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Ephrin-B2 controls VEGF-induced angiogenesis and lymphangiogenesis

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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Figure 1: Ephrin-B2 regulates angiogenesis.
Figure 2: Ephrin-B2 modulates endothelial cell sprouting.
Figure 3: VEGFR3 internalization and signalling.
Figure 4: Ephrin-B2 controls VEGFR3 internalization in vivo.

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  • 27 May 2010

    Three duplicated references have been removed

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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

  1. Yingdi Wang, Masanori Nakayama, Mara E. Pitulescu and Tim S. Schmidt: These authors contributed equally to this work.

Authors and Affiliations

  1. 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

  2. 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

  3. Departments of Physiology & Pharmacology and Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS6 6BS, UK

    Magdalena L. Bochenek & Catherine D. Nobes

  4. Centre de Biologie du Développement, Université de Toulouse, CNRS, CBD UMR 5547, F-31062 Toulouse cedex 9, France ,

    Alice Davy

  5. Theodor Kocher Institute, University of Berne, CH-3012 Bern, Switzerland

    Urban Deutsch

  6. Oncalis AG, Schlieren, Switzerland

    Urs Lüthi & Alcide Barberis

  7. Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215-5501, USA ,

    Laura E. Benjamin

  8. 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 to Ralf 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

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