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Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2
Transforming growth factor β (TGFβ) induces hepatic stellate cell (HSC) differentiation into tumor‐promoting myofibroblast, although underlying mechanism remains incompletely understood. Focal adhesion kinase (FAK) is activated in response to TGFβ stimulation, so it transmits TGFβ stimulus to extrac...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6996408/ https://www.ncbi.nlm.nih.gov/pubmed/32025610 http://dx.doi.org/10.1002/hep4.1452 |
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author | Chen, Yunru Li, Qing Tu, Kangsheng Wang, Yuanguo Wang, Xianghu Liu, Dandan Chen, Chen Liu, Donglian Yang, Rendong Qiu, Wei Kang, Ningling |
author_facet | Chen, Yunru Li, Qing Tu, Kangsheng Wang, Yuanguo Wang, Xianghu Liu, Dandan Chen, Chen Liu, Donglian Yang, Rendong Qiu, Wei Kang, Ningling |
author_sort | Chen, Yunru |
collection | PubMed |
description | Transforming growth factor β (TGFβ) induces hepatic stellate cell (HSC) differentiation into tumor‐promoting myofibroblast, although underlying mechanism remains incompletely understood. Focal adhesion kinase (FAK) is activated in response to TGFβ stimulation, so it transmits TGFβ stimulus to extracellular signal‐regulated kinase and P38 mitogen‐activated protein kinase signaling. However, it is unknown whether FAK can, in return, modulate TGFβ receptors. In this study, we tested whether FAK phosphorylated TGFβ receptor 2 (TGFβR2) and regulated TGFβR2 intracellular trafficking in HSCs. The FAKY397F mutant and PF‐573,228 were used to inhibit the kinase activity of FAK, the TGFβR2 protein level was quantitated by immunoblotting, and HSC differentiation into myofibroblast was assessed by expression of HSC activation markers, alpha‐smooth muscle actin, fibronectin, or connective tissue growth factor. We found that targeting FAK kinase activity suppressed the TGFβR2 protein level, TGFβ1‐induced mothers against decapentaplegic homolog phosphorylation, and myofibroblastic activation of HSCs. At the molecular and cellular level, active FAK (phosphorylated FAK at tyrosine 397) bound to TGFβR2 and kept TGFβR2 at the peripheral plasma membrane of HSCs, and it induced TGFβR2 phosphorylation at tyrosine 336. In contrast, targeting FAK or mutating Y336 to F on TGFβR2 led to lysosomal sorting and degradation of TGFβR2. Using RNA sequencing, we identified that the transcripts of 764 TGFβ target genes were influenced by FAK inhibition, and that through FAK, TGFβ1 stimulated HSCs to produce a panel of tumor‐promoting factors, including extracellular matrix remodeling proteins, growth factors and cytokines, and immune checkpoint molecule PD‐L1. Functionally, targeting FAK inhibited tumor‐promoting effects of HSCs in vitro and in a tumor implantation mouse model. Conclusion: FAK targets TGFβR2 to the plasma membrane and protects TGFβR2 from lysosome‐mediated degradation, thereby promoting TGFβ‐mediated HSC activation. FAK is a target for suppressing HSC activation and the hepatic tumor microenvironment. |
format | Online Article Text |
id | pubmed-6996408 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-69964082020-02-05 Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 Chen, Yunru Li, Qing Tu, Kangsheng Wang, Yuanguo Wang, Xianghu Liu, Dandan Chen, Chen Liu, Donglian Yang, Rendong Qiu, Wei Kang, Ningling Hepatol Commun Original Articles Transforming growth factor β (TGFβ) induces hepatic stellate cell (HSC) differentiation into tumor‐promoting myofibroblast, although underlying mechanism remains incompletely understood. Focal adhesion kinase (FAK) is activated in response to TGFβ stimulation, so it transmits TGFβ stimulus to extracellular signal‐regulated kinase and P38 mitogen‐activated protein kinase signaling. However, it is unknown whether FAK can, in return, modulate TGFβ receptors. In this study, we tested whether FAK phosphorylated TGFβ receptor 2 (TGFβR2) and regulated TGFβR2 intracellular trafficking in HSCs. The FAKY397F mutant and PF‐573,228 were used to inhibit the kinase activity of FAK, the TGFβR2 protein level was quantitated by immunoblotting, and HSC differentiation into myofibroblast was assessed by expression of HSC activation markers, alpha‐smooth muscle actin, fibronectin, or connective tissue growth factor. We found that targeting FAK kinase activity suppressed the TGFβR2 protein level, TGFβ1‐induced mothers against decapentaplegic homolog phosphorylation, and myofibroblastic activation of HSCs. At the molecular and cellular level, active FAK (phosphorylated FAK at tyrosine 397) bound to TGFβR2 and kept TGFβR2 at the peripheral plasma membrane of HSCs, and it induced TGFβR2 phosphorylation at tyrosine 336. In contrast, targeting FAK or mutating Y336 to F on TGFβR2 led to lysosomal sorting and degradation of TGFβR2. Using RNA sequencing, we identified that the transcripts of 764 TGFβ target genes were influenced by FAK inhibition, and that through FAK, TGFβ1 stimulated HSCs to produce a panel of tumor‐promoting factors, including extracellular matrix remodeling proteins, growth factors and cytokines, and immune checkpoint molecule PD‐L1. Functionally, targeting FAK inhibited tumor‐promoting effects of HSCs in vitro and in a tumor implantation mouse model. Conclusion: FAK targets TGFβR2 to the plasma membrane and protects TGFβR2 from lysosome‐mediated degradation, thereby promoting TGFβ‐mediated HSC activation. FAK is a target for suppressing HSC activation and the hepatic tumor microenvironment. John Wiley and Sons Inc. 2019-12-20 /pmc/articles/PMC6996408/ /pubmed/32025610 http://dx.doi.org/10.1002/hep4.1452 Text en © 2019 The Authors. Hepatology Communications published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Original Articles Chen, Yunru Li, Qing Tu, Kangsheng Wang, Yuanguo Wang, Xianghu Liu, Dandan Chen, Chen Liu, Donglian Yang, Rendong Qiu, Wei Kang, Ningling Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 |
title | Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 |
title_full | Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 |
title_fullStr | Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 |
title_full_unstemmed | Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 |
title_short | Focal Adhesion Kinase Promotes Hepatic Stellate Cell Activation by Regulating Plasma Membrane Localization of TGFβ Receptor 2 |
title_sort | focal adhesion kinase promotes hepatic stellate cell activation by regulating plasma membrane localization of tgfβ receptor 2 |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6996408/ https://www.ncbi.nlm.nih.gov/pubmed/32025610 http://dx.doi.org/10.1002/hep4.1452 |
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