Abstract
The BCR–ABL fusion oncoprotein accelerates differentiation and proliferation of myeloid cells during the chronic phase of chronic myeloid leukemia (CP-CML). Here, the role of CCAAT/enhancer binding protein β (C/EBPβ), a regulator for ‘emergency granulopoiesis,’ in the pathogenesis of CP-CML was examined. C/EBPβ expression was upregulated in Lineage− CD34+ CD38− hematopoietic stem cells (HSCs) and myeloid progenitors isolated from bone marrow of patients with CP-CML. In EML cells, a mouse HSC line, BCR–ABL upregulated C/EBPβ, at least in part, through the activation of STAT5. Myeloid differentiation and proliferation induced by BCR–ABL was significantly impaired in C/EBPβ-deficient bone marrow cells in vitro. Mice that were transplanted with BCR–ABL-transduced C/EBPβ knockout bone marrow cells survived longer than mice that received BCR–ABL-transduced wild-type (WT) bone marrow cells. Significantly higher levels of leukemic stem cells were maintained in BCR–ABL-transduced C/EBPβ-deficient cells than in BCR–ABL-transduced WT cells. These results suggest that C/EBPβ is involved in BCR–ABL-mediated myeloid expansion. Further elucidation of the molecular mechanisms underlying the C/EBPβ-mediated stem cell loss might reveal a novel therapeutic strategy for eradication of CML stem cells.
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
Cotta CV, Bueso-Ramos CE . New insights into the pathobiology and treatment of chronic myelogenous leukemia. Ann Diagn Pathol 2007; 11: 68–78.
Ren R . Mechanisms of BCR-ABL in the pathogenesis of chronic myelogenous leukaemia. Nat Rev Cancer 2005; 5: 172–183.
Sawyers CL . Chronic myeloid leukemia. N Engl J Med 1999; 340: 1330–1340.
Melo JV, Barnes DJ . Chronic myeloid leukaemia as a model of disease evolution in human cancer. Nat Rev Cancer 2007; 7: 441–453.
Huettner CS, Zhang P, Van Etten RA, Tenen DG . Reversibility of acute B-cell leukaemia induced by BCR-ABL1. Nat Genet 2000; 24: 57–60.
Huettner CS, Koschmieder S, Iwasaki H, Iwasaki-Arai J, Radomska HS, Akashi K et al. Inducible expression of BCR/ABL using human CD34 regulatory elements results in a megakaryocytic myeloproliferative syndrome. Blood 2003; 102: 3363–3370.
Koschmieder S, Gottgens B, Zhang P, Iwasaki-Arai J, Akashi K, Kutok JL et al. Inducible chronic phase of myeloid leukemia with expansion of hematopoietic stem cells in a transgenic model of BCR-ABL leukemogenesis. Blood 2005; 105: 324–334.
Goldman JM, Melo JV . Chronic myeloid leukemia--advances in biology and new approaches to treatment. N Engl J Med 2003; 349: 1451–1464.
Kimura S, Naito H, Segawa H, Kuroda J, Yuasa T, Sato K et al. NS-187, a potent and selective dual Bcr-Abl/Lyn tyrosine kinase inhibitor, is a novel agent for imatinib-resistant leukemia. Blood 2005; 106: 3948–3954.
Druker BJ, Guilhot F, O’Brien SG, Gathmann I, Kantarjian H, Gattermann N et al. Five-year follow-up of patients receiving imatinib for chronic myeloid leukemia. N Engl J Med 2006; 355: 2408–2417.
Kantarjian HM, Giles F, Gattermann N, Bhalla K, Alimena G, Palandri F et al. Nilotinib (formerly AMN107), a highly selective BCR-ABL tyrosine kinase inhibitor, is effective in patients with Philadelphia chromosome-positive chronic myelogenous leukemia in chronic phase following imatinib resistance and intolerance. Blood 2007; 110: 3540–3546.
Hochhaus A, Baccarani M, Deininger M, Apperley JF, Lipton JH, Goldberg SL et al. Dasatinib induces durable cytogenetic responses in patients with chronic myelogenous leukemia in chronic phase with resistance or intolerance to imatinib. Leukemia 2008; 22: 1200–1206.
Jamieson CH, Ailles LE, Dylla SJ, Muijtjens M, Jones C, Zehnder JL et al. Granulocyte-macrophage progenitors as candidate leukemic stem cells in blast-crisis CML. N Engl J Med 2004; 351: 657–667.
Passegue E, Weisman IL . Leukemic stem cells: where do they come from? Stem Cell Rev 2005; 1: 181–188.
Bruns I, Czibere A, Fischer JC, Roels F, Cadeddu RP, Buest S et al. The hematopoietic stem cell in chronic phase CML is characterized by a transcriptional profile resembling normal myeloid progenitor cells and reflecting loss of quiescence. Leukemia 2009; 23: 892–899.
Michor F, Hughes TP, Iwasa Y, Branford S, Shah NP, Sawyers CL et al. Dynamics of chronic myeloid leukaemia. Nature 2005; 435: 1267–1270.
Copland M, Hamilton A, Elrick LJ, Baird JW, Allan EK, Jordanides N et al. Dasatinib (BMS-354825) targets an earlier progenitor population than imatinib in primary CML but does not eliminate the quiescent fraction. Blood 2006; 107: 4532–4539.
Mahon FX, Rea D, Guilhot J, Guilhot F, Huguet F, Nicolini F et al. Discontinuation of imatinib in patients with chronic myeloid leukaemia who have maintained complete molecular remission for at least 2 years: the prospective, multicentre Stop Imatinib (STIM) trial. Lancet Oncol 2010; 11: 1029–1035.
Corbin AS, Agarwal A, Loriaux M, Cortes J, Deininger MW, Druker BJ . Human chronic myeloid leukemia stem cells are insensitive to imatinib despite inhibition of BCR-ABL activity. J Clin Invest 2011; 121: 396–409.
Gorre ME, Mohammed M, Ellwood K, Hsu N, Paquette R, Rao PN et al. Clinical resistance to STI-571 cancer therapy caused by BCR-ABL gene mutation or amplification. Science 2001; 293: 876–880.
Branford S, Rudzki Z, Walsh S, Grigg A, Arthur C, Taylor K et al. High frequency of point mutations clustered within the adenosine triphosphate-binding region of BCR/ABL in patients with chronic myeloid leukemia or Ph-positive acute lymphoblastic leukemia who develop imatinib (STI571) resistance. Blood 2002; 99: 3472–3475.
Hirai H, Zhang P, Dayaram T, Hetherington CJ, Mizuno S, Imanishi J et al. C/EBPbeta is required for ‘emergency’ granulopoiesis. Nat Immunol 2006; 7: 732–739.
Tsai S, Bartelmez S, Sitnicka E, Collins S . Lymphohematopoietic progenitors immortalized by a retroviral vector harboring a dominant-negative retinoic acid receptor can recapitulate lymphoid, myeloid, and erythroid development. Genes Dev 1994; 8: 2831–2841.
Screpanti I, Romani L, Musiani P, Modesti A, Fattori E . Lazzaro D et al. Lymphoproliferative disorder and imbalanced T-helper response in C/EBP beta-deficient mice. EMBO J 1995; 14: 1932–1941.
Hawley RG, Lieu FH, Fong AZ, Hawley TS . Versatile retroviral vectors for potential use in gene therapy. Gene Ther 1994; 1: 136–138.
Onishi M, Nosaka T, Misawa K, Mui AL, Gorman D, McMahon M et al. Identification and characterization of a constitutively active STAT5 mutant that promotes cell proliferation. Mol Cell Biol 1998; 18: 3871–3879.
Moriggl R, Gouilleux-Gruart V, Jahne R, Berchtold S, Gartmann C, Liu X et al. Deletion of the carboxyl-terminal transactivation domain of MGF-Stat5 results in sustained DNA binding and a dominant negative phenotype. Mol Cell Biol 1996; 16: 5691–5700.
Morita S, Kojima T, Kitamura T . Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther 2000; 7: 1063–1066.
Manz MG, Miyamoto T, Akashi K, Weissman IL . Prospective isolation of human clonogenic common myeloid progenitors. Proc Natl Acad Sci USA 2002; 99: 11872–11877.
Diaz-Blanco E, Bruns I, Neumann F, Fischer JC, Graef T, Rosskopf M et al. Molecular signature of CD34(+) hematopoietic stem and progenitor cells of patients with CML in chronic phase. Leukemia 2007; 21: 494–504.
Shuai K, Halpern J, ten Hoeve J, Rao X, Sawyers CL . Constitutive activation of STAT5 by the BCR-ABL oncogene in chronic myelogenous leukemia. Oncogene 1996; 13: 247–254.
Ilaria RL, Van Etten RA . P210 and P190(BCR/ABL) induce the tyrosine phosphorylation and DNA binding activity of multiple specific STAT family members. J Biol Chem 1996; 271: 31704–31710.
Ye D, Wolff N, Li L, Zhang S, Ilaria RL . STAT5 signaling is required for the efficient induction and maintenance of CML in mice. Blood 2006; 107: 4917–4925.
Steelman LS, Pohnert SC, Shelton JG, Franklin RA, Bertrand FE, McCubrey JA . JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis. Leukemia 2004; 18: 189–218.
Kharas MG, Fruman DA . ABL oncogenes and phosphoinositide 3-kinase: mechanism of activation and downstream effectors. Cancer Res 2005; 65: 2047–2053.
Quintas-Cardama A, Cortes J . Molecular biology of bcr-abl1-positive chronic myeloid leukemia. Blood 2009; 113: 1619–1630.
Guerzoni C, Bardini M, Mariani SA, Ferrari-Amorotti G, Neviani P, Panno ML et al. Inducible activation of CEBPB, a gene negatively regulated by BCR/ABL, inhibits proliferation and promotes differentiation of BCR/ABL-expressing cells. Blood 2006; 107: 4080–4089.
Schuster C, Forster K, Dierks H, Elsasser A, Behre G, Simon N et al. The effects of Bcr-Abl on C/EBP transcription-factor regulation and neutrophilic differentiation are reversed by the Abl kinase inhibitor imatinib mesylate. Blood 2003; 101: 655–663.
Steinman RA, Tweardy DJ . Granulocyte colony-stimulating factor receptor mRNA upregulation is an immediate early marker of myeloid differentiation and exhibits dysfunctional regulation in leukemic cells. Blood 1994; 83: 119–127.
Du Y, Campbell JL, Nalbant D, Youn H, Bass AC, Cobos E et al. Mapping gene expression patterns during myeloid differentiation using the EML hematopoietic progenitor cell line. Exp Hematol 2002; 30: 649–658.
Bjerregaard MD, Jurlander J, Klausen P, Borregaard N, Cowland JB . The in vivo profile of transcription factors during neutrophil differentiation in human bone marrow. Blood 2003; 101: 4322–4332.
Minami Y, Stuart SA, Ikawa T, Jiang Y, Banno A, Hunton IC et al. BCR-ABL-transformed GMP as myeloid leukemic stem cells. Proc Natl Acad Sci USA 2008; 105: 17967–17972.
Xie S, Wang Y, Liu J, Sun T, Wilson MB, Smithgall TE et al. Involvement of Jak2 tyrosine phosphorylation in Bcr-Abl transformation. Oncogene 2001; 20: 6188–6195.
Hoelbl A, Schuster C, Kovacic B, Zhu B, Wickre M, Hoelzl MA et al. Stat5 is indispensable for the maintenance of bcr/abl-positive leukaemia. EMBO Mol Med 2010; 2: 98–110.
Feldman GM, Rosenthal LA, Liu X, Hayes MP, Wynshaw-Boris A, Leonard WJ et al. STAT5A-deficient mice demonstrate a defect in granulocyte-macrophage colony-stimulating factor-induced proliferation and gene expression. Blood 1997; 90: 1768–1776.
Kimura A, Rieger MA, Simone JM, Chen W, Wickre MC, Zhu BM et al. The transcription factors STAT5A/B regulate GM-CSF-mediated granulopoiesis. Blood 2009; 114: 4721–4728.
Akagi T, Saitoh T, O’Kelly J, Akira S, Gombart AF, Koeffler HP . Impaired response to GM-CSF and G-CSF, and enhanced apoptosis in C/EBPbeta-deficient hematopoietic cells. Blood 2008; 111: 2999–3004.
Zhang H, Nguyen-Jackson H, Panopoulos AD, Li HS, Murray PJ, Watowich SS . STAT3 controls myeloid progenitor growth during emergency granulopoiesis. Blood 2010; 116: 2462–2471.
Liao W, Schones DE, Oh J, Cui Y, Cui K, Roh TY et al. Priming for T helper type 2 differentiation by interleukin 2-mediated induction of interleukin 4 receptor alpha-chain expression. Nat Immunol 2008; 9: 1288–1296.
Yang XP, Ghoreschi K, Steward-Tharp SM, Rodriguez-Canales J, Zhu J, Grainger JR et al. Opposing regulation of the locus encoding IL-17 through direct, reciprocal actions of STAT3 and STAT5. Nat Immunol 2011; 12: 247–254.
Schemionek M, Elling C, Steidl U, Baumer N, Hamilton A, Spieker T et al. BCR-ABL enhances differentiation of long-term repopulating hematopoietic stem cells. Blood 2010; 115: 3185–3195.
Guerzoni C, Ferrari-Amorotti G, Bardini M, Mariani SA, Calabretta B . Effects of C/EBPalpha and C/EBPbeta in BCR/ABL-expressing cells: differences and similarities. Cell Cycle 2006; 5: 1254–1257.
Sakka V, Tsiodras S, Giamarellos-Bourboulis EJ, Giamarellou H . An update on the etiology and diagnostic evaluation of a leukemoid reaction. Eur J Intern Med 2006; 17: 394–398.
Acknowledgements
We thank Dr Toshio Kitamura (University of Tokyo, Tokyo, Japan) and Dr Keiko Okuda (Kyoto Prefectural University of Medicine, Kyoto, Japan) for providing us with STAT5 mutant vectors and BCR–ABL-expressing vectors, respectively. We are grateful to Mikiko Katakami, Yoko Nakagawa and Yoshiko Manabe for their excellent technical assistance. This work was partly supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science and the Global COE Program ‘Center for Frontier Medicine’ from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, Takeda Science Foundation, the Kobayashi Foundation for Cancer Research, and Senshin Medical Research Foundation.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Leukemia website
Rights and permissions
About this article
Cite this article
Hayashi, Y., Hirai, H., Kamio, N. et al. C/EBPβ promotes BCR–ABL-mediated myeloid expansion and leukemic stem cell exhaustion. Leukemia 27, 619–628 (2013). https://doi.org/10.1038/leu.2012.258
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/leu.2012.258
Keywords
This article is cited by
-
Early induction of C/EBPβ expression as a potential marker of steroid responsive colitis
Scientific Reports (2019)
-
JMJD3 facilitates C/EBPβ-centered transcriptional program to exert oncorepressor activity in AML
Nature Communications (2018)
-
Constitutively active ABL family kinases, TEL/ABL and TEL/ARG, harbor distinct leukemogenic activities in vivo
Leukemia (2017)