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Am J Physiol Lung Cell Mol Physiol. 2019 Apr 10. doi: 10.1152/ajplung.00400.2018. [Epub ahead of print]

TGF-β activation impairs fibroblast ability to support adult lung epithelial progenitor cell organoid formation.

Author information

1
Division of Cell Biology, MRC Laboratory of Molecular Biology, United Kingdom.
2
ERIBA, University of Groningen, UMC Groningen.
3
Department of Molecular Pharmacology, University of Groningen.
4
Molecular Pharmacology, University of Groningen, Netherlands.
5
Center of Thoracic Surgery, Asklepios Fachkliniken München-Gauting.
6
ERIBA, University of Groningen, UMC Groningen, Netherlands.
7
Pulmonology, Leiden University Medical Center, Netherlands.
8
Pulmonology, LUMC, Netherlands.
9
University of Colorado Denver.

Abstract

Transforming growth factor-β (TGF-β)-induced fibroblast-to-myofibroblast differentiation contributes to remodeling in chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis, but whether this impacts the ability of fibroblasts to support lung epithelial repair remains little explored. We pre-treated human lung fibroblasts (primary [phFB] or MRC5 cells) with recombinant human TGF-β to induce myofibroblast differentiation, then co-cultured them with adult mouse lung EpCAM+ cells to investigate their capacity to support epithelial organoid formation in vitro. While control phFB and MRC5 lung fibroblasts supported organoid formation of mouse EpCAM+ cells, TGF-β-pre-treatment of both phFB and MRC5 impaired organoid-supporting ability. We performed RNA sequencing of TGF-β treated phFB, which revealed altered expression of key Wnt signaling pathway components and Wnt/β-catenin target genes, and modulated expression of secreted factors involved in mesenchymal-epithelial signaling. TGF-β profoundly skewed the transcriptional program induced by the Wnt/β-catenin activator CHIR99021 (CHIR). Supplementing organoid culture media recombinant hepatocyte growth factor (HGF) or fibroblast growth factor 7 (FGF7) promoted organoid formation when using TGF-β pre-treated fibroblasts. In conclusion, TGF-β-induced myofibroblast differentiation results in Wnt/β-catenin pathway skewing, and impairs fibroblast ability to support epithelial repair likely through multiple mechanisms including modulation of secreted growth factors.

KEYWORDS:

Lung regeneration/repair; Lung stem cells; Mesenchymal-epithelial signaling; TGF-β signaling; Wnt/β-catenin signaling

PMID:
30969812
DOI:
10.1152/ajplung.00400.2018

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