Abstract
Tyrosine kinase inhibitor (TKI) resistance and progression to blast crisis (BC), both related to persistent β-catenin activation, remain formidable challenges for chronic myeloid leukemia (CML). We observed overexpression of β-catenin in BC-CML stem/progenitor cells, particularly in granulocyte–macrophage progenitors, and highest among a novel CD34+CD38+CD123hiTim-3hi subset as determined by CyTOF analysis. Co-culture with mesenchymal stromal cells (MSCs) induced the expression of β-catenin and its target CD44 in CML cells. A novel Wnt/β-catenin signaling modulator, C82, and nilotinib synergistically killed KBM5T315I and TKI-resistant primary BC-CML cells with or without BCR–ABL kinase mutations even under leukemia/MSC co-culture conditions. Silencing of β-catenin by short interfering RNA restored sensitivity of primary BCR–ABLT315I/E255V BC-CML cells to nilotinib. Combining the C82 pro-drug, PRI-724, with nilotinib significantly prolonged the survival of NOD/SCID/IL2Rγ null mice injected with primary BCR–ABLT315I/E255V BC-CML cells. The combined treatment selectively targeted CML progenitors and inhibited CD44, c-Myc, survivin, p-CRKL and p-STAT5 expression. In addition, pretreating primary BC-CML cells with C82, or the combination, but not with nilotinib alone, significantly impaired their engraftment potential in NOD/SCID/IL2Rγ-null-3/GM/SF mice and significantly prolonged survival. Our data suggest potential benefit of concomitant β-catenin and Bcr–Abl inhibition to prevent or overcome Bcr–Abl kinase-dependent or -independent TKI resistance in BC-CML.
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References
De Klein A, van Kessel AG, Grosveld G, Bartram CR, Hagemeijer A, Bootsma D et al. A cellular oncogene is translocated to the Philadelphia chromosome in chronic myelocytic leukaemia. Nature 1982; 300: 765–767.
Apperley JF . Chronic myeloid leukaemia. Lancet 2015; 385: 1447–1459.
Goldman JM, Melo JV . Targeting the BCR-ABL tyrosine kinase in chronic myeloid leukemia. N Engl J Med 2001; 344: 1084–1086.
Hochhaus A, Kantarjian HM, Baccarani M, Lipton JH, Apperley JF, Druker BJ et al. Dasatinib induces notable hematologic and cytogenetic responses in chronic-phase chronic myeloid leukemia after failure of imatinib therapy. Blood 2007; 109: 2303–2309.
Mahon FX, Réa 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.
Saußele S, Richter J, Hochhaus A, Mahon FX . The concept of treatment-free remission in chronic myeloid leukemia. Leukemia 2016; 30: 1638–1647.
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.
Schemionek M, Elling C, Steidl U, Bäumer N, Hamilton A, Spieker T et al. BCR-ABL enhances differentiation of long-term repopulating hematopoietic stem cells. Blood 2010; 115: 3185–3195.
Clevers H . Wnt/beta-catenin signaling in development and disease. Cell 2006; 127: 469–480.
MacDonald BT, Tamai K, He X . Wnt/beta-catenin signaling: components, mechanisms, and diseases. Dev Cell 2009; 17: 9–26.
Moon RT, Bowerman B, Boutros M, Perrimon N . The promise and perils of Wnt signaling through beta-catenin. Science 2002; 296: 1644–1646.
Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K et al. A role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 2003; 423: 409–414.
Scheller M, Huelsken J, Rosenbauer F, Taketo MM, Birchmeier W, Tenen DG et al. Hematopoietic stem cell and multilineage defects generated by constitutive beta-catenin activation. Nat Immunol 2006; 7: 1037–1047.
Jeannet G, Scheller M, Scarpellino L, Duboux S, Gardiol N, Back J et al. Long-term, multilineage hematopoiesis occurs in the combined absence of beta-catenin and gamma-catenin. Blood 2008; 111: 142–149.
Abrahamsson AE, Geron I, Gotlib J, Dao KH, Barroga CF, Newton IG et al. Glycogen synthase kinase 3beta missplicing contributes to leukemia stem cell generation. Proc Natl Acad Sci USA 2009; 106: 3925–3929.
Wang Y, Krivtsov AV, Sinha AU, North TE, Goessling W, Feng Z et al. The Wnt/beta-catenin pathway is required for the development of leukemia stem cells in AML. Science 2010; 327: 1650–1653.
Zhao C, Blum J, Chen A, Kwon HY, Jung SH, Cook JM et al. Loss of beta-catenin impairs the renewal of normal and CML stem cells in vivo. Cancer Cell 2007; 12: 528–541.
Heidel FH, Bullinger L, Feng Z, Wang Z, Neff TA, Stein L et al. Genetic and pharmacologic inhibition of beta-catenin targets imatinib-resistant leukemia stem cells in CML. Cell Stem Cell 2012; 10: 412–424.
Hamilton A, Helgason GV, Schemionek M, Zhang B, Myssina S, Allan EK et al. Chronic myeloid leukemia stem cells are not dependent on Bcr-Abl kinase activity for their survival. Blood 2012; 119: 1501–1510.
Kavalerchik E, Goff D, Jamieson CH . Chronic myeloid leukemia stem cells. J Clin Oncol 2008; 26: 2911–2915.
Perrotti D, Jamieson C, Goldman J, Skorski T . Chronic myeloid leukemia: mechanisms of blastic transformation. J Clin Invest 2010; 120: 2254–2264.
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.
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.
Giotopoulos G, van der Weyden L, Osaki H, Rust AG, Gallipoli P, Meduri E et al. A novel mouse model identifies cooperating mutations and therapeutic targets critical for chronic myeloid leukemia progression. J Exp Med 2015; 212: 1551–1569.
Neviani P, Harb JG, Oaks JJ, Santhanam R, Walker CJ, Ellis JJ et al. PP2A-activating drugs selectively eradicate TKI-resistant chronic myeloid leukemic stem cells. J Clin Invest 2013; 123: 4144–4157.
Zhang B, Li M, McDonald T, Holyoake TL, Moon RT, Campana D et al. Microenvironmental protection of CML stem and progenitor cells from tyrosine kinase inhibitors through N-cadherin and Wnt-beta-catenin signaling. Blood 2013; 121: 1824–1838.
Kleppe M, Levine RL . Targeting beta-catenin in CML: leukemia stem cells beware!. Cell Stem Cell 2012; 10: 351–353.
El-Khoueiry AB, Ning Y, Yang D, Cole S, Kahn M, Zoghbi M et al. A phase I first-in-human study of PRI-724 in patients with advanced solid tumors. J Clin Oncol 2013; 31 (Suppl); Abstract 2501).
Eguchi M, Nguyen C, Lee SC, Kahn M . ICG-001, a novel small molecule regulator of TCF/beta-catenin transcription. Med Chem 2005; 1: 467–472.
Bendall SC, Simonds EF, Qiu P, Amir el-AD, Krutzik PO, Finck R et al. Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science 2011; 332: 687–696.
Qiu P, Simonds EF, Bendall SC, Gibbs Jr KD, Bruggner RV, Linderman MD et al. Extracting a cellular hierarchy from high-dimensional cytometry data with SPADE. Nat Biotechnol 2011; 29: 886–891.
Beran M, Pisa P, O’Brien S, Kurzrock R, Siciliano M, Cork A et al. Biological properties and growth in SCID mice of a new myelogenous leukemia cell line (KBM-5) derived from chronic myelogenous leukemia cells in the blastic phase. Cancer Res 1993; 53: 3603–3610.
Ricci C, Scappini B, Divoky V, Gatto S, Onida F, Verstovsek S et al. Mutation in the ATP-binding pocket of the ABL kinase domain in an STI571-resistant BCR/ABL-positive cell line. Cancer Res 2002; 62: 5995–5998.
Studeny M, Marini FC, Champlin RE, Zompetta C, Fidler IJ, Andreeff M . Bone marrow-derived mesenchymal stem cells as vehicles for interferon-beta delivery into tumors. Cancer Res 2002; 62: 3603–3608.
Konopleva M, Konoplev S, Hu W, Zaritskey AY, Afanasiev BV, Andreeff M . Stromal cells prevent apoptosis of AML cells by up-regulation of anti-apoptotic proteins. Leukemia 2002; 16: 1713–1724.
Holtz MS, Slovak ML, Zhang F, Sawyers CL, Forman SJ, Bhatia R . Imatinib mesylate (STI571) inhibits growth of primitive malignant progenitors in chronic myelogenous leukemia through reversal of abnormally increased proliferation. Blood 2002; 99: 3792–3800.
Mak DH, Schober WD, Chen W, Konopleva M, Cortes J, Kantarjian HM et al. Triptolide induces cell death independent of cellular responses to imatinib in blast crisis chronic myelogenous leukemia cells including quiescent CD34+ primitive progenitor cells. Mol Cancer Ther 2009; 8: 2509–2516.
Han L, Qiu P, Zeng Z, Jorgensen JL, Mak DH, Burks JK et al. Single-cell mass cytometry reveals intracellular survival/proliferative signaling in FLT3-ITD-mutated AML stem/progenitor cells. Cytometry A 2015; 87: 346–356.
Carter BZ, Mak PY, Mu H, Zhou HS, Mak DH, Schober W et al. Combined targeting of BCL-2 and BCR-ABL tyrosine kinase eradicates chronic myeloid leukemia stem cells. Sci Transl Med 2016; 8: 355ra117.
Chou TC, Talalay P . Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 1984; 22: 27–55.
Eiring AM, Khorashad JS, Anderson DJ, Yu F, Redwine HM, Mason CC et al. β-Catenin is required for intrinsic but not extrinsic BCR-ABL1 kinase-independent resistance to tyrosine kinase inhibitors in chronic myeloid leukemia. Leukemia 2015; 29: 2328–2337.
Krause DS, Lazarides K, von Andrian UH, Van Etten RA . Requirement for CD44 in homing and engraftment of BCR-ABL-expressing leukemic stem cells. Nat Med 2006; 12: 1175–1180.
Holm FL, Hellqvist E, Mason C . The niche specific role of CD44 splice isoform expression in blast crisis leukemia stem cell generation. Cancer Res 2014; 74 (19 Suppl); Abstract 4798).
Konopleva MY, Jordan CT . Leukemia stem cells and microenvironment: biology and therapeutic targeting. J Clin Oncol 2011; 29: 591–599.
Carter BZ, Mak DH, Cortes J, Andreeff M . The elusive chronic myeloid leukemia stem cell: does it matter and how do we eliminate it? Semin Hematol 2010; 47: 362–370.
Burchert A . Roots of imatinib resistance: a question of self-renewal? Drug Resist Updat 2007; 10: 152–161.
Savona M, Talpaz M . Getting to the stem of chronic myeloid leukaemia. Nat Rev Cancer 2008; 8: 341–350.
Haeno H, Levine RL, Gilliland DG, Michor F . A progenitor cell origin of myeloid malignancies. Proc Natl Acad Sci USA 2009; 106: 16616–16621.
Melkus M, Bennaceur-Griscelli A, Valogne Y, Flamant S, Chomel JC, Sorel N et al. Biological effects of T315I-mutated BCR-ABL in an embryonic stem cell-derived hematopoiesis model. Exp Hematol 2013; 4: 335–345.
Coluccia AM, Vacca A, Duñach M, Mologni L, Redaelli S, Bustos VH et al. Bcr-Abl stabilizes beta-catenin in chronic myeloid leukemia through its tyrosine phosphorylation. EMBO J 2007; 26: 1456–1466.
McCubrey JA, Steelman LS, Bertrand FE, Davis NM, Abrams SL, Montalto G et al. Multifaceted roles of GSK-3 and Wnt/β-catenin in hematopoiesis and leukemogenesis: opportunities for therapeutic intervention. Leukemia 2014; 28: 15–33.
Raaijmakers MH, Mukherjee S, Guo S, Zhang S, Kobayashi T, Schoonmaker JA et al. Bone progenitor dysfunction induces myelodysplasia and secondary leukaemia. Nature 2010; 464: 852–857.
Kode A, Manavalan JS, Mosialou I, Bhagat G, Rathinam CV, Luo N et al. Leukaemogenesis induced by an activating β-catenin mutation in osteoblasts. Nature 2014; 506: 240–244.
Eaves CJ, Humphries RK . Acute myeloid leukemia and the Wnt pathway. N Engl J Med 2010; 362: 2326–2327.
Ornatsky O, Bandura D, Baranov V, Nitz M, Winnik MA, Tanner S . Highly multiparametric analysis by mass cytometry. J Immunol Methods 2010; 361: 1–20.
Kikushige Y, Yuda J, Shima T, Miyamoto T, Akashi K . TIM-3 and its ligand, Galectin-9 constitute a pan-myeloid autocrine loop to develop and maintain malignant stem cells in most human myeloid leukemia. Blood 2014; 124: 835.
Kikushige Y, Miyamoto T, Yuda J, Jabbarzadeh-Tabrizi S, Shima T, Takayanagi S et al. A TIM-3/Gal-9 autocrine stimulatory loop drives self-renewal of human myeloid leukemia stem cells and leukemic progression. Cell Stem Cell 2015; 17: 341–352.
Tang D, Lotze MT . Tumor immunity times out: TIM-3 and HMGB1. Nat Immunol 2012; 13: 808–810.
Kabiri Z, Numata A, Kawasaki A, Edison, Tenen DG, Virshup DM . Wnts are dispensable for differentiation and self-renewal of adult murine hematopoietic stem cells. Blood 2015; 126: 1086–1094.
Acknowledgements
We thank Drs Michael Kahn for helping with technical questions and Numsen Hail for editing the manuscript. This work was supported in part by the research funding from PRISM Pharma/Eisai (to BZC) and grants from the National Institutes of Health (P01CA49639 and MD Anderson’s Cancer Center Support Grant CA016672), Cancer Prevention Research Institute of Texas (CPRIT, RP121010), MDACC moonshot program, and the Paul and Mary Haas Chair in Genetics (to MA) and grant from Science and Technology Planning Project of Guangdong Province, China (2016A020215112 to HSZ).
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Zhou, H., Mak, P., Mu, H. et al. Combined inhibition of β-catenin and Bcr–Abl synergistically targets tyrosine kinase inhibitor-resistant blast crisis chronic myeloid leukemia blasts and progenitors in vitro and in vivo. Leukemia 31, 2065–2074 (2017). https://doi.org/10.1038/leu.2017.87
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DOI: https://doi.org/10.1038/leu.2017.87
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