Abstract
Acute myeloid leukemia (AML) is an aggressive and lethal blood cancer originating from rare populations of leukemia stem cells (LSCs). AML relapse after conventional chemotherapy is caused by a remaining population of drug-resistant LSCs. Selective targeting of the chemoresistant population is a promising strategy for preventing and treating AML relapse. Polycomb repressive complex 2 (PRC2) trimethylates histone H3 at lysine 27 to maintain the stemness of LSCs. Here, we show that quiescent LSCs expressed the highest levels of enhancer of zeste (EZH) 1 and EZH2, the PRC2 catalytic subunits, in the AML hierarchy, and that dual inactivation of EZH1/2 eradicated quiescent LSCs to cure AML. Genetic deletion of Ezh1/2 in a mouse AML model induced cell cycle progression of quiescent LSCs and differentiation to LSCs, eventually eradicating AML LSCs. Quiescent LSCs showed PRC2-mediated suppression of Cyclin D, and Cyclin D-overexpressing AML was more sensitive to chemotherapy. We have developed a novel EZH1/2 dual inhibitor with potent inhibitory activity against both EZH1/2. In AML mouse models and patient-derived xenograft models, the inhibitor reduced the number of LSCs, impaired leukemia progression, and prolonged survival. Taken together, these results show that dual inhibition of EZH1/2 is an effective strategy for eliminating AML LSCs.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Kreso A, Dick JE . Evolution of the cancer stem cell model. Cell Stem Cell 2014; 14: 275–291.
Szer J . The prevalent predicament of relapsed acute myeloid leukemia. Hematology Am Soc Hematol Educ Program 2012; 2012: 43–48.
Ishikawa F, Yoshida S, Saito Y, Hijikata A, Kitamura H, Tanaka S et al. Chemotherapy-resistant human AML stem cells home to and engraft within the bone-marrow endosteal region. Nat Biotechnol 2007; 25: 1315–1321.
Gentles AJ, Plevritis SK, Majeti R, Alizadeh AA . Association of a leukemic stem cell gene expression signature with clinical outcomes in acute myeloid leukemia. JAMA 2012; 304: 2706–2715.
Takeishi S, Matsumoto A, Onoyama I, Naka K, Hirao A, Nakayama KI . Ablation of Fbxw7 eliminates leukemia-initiating cells by preventing quiescence. Cancer Cell [Internet].; 2013; 23: 347–361.
Lechman ER, Gentner B, Ng SWK, Schoof EM, van Galen P, Kennedy JA et al. MiR-126 regulates distinct self-renewal outcomes in normal and malignant hematopoietic stem cells. Cancer Cell 2016; 29: 214–228.
Bruedigam C, Bagger FO, Heidel FH, Paine Kuhn C, Guignes S, Song A et al. Telomerase inhibition effectively targets mouse and human aml stem cells and delays relapse following chemotherapy. Cell Stem Cell.; 2014; 15: 775–790.
Boyer La, Plath K, Zeitlinger J, Brambrink T, Medeiros La, Lee TI et al. Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature 2006; 441: 349–353.
Lee TI, Jenner RG, Boyer LA, Guenther MG, Levine SS, Kumar RM et al. Control of developmental regulators by polycomb in human embryonic stem cells. Cell 2006; 125: 301–313.
Cao R, Zhang Y . SUZ12 is required for both the histone methyltransferase activity and the silencing function of the EED-EZH2 complex. Mol Cell 2004; 15: 57–67.
Margueron R, Justin N, Ohno K, Sharpe ML, Son J, Drury WJ et al. Role of the polycomb protein EED in the propagation of repressive histone marks. Nature 2009; 461: 762–767.
Czermin B, Melfi R, McCabe D, Seitz V, Imhof A, Pirrotta V . Drosophila enhancer of Zeste/ESC complexes have a histone H3 methyltransferase activity that marks chromosomal Polycomb sites. Cell 2002; 111: 185–196.
Sparmann A, van Lohuizen M . Polycomb silencers control cell fate, development and cancer. Nat Rev Cancer 2006; 6: 846–856.
Xu F, Li X, Wu L, Zhang Q, Yang R, Yang Y 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 2011; 90: 643–653.
Raaphorst FM, van Kemenade FJ, Blokzijl T, Fieret E, Hamer KM, Satijn DP et al. Coexpression of BMI-1 and EZH2 polycomb group genes in Reed-Sternberg cells of Hodgkin’s disease. Am J Pathol 2000; 157: 709–715.
Van Kemenade FJ, Raaphorst FM, Blokzijl T, Fieret E, Hamer KM, Satijn DPE et al. Coexpression of BMI-1 and EZH2 polycomb-group proteins is associated with cycling cells and degree of malignancy in B-cell non-Hodgkin lymphoma. Blood 2001; 97: 3896–3901.
Sasaki D, Imaizumi Y, Hasegawa H, Osaka A, Tsukasaki K, Lim Choi Y et al. Overexpression of enhancer of zeste homolog 2 with trimethylation of lysine 27 on histone H3 in adult T-cell leukemia/lymphoma as a target for epigenetic therapy. Haematologica 2011; 96: 712–719.
Visser HPJ, Gunster MJ, Kluin-Nelemans HC, Manders EMM, Raaphorst FM, CJLM Meijer et al. The Polycomb group protein EZH2 is upregulated in proliferating, cultured human mantle cell lymphoma. Br J Haematol 2001; 112: 950–958.
Berg T, Thoene S, Yap D, Wee T, Schoeler N, Rosten P et al. A transgenic mouse model demonstrating the oncogenic role of mutations in the polycomb-group gene EZH2 in lymphomagenesis. Blood 2014; 123: 3914–3924.
Morin RD, Johnson NA, Severson TM, Mungall AJ, An J, Goya R et al. Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nat Genet 2010; 42: 181–185.
Sneeringer CJ, Scott MP, Kuntz KW, Knutson SK, Pollock RM, Richon VM 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 USA 2010; 107: 20980–20985.
McCabe MT, Graves AP, Ganji G, Diaz E, Halsey WS, Jiang Y et al. Mutation of A677 in histone methyltransferase EZH2 in human B-cell lymphoma promotes hypertrimethylation of histone H3 on lysine 27 (H3K27). Proc Natl Acad Sci USA 2012; 109: 2989–2994.
Majer CR, Jin L, Scott MP, Knutson SK, Kuntz KW, Keilhack H et al. A687V EZH2 is a gain-of-function mutation found in lymphoma patients. FEBS Lett 2012; 586: 3448–3451.
Neff T, Sinha AU, Kluk MJ, Zhu N, Khattab MH, Stein L et al. Polycomb repressive complex 2 is required for MLL-AF9 leukemia. Proc Natl Acad Sci USA 2012; 109: 5028–5033.
Tanaka S, Miyagi S, Sashida G, Chiba T, Yuan J, Mochizuki-Kashio M et al. Ezh2 augments leukemogenicity by reinforcing differentiation blockage in acute myeloid leukemia. Blood 2012; 120: 1107–1117.
Shi J, Wang E, Zuber J, Rappaport A, Taylor M, Johns C et al. The Polycomb complex PRC2 supports aberrant self-renewal in a mouse model of MLL-AF9;Nras(G12D) acute myeloid leukemia. Oncogene 2013; 32: 930–938.
Shen X, Liu Y, Hsu Y-J, Fujiwara Y, Kim J, Mao X et al. EZH1 mediates methylation on histone H3 Lysine 27 and complements EZH2 in maintaining stem cell identity and executing pluripotency. Mol Cell 2008; 32: 491–502.
Margueron R, Li G, Sarma K, Blais A, Zavadil J, Woodcock CL et al. Ezh1 and Ezh2 maintain repressive chromatin through different mechanisms. Mol Cell 2008; 32: 503–518.
Krivtsov AV, Twomey D, Feng Z, Stubbs MC, Wang Y, Faber J et al. Transformation from committed progenitor to leukaemia stem cell initiated by MLL-AF9. Nature 2006; 442: 818–822.
Huntly BJP, Shigematsu H, Deguchi K, Lee BH, Mizuno S, Duclos N et al. MOZ-TIF2, but not BCR-ABL, confers properties of leukemic stem cells to committed murine hematopoietic progenitors. Cancer Cell 2004; 6: 587–596.
Somervaille TCP, Matheny CJ, Spencer GJ, Iwasaki M, Rinn JL, Witten DM et al. Hierarchical maintenance of MLL myeloid leukemia stem cells employs a transcriptional program shared with embryonic rather than adult stem cells. Cell Stem Cell 2009; 4: 129–140.
Xie H, Xu J, Hsu JH, Nguyen M, Fujiwara Y, Peng C et al. Polycomb repressive complex 2 regulates normal hematopoietic stem cell function in a developmental-stage-specific manner. Cell Stem Cell 2014; 14: 68–80.
Caganova M, Carrisi C, Varano G, Mainoldi F, Zanardi F, Germain PL et al. Germinal center dysregulation by histone methyltransferase EZH2 promotes lymphomagenesis. J Clin Invest 2013; 123: 5009–5022.
Ghisi M, Kats L, Masson F, Li J, Kratina T, Vidacs E et al. Id2 and E proteins orchestrate the initiation and maintenance of MLL-rearranged acute myeloid leukemia. Cancer Cell 2016; 30: 59–74.
Ren S, Rollins BJ . Cyclin C/Cdk3 promotes Rb-dependent G0 exit. Cell 2004; 117: 239–251.
Mende N, Kuchen EE, Lesche M, Grinenko T, Kokkaliaris KD, Hanenberg H et al. CCND1-CDK4-mediated cell cycle progression provides a competitive advantage for human hematopoietic stem cells in vivo. J Exp Med 2015; 212: 1171–1183.
Laurenti E, Frelin C, Xie S, Ferrari R, Dunant CF, Zandi S et al. CDK6 levels regulate quiescence exit in human hematopoietic stem cells. Cell Stem Cell 2015; 16: 302–313.
Bernstein BE, Mikkelsen TS, Xie X, Kamal M, Huebert DJ, Cuff J et al. A bivalent chromatin structure marks key developmental genes in embryonic stem cells. Cell 2006; 125: 315–326.
Cui K, Zang C, Roh TY, Schones DE, Childs RW, Peng W et al. Chromatin signatures in multipotent human hematopoietic stem cells indicate the fate of bivalent genes during differentiation. Cell Stem Cell 2009; 4: 80–93.
Honma D, Kanno O, Watanabe J, Kinoshita J, Hirasawa M, Nosaka E et al. Novel orally bioavailable EZH1/2 dual inhibitors with greater antitumor efficacy than an EZH2 selective inhibitor. Cancer Sci 2017; 108: 2069–2078.
McCabe MT, Ott HM, Ganji G, Korenchuk S, Thompson C, Van Aller GS et al. EZH2 inhibition as a therapeutic strategy for lymphoma with EZH2-activating mutations. Nature 2012; 492: 108–112.
Xu B, On DM, Ma A, Parton T, Konze KD, Pattenden SG et al. Selective inhibition of EZH2 and EZH1 enzymatic activity by a small molecule suppresses MLL- rearranged leukemia. Blood 2015; 125: 346–357.
Acknowledgements
This work was supported in part by Acceleration Transformative research for Medical innovation from Japan Agency for Medical Research and Development; and the National Cancer Center Research and Development Fund (IK).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
DH, NA and KA are employees of Daiichi Sankyo, Co., Ltd. The remaining authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Leukemia website
Supplementary information
Rights and permissions
About this article
Cite this article
Fujita, S., Honma, D., Adachi, N. et al. Dual inhibition of EZH1/2 breaks the quiescence of leukemia stem cells in acute myeloid leukemia. Leukemia 32, 855–864 (2018). https://doi.org/10.1038/leu.2017.300
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/leu.2017.300
This article is cited by
-
FLT3 tyrosine kinase inhibition modulates PRC2 and promotes differentiation in acute myeloid leukemia
Leukemia (2024)
-
EZH1/2 as targets for cancer therapy
Cancer Gene Therapy (2023)
-
Dual inhibition of EZH1/2 induces cell cycle arrest of B cell acute lymphoblastic leukemia cells through upregulation of CDKN1C and TP53INP1
International Journal of Hematology (2023)
-
Valemetostat Tosilate: First Approval
Drugs (2022)
-
Integrative study of EZH2 mutational status, copy number, protein expression and H3K27 trimethylation in AML/MDS patients
Clinical Epigenetics (2021)