Letter | Published:

Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin

Nature Cell Biology volume 10, pages 329337 (2008) | Download Citation

Subjects

Abstract

The serine/threonine protein kinase Akt is involved in a variety of cellular processes including cell proliferation, survival, metabolism and gene expression. It is essential in vascular endothelial growth factor (VEGF)-mediated angiogenesis; however, it is not known how Akt regulates the migration of endothelial cells, a crucial process for vessel sprouting, branching and the formation of networks during angiogenesis. Here we report that Akt-mediated phosphorylation of Girdin, an actin-binding protein, promotes VEGF-dependent migration of endothelial cells and tube formation by these cells. We found that exogenously delivered adenovirus harbouring Girdin short interfering RNA in Matrigel embedded in mice, markedly inhibited VEGF-mediated angiogenesis. Targeted disruption of the Girdin gene in mice impaired vessel remodelling in the retina and angiogenesis from aortic rings, whereas Girdin was dispensable for embryonic vasculogenesis. These findings demonstrate that the Akt/Girdin signalling pathway is essential in VEGF-mediated postneonatal angiogenesis.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , & Akt1 in endothelial cell and angiogenesis. Cell Cycle 5, 512–518 (2006).

  2. 2.

    & Role of Akt signaling in vascular homeostasis and angiogenesis. Circ. Res. 90, 1243–1250 (2002).

  3. 3.

    , , & VEGF receptor signalling in control of vascular function. Nature Rev. Mol. Cell Biol. 7, 359–371 (2006).

  4. 4.

    et al. Protein kinase Bα/Akt1 regulates placental development and fetal growth. J. Biol. Chem. 278, 32124–32131 (2003).

  5. 5.

    et al. Microvascular patterning is controlled by fine-tuning the Akt signal. Proc. Natl Acad. Sci. USA 102, 128–133 (2005).

  6. 6.

    et al. Akt1/protein kinase Bα is critical for ischemic and VEGF-mediated angiogenesis. J. Clin. Invest. 115, 2119–2127 (2005).

  7. 7.

    et al. Akt1 regulates pathological angiogenesis, vascular maturation and permeability in vivo. Nature Med. 11, 1188–1196 (2005).

  8. 8.

    et al. Pathological angiogenesis is induced by sustained Akt signaling and inhibited by rapamycin. Cancer Cell 10, 159–170 (2006).

  9. 9.

    et al. Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3-kinase/Akt signal transduction pathway. Requirement for Flk-1/KDR activation. J. Biol. Chem. 273, 30336–30343 (1998).

  10. 10.

    & Regulation of cardiac growth and coronary angiogenesis by the Akt/PKB signaling pathway. Genes Dev. 20, 3347–3365 (2006).

  11. 11.

    et al. Akt/PKB regulates actin organization and cell motility via Girdin/APE. Dev. Cell 9, 389–402 (2005).

  12. 12.

    et al. A novel protein kinase B (PKB)/AKT-binding protein enhances PKB kinase activity and regulates DNA synthesis. J. Biol. Chem. 280, 18525–18535 (2005).

  13. 13.

    , , , & Identification and characterization of GIV, a novel Gαi/s-interacting protein found on COPI, endoplasmic reticulum-Golgi transport vesicles. J. Biol. Chem. 280, 22012–22020 (2005).

  14. 14.

    et al. A novel hook-related protein family and the characterization of hook-related protein 1. Traffic 6, 442–458 (2005).

  15. 15.

    The production of 'cell cortices' for light and electron microscopy. Traffic 1, 545–552 (2000).

  16. 16.

    & Vascular endothelial growth factor stimulates tyrosine phosphorylation and recruitment to new focal adhesions of focal adhesion kinase and paxillin in endothelial cells. J. Biol. Chem. 272, 15442–15451 (1997).

  17. 17.

    et al. WAVE2 is required for directed cell migration and cardiovascular development. Nature 424, 452–456 (2003).

  18. 18.

    et al. Enhanced pathological angiogenesis in mice lacking β3 integrin or β3 and β5 integrins. Nature Med. 8, 27–34 (2002).

  19. 19.

    & Retinal angiogenesis in development and disease. Nature 438, 960–966 (2005).

  20. 20.

    & Mechanisms of endothelial cell guidance and vascular patterning in the developing mouse retina. Prog. Retin. Eye Res. 25, 277–295 (2006).

  21. 21.

    et al. VEGF guides angiogenic sprouting utilizing endothelial tip cell filopodia. J. Cell Biol. 161, 1163–1177 (2003).

  22. 22.

    et al. Mouse Aortic Ring Assay: A New Approach of the Molecular Genetics of Angiogenesis. Biol. Proced. Online 4, 24–31 (2002).

  23. 23.

    et al. Vascular endothelial growth factor-stimulated actin reorganization and migration of endothelial cells is regulated via the serine/threonine kinase Akt. Circ. Res. 86, 892–896 (2000).

  24. 24.

    & Rho, Rac, Pak and angiogenesis: old roles and newly identified responsibilities in endothelial cells. Cancer Lett. 229, 13–23 (2005).

  25. 25.

    , , & Rac regulates endothelial morphogenesis and capillary assembly. Mol. Biol. Cell 13, 2474–2485 (2002).

  26. 26.

    & The Cdc42 and Rac1 GTPases are required for capillary lumen formation in three-dimensional extracellular matrices. J. Cell Sci. 115, 1123–1136 (2002).

  27. 27.

    et al. Small GTP-binding protein Rac is an essential mediator of vascular endothelial growth factor-induced endothelial fenestrations and vascular permeability. Circulation 107, 1532–1538 (2003).

Download references

Acknowledgements

We thank Y. Iwata (Ogaki Municipal Hospital) for providing haemangioma specimens, M. Nakayama (Nagoya University) for helpful discussion, and N. Takakura (Osaka University) for discussions on angiogenesis assays. This work was supported by Grants-in-Aid for 21st century Center of Excellence (COE) Research, Scientific Research (A), and Scientific Research on Priority Area 'Cancer' (to M.T.), Grant-in-Aid for Exploratory Research (to N.A.) and Program for Improvement of Research Environment for Young Researchers from Special Coordination Funds for Promoting Science and Technology (SCF) (to A.E.) commissioned by the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.

Author information

Author notes

    • Tomoya Kitamura
    • , Naoya Asai
    •  & Atsushi Enomoto

    These authors equally contributed to this work.

Affiliations

  1. Department of Cardiology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.

    • Tomoya Kitamura
    • , Kengo Maeda
    • , Takahisa Kondo
    •  & Toyoaki Murohara
  2. Department of Pathology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.

    • Naoya Asai
    • , Atsushi Enomoto
    • , Takuya Kato
    • , Maki Ishida
    • , Ping Jiang
    •  & Masahide Takahashi
  3. Department of Cell Pharmacology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.

    • Takashi Watanabe
  4. Institute for Advanced Research, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.

    • Atsushi Enomoto
    •  & Takashi Watanabe
  5. Division of Associated Research Projects, EcoTopia Science Institute, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.

    • Jiro Usukura
  6. Department of Genetics and Development, Columbia University, 701 West 168th Street, New York, NY 10032, USA.

    • Frank Costantini
  7. Division of Molecular Pathology, Center for Neurological Disease and Cancer, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.

    • Masahide Takahashi

Authors

  1. Search for Tomoya Kitamura in:

  2. Search for Naoya Asai in:

  3. Search for Atsushi Enomoto in:

  4. Search for Kengo Maeda in:

  5. Search for Takuya Kato in:

  6. Search for Maki Ishida in:

  7. Search for Ping Jiang in:

  8. Search for Takashi Watanabe in:

  9. Search for Jiro Usukura in:

  10. Search for Takahisa Kondo in:

  11. Search for Frank Costantini in:

  12. Search for Toyoaki Murohara in:

  13. Search for Masahide Takahashi in:

Contributions

To.Ki., A.E., K.M., Ta.Ka. and M.I. performed biochemical and cell biological experiments. N.A. generated Girdin-deficient mice with assistance from F.C. and analysed their phenotype. P.J. contributed to the immunohistochemistry experiments. T.W. and J.U. performed the freeze-replica electron microscopic analyses. Ta. Ko., T.M and M.T provided team leadership and project management. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Masahide Takahashi.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary figures S1, S2, S3, S4, S5, S6 and S7

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb1695

Further reading