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Mesenchymal Stromal Cells Accelerate Epithelial Tight Junction Assembly via the AMP-Activated Protein Kinase Pathway, Independently of Liver Kinase B1

Authors :
Jean-Marie Krzesinski
Pauline Erpicum
François Jouret
Michael Sebbagh
Pascal Rowart
Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA-Research)
Université de Liège
Centre de Recherche en Cancérologie de Marseille (CRCM)
Aix Marseille Université (AMU)-Institut Paoli-Calmettes
Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
MITOYAN, Louciné
Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Paoli-Calmettes
Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Aix Marseille Université (AMU)
Source :
Stem Cells International, Stem Cells International, 2017, Stem Cells International, Hindawi Publishing Corporation, 2017, Stem Cells International, Vol 2017 (2017)
Publication Year :
2017
Publisher :
HAL CCSD, 2017.

Abstract

Background. Mesenchymal stromal cells (MSC) are fibroblast-like multipotent cells capable of tissue-repair properties. Given the essentiality of tight junctions (TJ) in epithelial integrity, we hypothesized that MSC modulate TJ formation, via the AMP-activated kinase (AMPK) pathway. Liver kinase-β1 (LKB1) and Ca2+-calmodulin-dependent protein kinase kinase (CaMKK) represent the main kinases that activate AMPK.Methods. The in vitro Ca2+switch from 5 μM to 1.8 mM was performed using epithelial Madin-Darby canine kidney (MDCK) cells cultured alone or cocultured with rat bone marrow-derived MSC or preexposed to MSC-conditioned medium. TJ assembly was measured by assessing ZO-1 relocation to cell-cell contacts. Experiments were conducted using MDCK stably expressing short-hairpin-RNA (shRNA) against LKB1 or luciferase (LUC, as controls). Compound STO-609 (50 μM) was used as CaMKK inhibitor.Results. Following Ca2+switch, ZO-1 relocation and phosphorylation/activation of AMPK were significantly higher in MDCK/MSC compared to MDCK. No difference in AMPK phosphorylation was observed between LKB1-shRNA and Luc-shRNA MDCK following Ca2+switch. Conversely, incubation with STO-609 prior to Ca2+switch prevented AMPK phosphorylation and ZO-1 relocation. MSC-conditioned medium slightly but significantly increased AMPK activation and accelerated TJ-associated distribution of ZO-1 post Ca2+switch in comparison to regular medium.Conclusions. MSC modulate the assembly of epithelial TJ, via the CaMKK/AMPK pathway independently of LKB1.

Details

Language :
English
ISSN :
1687966X and 16879678
Database :
OpenAIRE
Journal :
Stem Cells International, Stem Cells International, 2017, Stem Cells International, Hindawi Publishing Corporation, 2017, Stem Cells International, Vol 2017 (2017)
Accession number :
edsair.doi.dedup.....2a0f3aa449f57d4fc6c2d01a9ed4a472