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Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells

Nature volume 441, pages 475–482 (2006)Cite this article

Abstract

Recent advances have highlighted extensive phenotypic and functional similarities between normal stem cells and cancer stem cells. This raises the question of whether disease therapies can be developed that eliminate cancer stem cells without eliminating normal stem cells. Here we address this issue by conditionally deleting the Pten tumour suppressor gene in adult haematopoietic cells. This led to myeloproliferative disease within days and transplantable leukaemias within weeks. Pten deletion also promoted haematopoietic stem cell (HSC) proliferation. However, this led to HSC depletion via a cell-autonomous mechanism, preventing these cells from stably reconstituting irradiated mice. In contrast to leukaemia-initiating cells, HSCs were therefore unable to maintain themselves without Pten. These effects were mostly mediated by mTOR as they were inhibited by rapamycin. Rapamycin not only depleted leukaemia-initiating cells but also restored normal HSC function. Mechanistic differences between normal stem cells and cancer stem cells can thus be targeted to deplete cancer stem cells without damaging normal stem cells.

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Figure 1: Pten deletion from adult haematopoietic cells leads to myeloproliferative disease that progresses to AML and ALL.
Figure 2: HSCs proliferate after Pten deletion, transiently expanding in number before becoming depleted.
Figure 3: Pten is required cell-autonomously for HSC maintenance.
Figure 4: AML- and ALL-initiating cells are rare in Pten -deleted mice, but are transplantable, are contained within multiple distinct populations, and are highly enriched among cells that express HSC markers.
Figure 5: Rapamycin depletes leukaemia-initiating cells.
Figure 6: Rapamycin rescues normal HSC function after Pten deletion.

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Acknowledgements

This work was supported by the Howard Hughes Medical Institute. O.H.Y. was supported by a predoctoral fellowship from the University of Michigan (UM) Institute of Gerontology. We thank the UM Flow-cytometry Core Facility, which was supported by the UM-Comprehensive Cancer Center. We also thank E. Smith in the Hybridoma Core Facility for antibody production, supported in part through the Rheumatic Disease Core Center; A. Burgess and N. McAnsh of the UM Comprehensive Cancer Center Tissue Core; and C. Mountford for excellent mouse colony management. Author Contributions O.H.Y. performed all experiments and participated in the design and interpretation of experiments. R.V. performed all pathology on the mice with help from O.H.Y. B.K.T. and D.O.F. performed spectral karyotype analysis with help from O.H.Y. W.G. and H.W. provided the Ptenfl/fl mice and discussed pre-publication results. S.J.M. participated in the design and interpretation of experiments, and wrote the paper with O.H.Y.

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Authors and Affiliations

  1. Howard Hughes Medical Institute, Life Sciences Institute, Department of Internal Medicine, and Center for Stem Cell Biology, University of Michigan, Ann Arbor, Michigan, 48109-2216, USA

    Ömer H. Yilmaz & Sean J. Morrison

  2. Department of Pathology, University of Michigan, Ann Arbor, Michigan, 48109-2216, USA

    Riccardo Valdez, Brian K. Theisen & David O. Ferguson

  3. Molecular and Medical Pharmacology, UCLA School of Medicine, Los Angeles, California, 90095-1735, USA

    Wei Guo & Hong Wu

Authors
  1. Ömer H. Yilmaz
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Correspondence to Sean J. Morrison.

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Supplementary information

Supplementary Figure 1

Seven injections of pIpC over a 14 day period led to complete deletion of Pten in mature myeloid cells and in HSCs of adult Pten fl/fl Mx-1-Cre mice. (PDF 129 kb)

Supplementary Figure 2

The frequencies of HSCs and hematopoietic progenitors increases significantly in the spleen within days of Pten deletion. (PDF 277 kb)

Supplementary Figure 3

Myeloid leukemias acquired significant aneuploidy and/or chromosomal translocations after Pten deletion and were clonal or oligoclonal. (PDF 232 kb)

Supplementary Figure 4

No effect of Pten deletion on the rate of cell death observed within whole bone marrow or within the Flk2-Sca-1+Lin-c-kit+CD48- HSC population. (PDF 265 kb)

Supplementary Figure 5

Pten deletion did not affect the clonogenicity or differentiation of HSCs in culture. (PDF 92 kb)

Supplementary Figure 6

Rapamycin reduced the frequency of AML blasts cells that formed colonies in methylcellulose, the size of those colonies, and the percentage of cultured blast cells in S phase of the cell cycle. (PDF 179 kb)

Supplementary Methods

This file contains additional details of the methods used in this study. (DOC 104 kb)

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Yilmaz, Ö., Valdez, R., Theisen, B. et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature 441, 475–482 (2006). https://doi.org/10.1038/nature04703

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