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Glioma stem cells promote radioresistance by preferential activation of the DNA damage response

Abstract

Ionizing radiation represents the most effective therapy for glioblastoma (World Health Organization grade IV glioma), one of the most lethal human malignancies1, but radiotherapy remains only palliative2 because of radioresistance. The mechanisms underlying tumour radioresistance have remained elusive. Here we show that cancer stem cells contribute to glioma radioresistance through preferential activation of the DNA damage checkpoint response and an increase in DNA repair capacity. The fraction of tumour cells expressing CD133 (Prominin-1), a marker for both neural stem cells and brain cancer stem cells3,4,5,6, is enriched after radiation in gliomas. In both cell culture and the brains of immunocompromised mice, CD133-expressing glioma cells survive ionizing radiation in increased proportions relative to most tumour cells, which lack CD133. CD133-expressing tumour cells isolated from both human glioma xenografts and primary patient glioblastoma specimens preferentially activate the DNA damage checkpoint in response to radiation, and repair radiation-induced DNA damage more effectively than CD133-negative tumour cells. In addition, the radioresistance of CD133-positive glioma stem cells can be reversed with a specific inhibitor of the Chk1 and Chk2 checkpoint kinases. Our results suggest that CD133-positive tumour cells represent the cellular population that confers glioma radioresistance and could be the source of tumour recurrence after radiation. Targeting DNA damage checkpoint response in cancer stem cells may overcome this radioresistance and provide a therapeutic model for malignant brain cancers.

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Figure 1: Enrichment of CD133 + tumour subpopulations after irradiation in vitro and in vivo , and enhancement of intracranial tumour formation by increased CD133 + fraction.
Figure 2: Characterization of CD133 + and CD133 - cells from human glioma xenografts and primary glioblastoma specimens.
Figure 3: CD133 + tumour cells show radioresistance and lower sensitivity to radiation-induced apoptosis than CD133 - tumour cells dependent on checkpoint kinase activity.
Figure 4: CD133 + glioma cells preferentially activate the DNA damage checkpoint and repair IR-induced DNA damage more efficiently than CD133 - cells.

References

  1. Legler, J. M. et al. Brain and other central nervous system cancers: recent trends in incidence and mortality. J. Natl Cancer Inst. 91, 1382–1390 (1999)

    Article  CAS  PubMed  Google Scholar 

  2. Garden, A. S. et al. Outcome and patterns of failure following limited-volume irradiation for malignant astrocytomas. Radiother. Oncol. 20, 99–110 (1991)

    Article  CAS  PubMed  Google Scholar 

  3. Singh, S. K. et al. Identification of human brain tumour initiating cells. Nature 432, 396–401 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  4. Hemmati, H. D. et al. Cancerous stem cells can arise from pediatric brain tumours. Proc. Natl Acad. Sci. USA 100, 15178–15183 (2003)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  5. Singh, S. K. et al. Identification of a cancer stem cell in human brain tumours. Cancer Res. 63, 5821–5828 (2003)

    CAS  PubMed  Google Scholar 

  6. Uchida, N. et al. Direct isolation of human central nervous system stem cells. Proc. Natl Acad. Sci. USA 97, 14720–14725 (2000)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  7. Galli, R. et al. Isolation and characterization of tumourigenic, stem-like neural precursors from human glioblastoma. Cancer Res. 64, 7011–7021 (2004)

    Article  CAS  PubMed  Google Scholar 

  8. Izant, J. G., Weatherbee, J. A. & McIntosh, J. R. A microtubule-associated protein antigen unique to mitotic spindle microtubules in PtK1 cells. J. Cell Biol. 96, 424–434 (1983)

    Article  CAS  PubMed  Google Scholar 

  9. Ignatova, T. N. et al. Human cortical glial tumours contain neural stem-like cells expressing astroglial and neuronal markers in vitro.. Glia 39, 193–206 (2002)

    Article  PubMed  Google Scholar 

  10. Sanai, N. et al. Unique astrocyte ribbon in adult human brain contains neural stem cells but lacks chain migration. Nature 427, 740–744 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  11. Reynolds, B. A. & Weiss, S. Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255, 1707–1710 (1992)

    Article  ADS  CAS  PubMed  Google Scholar 

  12. Oostendorp, R. A., Audet, J., Miller, C. & Eaves, C. J. Cell division tracking and expansion of hematopoietic long-term repopulating cells. Leukemia 13, 499–501 (1999)

    Article  CAS  PubMed  Google Scholar 

  13. Marin, L. et al. Flow cytometric quantification of apoptosis and proliferation in mixed lymphocyte culture. Cytometry A 51, 107–118 (2003)

    Article  PubMed  Google Scholar 

  14. Abraham, R. T. Cell cycle checkpoint signaling through the ATM and ATR kinases. Genes Dev. 15, 2177–2196 (2001)

    Article  CAS  PubMed  Google Scholar 

  15. Carr, A. M. DNA structure dependent checkpoints as regulators of DNA repair. DNA Repair 1, 983–994 (2002)

    Article  CAS  PubMed  Google Scholar 

  16. Zhou, B. B. & Elledge, S. J. The DNA damage response: putting checkpoints in perspective. Nature 408, 433–439 (2000)

    Article  ADS  CAS  PubMed  Google Scholar 

  17. Kastan, M. B. & Bartek, J. Cell-cycle checkpoints and cancer. Nature 432, 316–323 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  18. Ali, A. et al. Requirement of protein phosphatase 5 in DNA-damage-induced ATM activation. Genes Dev. 18, 249–254 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Bao, S. et al. ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses. Nature 411, 969–974 (2001)

    Article  ADS  CAS  PubMed  Google Scholar 

  20. Sancar, A., Lindsey-Boltz, L. A., Unsal-Kacmaz, K. & Linn, S. Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu. Rev. Biochem. 73, 39–85 (2004)

    Article  CAS  PubMed  Google Scholar 

  21. Tice, R. R. et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen. 35, 206–221 (2000)

    Article  CAS  PubMed  Google Scholar 

  22. Balajee, A. S. & Geard, C. R. Replication protein A and γ-H2AX foci assembly is triggered by cellular response to DNA double-strand breaks. Exp. Cell Res. 300, 320–334 (2004)

    Article  CAS  PubMed  Google Scholar 

  23. Curman, D. et al. Inhibition of the G2 DNA damage checkpoint and of protein kinases Chk1 and Chk2 by the marine sponge alkaloid debromohymenialdisine. J. Biol. Chem. 276, 17914–17919 (2001)

    Article  CAS  PubMed  Google Scholar 

  24. Gorgoulis, V. G. et al. Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature 434, 907–913 (2005)

    Article  ADS  CAS  PubMed  Google Scholar 

  25. Bartkova, J. et al. DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434, 864–870 (2005)

    Article  ADS  CAS  PubMed  Google Scholar 

  26. Bao, S. et al. Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor. Cancer Res. 66, 7843–7848 (2006)

    Article  CAS  PubMed  Google Scholar 

  27. Lee, A. et al. Isolation of neural stem cells from the postnatal cerebellum. Nature Neurosci. 8, 723–729 (2005)

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Y. H. Sun, S. Keir, D. Satterfield, L. Ehinger and J. Faison for technical assistance; M. Cook and T. R. Dissanayake for assistance with flow cytometry; Z. Lu for assistance with fluorescent microscopy; and X.-F. Wang, H. Lin, T.P. Yao, H. Friedman and R. Wechsler-Reya for discussions. Financial support was provided by the Childhood Brain Tumor Foundation, the Pediatric Brain Tumor Foundation of the United States, Accelerate Brain Cancer Cure, a grant from the Duke Comprehensive Cancer Center Kislak–Fields Family Fund (to J.N.R.), and grants from the NIH (to J.N.R. and to D.D.B.). J.N.R. is a Damon Runyon-Lilly Clinical Investigator supported by the Damon Runyon Cancer Research Foundation and a Sidney Kimmel Foundation for Cancer Research Scholar. A.B.H. is a Paul Brazen/American Brain Tumor Association Fellow. Author Contributions Q.W., S.B., Y.H. and Q.S. did the experimental work. R.E.M. performed pathological analysis and assisted in human tumour specimen acquisition. S.B. and J.N.R. wrote the paper and designed the experiments. A.B.H., M.W.D. and D.D.B. provided intellectual input and helped with experimental design.

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Correspondence to Jeremy N. Rich.

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Supplementary Notes

This file contains Supplementary Tables and Supplementary Figures 1–17. (PDF 4052 kb)

Supplementary Methods

This file contains additional details of the methods used in this study. (DOC 36 kb)

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Bao, S., Wu, Q., McLendon, R. et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 444, 756–760 (2006). https://doi.org/10.1038/nature05236

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