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Histone H2AX is integral to hypoxia-driven neovascularization

Nature Medicine volume 15, pages 553–558 (2009)Cite this article

Abstract

H2A histone family member X (H2AX, encoded by H2AFX) and its C-terminal phosphorylation (γ-H2AX) participates in the DNA damage response and mediates DNA repair1,2,3,4,5,6. Hypoxia is a physiological stress that induces a replication-associated DNA damage response7. Moreover, hypoxia is the major driving force for neovascularization8, as the hypoxia-mediated induction of vascular growth factors triggers endothelial cell proliferation8. Here we studied the role of the hypoxia-induced DNA damage response in endothelial cell function and in hypoxia-driven neovascularization in vivo. Hypoxia induced replication-associated generation of γ-H2AX in endothelial cells in vitro and in mice. Both in cultured cells and in mice, endothelial cell proliferation under hypoxic conditions was reduced by H2AX deficiency. Whereas developmental angiogenesis was not affected in H2afx−/− mice, hypoxia-induced neovascularization during pathologic proliferative retinopathy, in response to hind limb ischemia or during tumor angiogenesis was substantially lower in H2afx−/− mice. Moreover, endothelial-specific H2afx deletion resulted in reduced hypoxia-driven retina neovascularization and tumor neovascularization. Our findings establish that H2AX, and hence activation of the DNA repair response, is needed for endothelial cells to maintain their proliferation under hypoxic conditions and is crucial for hypoxia-driven neovascularization.

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Figure 1: Generation of γ-H2AX in response to hypoxia in endothelial cells and the importance of H2AX for endothelial cell proliferation under hypoxia.
Figure 2: A role for H2AX in retinal neovascularization under hypoxia.
Figure 3: H2AX deficiency decreases neovascularization and blood flow after hind limb ischemia.
Figure 4: Tumor angiogenesis and tumor growth are reduced due to H2AX deficiency.

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References

  1. Rouse, J. & Jackson, S.P. Interfaces between the detection, signaling and repair of DNA damage. Science 297, 547–551 (2002).

    Article  CAS  Google Scholar 

  2. McGowan, C.H. & Russell, P. The DNA damage response: sensing and signaling. Curr. Opin. Cell Biol. 16, 629–633 (2004).

    Article  CAS  Google Scholar 

  3. Fernandez-Capetillo, O., Lee, A., Nussenzweig, M. & Nussenzweig, A. H2AX: the histone guardian of the genome. DNA Repair (Amst.) 3, 959–967 (2004).

    Article  CAS  Google Scholar 

  4. Celeste, A. et al. Histone H2AX phosphorylation is dispensable for the initial recognition of DNA breaks. Nat. Cell Biol. 5, 675–679 (2003).

    Article  CAS  Google Scholar 

  5. Bassing, C.H. & Alt, F.W. H2AX may function as an anchor to hold broken chromosomal DNA ends in close proximity. Cell Cycle 3, 149–153 (2004).

    Article  CAS  Google Scholar 

  6. Sedelnikova, O.A., Pilch, D.R., Redon, C. & Bonner, W.M. Histone H2AX in DNA damage and repair. Cancer Biol. Ther. 2, 233–235 (2003).

    Article  CAS  Google Scholar 

  7. Hammond, E.M. & Giaccia, A.J. The role of ATM and ATR in the cellular response to hypoxia and re-oxygenation. DNA Repair (Amst.) 3, 1117–1122 (2004).

    Article  CAS  Google Scholar 

  8. Carmeliet, P. Angiogenesis in life, disease and medicine. Nature 438, 932–936 (2005).

    Article  CAS  Google Scholar 

  9. Hurley, P.J. & Bunz, F. ATM and ATR: components of an integrated circuit. Cell Cycle 6, 414–417 (2007).

    Article  CAS  Google Scholar 

  10. Zhang, Y.W., Hunter, T. & Abraham, R.T. Turning the replication checkpoint on and off. Cell Cycle 5, 125–128 (2006).

    Article  CAS  Google Scholar 

  11. Shiloh, Y. ATM and ATR: networking cellular responses to DNA damage. Curr. Opin. Genet. Dev. 11, 71–77 (2001).

    Article  CAS  Google Scholar 

  12. Lopes, M. et al. The DNA replication checkpoint response stabilizes stalled replication forks. Nature 412, 557–561 (2001).

    Article  CAS  Google Scholar 

  13. Hammond, E.M., Dorie, M.J. & Giaccia, A.J. ATR/ATM targets are phosphorylated by ATR in response to hypoxia and ATM in response to reoxygenation. J. Biol. Chem. 278, 12207–12213 (2003).

    Article  CAS  Google Scholar 

  14. Hammond, E.M., Dorie, M.J. & Giaccia, A.J. Inhibition of ATR leads to increased sensitivity to hypoxia/reoxygenation. Cancer Res. 64, 6556–6562 (2004).

    Article  CAS  Google Scholar 

  15. Gibson, S.L., Bindra, R.S. & Glazer, P.M. Hypoxia-induced phosphorylation of Chk2 in an ataxia telangiectasia mutated-dependent manner. Cancer Res. 65, 10734–10741 (2005).

    Article  CAS  Google Scholar 

  16. Arjamaa, O. & Nikinmaa, M. Oxygen-dependent diseases in the retina: role of hypoxia-inducible factors. Exp. Eye Res. 83, 473–483 (2006).

    Article  CAS  Google Scholar 

  17. Campochiaro, P.A. & Hackett, S.F. Ocular neovascularization: a valuable model system. Oncogene 22, 6537–6548 (2003).

    Article  CAS  Google Scholar 

  18. Noguera-Troise, I. et al. Blockade of Dll4 inhibits tumour growth by promoting non-productive angiogenesis. Nature 444, 1032–1037 (2006).

    Article  CAS  Google Scholar 

  19. Celeste, A. et al. Genomic instability in mice lacking histone H2AX. Science 296, 922–927 (2002).

    Article  CAS  Google Scholar 

  20. Celis, J.E. & Celis, A. Cell cycle-dependent variations in the distribution of the nuclear protein cyclin proliferating cell nuclear antigen in cultured cells: subdivision of S phase. Proc. Natl. Acad. Sci. USA 82, 3262–3266 (1985).

    Article  CAS  Google Scholar 

  21. Zou, L. & Elledge, S.J. Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300, 1542–1548 (2003).

    Article  CAS  Google Scholar 

  22. Economopoulou, M. et al. Inhibition of pathologic retinal neovascularization by alpha-defensins. Blood 106, 3831–3838 (2005).

    Article  CAS  Google Scholar 

  23. Smith, L.E. et al. Oxygen-induced retinopathy in the mouse. Invest. Ophthalmol. Vis. Sci. 35, 101–111 (1994).

    CAS  Google Scholar 

  24. Alva, J.A. et al. VE-Cadherin-Cre-recombinase transgenic mouse: a tool for lineage analysis and gene deletionin endothelial cells. Dev. Dyn. 235, 759–767 (2006).

    Article  CAS  Google Scholar 

  25. Bassing, C.H. et al. Increased ionizing radiation sensitivity and genomic instability in the absence of histone H2AX. Proc. Natl. Acad. Sci. USA 99, 8173–8178 (2002).

    Article  CAS  Google Scholar 

  26. Luttun, A. et al. Revascularization of ischemic tissues by PlGF treatment, and inhibition of tumor angiogenesis, arthritis and atherosclerosis by anti-Flt1. Nat. Med. 8, 831–840 (2002).

    Article  CAS  Google Scholar 

  27. Hammond, E.M., Denko, N.C., Dorie, M.J., Abraham, R.T. & Giaccia, A.J. Hypoxia links ATR and p53 through replication arrest. Mol. Cell. Biol. 22, 1834–1843 (2002).

    Article  CAS  Google Scholar 

  28. Hammond, E.M., Kaufmann, M.R. & Giaccia, A.J. Oxygen-sending and the DNA damage response. Curr. Opin. Cell Biol. 19, 680–684 (2007).

    Article  CAS  Google Scholar 

  29. Huang, L.E., Bindra, R.S., Glazer, P.M. & Harris, A.L. Hypoxia-induced genetic instability—a calculated mechanism underlying tumor progression. J. Mol. Med. 85, 139–148 (2007).

    Article  CAS  Google Scholar 

  30. To, K.K.W., Sedelnikova, O.A., Samons, M., Bonner, W.M. & Huang, L.E. The phosphorylation status of PAS-B distinguishes HIF-1α from HIF-2α in NBS1 repression. EMBO J. 25, 4784–4794 (2006).

    Article  CAS  Google Scholar 

  31. Orlova, V.V., Economopoulou, M., Lupu, F., Santoso, S. & Chavakis, T. Junctional adhesion molecule-C regulates vascular endothelial permeability by modulating VE-cadherin–mediated cell-cell contacts. J. Exp. Med. 203, 2703–2714 (2006).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  33. Lin, M.I., Yu, J., Murata, T. & Sessa, W.C. Caveolin-1–deficient mice have increased tumor microvascular permeability, angiogenesis and growth. Cancer Res. 67, 2849–2856 (2007).

    Article  CAS  Google Scholar 

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Acknowledgements

This research was supported by the Intramural Research Program of the US National Institutes of Health National Cancer Institute. H.F.L. was supported by the German Academy of Sciences (Leopoldina). We would like to acknowledge M.E. Kruhlak for the help with microscopy, D. Winkler and S. Kaul for help with genotyping, G. Tosato for help with Matrigel experiments, F. Alt (Harvard University) for providing the H2afx floxed mice and A. Singer and D.S. Singer for critically reading the manuscript.

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Author notes

  1. Matina Economopoulou and Harald F Langer: These authors contributed equally to this work.

Authors and Affiliations

  1. Experimental Immunology Branch, National Cancer Institute, Bethesda, Maryland, USA

    Matina Economopoulou, Harald F Langer, Arkady Celeste, Valeria V Orlova, Eun Young Choi, Elsa Callen, Andre Nussenzweig & Triantafyllos Chavakis

  2. Translational Medicine Branch, National Heart, Lung and Blood Institute, Bethesda, Maryland, USA

    Mingchao Ma & Manfred Boehm

  3. Genetics of Development and Disease Branch, National Institute for Diabetes and Digestive and Kidney Diseases, US National Institutes of Health, Bethesda, Maryland, USA

    Athanassios Vassilopoulos & Chuxia Deng

  4. Department of Pathology and Laboratory Medicine, Center for Childhood Cancer Research, Children's Hospital of Philadelphia, Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, USA

    Craig H Bassing

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Contributions

M.E. designed and conducted experiments and wrote the manuscript, H.F.L. conducted experiments, A.C. designed experiments, V.V.O. conducted experiments, E.Y.C., M.M., A.V. and E.C. conducted experiments, C.D. supervised experiments, C.H.B. generated H2afx floxed mice, M.B. designed, conducted and supervised experiments, A.N. supervised and designed experiments, and T.C. designed, conducted and supervised experiments and wrote the manuscript.

Corresponding author

Correspondence to Triantafyllos Chavakis.

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Economopoulou, M., Langer, H., Celeste, A. et al. Histone H2AX is integral to hypoxia-driven neovascularization. Nat Med 15, 553–558 (2009). https://doi.org/10.1038/nm.1947

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