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Acute Leukemias

EVI-1 modulates leukemogenic potential and apoptosis sensitivity in human acute lymphoblastic leukemia

Leukemia volume 27, pages 56–65 (2013)Cite this article

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Abstract

The transcriptional regulator ecotropic viral integration site-1 (EVI-1) has mainly been studied for its role in myeloid malignancies, in which high EVI-1 levels are associated with particularly aggressive disease. The role of EVI-1 in lymphoid cells, however, is largely unknown. Here we show that EVI-1 is indeed expressed in lymphoid malignancies such as acute lymphoblastic leukemia (ALL) and a subset of chronic lymphocytic leukemia. Expression data from pediatric ALL further suggest that high EVI-1 levels are associated with poor prognosis. Suppression of EVI-1 expression by RNA interference reduces cell growth and enhances apoptosis sensitivity in response to various stimuli in lymphoblastic leukemia cells. At the molecular level, EVI-1 modulates expression of several apoptosis-related genes (such as BCL2, BCL-x, XIAP, NOXA, PUMA, TRAIL-R1). Furthermore, EVI-1 knockdown strongly impairs in vivo engraftment of lymphoblastic leukemia cells upon transplantation in immune-permissive NOD/SCID/IL2Rγnull mice, conferring a survival benefit when compared with mice transplanted with control cells. Thus, our data show that EVI-1 is expressed not only in myeloid but also in lymphoid leukemias, and contributes to the leukemogenic potential and apoptosis resistance of ALL cells.

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References

  1. Mucenski ML, Taylor BA, Ihle JN, Hartley JW, Morse HC, Jenkins NA et al. Identification of a common ecotropic viral integration site, Evi-1, in the DNA of AKXD murine myeloid tumors. Mol Cell Biol 1988; 8: 301–308.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Ogawa S, Kurokawa M, Tanaka T, Mitani K, Inazawa J, Hangaishi A et al. Structurally altered Evi-1 protein generated in the 3q21q26 syndrome. Oncogene 1996; 13: 183–191.

    CAS  PubMed  Google Scholar 

  3. Suzukawa K, Parganas E, Gajjar A, Abe T, Takahashi S, Tani K et al. Identification of a breakpoint cluster region 3′ of the ribophorin I gene at 3q21 associated with the transcriptional activation of the EVI1 gene in acute myelogenous leukemias with inv(3)(q21q26). Blood 1994; 84: 2681–2688.

    CAS  PubMed  Google Scholar 

  4. Buonamici S, Li D, Chi Y, Zhao R, Wang X, Brace L et al. EVI1 induces myelodysplastic syndrome in mice. J Clin Invest 2004; 114: 713–719.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Jin G, Yamazaki Y, Takuwa M, Takahara T, Kaneko K, Kuwata T et al. Trib1 and Evi1 cooperate with Hoxa and Meis1 in myeloid leukemogenesis. Blood 2007; 109: 3998–4005.

    Article  CAS  PubMed  Google Scholar 

  6. Barjesteh van Waalwijk van Doorn-Khosrovani S, Erpelinck C, van Putten WL, Valk PJ, van der Poel-van de Luytgaarde S, Hack R et al. High EVI1 expression predicts poor survival in acute myeloid leukemia: a study of 319 de novo AML patients. Blood 2003; 101: 837–845.

    Article  PubMed  Google Scholar 

  7. Groschel S, Lugthart S, Schlenk RF, Valk PJ, Eiwen K, Goudswaard C et al. High EVI1 expression predicts outcome in younger adult patients with acute myeloid leukemia and is associated with distinct cytogenetic abnormalities. J Clin Oncol 2010; 28: 2101–2107.

    Article  PubMed  Google Scholar 

  8. Daghistani M, Marin D, Khorashad JS, Wang L, May PC, Paliompeis C et al. EVI-1 oncogene expression predicts survival in chronic-phase CML patients resistant to imatinib treated with second-generation tyrosine kinase inhibitors. Blood 2010; 116: 6014–6017.

    Article  CAS  PubMed  Google Scholar 

  9. Valk PJ, Verhaak RG, Beijen MA, Erpelinck CA, Barjesteh van Waalwijk van Doorn-Khosrovani S, Boer JM et al. Prognostically useful gene-expression profiles in acute myeloid leukemia. N Engl J Med 2004; 350: 1617–1628.

    Article  CAS  PubMed  Google Scholar 

  10. Hirai H . The transcription factor Evi-1. Int J Biochem Cell Biol 1999; 31: 1367–1371.

    Article  CAS  PubMed  Google Scholar 

  11. Wieser R . The oncogene and developmental regulator EVI1: expression, biochemical properties, and biological functions. Gene 2007; 396: 346–357.

    Article  CAS  PubMed  Google Scholar 

  12. Hoyt PR, Bartholomew C, Davis AJ, Yutzey K, Gamer LW, Potter SS et al. The Evi1 proto-oncogene is required at midgestation for neural, heart, and paraxial mesenchyme development. Mech Dev 1997; 65: 55–70.

    Article  CAS  PubMed  Google Scholar 

  13. Goyama S, Yamamoto G, Shimabe M, Sato T, Ichikawa M, Ogawa S et al. Evi-1 Is a critical regulator for hematopoietic stem cells and transformed leukemic cells. Cell Stem Cell 2008; 3: 207–220.

    Article  CAS  PubMed  Google Scholar 

  14. Yuasa H, Oike Y, Iwama A, Nishikata I, Sugiyama D, Perkins A et al. Oncogenic transcription factor Evi1 regulates hematopoietic stem cell proliferation through GATA-2 expression. EMBO J 2005; 24: 1976–1987.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Gerhardt TM, Schmahl GE, Flotho C, Rath AV, Niemeyer CM . Expression of the Evi-1 gene in haemopoietic cells of children with juvenile myelomonocytic leukaemia and normal donors. Br J Haematol 1997; 99: 882–887.

    Article  CAS  PubMed  Google Scholar 

  16. Privitera E, Longoni D, Brambillasca F, Biondi A . EVI-1 gene expression in myeloid clonogenic cells from juvenile myelomonocytic leukemia (JMML). Leukemia 1997; 11: 2045–2048.

    Article  CAS  PubMed  Google Scholar 

  17. Delwel R, Funabiki T, Kreider BL, Morishita K, Ihle JN . Four of the seven zinc fingers of the Evi-1 myeloid-transforming gene are required for sequence-specific binding to GA(C/T)AAGA(T/C)AAGATAA. Mol Cell Biol 1993; 13: 4291–4300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Perkins AS, Fishel R, Jenkins NA, Copeland NG . Evi-1 a murine zinc finger proto-oncogene, encodes a sequence-specific DNA-binding protein. Mol Cell Biol 1991; 11: 2665–2674.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Izutsu K, Kurokawa M, Imai Y, Maki K, Mitani K, Hirai H . The corepressor CtBP interacts with Evi-1 to repress transforming growth factor beta signaling. Blood 2001; 97: 2815–2822.

    Article  CAS  PubMed  Google Scholar 

  20. Kurokawa M, Mitani K, Irie K, Matsuyama T, Takahashi T, Chiba S et al. The oncoprotein Evi-1 represses TGF-beta signalling by inhibiting Smad3. Nature 1998; 394: 92–96.

    Article  CAS  PubMed  Google Scholar 

  21. Cattaneo F, Nucifora G . EVI1 recruits the histone methyltransferase SUV39H1 for transcription repression. J Cell Biochem 2008; 105: 344–352.

    Article  CAS  PubMed  Google Scholar 

  22. Spensberger D, Delwel R . A novel interaction between the proto-oncogene Evi1 and histone methyltransferases, SUV39H1 and G9a. FEBS Lett 2008; 582: 2761–2767.

    Article  CAS  PubMed  Google Scholar 

  23. Vinatzer U, Taplick J, Seiser C, Fonatsch C, Wieser R . The leukaemia-associated transcription factors EVI-1 and MDS1/EVI1 repress transcription and interact with histone deacetylase. Br J Haematol 2001; 114: 566–573.

    Article  CAS  PubMed  Google Scholar 

  24. Goyama S, Nitta E, Yoshino T, Kako S, Watanabe-Okochi N, Shimabe M et al. EVI-1 interacts with histone methyltransferases SUV39H1 and G9a for transcriptional repression and bone marrow immortalization. Leukemia 2010; 24: 81–88.

    Article  CAS  PubMed  Google Scholar 

  25. Poppe B, Dastugue N, Vandesompele J, Cauwelier B, De Smet B, Yigit N et al. EVI1 is consistently expressed as principal transcript in common and rare recurrent 3q26 rearrangements. Genes Chromosomes Cancer 2006; 45: 349–356.

    Article  CAS  PubMed  Google Scholar 

  26. De Weer A, Poppe B, Cauwelier B, Van Roy N, Dastugue N, Hagemeijer A et al. Screening for EVI1: ectopic expression absent in T-cell acute lymphoblastic leukemia patients and cell lines. Cancer Genet Cytogenet 2006; 171: 79–80.

    Article  CAS  PubMed  Google Scholar 

  27. Andersson A, Ritz C, Lindgren D, Eden P, Lassen C, Heldrup J et al. Microarray-based classification of a consecutive series of 121 childhood acute leukemias: prediction of leukemic and genetic subtype as well as of minimal residual disease status. Leukemia 2007; 21: 1198–1203.

    Article  CAS  PubMed  Google Scholar 

  28. Cario G, Stanulla M, Fine BM, Teuffel O, Neuhoff NV, Schrauder A et al. Distinct gene expression profiles determine molecular treatment response in childhood acute lymphoblastic leukemia. Blood 2005; 105: 821–826.

    Article  CAS  PubMed  Google Scholar 

  29. Gunnarsson R, Isaksson A, Mansouri M, Goransson H, Jansson M, Cahill N et al. Large but not small copy-number alterations correlate to high-risk genomic aberrations and survival in chronic lymphocytic leukemia: a high-resolution genomic screening of newly diagnosed patients. Leukemia 2010; 24: 211–215.

    Article  CAS  PubMed  Google Scholar 

  30. Riedt T, Ebinger M, Salih HR, Tomiuk J, Handgretinger R, Kanz L et al. Aberrant expression of the homeobox gene CDX2 in pediatric acute lymphoblastic leukemia. Blood 2009; 113: 4049–4051.

    Article  CAS  PubMed  Google Scholar 

  31. Conter V, Bartram CR, Valsecchi MG, Schrauder A, Panzer-Grumayer R, Moricke A et al. Molecular response to treatment redefines all prognostic factors in children and adolescents with B-cell precursor acute lymphoblastic leukemia: results in 3184 patients of the AIEOP-BFM ALL 2000 study. Blood 2010; 115: 3206–3214.

    Article  CAS  PubMed  Google Scholar 

  32. Anastasov N, Klier M, Koch I, Angermeier D, Hofler H, Fend F et al. Efficient shRNA delivery into B and T lymphoma cells using lentiviral vector-mediated transfer. J Hematop 2009; 2: 9–19.

    Article  PubMed  Google Scholar 

  33. Maetzig T, Brugman MH, Bartels S, Heinz N, Kustikova OS, Modlich U et al. Polyclonal fluctuation of lentiviral vector-transduced and expanded murine hematopoietic stem cells. Blood 2011; 117: 3053–3064.

    Article  CAS  PubMed  Google Scholar 

  34. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C . A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 1991; 139: 271–279.

    Article  CAS  PubMed  Google Scholar 

  35. Kunder S, Calzada-Wack J, Holzlwimmer G, Muller J, Kloss C, Howat W et al. A comprehensive antibody panel for immunohistochemical analysis of formalin-fixed, paraffin-embedded hematopoietic neoplasms of mice: analysis of mouse specific and human antibodies cross-reactive with murine tissue. Toxicol Pathol 2007; 35: 366–375.

    Article  CAS  PubMed  Google Scholar 

  36. Barjesteh van Waalwijk van Doorn-Kh S, Erpelinck C, Löwenberg B, Delwal R . Low expression of MDS1-EVI1–like-1 (MEL1) and EVI1-like-1 (EL1) genes in favorable-risk acute myeloid leukemia. Exp Hematol 2003; 31: 1066–1072.

    Article  Google Scholar 

  37. Vinatzer U, Mannhalter C, Mitterbauer M, Gruener H, Greinix H, Schmidt HH et al. Quantitative comparison of the expression of EVI1 and its presumptive antagonist, MDS1/EVI1, in patients with myeloid leukemia. Genes Chromosomes Cancer 2003; 36: 80–89.

    Article  CAS  PubMed  Google Scholar 

  38. Srinivasula SM, Ashwell JD . IAPs: what’s in a name? Mol Cell 2008; 30: 123–135.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Zhang XD, Franco A, Myers K, Gray C, Nguyen T, Hersey P . Relation of TNF-related apoptosis-inducing ligand (TRAIL) receptor and FLICE-inhibitory protein expression to TRAIL-induced apoptosis of melanoma. Cancer Res 1999; 59: 2747–2753.

    CAS  PubMed  Google Scholar 

  40. Leverkus M, Neumann M, Mengling T, Rauch CT, Brocker EB, Krammer PH et al. Regulation of tumor necrosis factor-related apoptosis-inducing ligand sensitivity in primary and transformed human keratinocytes. Cancer Res 2000; 60: 553–559.

    CAS  PubMed  Google Scholar 

  41. Eggert A, Grotzer MA, Zuzak TJ, Wiewrodt BR, Ho R, Ikegaki N et al. Resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in neuroblastoma cells correlates with a loss of caspase-8 expression. Cancer Res 2001; 61: 1314–1319.

    CAS  PubMed  Google Scholar 

  42. Shultz LD, Lyons BL, Burzenski LM, Gott B, Chen X, Chaleff S et al. Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells. J Immunol 2005; 174: 6477–6489.

    Article  CAS  PubMed  Google Scholar 

  43. McDermott SP, Eppert K, Lechman ER, Doedens M, Dick JE . Comparison of human cord blood engraftment between immunocompromised mouse strains. Blood 2010; 116: 193–200.

    Article  CAS  PubMed  Google Scholar 

  44. Notta F, Doulatov S, Dick JE . Engraftment of human hematopoietic stem cells is more efficient in female NOD/SCID/IL-2Rgc-null recipients. Blood 2010; 115: 3704–3707.

    Article  CAS  PubMed  Google Scholar 

  45. Russell M, List A, Greenberg P, Woodward S, Glinsmann B, Parganas E et al. Expression of EVI1 in myelodysplastic syndromes and other hematologic malignancies without 3q26 translocations. Blood 1994; 84: 1243–1248.

    CAS  PubMed  Google Scholar 

  46. Arai S, Yoshimi A, Shimabe M, Ichikawa M, Nakagawa M, Imai Y et al. Evi-1 is a transcriptional target of MLL oncoproteins in hematopoietic stem cells. Blood 2010; 117: 6304–6314.

    Article  PubMed  Google Scholar 

  47. Modlich U, Schambach A, Brugman MH, Wicke DC, Knoess S, Li Z et al. Leukemia induction after a single retroviral vector insertion in Evi1 or Prdm16. Leukemia 2008; 22: 1519–1528.

    Article  CAS  PubMed  Google Scholar 

  48. Chiu PP, Jiang H, Dick JE . Leukemia-initiating cells in human T-lymphoblastic leukemia exhibit glucocorticoid resistance. Blood 2010; 116: 5268–5279.

    Article  CAS  PubMed  Google Scholar 

  49. Bonnet D . Humanized model to study leukemic stem cells. Methods Mol Biol 2009; 538: 247–262.

    Article  CAS  PubMed  Google Scholar 

  50. Morisot S, Wayne AS, Bohana-Kashtan O, Kaplan IM, Gocke CD, Hildreth R et al. High frequencies of leukemia stem cells in poor-outcome childhood precursor-B acute lymphoblastic leukemias. Leukemia 2010; 24: 1859–1866.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Meyer LH, Eckhoff SM, Queudeville M, Kraus JM, Giordan M, Stursberg J et al. Early relapse in all is identified by time to leukemia in NOD/SCID mice and is characterized by a gene signature involving survival pathways. Cancer Cell 2011; 19: 206–217.

    Article  CAS  PubMed  Google Scholar 

  52. Kurokawa M, Mitani K, Yamagata T, Takahashi T, Izutsu K, Ogawa S et al. The evi-1 oncoprotein inhibits c-Jun N-terminal kinase and prevents stress-induced cell death. EMBO J 2000; 19: 2958–2968.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Liu Y, Chen L, Ko TC, Fields AP, Thompson EA . Evi1 is a survival factor which conveys resistance to both TGFbeta- and taxol-mediated cell death via PI3K/AKT. Oncogene 2006; 25: 3565–3575.

    Article  CAS  PubMed  Google Scholar 

  54. Yoshimi A, Goyama S, Watanabe-Okochi N, Yoshiki Y, Nannya Y, Nitta E et al. Evi1 represses PTEN expression and activates PI3K/AKT/mTOR via interactions with polycomb proteins. Blood 2011; 117: 3617–3628.

    Article  CAS  PubMed  Google Scholar 

  55. Jazaeri AA, Ferriss JS, Bryant JL, Dalton MS, Dutta A . Evaluation of EVI1 and EVI1s (Delta324) as potential therapeutic targets in ovarian cancer. Gynecol Oncol 2010; 118: 189–195.

    Article  CAS  PubMed  Google Scholar 

  56. Roos WP, Kaina B . DNA damage-induced cell death by apoptosis. Trends Mol Med 2006; 12: 440–450.

    Article  CAS  PubMed  Google Scholar 

  57. Pradhan AK, Mohapatra AD, Nayak KB, Chakraborty S . Acetylation of the proto-oncogene EVI1 abrogates Bcl-xL promoter binding and induces apoptosis. PLoS One 2011; 6: e25370.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Shackelford D, Kenific C, Blusztajn A, Waxman S, Ren R . Targeted degradation of the AML1/MDS1/EVI1 oncoprotein by arsenic trioxide. Cancer Res 2006; 66: 11360–11369.

    Article  CAS  PubMed  Google Scholar 

  59. Raza A, Buonamici S, Lisak L, Tahir S, Li D, Imran M et al. Arsenic trioxide and thalidomide combination produces multi-lineage hematological responses in myelodysplastic syndromes patients, particularly in those with high pre-therapy EVI1 expression. Leuk Res 2004; 28: 791–803.

    Article  CAS  PubMed  Google Scholar 

  60. Zhang Y, Sicot G, Cui X, Vogel M, Wuertzer CA, Lezon-Geyda K et al. Targeting a DNA binding motif of the EVI1 protein by a pyrrole-imidazole polyamide. Biochemistry 2011; 50: 10431–10441.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This study was supported by the Max-Eder Program of the Deutsche Krebshilfe, the Deutsche Forschungsgemeinschaft (SFB773, SFB685, KFO 183/TP4) and the Fortüne Programme of the University of Tuebingen. We thank Claudia Kloss, Sascha Bjoern Fischer and Michael Urschitz for help with histological analyses, FACS analyses and statistical analysis, and Axel Schambach, Maike Stahlhut and Teng Cheong Ha for expression vectors.

Authors contributions

MK, MCA, MG, HW, SG, LQM, OCR and SL designed and performed experiments. CL, MK, MCA, LQM, FE and KSO analyzed results. ME performed the analysis of the pediatric ALL patient cohort. RH, ME, OSK and HRS contributed critical reagents. CL and MK wrote the paper. KSO, MCA, RH, LK, FE, FF and HRS contributed to writing the paper. CL designed the research. All authors contributed to editing of the manuscript.

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Authors and Affiliations

  1. Department of Hematology and Oncology, University of Tuebingen Medical Center II, Tuebingen, Germany

    M Konantz, M Grauer, H Wang, S Grzywna, H R Salih, L Kanz & C Lengerke

  2. Department of Pediatric Hematology and Oncology, University of Tuebingen Children’s Hospital, Tuebingen, Germany

    M C André, M Ebinger & R Handgretinger

  3. Department of Pediatric Intensive Care Medicine, University Children's Hospital (UKBB), Basel, Switzerland

    M C André

  4. Interfaculty Institute of Biochemistry, University of Tuebingen, Tuebingen, Germany

    O C Rothfuss, S Lehle, K Schulze-Osthoff & F Essmann

  5. Department of Experimental Hematology, Hannover Medical School, Hannover, Germany

    O S Kustikova & C Baum

  6. Institute of Pathology, University of Tuebingen, Tuebingen, Germany

    F Fend & L Quintanilla-Martinez

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  1. M Konantz
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Correspondence to C Lengerke.

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Konantz, M., André, M., Ebinger, M. et al. EVI-1 modulates leukemogenic potential and apoptosis sensitivity in human acute lymphoblastic leukemia. Leukemia 27, 56–65 (2013). https://doi.org/10.1038/leu.2012.211

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