J 2020

3D Cell Culture Models Demonstrate a Role for FGF and WNT Signaling in Regulation of Lung Epithelial Cell Fate and Morphogenesis

RABATA, Anas, Radek FEDR, Karel SOUČEK, Aleš HAMPL, Zuzana KOLEDOVÁ et. al.

Basic information

Original name

3D Cell Culture Models Demonstrate a Role for FGF and WNT Signaling in Regulation of Lung Epithelial Cell Fate and Morphogenesis

Authors

RABATA, Anas (760 Syrian Arab Republic, belonging to the institution), Radek FEDR (203 Czech Republic), Karel SOUČEK (203 Czech Republic), Aleš HAMPL (203 Czech Republic, belonging to the institution) and Zuzana KOLEDOVÁ (703 Slovakia, guarantor, belonging to the institution)

Edition

Frontiers in Cell and Developmental Biology, Lausanne, Frontiers Media S.A. 2020, 2296-634X

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10601 Cell biology

Country of publisher

Switzerland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 6.684

RIV identification code

RIV/00216224:14110/20:00116303

Organization unit

Faculty of Medicine

UT WoS

000558854700001

Keywords in English

3D cell culture; epithelial cell; FGF signaling; lung; morphogenesis; organoid; WNT signaling

Tags

International impact, Reviewed
Změněno: 7/2/2022 12:54, Mgr. Tereza Miškechová

Abstract

V originále

FGF signaling plays an essential role in lung development, homeostasis, and regeneration. We employed mouse 3D cell culture models and imaging to studyex vivothe role of FGF ligands and the interplay of FGF signaling with epithelial growth factor (EGF) and WNT signaling pathways in lung epithelial morphogenesis and differentiation. In non-adherent conditions, FGF signaling promoted formation of lungospheres from lung epithelial stem/progenitor cells (LSPCs). Ultrastructural and immunohistochemical analyses showed that LSPCs produced more differentiated lung cell progeny. In a 3D extracellular matrix, FGF2, FGF7, FGF9, and FGF10 promoted lung organoid formation. FGF9 showed reduced capacity to promote lung organoid formation, suggesting that FGF9 has a reduced ability to sustain LSPC survival and/or initial divisions. FGF7 and FGF10 produced bigger organoids and induced organoid branching with higher frequency than FGF2 or FGF9. Higher FGF concentration and/or the use of FGF2 with increased stability and affinity to FGF receptors both increased lung organoid and lungosphere formation efficiency, respectively, suggesting that the level of FGF signaling is a crucial driver of LSPC survival and differentiation, and also lung epithelial morphogenesis. EGF signaling played a supportive but non-essential role in FGF-induced lung organoid formation. Analysis of tissue architecture and cell type composition confirmed that the lung organoids contained alveolar-like regions with cells expressing alveolar type I and type II cell markers, as well as airway-like structures with club cells and ciliated cells. FGF ligands showed differences in promoting distinct lung epithelial cell types. FGF9 was a potent inducer of more proximal cell types, including ciliated and basal cells. FGF7 and FGF10 directed the differentiation toward distal lung lineages. WNT signaling enhanced the efficiency of lung organoid formation, but in the absence of FGF10 signaling, the organoids displayed limited branching and less differentiated phenotype. In summary, we present lung 3D cell culture models as useful tools to study the role and interplay of signaling pathways in postnatal lung development and homeostasis, and we reveal distinct roles for FGF ligands in regulation of mouse lung morphogenesis and differentiationex vivo.

Links

LM2015062, research and development project
Name: Národní infrastruktura pro biologické a medicínské zobrazování
Investor: Ministry of Education, Youth and Sports of the CR
MUNI/A/1382/2019, interní kód MU
Name: Zdroje pro tkáňové inženýrství 10 (Acronym: TissueEng 10)
Investor: Masaryk University, Category A
ROZV/28/LF19/2020, interní kód MU
Name: Regulace morfogeneze epitelu mléčné žlázy pomocí mechanických sil a dynamiky signalizace
Investor: Ministry of Education, Youth and Sports of the CR, Internal development projects