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Angiopoietins assemble distinct Tie2 signalling complexes in endothelial cell–cell and cell–matrix contacts

Nature Cell Biology volume 10, pages 527–537 (2008)Cite this article

Abstract

The receptor tyrosine kinase Tie2, and its activating ligand Angiopoietin-1 (Ang1), are required for vascular remodelling and vessel integrity, whereas Ang2 may counteract these functions. However, it is not known how Tie2 transduces these different signals. Here, we show that Ang1 induces unique Tie2 complexes in mobile and confluent endothelial cells. Matrix-bound Ang1 induced cell adhesion, motility and Tie2 activation in cell–matrix contacts that became translocated to the trailing edge in migrating endothelial cells. In contrast, in contacting cells Ang1 induced Tie2 translocation to cell–cell contacts and the formation of homotypic Tie2–Tie2 trans-associated complexes that included the vascular endothelial phosphotyrosine phosphatase, leading to inhibition of paracellular permeability. Distinct signalling proteins were preferentially activated by Tie2 in the cell–matrix and cell–cell contacts, where Ang2 inhibited Ang1-induced Tie2 activation. This novel type of cellular microenvironment-dependent receptor tyrosine kinase activation may explain some of the effects of angiopoietins in angiogenesis and vessel stabilization.

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Figure 1: Ang1 induces translocation of Tie1 and Tie2 to endothelial cell–cell contacts.
Figure 2: Soluble and substrate bound Ang1 induce Tie2 translocation to cell–matrix contacts during cell spreading and migration.
Figure 3: The extracellular and transmembrane domains of Tie2 suffice for translocation to cell–cell contacts in response to Ang1.
Figure 4: Ang1 induces activation of Tie2 in trans.
Figure 5: Distinct downstream signalling by Ang1 in the presence and absence of cell–cell contacts.
Figure 6: VE–PTP and Tie2 interact in endothelial cell–cell contacts.
Figure 7: A schematic representation of a model of Ang1–Tie2 interactions at cell–matrix and cell–cell contacts.

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Acknowledgements

We thank: D. Dumont for the Dok-R construct; M. Jeltsch for VEGF-C; J. Partanen and H. Augustin for constructive criticism on the manuscript; N. Ihalainen, T. LaakkonenV. Pihlajaniemi, K. Pulkki and J. Träskelin for excellent technical assistance; S. Eskelinen, A. Manninen and the Molecular Imaging Unit of Biomedicum Helsinki for advice. This work was supported by grants from the the Academy of Finland (213327 (P. S, 116138 (L.E.), 202852, 204312 (K. A.)), the Louis Jeantet Foundation, the Human Frontier Science Program, the Finnish Cancer Organizations, the Novo Nordisk Foundation, National Institute of Health grants HL075183-02 (K. A.), AR36820 and AR48564 (B. R. O.) and by the KOSEF through the NRL (2004-02376, G. Y. K.) funded by the MOST.

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Author notes

  1. Pipsa Saharinen and Lauri Eklund: These authors contributed equally to this work.

Authors and Affiliations

  1. Molecular/Cancer Biology Laboratory and Ludwig Institute for Cancer Research, Biomedicum Helsinki, University of Helsinki, Haartmaninkatu 8, P.O.B. 63, Helsinki, 00014, Finland

    Pipsa Saharinen, Juho Miettinen, Andrey Anisimov & Kari Alitalo

  2. Biocenter Oulu and Department of Medical Biochemistry and Molecular Biology, Oulu Centre for Cell-Matrix Research, University of Oulu, P.O.B. 5000, 90014 University of Oulu, Finland

    Lauri Eklund & Riikka Wirkkala

  3. Max-Planck-Institute of Molecular Biomedicine, Roentgenstr, D-48149, Muenster, Germany

    Mark Winderlich, Astrid Nottebaum & Dietmar Vestweber

  4. Theodor Kocher Institute, University of Berne, Freiestrasse 1, Bern, CH-3012, Switzerland

    Urban Deutsch

  5. National Research Laboratory of Vascular Biology and Biomedical Center and Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, Guseong-dong, 305-701, Daejeon, Republic of Korea

    Gou Young Koh

  6. Department of Developmental Biology, Harvard School of Dental Medicine, 188 Longwood Avenue, Boston, 02115, MA, USA

    Lauri Eklund & Bjorn R. Olsen

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Contributions

P. S., L. E. and K. A. designed most of the experiments; P. S. and L. E. participated in the experimental work, performed most of the data collection, data analysis. writing and processing of figures; J. M. and R. W. performed a significant amount of the experimental work and some planning of experiments; A. A. participated in the S2 cell experiments; M. W., A. N. and D. W. performed VE-PTP staining of mouse endothelioma cells and produced anti-VE-PTP anti-serum; U. D. provided Tie2−/− and Tie2+/+ endothelioma cells; G. Y. K. provided COMP–Ang1; U. D. and G. Y. K. provided advice. Most of the work was carried out in the laboratories of L. E. and K. A., and some experiments in the laboratory of B. R. O.

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Correspondence to Kari Alitalo.

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Saharinen, P., Eklund, L., Miettinen, J. et al. Angiopoietins assemble distinct Tie2 signalling complexes in endothelial cell–cell and cell–matrix contacts. Nat Cell Biol 10, 527–537 (2008). https://doi.org/10.1038/ncb1715

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