Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Enhanced pathological angiogenesis in mice lacking β3 integrin or β3 and β5 integrins

Nature Medicine volume 8, pages 27–34 (2002)Cite this article

Abstract

Inhibition of αvβ3 or αvβ5 integrin function has been reported to suppress neovascularization and tumor growth, suggesting that these integrins are critical modulators of angiogenesis. Here we report that mice lacking β3 integrins or both β3 and β5 integrins not only support tumorigenesis, but have enhanced tumor growth as well. Moreover, the tumors in these integrin-deficient mice display enhanced angiogenesis, strongly suggesting that neither β3 nor β5 integrins are essential for neovascularization. We also observed that angiogenic responses to hypoxia and vascular endothelial growth factor (VEGF) are augmented significantly in the absence of β3 integrins. We found no evidence that the expression or functions of other integrins were altered as a consequence of the β3 deficiency, but we did observe elevated levels of VEGF receptor-2 (also called Flk-1) in β3-null endothelial cells. These data indicate that αvβ3 and αvβ5 integrins are not essential for vascular development or pathological angiogenesis and highlight the need for further evaluation of the mechanisms of action of αv-integrin antagonists in anti-angiogenic therapeutics.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Tumor growth and angiogenesis are enhanced in β3-deficient mice.
Figure 2: β3/β5-null mice show enhanced tumor growth and angiogenesis.
Figure 3: β3 deficiency does not affect the expression or function of other integrins.
Figure 4: Hypoxia-induced retinal angiogenesis is enhanced in β3-null mice.
Figure 5: β3 deficiency enhances VEGF-induced vessel growth in vivo and in vitro.
Figure 6: Enhanced VEGFR-2 expression on β3-null endothelial cells.

Similar content being viewed by others

References

  1. Ferrara, N. & Alitalo, K. Clinical applications of angiogenic growth factors and their inhibitors. Nature Med. 5, 1359–1364 (1999).

    Article  CAS  Google Scholar 

  2. Hynes, R.O., Bader, B.L. & Hodivala-Dilke, K. Integrins in vascular development. Braz. J. Med. Biol. Res. 32, 501–510 (1999).

    Article  CAS  Google Scholar 

  3. Carmeliet, P. & Jain, R.K. Angiogenesis in cancer and other diseases. Nature 407, 249–257 (2000).

    Article  CAS  Google Scholar 

  4. Carmeliet, P. et al. Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele. Nature 380, 435–439 (1996).

    Article  CAS  Google Scholar 

  5. Shalaby, F. et al. Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 376, 62–66 (1995).

    Article  CAS  Google Scholar 

  6. Fong, G.H., Rossant, J., Gertsenstein, M. & Breitman, M.L. Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium. Nature 376, 66–70 (1995).

    Article  CAS  Google Scholar 

  7. Brekken, R.A. et al. Selective inhibition of vascular endothelial growth factor (VEGF) receptor 2 (KDR/Flk-1) activity by a monoclonal anti-VEGF antibody blocks tumor growth in mice. Cancer Res. 60, 5117–5124 (2000).

    CAS  Google Scholar 

  8. Saaristo, A., Karpanen, T. & Alitalo, K. Mechanisms of angiogenesis and their use in the inhibition of tumor growth and metastasis. Oncogene 19, 6122–6129 (2000).

    Article  CAS  Google Scholar 

  9. Senger, D.R. et al. Angiogenesis promoted by vascular endothelial growth factor: Regulation through α1β1 and α2β1 integrins. Proc. Natl. Acad. Sci. USA 94, 13612–13617 (1997).

    Article  CAS  Google Scholar 

  10. Kim, S., Harris, M. & Varner, J.A. Regulation of Integrin αvβ3-mediated Endothelial Cell Migration and Angiogenesis by Integrin α5β1 and protein kinase A. J. Biol. Chem. 275, 33920–33928 (2000a).

    Article  CAS  Google Scholar 

  11. Brooks, P.C., Clark, R.A.F. & Cheresh, D.A. Requirement of vascular integrin αvβ3 for angiogenesis. Science 264, 569–571 (1994).

    Article  CAS  Google Scholar 

  12. Brooks, P.C. et al. Integrin αvβ3 antagonists promote tumor regression by inducing apoptosis angiogenic blood vessels. Cell 79, 1157–1164 (1994).

    Article  CAS  Google Scholar 

  13. Brooks, P.C. et al. Antiintegrin αvβ3 blocks human breast-cancer growth and angiogenesis in human skin. J. Clin. Invest. 96, 1815–1822 (1995).

    Article  CAS  Google Scholar 

  14. Drake, C.J., Cheresh, D.A. & Little, C.D. An antagonist of integrin αvβ3 prevents maturation of blood vessels during embryonic neovascularization. J. Cell Sci. 108, 2655–2661 (1995).

    CAS  Google Scholar 

  15. Friedlander, M. et al. Definition of two angiogenic pathways by distinct αv integrins. Science 270, 1500–1502 (1995).

    Article  CAS  Google Scholar 

  16. Friedlander, M. et al. Involvement of integrins αvβ3 and αvβ5 in ocular neovascular diseases. Proc. Natl. Acad. Sci. USA 93, 9764–9769 (1996).

    Article  CAS  Google Scholar 

  17. Eliceiri, B.P. & Cheresh, D.A. The role of αv integrins during angiogenesis: Insights into potential mechanisms of action and clinical development. J. Clin. Invest. 103, 1227–1230 (1999).

    Article  CAS  Google Scholar 

  18. Varner, J.A. & Cheresh, D.A. Integrins and cancer. Curr. Opin. Cell Biol. 8, 724–730 (1996).

    Article  CAS  Google Scholar 

  19. Hammes, H.P., Brownlee, M., Jonczyk, A., Sutter, A. & Preissner, K.T. Subcutaneous injection of a cyclic peptide antagonist of vitronectin receptor-type integrins inhibits retinal neovascularization. Nature Med. 2, 820–820 (1996).

    Article  CAS  Google Scholar 

  20. Kumar, C.C. et al. Inhibition of angiogenesis and tumor growth by SCH221153, a dual αvβ3 and αvβ5 integrin receptor antagonist. Cancer Res. 61, 2232–2238 (2001).

    CAS  Google Scholar 

  21. Gutheil, J.C. et al. Targeted antiangiogenic therapy for cancer using vitaxin: A humanized monoclonal antibody to the integrin αvβ3 . Clin. Cancer Res. 6, 3056–3061 (2000).

    CAS  PubMed Central  Google Scholar 

  22. Bader, B.L., Rayburn, H., Crowley, D. & Hynes, R.O. Extensive vasculogenesis, angiogensis, and organogenesis precede lethality in mice lacking all αv integrins. Cell 95, 507–519 (1998).

    Article  CAS  Google Scholar 

  23. Hodivala-Dilke, K.M. et al. β3-integrin-deficient mice are a model for Glanzmann thrombasthenia showing placental defects and reduced survival. J. Clin. Invest. 103, 229–238 (1999).

    Article  CAS  Google Scholar 

  24. Huang, X.Z., Griffiths, M., Wu, J.F., Farese, R.V. & Sheppard, D. Normal development, wound healing, and adenovirus susceptibility in β5-deficient mice. Mol. Cell. Biol. 20, 755–759 (2000).

    Article  CAS  Google Scholar 

  25. Kim, S., Bell, K., Mouse, S.A. & Varner, J.A. Regulation of angiogenesis in vivo by ligation of integrin α5β1 with the central cell-binding domain of fibronectin. Am. J. Pathol. 156, 1345–1362 (2000b).

    Article  CAS  Google Scholar 

  26. Pierce, E.A. Regulation of vascular endothelial growth factor by oxygen in a model of retinopathy of prematurity (vol 114, pg 1219, 1996). Arch. Ophthalmol. 115, 427–427 (1997).

    Article  Google Scholar 

  27. Stone, E.M. & Sheffield, V.C. Genetic approaches to human retinal disorders. Invest. Ophthalmol. Vis. Sci. 37, 3100–3100 (1996).

    Google Scholar 

  28. Kroon, M.E., Koolwijk, P., van der Vecht, B. & van Hinsbergh, V.W.M. Urokinase receptor expression on human microvascular endothelial cells is increased by hypoxia: Implications for capillary-like tube formation in a fibrin matrix. Blood 96, 2775–2783 (2000).

    CAS  Google Scholar 

  29. DiazGonzalez, F., Forsyth, J., Steiner, B. & Ginsberg, M.H. Trans-dominant inhibition of integrin function. Mol. Biol. Cell 7, 1939–1951 (1996).

    Article  CAS  Google Scholar 

  30. Blystone, S.D., Graham, I.L., Lindberg, F.P. & Brown, E.J. Integrin αvβ3 differentially regulates adhesive and phagocytic functions of the fibronectin receptor α5β1 . J. Cell Biol 127, 1129–1137 (1994).

    Article  CAS  Google Scholar 

  31. Huhtala, P. et al. Cooperative signalling by α5β1 and α4β1 integrins regulates metalloproteinase gene expression in fibroblasts adhering to fibronectin. J. Cell Biol 129, 876–879 (1995).

    Article  Google Scholar 

  32. Chen, Y. et al. “Inside out” signal transduction inhibited by isolated integrin cytoplasmic domains. J. Biol. Chem. 269, 18307–18310 (1994).

    CAS  Google Scholar 

  33. LaFlamme, S.E., Thomas, L.A., Yamada, S.S. & Yamada, K.M. Single subunit chimeric integrins as mimics and inhibitors of endogenous integrin functions in receptor localization, cell spreading and migration, and matrix assembly. J. Cell Biol. 126, 1287–1298 (1994).

    Article  CAS  Google Scholar 

  34. Taverna, D. & Hynes, R.O. Reduced blood vessel formation and tumor growth in α5-integrin-negative teratocarcinomas and embryoid bodies. Cancer Res. 61, 5255–5261 (2001).

    CAS  Google Scholar 

  35. Asahara, T. et al. VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J 18, 3964–3972 (1999).

    Article  CAS  Google Scholar 

  36. Isner, J.M. & Asahara, T. Angiogenesis and vasculogenesis as therapeutic strategies for postnatal neovascularization. J. Clin. Invest. 103, 1231–1236 (1999).

    Article  CAS  Google Scholar 

  37. Kalka, C. et al. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc. Natl. Acad. Sci. USA 97, 3422–3427 (2000).

    Article  CAS  Google Scholar 

  38. Takahashi, T. et al. Ischemia- and cytokine-induced mobilization of bone marrow–derived endothelial progenitor cells for neovascularization. Nature Med. 5, 434–438 (1999).

    Article  CAS  Google Scholar 

  39. Carmeliet, P. et al. Synergism between vascular endothelial growth factor and placental growth factor contributs to angiogenesis and plasma extravasation in pathological conditions. Nature Med. 7, 575–583 (2001).

    Article  CAS  Google Scholar 

  40. Soldi, R. et al. Role of αvβ3 integrin in the activation of vascular endothelial growth factor receptor-2. EMBO J. 18, 882–892 (1999).

    Article  CAS  Google Scholar 

  41. Byzova, T.V. et al. A mechanism for modulation of cellular responses to VEGF: Activation of the integrins. Mol. Cell 6, 851–860 (2000).

    CAS  Google Scholar 

  42. Marcantonio, E. & Hynes, R. Antibodies to the conserved cytoplasmic domain of the integrin β1 subunit react with proteins in vertebrates, invertebrates, and fungi. J. Cell Biol. 106, 1765–1772 (1988).

    Article  CAS  Google Scholar 

  43. Albrecht-Buehler, G. The phagokinectic tracks of 3T3 Cells. Cell 11, 395–404 (1977).

    Article  CAS  Google Scholar 

  44. Passaniti, A. et al. A simple, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor. Lab. Invest. 67, 519–528 (1992).

    CAS  Google Scholar 

  45. Nicosia, R.F. & Ottinetti, A. Modulation of microvascular growth and morphogenesis by reconstituted basement-membrane gel in 3-dimensional cultures of rat aorta—a comparative-study of angiogenesis in Matrigel, collagen, fibrin, and plasma clot. In Vitro Cell. Dev. Biol. 26, 119–128 (1990).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B. Coller, E. Ruoslahti and L. Reichardt for antibodies against mouse β3-, αv and β5 integrin antibodies, respectively; G. Saunders, S. Watling and C. Wren for their technical assistance; G. Elias and colleagues for help with histology; J. Marshall for his gift of human β3-construct; F. Parkinson for her help in preparing the manuscript; and I. Hart for criticism during this study. This work was supported in part by grants from the NIH (PO1HL41484, PO1HL66105 and RO1CA17007 to R.O.H. and R01 HL64353, RO1 HL53949 to D.S.), and by the Howard Hughes Medical Institute. R.O.H. is an investigator and D.T. is an Associate of the Howard Hughes Medical Institute.

Author information

Author notes

  1. Louise E. Reynolds and Lorenza Wyder: L.E.R. and L.W. contributed equally to this study.

Authors and Affiliations

  1. Richard Dimbleby Department, Cell Adhesion and Disease Laboratory, Imperial Cancer Research Fund, St. Thomas' Hospital, London, UK

    Louise E. Reynolds, Lorenza Wyder, Stephen D. Robinson & Kairbaan M. Hodivala-Dilke

  2. Howard Hughes Medical Institute, Center for Cancer Research, MIT, Cambridge, 02139, Massachusetts, USA

    Julie C. Lively, Daniela Taverna & Richard O. Hynes

  3. Lung Biology Center, University of California, San Francisco, California, USA

    Xiaozhu Huang & Dean Sheppard

Authors
  1. Louise E. Reynolds
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lorenza Wyder
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Julie C. Lively
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Daniela Taverna
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stephen D. Robinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiaozhu Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Dean Sheppard
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Richard O. Hynes
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Kairbaan M. Hodivala-Dilke
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kairbaan M. Hodivala-Dilke.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Reynolds, L., Wyder, L., Lively, J. et al. Enhanced pathological angiogenesis in mice lacking β3 integrin or β3 and β5 integrins. Nat Med 8, 27–34 (2002). https://doi.org/10.1038/nm0102-27

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm0102-27

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing