investigate direct effects of endogenous non-neuronal acetylcholine on epithelial cell differentiation. Human airway epithelial cells from healthy donors were cultured at an air-liquid interface (ALI). Cells were exposed to the muscarinic antagonist tiotropium (10 nM), interleukin (IL)-13 (1, 2 and 5 ng/mL), or a combination of IL-13 and tiotropium, during or after differentiation at the ALI. Human airway epithelial cells expressed all components of the non-neuronal cholinergic system, suggesting acetylcholine production. Tiotropium had no effects on epithelial cell differentiation after air exposure. Differentiation into goblet cells was barely induced after air exposure. Therefore, IL-13 (1 ng/mL) was used to induce goblet cell metaplasia. IL-13 induced MUC5AC-positive cells (5-fold) and goblet cells (14-fold), as assessed by histochemistry, and MUC5AC gene expression (105-fold). These effects were partly prevented by tiotropium (47-92%). Goblet cell metaplasia was induced by IL-13 in a dose-dependent manner, which was inhibited by tiotropium. In addition, tiotropium reversed goblet cell metaplasia induced by 2 weeks of IL-13 exposure. IL-13 decreased forkhead box protein A2 (FoxA2) expression (1.6-fold) and increased FoxA3 (3.6-fold) and SAM-pointed domain-containing ETS transcription factor (SPDEF) (5.2-fold) expression. Tiotropium prevented the effects on FoxA2 and FoxA3, but not on SPDEF. We demonstrate that tiotropium has no effects on epithelial cell differentiation after air exposure, but inhibits and reverses IL-13-induced goblet cell metaplasia, possibly via FoxA2 and FoxA3. This indicates that non-neuronal acetylcholine contributes to goblet cell differentiation by a direct effect on epithelial cells.
ISSN:	1468-3296
DOI:	10.1136/thoraxjnl-2014-205731