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CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions

Vascular endothelial (VE) cadherin is a key component of endothelial adherens junctions (AJs) and plays an important role in maintaining vascular integrity. Endocytosis of VE-cadherin regulates junctional strength and a decrease of surface VE-cadherin reduces vascular stability. However, disruption...

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Autores principales: Chrifi, Ihsan, Louzao-Martinez, Laura, Brandt, Maarten M., van Dijk, Christian G. M., Bürgisser, Petra E., Zhu, Changbin, Kros, Johan M., Verhaar, Marianne C., Duncker, Dirk J., Cheng, Caroline
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6510885/
https://www.ncbi.nlm.nih.gov/pubmed/30097810
http://dx.doi.org/10.1007/s10456-018-9638-1
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author Chrifi, Ihsan
Louzao-Martinez, Laura
Brandt, Maarten M.
van Dijk, Christian G. M.
Bürgisser, Petra E.
Zhu, Changbin
Kros, Johan M.
Verhaar, Marianne C.
Duncker, Dirk J.
Cheng, Caroline
author_facet Chrifi, Ihsan
Louzao-Martinez, Laura
Brandt, Maarten M.
van Dijk, Christian G. M.
Bürgisser, Petra E.
Zhu, Changbin
Kros, Johan M.
Verhaar, Marianne C.
Duncker, Dirk J.
Cheng, Caroline
author_sort Chrifi, Ihsan
collection PubMed
description Vascular endothelial (VE) cadherin is a key component of endothelial adherens junctions (AJs) and plays an important role in maintaining vascular integrity. Endocytosis of VE-cadherin regulates junctional strength and a decrease of surface VE-cadherin reduces vascular stability. However, disruption of AJs is also a requirement for vascular sprouting. Identifying novel regulators of endothelial endocytosis could enhance our understanding of angiogenesis. Here, we evaluated the angiogenic potential of (CKLF-like MARVEL transmembrane domain 4) CMTM4 and assessed in which molecular pathway CMTM4 is involved during angiogenesis. Using a 3D vascular assay composed of GFP-labeled HUVECs and dsRED-labeled pericytes, we demonstrated in vitro that siRNA-mediated CMTM4 silencing impairs vascular sprouting. In vivo, CMTM4 silencing by morpholino injection in zebrafish larvae inhibits intersomitic vessel growth. Intracellular staining revealed that CMTM4 colocalizes with Rab4(+) and Rab7(+) vesicles, both markers of the endocytic trafficking pathway. CMTM4 colocalizes with both membrane-bound and internalized VE-cadherin. Adenovirus-mediated CMTM4 overexpression enhances the endothelial endocytic pathway, in particular the rapid recycling pathway, shown by an increase in early endosomal antigen-1 positive (EEA1(+)), Rab4(+), Rab11(+) , and Rab7(+) vesicles. CMTM4 overexpression enhances membrane-bound VE-cadherin internalization, whereas CMTM4 knockdown decreases internalization of VE-cadherin. CMTM4 overexpression promotes endothelial barrier function, shown by an increase in recovery of transendothelial electrical resistance (TEER) after thrombin stimulation. We have identified in this study a novel regulatory function for CMTM4 in angiogenesis. CMTM4 plays an important role in the turnover of membrane-bound VE-cadherin at AJs, mediating endothelial barrier function and controlling vascular sprouting. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10456-018-9638-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-65108852019-05-28 CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions Chrifi, Ihsan Louzao-Martinez, Laura Brandt, Maarten M. van Dijk, Christian G. M. Bürgisser, Petra E. Zhu, Changbin Kros, Johan M. Verhaar, Marianne C. Duncker, Dirk J. Cheng, Caroline Angiogenesis Original Paper Vascular endothelial (VE) cadherin is a key component of endothelial adherens junctions (AJs) and plays an important role in maintaining vascular integrity. Endocytosis of VE-cadherin regulates junctional strength and a decrease of surface VE-cadherin reduces vascular stability. However, disruption of AJs is also a requirement for vascular sprouting. Identifying novel regulators of endothelial endocytosis could enhance our understanding of angiogenesis. Here, we evaluated the angiogenic potential of (CKLF-like MARVEL transmembrane domain 4) CMTM4 and assessed in which molecular pathway CMTM4 is involved during angiogenesis. Using a 3D vascular assay composed of GFP-labeled HUVECs and dsRED-labeled pericytes, we demonstrated in vitro that siRNA-mediated CMTM4 silencing impairs vascular sprouting. In vivo, CMTM4 silencing by morpholino injection in zebrafish larvae inhibits intersomitic vessel growth. Intracellular staining revealed that CMTM4 colocalizes with Rab4(+) and Rab7(+) vesicles, both markers of the endocytic trafficking pathway. CMTM4 colocalizes with both membrane-bound and internalized VE-cadherin. Adenovirus-mediated CMTM4 overexpression enhances the endothelial endocytic pathway, in particular the rapid recycling pathway, shown by an increase in early endosomal antigen-1 positive (EEA1(+)), Rab4(+), Rab11(+) , and Rab7(+) vesicles. CMTM4 overexpression enhances membrane-bound VE-cadherin internalization, whereas CMTM4 knockdown decreases internalization of VE-cadherin. CMTM4 overexpression promotes endothelial barrier function, shown by an increase in recovery of transendothelial electrical resistance (TEER) after thrombin stimulation. We have identified in this study a novel regulatory function for CMTM4 in angiogenesis. CMTM4 plays an important role in the turnover of membrane-bound VE-cadherin at AJs, mediating endothelial barrier function and controlling vascular sprouting. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10456-018-9638-1) contains supplementary material, which is available to authorized users. Springer Netherlands 2018-08-10 2019 /pmc/articles/PMC6510885/ /pubmed/30097810 http://dx.doi.org/10.1007/s10456-018-9638-1 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Paper
Chrifi, Ihsan
Louzao-Martinez, Laura
Brandt, Maarten M.
van Dijk, Christian G. M.
Bürgisser, Petra E.
Zhu, Changbin
Kros, Johan M.
Verhaar, Marianne C.
Duncker, Dirk J.
Cheng, Caroline
CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions
title CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions
title_full CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions
title_fullStr CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions
title_full_unstemmed CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions
title_short CMTM4 regulates angiogenesis by promoting cell surface recycling of VE-cadherin to endothelial adherens junctions
title_sort cmtm4 regulates angiogenesis by promoting cell surface recycling of ve-cadherin to endothelial adherens junctions
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6510885/
https://www.ncbi.nlm.nih.gov/pubmed/30097810
http://dx.doi.org/10.1007/s10456-018-9638-1
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