PUBLICATION

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

Authors
Chrifi, I., Louzao-Martinez, L., Brandt, M.M., van Dijk, C.G.M., Bürgisser, P.E., Zhu, C., Kros, J.M., Verhaar, M.C., Duncker, D.J., Cheng, C.
ID
ZDB-PUB-180814-2
Date
2018
Source
Angiogenesis   22(1): 75-93 (Journal)
Registered Authors
Keywords
Adherens junctions, Angiogenesis, CMTM4, Endothelial cells, VE-cadherin
MeSH Terms
  • Adherens Junctions/genetics
  • Adherens Junctions/metabolism*
  • Antigens, CD/genetics
  • Cadherins/genetics
  • Endocytosis*
  • Gene Silencing
  • Human Umbilical Vein Endothelial Cells/cytology
  • Human Umbilical Vein Endothelial Cells/metabolism*
  • Humans
  • MARVEL Domain-Containing Proteins/genetics
  • MARVEL Domain-Containing Proteins/metabolism*
  • Neovascularization, Physiologic*
PubMed
30097810 Full text @ Angiogenesis
Abstract
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.
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