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Identification of human brain tumour initiating cells

Nature volume 432, pages 396–401 (2004)Cite this article

Abstract

The cancer stem cell (CSC) hypothesis suggests that neoplastic clones are maintained exclusively by a rare fraction of cells with stem cell properties1,2. Although the existence of CSCs in human leukaemia is established3,4, little evidence exists for CSCs in solid tumours, except for breast cancer5. Recently, we prospectively isolated a CD133+ cell subpopulation from human brain tumours that exhibited stem cell properties in vitro6. However, the true measures of CSCs are their capacity for self renewal and exact recapitulation of the original tumour1,2,7. Here we report the development of a xenograft assay that identified human brain tumour initiating cells that initiate tumours in vivo. Only the CD133+ brain tumour fraction contains cells that are capable of tumour initiation in NOD-SCID (non-obese diabetic, severe combined immunodeficient) mouse brains. Injection of as few as 100 CD133+ cells produced a tumour that could be serially transplanted and was a phenocopy of the patient's original tumour, whereas injection of 105 CD133- cells engrafted but did not cause a tumour. Thus, the identification of brain tumour initiating cells provides insights into human brain tumour pathogenesis, giving strong support for the CSC hypothesis as the basis for many solid tumours5, and establishes a previously unidentified cellular target for more effective cancer therapies.

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Figure 1: CD133+ tumour cells initiate tumours upon intracranial transplantation into the adult NOD-SCID mouse forebrain.
Figure 2: CD133+ xenograft from a medulloblastoma (right side) resembles the original patient tumour (left side).
Figure 3: CD133+ xenograft from a GBM (right) resembles the original patient tumour (left).
Figure 4: Only the CD133+ BTIC, and not the CD133- tumour cell, is capable of tumour initiation.

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Acknowledgements

We thank M. Borden, J. Ma, I. Diplock, M. Ho and C. Gibson for technical assistance, and we are grateful to V. Bonn, L. Davidson, N. Lifshitz and J. Chen of the Mouse Imaging Centre for help with neuroimaging. We thank J. Dick for discussions. S. Singh was supported by a a Terry Fox Foundation fellowship from the Canadian Cancer Society, the Neurosurgical Research and Education Foundation and the American Brain Tumor Association. This work was supported by the Canadian Cancer Society, the Canadian Institutes of Health Research, the Foundation of The Hospital for Sick Children, BrainChild and the Jack Baker and Jessica Durigon family funds.

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Authors and Affiliations

  1. The Arthur and Sonia Labatt Brain Tumor Research Centre, University of Toronto, 555 University Avenue, Toronto, M5G 1X8, Canada

    Sheila K. Singh, Cynthia Hawkins, Ian D. Clarke, Takuichiro Hide & Peter B. Dirks

  2. Program in Developmental Biology, University of Toronto, 555 University Avenue, Toronto, M5G 1X8, Canada

    Sheila K. Singh, Ian D. Clarke, Takuichiro Hide & Peter B. Dirks

  3. Division of Neurosurgery, University of Toronto, 555 University Avenue, M5G 1X8, Toronto, Canada

    Sheila K. Singh, Michael D. Cusimano & Peter B. Dirks

  4. Department of Pediatric Laboratory Medicine, University of Toronto, 555 University Avenue, M5G 1X8, Toronto, Canada

    Cynthia Hawkins

  5. Integrative Biology Program, The Hospital for Sick Children and University of Toronto, 555 University Avenue, M5G 1X8, Toronto, Canada

    R. Mark Henkelman

  6. Ontario Cancer Institute and University of Toronto, 610 University Avenue, M5G 2M9, Toronto, Canada

    Jeremy A. Squire & Jane Bayani

  7. Division of Neurosurgery, St Michael's Hospital and University of Toronto, 30 Bond Street, M5B 1W8, Toronto, Canada

    Michael D. Cusimano

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Corresponding author

Correspondence to Peter B. Dirks.

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Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Table 1

Patient information. (DOC 23 kb)

Supplementary Table 2

CD133 frequency and primary sphere formation rate of sorted tumours. (DOC 24 kb)

Supplementary Table 3

Summary of tumour cell injections into NOD–SCID mice. (DOC 28 kb)

Supplementary Table 4

Comparison of histology and immunohistochemistry of CD133+ xenotransplanted tumours and original patient tumours. (DOC 39 kb)

Supplementary Figure 1

Phenotypic characteristics of glioblastoma and medulloblastoma xenografts. (DOC 273 kb)

Supplementary Figure 2

CD133+ and CD133- cells from an original patient medulloblastoma show clonal karyotype abnormalities. (DOC 248 kb)

Supplementary Figure 3

CD133+ and CD133- cells from patient GBMs show clonal karyotype abnormalities. (DOC 730 kb)

Supplementary Figure 4

EGFR Amplification in GBM and resulting xenografts. (DOC 758 kb)

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Singh, S., Hawkins, C., Clarke, I. et al. Identification of human brain tumour initiating cells. Nature 432, 396–401 (2004). https://doi.org/10.1038/nature03128

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