Ectopic expression of the histone methyltransferase Ezh2 in haematopoietic stem cells causes myeloproliferative disease

  • Nature Communications 3, Article number: 623 (2012)
  • doi:10.1038/ncomms1623
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Recent evidence shows increased and decreased expression of Ezh2 in cancer, suggesting a dual role as an oncogene or tumour suppressor. To investigate the mechanism by which Ezh2-mediated H3K27 methylation leads to cancer, we generated conditional Ezh2 knock-in (Ezh2-KI) mice. Here we show that induced Ezh2 haematopoietic expression increases the number and proliferation of repopulating haematopoietic stem cells. Ezh2-KI mice develop myeloproliferative disorder, featuring excessive myeloid expansion in bone marrow and spleen, leukocytosis and splenomegaly. Competitive and serial transplantations demonstrate progressive myeloid commitment of Ezh2-KI haematopoietic stem cells. Transplanted self-renewing haematopoietic stem cells from Ezh2-KI mice induce myeloproliferative disorder, suggesting that the Ezh2 gain-of-function arises in the haematopoietic stem cell pool, and not at later stages of myelopoiesis. At the molecular level, Ezh2 regulates haematopoietic stem cell-specific genes such as Evi-1 and Ntrk3, aberrantly found in haematologic malignancies. These results demonstrate a stem cell-specific Ezh2 oncogenic role in myeloid disorders, and suggest possible therapeutic applications in Ezh2-related haematological malignancies.

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Change history

  • Updated online 07 March 2017

    This paper has been retracted at the request of the authors.


  1. 1.

    & Hematopoietic stem cell self-renewal. Curr. Opin. Genet. Dev. 16, 496–501 (2006).

  2. 2.

    & Polycomb group proteins: navigators of lineage pathways led astray in cancer. Nat. Rev. Cancer 9, 773–784 (2009).

  3. 3.

    & The Polycomb complex PRC2 and its mark in life. Nature 469, 343–349 (2011).

  4. 4.

    et al. An embryonic stem cell-like gene expression signature in poorly differentiated aggressive human tumors. Nat. Genet. 40, 499–507 (2008).

  5. 5.

    et al. Polycomb-mediated methylation on Lys27 of histone H3 pre-marks genes for de novo methylation in cancer. Nat. Genet. 39, 232–236 (2007).

  6. 6.

    , & Polycomb proteins in hematologic malignancies. Blood 116, 5465–5475 (2010).

  7. 7.

    et al. Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nat. Genet. 42, 181–185 (2010).

  8. 8.

    et al. Coordinated activities of wild-type plus mutant EZH2 drive tumor-associated hypertrimethylation of lysine 27 on histone H3 (H3K27) in human B-cell lymphomas. Proc. Natl Acad. Sci. U S A 107, 20980–20985 (2011).

  9. 9.

    et al. Somatic mutations at EZH2 Y641 act dominantly through a mechanism of selectively altered PRC2 catalytic activity, to increase H3K27 trimethylation. Blood 117, 2451–2459 (2010).

  10. 10.

    et al. Overexpression of the EZH2, RING1 and BMI1 genes is common in myelodysplastic syndromes: relation to adverse epigenetic alteration and poor prognostic scoring. Ann. Hematol. 90, 643–653 (2010).

  11. 11.

    et al. Signatures of polycomb repression and reduced H3K4 trimethylation are associated with p15INK4b DNA methylation in AML. Blood 115, 3098–3108 (2010).

  12. 12.

    et al. Combined epigenetic therapy with the histone methyltransferase EZH2 inhibitor 3-deazaneplanocin A and the histone deacetylase inhibitor panobinostat against human AML cells. Blood 114, 2733–2743 (2009).

  13. 13.

    et al. Somatic mutations of the histone methyltransferase gene EZH2 in myelodysplastic syndromes. Nat. Genet. 42, 665–667 (2010).

  14. 14.

    et al. Inactivating mutations of the histone methyltransferase gene EZH2 in myeloid disorders. Nat. Genet. 42, 722–726 (2010).

  15. 15.

    et al. Transgenic mice with hematopoietic and lymphoid specific expression of Cre. Eur. J. Immunol. 33, 314–325 (2003).

  16. 16.

    et al. SOCS3 is a critical physiological negative regulator of G-CSF signaling and emergency granulopoiesis. Immunity 20, 153–165 (2004).

  17. 17.

    et al. Promoter elements of vav drive transgene expression in vivo throughout the hematopoietic compartment. Blood 94, 1855–1863 (1999).

  18. 18.

    et al. Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J. Exp. Med. 183, 1797–1806 (1996).

  19. 19.

    et al. Restoration of p53 function leads to tumour regression in vivo. Nature 445, 661–665 (2007).

  20. 20.

    et al. Conditional gene targeting in macrophages and granulocytes using LysMcre mice. Transgenic Res. 8, 265–277 (1999).

  21. 21.

    et al. Global analysis of proliferation and cell cycle gene expression in the regulation of hematopoietic stem and progenitor cell fates. J. Exp. Med. 202, 1599–1611 (2005).

  22. 22.

    et al. Gene ontology: tool for the unification of biology. The gene ontology consortium. Nat. Genet. 25, 25–29 (2000).

  23. 23.

    , & Lack of evidence that hematopoietic stem cells depend on N-cadherin-mediated adhesion to osteoblasts for their maintenance. Cell Stem Cell 1, 204–217 (2007).

  24. 24.

    et al. Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell 118, 149–161 (2004).

  25. 25.

    et al. Id1 is a common downstream target of oncogenic tyrosine kinases in leukemic cells. Blood 112, 1981–1992 (2008).

  26. 26.

    et al. Molecular signatures of quiescent, mobilized and leukemia-initiating hematopoietic stem cells. PLoS One 5, e8785 (2010).

  27. 27.

    et al. EVI1 induces myelodysplastic syndrome in mice. J. Clin. Invest. 114, 713–719 (2004).

  28. 28.

    et al. Evi-1 is a critical regulator for hematopoietic stem cells and transformed leukemic cells. Cell Stem Cell 3, 207–220 (2008).

  29. 29.

    et al. EVI-1 interacts with histone methyltransferases SUV39H1 and G9a for transcriptional repression and bone marrow immortalization. Leukemia 24, 81–88 (2010).

  30. 30.

    et al. The Polycomb group proteins bind throughout the INK4A-ARF locus and are disassociated in senescent cells. Genes Dev. 21, 525–530 (2007).

  31. 31.

    & Polycomb group proteins: multi-faceted regulators of somatic stem cells and cancer. Cell Stem Cell 7, 299–313 (2010).

  32. 32.

    & Oncogenes in myeloproliferative disorders. Cell Cycle 6, 550–566 (2007).

  33. 33.

    et al. Bethesda proposals for classification of nonlymphoid hematopoietic neoplasms in mice. Blood 100, 238–245 (2002).

  34. 34.

    et al. The Polycomb group protein EZH2 directly controls DNA methylation. Nature 439, 871–874 (2006).

  35. 35.

    et al. A stem cell-like chromatin pattern may predispose tumor suppressor genes to DNA hypermethylation and heritable silencing. Nat. Genet. 39, 237–242 (2007).

  36. 36.

    et al. Epigenetic stem cell signature in cancer. Nat. Genet. 39, 157–158 (2007).

  37. 37.

    & Inflammation and cancer. Nature 420, 860–867 (2002).

  38. 38.

    & The epigenomics of cancer. Cell 128, 683–692 (2007).

  39. 39.

    et al. Clinical effect of point mutations in myelodysplastic syndromes. N. Engl. J. Med. 364, 2496–2506 (2011).

  40. 40.

    & Polycomb complexes and silencing mechanisms. Curr. Opin. Cell Biol. 16, 239–246 (2004).

  41. 41.

    & Regulating quiescence: new insights into hematopoietic stem cell biology. Dev. Cell 10, 415–417 (2006).

  42. 42.

    et al. Hematopoiesis and leukemogenesis in mice expressing oncogenic NrasG12D from the endogenous locus. Blood 117, 2022–2032 (2011).

  43. 43.

    Regulating the leukaemia stem cell. Best Pract. Res. Clin. Haematol. 22, 483–487 (2009).

  44. 44.

    et al. Somatic activation of oncogenic Kras in hematopoietic cells initiates a rapidly fatal myeloproliferative disorder. Proc. Natl Acad. Sci. U S A 101, 597–602 (2004).

  45. 45.

    et al. Physiological Jak2V617F expression causes a lethal myeloproliferative neoplasm with differential effects on hematopoietic stem and progenitor cells. Cancer Cell 17, 584–596 (2010).

  46. 46.

    , & Mechanisms controlling hematopoietic stem cell functions during normal hematopoiesis and hematological malignancies. Wiley Interdiscip. Rev. Syst. Biol. Med. 3, 681–701 (2011).

  47. 47.

    et al. miR-33-mediated downregulation of p53 controls hematopoietic stem cell self-renewal. Cell Cycle 9, 3277–3285 (2010).

  48. 48.

    , & Mechanistic principles of chromatin remodeling guided by siRNAs and miRNAs. Cell Cycle 7, 2601–2608 (2008).

  49. 49.

    et al. Oncogenic activity of Cdc6 through repression of the INK4/ARF locus. Nature 440, 702–706 (2006).

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We are grateful to Giovanna Giovinazzo, Luis-Miguel Criado, Maria Montoya and their teams (CNIC) for technical support. We also thank Simon Bartlett (CNIC) for English editing, Dr Serrano for helpful discussion, and Purificacion Arribas for her wonderful technical assistance. S. Gonzalez is funded by the Human Frontiers Science Program Organization, the Spanish Ministries of Science and Innovation (SAF2010-15386) and Health (FIS PI06/0627) The CNIC is supported by the Ministry of Science and Innovation and the Pro-CNIC Foundation.

Author information


  1. Stem Cell Aging Group, Spanish National Cardiovascular Research Center (CNIC), E-28029 Madrid, Spain.

    • A. Herrera-Merchan
    • , L. Arranz
    •  & S. Gonzalez
  2. Cellomic Unit, Spanish National Cardiovascular Research Center (CNIC), E-28029 Madrid, Spain.

    • J.M. Ligos
  3. Animal Unit, Spanish National Cardiovascular Research Center (CNIC), E-28029 Madrid, Spain.

    • A. de Molina
  4. Genomics Unit, Spanish National Cancer Research Center (CNIO), E-28029 Madrid, Spain.

    • O. Dominguez


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A.H-M. performed most of the experiments, contributed to data analysis, and discussion of the paper. L.A. helps with technical issues regarding gene expression assays. J.M.L. supervised the cytometric analyses. A.M. performed all pathological analyses. O.D. designed and supervised the gene expression data. S.G. designed and supervised the study, secured funding, analysed the data and wrote the paper. All authors discussed the results and commented on the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to S. Gonzalez.

Supplementary information

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    Supplementary Information

    Supplementary Figures S1-S14, Supplementary Tables S1-S3, Supplementary Methods and Supplementary References


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