PUBLICATION
CMTM3 (CKLF-Like Marvel Transmembrane Domain 3) Mediates Angiogenesis by Regulating Cell Surface Availability of VE-Cadherin in Endothelial Adherens Junctions
- Authors
- Chrifi, I., Louzao-Martinez, L., Brandt, M., van Dijk, C.G.M., Burgisser, P., Zhu, C., Kros, J.M., Duncker, D.J., Cheng, C.
- ID
- ZDB-PUB-170422-9
- Date
- 2017
- Source
- Arteriosclerosis, Thrombosis, and Vascular Biology 37(6): 1098-1114 (Journal)
- Registered Authors
- Keywords
- adherens junctions, adhesive, cadherins, endocytosis, endothelial cells
- MeSH Terms
-
- Adherens Junctions/metabolism*
- Animals
- Antigens, CD/metabolism*
- Cadherins/metabolism*
- Capillary Permeability
- Cell Membrane/metabolism*
- Cells, Cultured
- Chemokines/genetics
- Chemokines/metabolism*
- Coculture Techniques
- Electric Impedance
- Endocytosis
- Endosomes/metabolism
- Gene Expression Regulation
- Human Umbilical Vein Endothelial Cells/metabolism*
- Humans
- MARVEL Domain-Containing Proteins/genetics
- MARVEL Domain-Containing Proteins/metabolism*
- Neovascularization, Physiologic*
- Pericytes/metabolism
- RNA Interference
- Signal Transduction
- Time Factors
- Transfection
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish/metabolism
- Zebrafish Proteins/genetics
- Zebrafish Proteins/metabolism
- PubMed
- 28428220 Full text @ Arterio., Thromb., and Vas. Bio.
Citation
Chrifi, I., Louzao-Martinez, L., Brandt, M., van Dijk, C.G.M., Burgisser, P., Zhu, C., Kros, J.M., Duncker, D.J., Cheng, C. (2017) CMTM3 (CKLF-Like Marvel Transmembrane Domain 3) Mediates Angiogenesis by Regulating Cell Surface Availability of VE-Cadherin in Endothelial Adherens Junctions. Arteriosclerosis, Thrombosis, and Vascular Biology. 37(6):1098-1114.
Abstract
Objective Decrease in VE-cadherin adherens junctions reduces vascular stability, whereas disruption of adherens junctions is a requirement for neovessel sprouting during angiogenesis. Endocytosis plays a key role in regulating junctional strength by altering bioavailability of cell surface proteins, including VE-cadherin. Identification of new mediators of endothelial endocytosis could enhance our understanding of angiogenesis. Here, we assessed the function of CMTM3 (CKLF-like MARVEL transmembrane domain 3), which we have previously identified as highly expressed in Flk1+ endothelial progenitor cells during embryonic development.
Approach and results Using a 3-dimensional coculture of human umbilical vein endothelial cells-GFP (green fluorescent protein) and pericytes-RFP (red fluorescent protein), we demonstrated that siRNA-mediated CMTM3 silencing in human umbilical vein endothelial cells impairs angiogenesis. In vivo CMTM3 inhibition by morpholino injection in developing zebrafish larvae confirmed that CMTM3 expression is required for vascular sprouting. CMTM3 knockdown in human umbilical vein endothelial cells does not affect proliferation or migration. Intracellular staining demonstrated that CMTM3 colocalizes with early endosome markers EEA1 (early endosome marker 1) and Clathrin+ vesicles and with cytosolic VE-cadherin in human umbilical vein endothelial cells. Adenovirus-mediated CMTM3 overexpression enhances endothelial endocytosis, shown by an increase in Clathrin+, EEA1+, Rab11+, Rab5+, and Rab7+ vesicles. CMTM3 overexpression enhances, whereas CMTM3 knockdown decreases internalization of cell surface VE-cadherin in vitro. CMTM3 promotes loss of endothelial barrier function in thrombin-induced responses, shown by transendothelial electric resistance measurements in vitro.
Conclusions In this study, we have identified a new regulatory function for CMTM3 in angiogenesis. CMTM3 is involved in VE-cadherin turnover and is a regulator of the cell surface pool of VE-cadherin. Therefore, CMTM3 mediates cell-cell adhesion at adherens junctions and contributes to the control of vascular sprouting.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping