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Differential function of Tie2 at cell–cell contacts and cell–substratum contacts regulated by angiopoietin-1

Nature Cell Biology volume 10, pages 513–526 (2008)Cite this article

Abstract

Tie2 belongs to the receptor tyrosine kinase family and functions as a receptor for Angiopoietin-1 (Ang1). Gene-targeting analyses of either Ang1 or Tie2 in mice reveal a critical role of Ang1–Tie2 signalling in developmental vascular formation. It remains elusive how the Tie2 signalling pathway plays distinct roles in both vascular quiescence and angiogenesis. We demonstrate here that Ang1 bridges Tie2 at cell–cell contacts, resulting in trans-association of Tie2 in the presence of cell–cell contacts. In clear contrast, in isolated cells, extracellular matrix-bound Ang1 locates Tie2 at cell–substratum contacts. Furthermore, Tie2 activated at cell–cell or cell–substratum contacts leads to preferential activation of Akt and Erk, respectively. Microarray analyses and real-time PCR validation clearly show the differential gene expression profile in vascular endothelial cells upon Ang1 stimulation in the presence or absence of cell–cell contacts, implying downstream signalling is dependent upon the spatial localization of Tie2.

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Figure 1: Tie2 is recruited to cell–cell contacts upon Ang1 stimulation in vascular endothelial cells.
Figure 2: Ang1 induces trans-association of Tie2 at cell–cell contacts.
Figure 3: ECM-bound Ang1 anchors Tie2 to cell–substratum contacts in the absence of cell–cell adhesions.
Figure 4: Trans-association of Tie2 leads to the preferential activation of Akt.
Figure 5: Activation of Erk by Tie2 at cell–substratum contacts partly depends upon FAK and is involved in endothelial cell migration.
Figure 6: The presence of cell–cell contacts determines the preferential activation of Akt and subsequent phosphorylation of Foxo1 and eNOS.
Figure 7: Ang1 stimulation leads to a distinct pattern of gene expression in HUVECs in the presence or absence of cell–cell contacts.
Figure 8: Schematic representation of a proposed model for how Ang1–Tie2 signalling is involved in both vascular quiescence and angiogenesis.

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Acknowledgements

We are grateful to T. Suda (Keio University, Tokyo, Japan) for the Tie2 cDNA, to A. Fukamizu (University of Tsukuba, Tsukuba, Japan) for the Foxo1 cDNA, to K.M. Yamada (National Institute of Health) for GFP–tensin, to J. Nakae (Kobe University Graduate School of Medicine, Kobe, Japan) for the adenovirus encoding Foxo1 mutant, to A. Mizushima, M. Sone, M. Maeoka, and Y. Matsuura for technical assistance, to M. Masuda, H. Hanada, and S. Yamanoto for helpful advice and to J.T. Pearson and J.S. Gutkind for critical reading of the manuscript. This work was supported in part by grants from the Ministry of Education, Science, Sports and Culture of Japan; the Ministry of Health, Labour, and Welfare of Japan; and the Program for the Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (to S.F., T.M, T.K. N.M.); the Naito Foundation (to S.F.); Takeda Medical Research Foundation (to N.M); and KOSEF through the NRL Program (2004-02376 to G.Y.K) funded by the MOST.

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Authors and Affiliations

  1. Department of Structural Analysis, National Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, 565-8565, Osaka, Japan

    Shigetomo Fukuhara, Keisuke Sako, Kazuomi Noda & Naoki Mochizuki

  2. The Research Center for Advanced Science and Technology, University of Tokyo, Laboratory for System Biology and Medicine, 4-6-1, Komaba, Meguro, 153-8904, Tokyo, Japan

    Takashi Minami & Tatsuhiko Kodama

  3. Biomedical Research Center and Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Guseong-dong, 305-701, Daejeon, Korea

    Hak Zoo Kim & Gou Young Koh

  4. Division of Genetics, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokane-dai, Minato-ku, 108-8639, Tokyo, Japan

    Masabumi Shibuya

  5. Department of Signal Transduction, Research Institute of Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, 565-0871, Osaka, Japan

    Nobuyuki Takakura

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Contributions

S. F. and N. M. designed and wrote the paper. S. F. performed the all cell biological and biochemical analysis. K. S. and K. N. helped with the experiments performed by S. F. T. M. and T. K. performed microarray analyses. M. S. and N. T. helped with VEGF-related and Tie2–BaF experiments. H. Z. K. and G. Y. K. prepared several forms of recombinant Ang1.

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Correspondence to Shigetomo Fukuhara or Naoki Mochizuki.

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Fukuhara, S., Sako, K., Minami, T. et al. Differential function of Tie2 at cell–cell contacts and cell–substratum contacts regulated by angiopoietin-1. Nat Cell Biol 10, 513–526 (2008). https://doi.org/10.1038/ncb1714

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