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Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells

Neural cell adhesion molecule (NCAM) and fibroblast growth factor receptor 1 (FGFR1) cross-talk have been involved in epithelial-to-mesenchymal transition (EMT) process during carcinogenesis. Since EMT also contributes to maladaptive repair and parenchymal damage during renal fibrosis, we became enc...

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Autores principales: Životić, Maja, Tampe, Björn, Müller, Gerhard, Müller, Claudia, Lipkovski, Aleksandar, Xu, Xingbo, Nyamsuren, Gunsmaa, Zeisberg, Michael, Marković-Lipkovski, Jasmina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6211750/
https://www.ncbi.nlm.nih.gov/pubmed/30383875
http://dx.doi.org/10.1371/journal.pone.0206786
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author Životić, Maja
Tampe, Björn
Müller, Gerhard
Müller, Claudia
Lipkovski, Aleksandar
Xu, Xingbo
Nyamsuren, Gunsmaa
Zeisberg, Michael
Marković-Lipkovski, Jasmina
author_facet Životić, Maja
Tampe, Björn
Müller, Gerhard
Müller, Claudia
Lipkovski, Aleksandar
Xu, Xingbo
Nyamsuren, Gunsmaa
Zeisberg, Michael
Marković-Lipkovski, Jasmina
author_sort Životić, Maja
collection PubMed
description Neural cell adhesion molecule (NCAM) and fibroblast growth factor receptor 1 (FGFR1) cross-talk have been involved in epithelial-to-mesenchymal transition (EMT) process during carcinogenesis. Since EMT also contributes to maladaptive repair and parenchymal damage during renal fibrosis, we became encouraged to explore the role of NCAM/FGFR1 signaling as initiating or driving forces of EMT program in cultured human proximal tubular epithelial cells (TECs). TECs stimulated with TGF-β1 (10ng/mL) was used as an established in vitro EMT model. TGF-β1 downstream effectors were detected in vitro, as well as in 50 biopsies of different human kidney diseases to explore their in vivo correlation. NCAM/FGFR1 signaling and its modulation by FGFR1 inhibitor PD173074 (100nM) were analyzed by light microscopy, immunolabeling, qRT-PCR and scratch assays. Morphological changes associated with EMT appeared 48h after TGF-ß1 treatment and was clearly apparent after 72 hours, followed by loss of CDH1 (encoding E-Cadherin) and transcriptional induction of SNAI1 (SNAIL), SNAI2 (SLUG), TWIST1, MMP2, MMP9, CDH2 (N-Cadherin), ITGA5 (integrin-α5), ITGB1 (integrin-β1), ACTA2 (α-SMA) and S100A4 (FSP1). Moreover, at the early stage of EMT program (24 hours upon TGF-β1 exposure), transcriptional induction of several NCAM isoforms along with FGFR1 was observed, implicating a mechanistic link between NCAM/FGFR1 signaling and induction of EMT. These assumptions were further supported by the inhibition of the EMT program after specific blocking of FGFR1 signaling by PD173074. Finally, there was evidence for an in vivo TGF-β1 pathway activation in diseased human kidneys and correlation with impaired renal excretory functions. Collectively, NCAM/FGFR1 signaling appears to be involved in the initial phase of TGF-ß1 initiated EMT which can be effectively suppressed by application of FGFR inhibitor.
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spelling pubmed-62117502018-11-19 Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells Životić, Maja Tampe, Björn Müller, Gerhard Müller, Claudia Lipkovski, Aleksandar Xu, Xingbo Nyamsuren, Gunsmaa Zeisberg, Michael Marković-Lipkovski, Jasmina PLoS One Research Article Neural cell adhesion molecule (NCAM) and fibroblast growth factor receptor 1 (FGFR1) cross-talk have been involved in epithelial-to-mesenchymal transition (EMT) process during carcinogenesis. Since EMT also contributes to maladaptive repair and parenchymal damage during renal fibrosis, we became encouraged to explore the role of NCAM/FGFR1 signaling as initiating or driving forces of EMT program in cultured human proximal tubular epithelial cells (TECs). TECs stimulated with TGF-β1 (10ng/mL) was used as an established in vitro EMT model. TGF-β1 downstream effectors were detected in vitro, as well as in 50 biopsies of different human kidney diseases to explore their in vivo correlation. NCAM/FGFR1 signaling and its modulation by FGFR1 inhibitor PD173074 (100nM) were analyzed by light microscopy, immunolabeling, qRT-PCR and scratch assays. Morphological changes associated with EMT appeared 48h after TGF-ß1 treatment and was clearly apparent after 72 hours, followed by loss of CDH1 (encoding E-Cadherin) and transcriptional induction of SNAI1 (SNAIL), SNAI2 (SLUG), TWIST1, MMP2, MMP9, CDH2 (N-Cadherin), ITGA5 (integrin-α5), ITGB1 (integrin-β1), ACTA2 (α-SMA) and S100A4 (FSP1). Moreover, at the early stage of EMT program (24 hours upon TGF-β1 exposure), transcriptional induction of several NCAM isoforms along with FGFR1 was observed, implicating a mechanistic link between NCAM/FGFR1 signaling and induction of EMT. These assumptions were further supported by the inhibition of the EMT program after specific blocking of FGFR1 signaling by PD173074. Finally, there was evidence for an in vivo TGF-β1 pathway activation in diseased human kidneys and correlation with impaired renal excretory functions. Collectively, NCAM/FGFR1 signaling appears to be involved in the initial phase of TGF-ß1 initiated EMT which can be effectively suppressed by application of FGFR inhibitor. Public Library of Science 2018-11-01 /pmc/articles/PMC6211750/ /pubmed/30383875 http://dx.doi.org/10.1371/journal.pone.0206786 Text en © 2018 Životić et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Životić, Maja
Tampe, Björn
Müller, Gerhard
Müller, Claudia
Lipkovski, Aleksandar
Xu, Xingbo
Nyamsuren, Gunsmaa
Zeisberg, Michael
Marković-Lipkovski, Jasmina
Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells
title Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells
title_full Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells
title_fullStr Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells
title_full_unstemmed Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells
title_short Modulation of NCAM/FGFR1 signaling suppresses EMT program in human proximal tubular epithelial cells
title_sort modulation of ncam/fgfr1 signaling suppresses emt program in human proximal tubular epithelial cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6211750/
https://www.ncbi.nlm.nih.gov/pubmed/30383875
http://dx.doi.org/10.1371/journal.pone.0206786
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