Multi-genetic events collaboratively contribute to Pten-null leukaemia stem-cell formation

Abstract

Cancer stem cells, which share many common properties and regulatory machineries with normal stem cells, have recently been proposed to be responsible for tumorigenesis and to contribute to cancer resistance1. The main challenges in cancer biology are to identify cancer stem cells and to define the molecular events required for transforming normal cells to cancer stem cells. Here we show that Pten deletion in mouse haematopoietic stem cells leads to a myeloproliferative disorder, followed by acute T-lymphoblastic leukaemia (T-ALL). Self-renewable leukaemia stem cells (LSCs) are enriched in the c-KitmidCD3+Lin- compartment, where unphosphorylated β-catenin is significantly increased. Conditional ablation of one allele of the β-catenin gene substantially decreases the incidence and delays the occurrence of T-ALL caused by Pten loss, indicating that activation of the β-catenin pathway may contribute to the formation or expansion of the LSC population. Moreover, a recurring chromosomal translocation, T(14;15), results in aberrant overexpression of the c-myc oncogene in c-KitmidCD3+Lin- LSCs and CD3+ leukaemic blasts, recapitulating a subset of human T-ALL. No alterations in Notch1 signalling are detected in this model, suggesting that Pten inactivation and c-myc overexpression may substitute functionally for Notch1 abnormalities2,3, leading to T-ALL development. Our study indicates that multiple genetic or molecular alterations contribute cooperatively to LSC transformation.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: VEC-Cre -mediated Pten loss leads to MPD and leukaemogenesis.
Figure 2: Self-renewing LSCs are enriched in the c-Kit mid CD3 + compartment.
Figure 3: β-Catenin activation in LSCs and its role in leukaemogenesis.
Figure 4: The recurring translocation T(14;15) involves the Tcra/Tcrd cluster and the c- myc gene and results in aberrant overexpression of c- myc in LSCs and T-ALL blasts.

References

  1. 1

    Reya, T., Morrison, S. J., Clarke, M. F. & Weissman, I. L. Stem cells, cancer, and cancer stem cells. Nature 414, 105–111 (2001)

  2. 2

    Maser, R. S. et al. Chromosomally unstable mouse tumours have genomic alterations similar to diverse human cancers. Nature 447, 966–971 (2007)

  3. 3

    Weng, A. P. et al. Activating mutations of NOTCH1 in human T cell acute lymphoblastic leukemia. Science 306, 269–271 (2004)

  4. 4

    Fukuda, R. et al. Alteration of phosphatidylinositol 3-kinase cascade in the multilobulated nuclear formation of adult T cell leukemia/lymphoma (ATLL). Proc. Natl Acad. Sci. USA 102, 15213–15218 (2005)

  5. 5

    Yilmaz, O. H. et al. Pten dependence distinguishes haematopoietic stem cells from leukaemia-initiating cells. Nature 441, 475–482 (2006)

  6. 6

    Zhang, J. et al. PTEN maintains haematopoietic stem cells and acts in lineage choice and leukaemia prevention. Nature 441, 518–522 (2006)

  7. 7

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

  8. 8

    Borowitz, M. J., Guenther, K. L., Shults, K. E. & Stelzer, G. T. Immunophenotyping of acute leukemia by flow cytometric analysis. Use of CD45 and right-angle light scatter to gate on leukemic blasts in three-color analysis. Am. J. Clin. Pathol. 100, 534–540 (1993)

  9. 9

    Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)

  10. 10

    Bonnet, D. & Dick, J. E. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nature Med. 3, 730–737 (1997)

  11. 11

    Passegue, E., Wagner, E. F. & Weissman Jun, I. L. B deficiency leads to a myeloproliferative disorder arising from hematopoietic stem cells. Cell 119, 431–443 (2004)

  12. 12

    Castor, A. et al. Distinct patterns of hematopoietic stem cell involvement in acute lymphoblastic leukemia. Nature Med. 11, 630–637 (2005)

  13. 13

    Jamieson, C. H. et al. Granulocyte–macrophage progenitors as candidate leukemic stem cells in blast-crisis CML. N. Engl. J. Med. 351, 657–667 (2004)

  14. 14

    Kelly, P. N. et al. Tumor growth need not be driven by rare cancer stem cells. Science 317, 337 (2007)

  15. 15

    Reya, T. et al. A role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 423, 409–414 (2003)

  16. 16

    Erikson, J. et al. Deregulation of c-myc by translocation of the alpha-locus of the T-cell receptor in T-cell leukemias. Science 232, 884–886 (1986)

  17. 17

    Boxer, L. M. & Dang, C. V. Translocations involving c-myc and c-myc function. Oncogene 20, 5595–5610 (2001)

  18. 18

    Weissman, I. Stem cell research: paths to cancer therapies and regenerative medicine. J. Am. Med. Assoc. 294, 1359–1366 (2005)

  19. 19

    Shen, W. H. et al. Essential role for nuclear PTEN in maintaining chromosomal integrity. Cell 128, 157–170 (2007)

  20. 20

    Palomero, T. et al. NOTCH1 directly regulates c-MYC and activates a feed-forward-loop transcriptional network promoting leukemic cell growth. Proc. Natl Acad. Sci. USA 103, 18261–18266 (2006)

  21. 21

    Sharma, V. M. et al. Notch1 contributes to mouse T-cell leukemia by directly inducing the expression of c-myc . Mol. Cell. Biol. 26, 8022–8031 (2006)

  22. 22

    Weng, A. P. et al. c-Myc is an important direct target of Notch1 in T-cell acute lymphoblastic leukemia/lymphoma. Genes Dev. 20, 2096–2109 (2006)

  23. 23

    Palomero, T. et al. Mutational loss of PTEN induces resistance to NOTCH1 inhibition in T-cell leukemia. Nature Med. 13, 1203–1210 (2007)

  24. 24

    Lesche, R. et al. Cre/loxP-mediated inactivation of the murine Pten tumor suppressor gene. Genesis 32, 148–149 (2002)

  25. 25

    Brault, V. et al. Inactivation of the β-catenin gene by Wnt1-Cre-mediated deletion results in dramatic brain malformation and failure of craniofacial development. Development 128, 1253–1264 (2001)

  26. 26

    Putti, M. C. et al. Expression of myeloid markers lacks prognostic impact in children treated for acute lymphoblastic leukemia: Italian experience in AIEOP-ALL 88–91 studies. Blood 92, 795–801 (1998)

  27. 27

    Boomer, T. et al. Detection of E2A translocations in leukemias via fluorescence in situ hybridization. Leukemia 15, 95–102 (2001)

Download references

Acknowledgements

We thank O. Witte, K. Dorshkind, J. Said, R. Gatti, K. Sakamoto, S. Schubbert, R. Hill and B. Valamehr for helpful comments; D. Cheng from O. Witte’s laboratory for cell sorting; J. Gao for retro-orbital bleeding and genotyping; and the Department of Pathology’s Tissue Procurement Core Laboratory (partly supported by National Cancer Institute grant CA16042) for tissue procurement. W.G. is supported by a California Institute of Regenerative Medicine (CIRM) training fellowship. J.E.Y. was supported by the University of California Los Angeles Amgen Scholar Program. J.Y.C. is supported by the Beckman Undergraduate Research Program.

Author Contributions W.G. and H.W. designed the experiments, and W.G. performed a majority of the work involved in this study. J.L.L. performed immunohistochemistry on tissue sections and cytospin slides (Fig. 3a), and participated in FACS-Gal analysis, PB profile analysis and histological analysis. C.-J.C. performed RT–PCR and western blotting for c-myc expression (Fig. 4e and data not shown). X.L. maintained mouse colonies and was responsible for intravenous injection; S.M. performed western blotting and TOPflash reporter assay for β-catenin activation by c-Myc (Supplementary Fig. 9c, d). J.E.Y. performed Notch1 mutation screening and RT–PCR analysis for Fbxw7 and Hes1 expression (Supplementary Fig. 10). J.Y.C. performed Pten and Ctnnb1 genotyping and FACS analysis; Y.X. and M.V.-G. performed SKY and FISH analysis (Fig. 4a, b, d, and Supplementary Fig. 8). M.L.I.-A. provided VE-Cadherin-Cre+ mice. W.G., J.L.L. and H.W. wrote the paper. All authors discussed the results and commented on the manuscript.

Author information

Correspondence to Hong Wu.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-11 with Legends. (PDF 4635 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Guo, W., Lasky, J., Chang, C. et al. Multi-genetic events collaboratively contribute to Pten-null leukaemia stem-cell formation. Nature 453, 529–533 (2008). https://doi.org/10.1038/nature06933

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.