Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis (original) (raw)
- Letter
- Published: 27 May 2010
- Masanori Nakayama2 na1,
- Mara E. Pitulescu2 na1,
- Tim S. Schmidt1 na1,
- Magdalena L. Bochenek2,3,
- Akira Sakakibara1,
- Susanne Adams1,2,
- Alice Davy4,
- Urban Deutsch5,
- Urs Lüthi6,
- Alcide Barberis6,
- Laura E. Benjamin7,
- Taija Mäkinen8,
- Catherine D. Nobes3 &
- …
- Ralf H. Adams1,2
Nature volume 465, pages 483–486 (2010)Cite this article
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Abstract
In development, tissue regeneration or certain diseases, angiogenic growth leads to the expansion of blood vessels and the lymphatic vasculature. This involves endothelial cell proliferation as well as angiogenic sprouting, in which a subset of cells, termed tip cells, acquires motile, invasive behaviour and extends filopodial protrusions1,2,3. Although it is already appreciated that angiogenesis is triggered by tissue-derived signals, such as vascular endothelial growth factor (VEGF) family growth factors, the resulting signalling processes in endothelial cells are only partly understood. Here we show with genetic experiments in mouse and zebrafish that ephrin-B2, a transmembrane ligand for Eph receptor tyrosine kinases, promotes sprouting behaviour and motility in the angiogenic endothelium. We link this pro-angiogenic function to a crucial role of ephrin-B2 in the VEGF signalling pathway, which we have studied in detail for VEGFR3, the receptor for VEGF-C. In the absence of ephrin-B2, the internalization of VEGFR3 in cultured cells and mutant mice is defective, which compromises downstream signal transduction by the small GTPase Rac1, Akt and the mitogen-activated protein kinase Erk. Our results show that full VEGFR3 signalling is coupled to receptor internalization. Ephrin-B2 is a key regulator of this process and thereby controls angiogenic and lymphangiogenic growth.
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27 May 2010
Three duplicated references have been removed
References
- Adams, R. H. & Alitalo, K. Molecular regulation of angiogenesis and lymphangiogenesis. Nature Rev. Mol. Cell Biol. 8, 464–478 (2007)
Article CAS Google Scholar - Gerhardt, H. & Betsholtz, C. How do endothelial cells orientate? Experientia Suppl. 94, 3–15 (2005)
- Klagsbrun, M. & Eichmann, A. A role for axon guidance receptors and ligands in blood vessel development and tumor angiogenesis. Cytokine Growth Factor Rev. 16, 535–548 (2005)
Article CAS PubMed Google Scholar - Arvanitis, D. & Davy, A. Eph/ephrin signaling: networks. Genes Dev. 22, 416–429 (2008)
Article CAS PubMed PubMed Central Google Scholar - Egea, J. & Klein, R. Bidirectional Eph-ephrin signaling during axon guidance. Trends Cell Biol. 17, 230–238 (2007)
Article CAS PubMed Google Scholar - Poliakov, A., Cotrina, M. & Wilkinson, D. G. Diverse roles of eph receptors and ephrins in the regulation of cell migration and tissue assembly. Dev. Cell 7, 465–480 (2004)
Article CAS PubMed Google Scholar - Pasquale, E. B. Eph receptor signalling casts a wide net on cell behaviour. Nature Rev. Mol. Cell Biol. 6, 462–475 (2005)
Article CAS Google Scholar - Lawson, N. D. & Weinstein, B. M. Arteries and veins: making a difference with zebrafish. Nature Rev. Genet. 3, 674–682 (2002)
Article CAS PubMed Google Scholar - Lamont, R. E. & Childs, S. MAPping out arteries and veins. Sci. STKE 2006, pe39 (2006)
Article PubMed Google Scholar - le Noble, F. et al. Flow regulates arterial–venous differentiation in the chick embryo yolk sac. Development 131, 361–375 (2004)
Article CAS PubMed Google Scholar - Wang, H. U., Chen, Z. F. & Anderson, D. J. Molecular distinction and angiogenic interaction between embryonic arteries and veins revealed by ephrin-B2 and its receptor Eph-B4. Cell 93, 741–753 (1998)
Article CAS PubMed Google Scholar - Gerety, S. S., Wang, H. U., Chen, Z. F. & Anderson, D. J. Symmetrical mutant phenotypes of the receptor EphB4 and its specific transmembrane ligand ephrin-B2 in cardiovascular development. Mol. Cell 4, 403–414 (1999)
Article CAS PubMed Google Scholar - Adams, R. H. et al. Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. Genes Dev. 13, 295–306 (1999)
Article MathSciNet CAS PubMed PubMed Central Google Scholar - Shin, D. et al. Expression of ephrinB2 identifies a stable genetic difference between arterial and venous vascular smooth muscle as well as endothelial cells, and marks subsets of microvessels at sites of adult neovascularization. Dev. Biol. 230, 139–150 (2001)
Article CAS PubMed Google Scholar - Gale, N. W. et al. Ephrin-B2 selectively marks arterial vessels and neovascularization sites in the adult, with expression in both endothelial and smooth-muscle cells. Dev. Biol. 230, 151–160 (2001)
Article CAS PubMed Google Scholar - Davy, A. & Soriano, P. Ephrin-B2 forward signaling regulates somite patterning and neural crest cell development. Dev. Biol. 304, 182–193 (2007)
Article CAS PubMed Google Scholar - Taylor, A. C., Murfee, W. L. & Peirce, S. M. EphB4 expression along adult rat microvascular networks: EphB4 is more than a venous specific marker. Microcirculation 14, 253–267 (2007)
Article CAS PubMed Google Scholar - Zhu, Z., Zheng, T., Lee, C. G., Homer, R. J. & Elias, J. A. Tetracycline-controlled transcriptional regulation systems: advances and application in transgenic animal modeling. Semin. Cell Dev. Biol. 13, 121–128 (2002)
Article CAS PubMed Google Scholar - Sun, J. F. et al. Microvascular patterning is controlled by fine-tuning the Akt signal. Proc. Natl Acad. Sci. USA 102, 128–133 (2005)
Article ADS CAS PubMed Google Scholar - Baluk, P., Hashizume, H. & McDonald, D. M. Cellular abnormalities of blood vessels as targets in cancer. Curr. Opin. Genet. Dev. 15, 102–111 (2005)
Article CAS PubMed Google Scholar - Makinen, T. et al. PDZ interaction site in ephrinB2 is required for the remodeling of lymphatic vasculature. Genes Dev. 19, 397–410 (2005)
Article CAS PubMed PubMed Central Google Scholar - Tammela, T. et al. Blocking VEGFR-3 suppresses angiogenic sprouting and vascular network formation. Nature 454, 656–660 (2008)
Article ADS CAS PubMed Google Scholar - Tammela, T., Enholm, B., Alitalo, K. & Paavonen, K. The biology of vascular endothelial growth factors. Cardiovasc. Res. 65, 550–563 (2005)
Article CAS PubMed Google Scholar - McColl, B. K., Stacker, S. A. & Achen, M. G. Molecular regulation of the VEGF family–inducers of angiogenesis and lymphangiogenesis. APMIS 112, 463–480 (2004)
Article CAS PubMed Google Scholar - Zachary, I. & Gliki, G. Signaling transduction mechanisms mediating biological actions of the vascular endothelial growth factor family. Cardiovasc. Res. 49, 568–581 (2001)
Article CAS PubMed Google Scholar - Olsson, A. K., Dimberg, A., Kreuger, J. & Claesson-Welsh, L. VEGF receptor signalling – in control of vascular function. Nature Rev. Mol. Cell Biol. 7, 359–371 (2006)
Article CAS Google Scholar - Macia, E. et al. Dynasore, a cell-permeable inhibitor of dynamin. Dev. Cell 10, 839–850 (2006)
Article CAS PubMed Google Scholar - Sawamiphak, S. et al. Ephrin-B2 regulates VEGF-R2 function in developmental and tumour angiogenesis. Nature 10.1038/nature08995 (this issue)
- Grunwald, I. C. et al. Hippocampal plasticity requires postsynaptic ephrinBs. Nature Neurosci. 7, 33–40 (2004)
Article CAS PubMed Google Scholar - Foo, S. S. et al. Ephrin-B2 controls cell motility and adhesion during blood-vessel-wall assembly. Cell 124, 161–173 (2006)
Article CAS PubMed Google Scholar - Deutsch, U. et al. Inducible endothelial cell-specific gene expression in transgenic mouse embryos and adult mice. Exp. Cell Res. 314, 1202–1216 (2008)
Article CAS PubMed Google Scholar - Osoegawa, K. et al. Bacterial artificial chromosome libraries for mouse sequencing and functional analysis. Genome Res. 10, 116–128 (2000)
PubMed PubMed Central CAS Google Scholar - Feil, R., Wagner, J., Metzger, D. & Chambon, P. Regulation of Cre recombinase activity by mutated estrogen receptor ligand-binding domains. Biochem. Biophys. Res. Commun. 237, 752–757 (1997)
Article CAS PubMed Google Scholar - Copeland, N. G., Jenkins, N. A. & Court, D. L. Recombineering: a powerful new tool for mouse functional genomics. Nature Rev. Genet. 2, 769–779 (2001)
Article CAS PubMed Google Scholar - Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)
Article CAS PubMed Google Scholar - Jat, P. S. et al. Direct derivation of conditionally immortal cell lines from an H-2Kb-tsA58 transgenic mouse. Proc. Natl Acad. Sci. USA 88, 5096–5100 (1991)
Article ADS CAS PubMed Google Scholar - Morgan, S. M., Samulowitz, U., Darley, L., Simmons, D. L. & Vestweber, D. Biochemical characterization and molecular cloning of a novel endothelial-specific sialomucin. Blood 93, 165–175 (1999)
Article CAS PubMed Google Scholar - Lawson, N. D. & Weinstein, B. M. In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev. Biol. 248, 307–318 (2002)
Article CAS PubMed Google Scholar
Acknowledgements
We thank R. Benedito, I. Schmidt, S. Hoffmann, I. Rosewell, S.M. Kuijper, F. Gisler and N. Hostettler for their help, N. Copeland and A. Eichmann for information and reagents, P. Chambon for the CreERT2 cDNA, A.L. Bermange, J.D. Leslie and J. Lewis for help with zebrafish experiments, and A. Acker-Palmer for discussions and for reading the manuscript. Cancer Research UK, the Max-Planck-Society, the German Research Foundation (programmes SFB 629 and SPP 1190) and the EMBO LTF programme provided funding.
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Author notes
- Yingdi Wang, Masanori Nakayama, Mara E. Pitulescu and Tim S. Schmidt: These authors contributed equally to this work.
Authors and Affiliations
- Vascular Development Laboratory, Cancer Research UK London Research Institute, London WC2A 3PX, UK
Yingdi Wang, Tim S. Schmidt, Akira Sakakibara, Susanne Adams & Ralf H. Adams - Department of Tissue Morphogenesis, Max-Planck-Institute for Molecular Biomedicine, and Faculty of Medicine, University of Münster, D-48149 Münster, Germany
Masanori Nakayama, Mara E. Pitulescu, Magdalena L. Bochenek, Susanne Adams & Ralf H. Adams - Departments of Physiology & Pharmacology and Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS6 6BS, UK
Magdalena L. Bochenek & Catherine D. Nobes - Centre de Biologie du Développement, Université de Toulouse, CNRS, CBD UMR 5547, F-31062 Toulouse cedex 9, France ,
Alice Davy - Theodor Kocher Institute, University of Berne, CH-3012 Bern, Switzerland
Urban Deutsch - Oncalis AG, Schlieren, Switzerland
Urs Lüthi & Alcide Barberis - Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215-5501, USA ,
Laura E. Benjamin - Cancer Research UK London Research Institute, Lymphatic Development Laboratory, London WC2A 3PX, UK
Taija Mäkinen
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Contributions
Y.W., M.E.P., M.N., C.D.N. and R.H.A. designed experiments. Y.W., M.E.P. and T.S.S. characterized mouse mutants. M.L.B. and A.S. performed zebrafish experiments, M.L.B. microinjection assays and M.N. all other cell culture experiments. A.D., U.D., L.E.B., S.A. and T.M. generated mouse mutants or lines, U.L. and A.B. the EphB4 inhibitors. Y.W., M.N., M.E.P. and R.H.A. wrote the manuscript.
Corresponding author
Correspondence toRalf H. Adams.
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Competing interests
U.L. and A.B. are employed by Oncalis, the company that has developed the inhibitors ONC-101 and ONC-102.
Supplementary information
Supplementary Information
This file contains Supplementary Methods and References, Supplementary Figures 1-16 with legends and full captions for Supplementary Movies S1-S3. (PDF 8879 kb)
Supplementary Movie S1
A fluorescent time-lapse movie showing dynamics of intersegmental vessels in 27 hpf _fli1_-EGFP embryo injected with control morpholino. (MOV 2136 kb)
Supplementary Movie S2
Intersegmental vessels in 27 hpf _efnb2a_-MO-injected _fli1_-EGFP embryo showed few filopodia and instead blunt, bleb-like protrusions were seen on the cell surface. (MOV 2298 kb)
Supplementary Movie S3
Ephrin-B2 overexpression in single cells within a confluent monolayer of (uninjected) HUVECs. (MOV 3090 kb)
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Wang, Y., Nakayama, M., Pitulescu, M. et al. Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis.Nature 465, 483–486 (2010). https://doi.org/10.1038/nature09002
- Received: 25 October 2009
- Revised: 02 March 2010
- Accepted: 05 May 2010
- Published: 27 May 2010
- Issue Date: 27 May 2010
- DOI: https://doi.org/10.1038/nature09002
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Editorial Summary
Ephrin-B2/VEGF in angiogenesis control
Ephrin-B ligands are well known as axon guidance molecules. Ephrin-B2 is also known to play a role in angiogenic remodelling. Two studies now show that signalling through ephrin-B2 controls vessel sprouting. Mechanistically, ephrin-B2 seems to function in part by regulating VEGFR internalization and signalling. The finding suggests that blocking ephrin-B2 signalling may be an alternative approach to blocking VEGFR function in angiogenesis.