The online version of this article (https://doi.org/10.1186/s12885-018-4346-1) contains supplementary material, which is available to authorized users.
The addition of high-dose cytarabine to the treatment of mantle cell lymphoma (MCL) has significantly prolonged survival of patients, but relapses are common and are normally associated with increased resistance. To elucidate the mechanisms responsible for cytarabine resistance, and to create a tool for drug discovery investigations, we established a unique and molecularly reproducible cytarabine resistant model from the Z138 MCL cell line.
Effects of different substances on cytarabine-sensitive and resistant cells were evaluated by assessment of cell proliferation using [methyl-14C]-thymidine incorporation and molecular changes were investigated by protein and gene expression analyses.
Gene expression profiling revealed that major transcriptional changes occur during the initial phase of adaptation to cellular growth in cytarabine containing media, and only few key genes, including SPIB, are deregulated upon the later development of resistance. Resistance was shown to be mediated by down-regulation of the deoxycytidine kinase (dCK) protein, responsible for activation of nucleoside analogue prodrugs. This key event, emphasized by cross-resistance to other nucleoside analogues, did not only effect resistance but also levels of SPIB and NF-κB, as assessed through forced overexpression in resistant cells. Thus, for the first time we show that regulation of drug resistance through prevention of conversion of pro-drug into active drug are closely linked to increased proliferation and resistance to apoptosis in MCL. Using drug libraries, we identify several substances with growth reducing effect on cytarabine resistant cells. We further hypothesized that co-treatment with bortezomib could prevent resistance development. This was confirmed and show that the dCK levels are retained upon co-treatment, indicating a clinical use for bortezomib treatment in combination with cytarabine to avoid development of resistance. The possibility to predict cytarabine resistance in diagnostic samples was assessed, but analysis show that a majority of patients have moderate to high expression of dCK at diagnosis, corresponding well to the initial clinical response to cytarabine treatment.
We show that cytarabine resistance potentially can be avoided or at least delayed through co-treatment with bortezomib, and that down-regulation of dCK and up-regulation of SPIB and NF-κB are the main molecular events driving cytarabine resistance development.
Levine PH, Hoover R. The emerging epidemic of non-Hodgkin's lymphoma: current knowledge regarding etiological factors. Cancer Epidemiol Biomarkers Prev. 1992;1:515–7. PubMed
Chaturvedi NK, Rajule RN, Shukla A, Radhakrishnan P, Todd GL, Natarajan A, Vose JM, Joshi SS. Novel treatment for mantle cell lymphoma including therapy-resistant tumor by NF-κB and mTOR dual-targeting approach. Mol Cancer Ther. 2013; https://doi.org/10.1158/1535-7163.MCT-13-0239.
Dreyling M, Kluin-Nelemans HC, Bea S, Klapper W, Vogt N, Delfau-Larue MH, Hutter G, Cheah C, Chiappella A, Cortelazzo S, et al. Update on the molecular pathogenesis and clinical treatment of mantle cell lymphoma: report of the 11th annual conference of the European mantle cell lymphoma network. Leuk Lymphoma. 2013;54:699–707. CrossRefPubMed
Geisler CH, Kolstad A, Laurell A, Jerkeman M, Raty R, Andersen NS, Pedersen LB, Eriksson M, Nordstrom M, Kimby E, et al. Nordic MCL2 trial update: six-year follow-up after intensive immunochemotherapy for untreated mantle cell lymphoma followed by BEAM or BEAC + autologous stem-cell support: still very long survival but late relapses do occur. Br J Haematol. 2012;158:355–62. CrossRefPubMed
Geisler CH, Kolstad A, Laurell A, Andersen NS, Pedersen LB, Jerkeman M, Eriksson M, Nordström M, Kimby E, Boesen AM, et al. Long-term progression-free survival of mantle cell lymphoma after intensive front-line immunochemotherapy with in vivo–purged stem cell rescue: a nonrandomized phase 2 multicenter study by the Nordic lymphoma group. Blood. 2008;112:2687–93. CrossRefPubMedPubMedCentral
Marcé S, Molina-Arcas M, Villamor N, Casado F, Campo E, Pastor-Anglada M, Colomer D. Expression of human equilibrative nucleoside transporter 1 (hENT1) and its correlation with gemcitabine uptake and cytotoxicity in mantle cell lymphoma. Hematol J. 2006;91:895–902.
Klanova M, Lorkova L, Vit O, Maswabi B, Molinsky J, Pospisilova J, Vockova P, Mavis C, Lateckova L, Kulvait V, et al. Downregulation of deoxycytidine kinase in cytarabine-resistant mantle cell lymphoma cells confers cross-resistance to nucleoside analogs gemcitabine, fludarabine and cladribine, but not to other classes of anti-lymphoma agents. Mol Cancer. 2014;13:159. CrossRefPubMedPubMedCentral
Dumontet C, Fabianowska-Majewska K, Mantincic D, Callet Bauchu E, Tigaud I, Gandhi V, Lepoivre M, Peters GJ, Rolland MO, Wyczechowska D, et al. Common resistance mechanisms to deoxynucleoside analogues in variants of the human erythroleukaemic line K562. Br J Haematol. 1999;106:78–85. CrossRefPubMed
Kuci V, Nordstrom L, Conrotto P, Ek S. SOX11 and HIG-2 are cross-regulated and affect growth in mantle cell lymphoma. Leuk Lymphoma. 2016;57(8):1883–92. https://doi.org/10.3109/10428194.2015.1121257.
Nordström L, Sernbo S, Eden P, Grønbæk K, Kolstad A, Räty R, Karjalainen M-L, Geisler C, Ralfkiær E, Sundström C, et al. SOX11 and TP53 add prognostic information to MIPI in a homogenously treated cohort of mantle cell lymphoma – a Nordic lymphoma group study. Br J Haematol. 2014;166:98–108. CrossRefPubMedPubMedCentral
Eskelund CW, Dahl C, Hansen JW, Westman M, Kolstad A, Pedersen LB, Montano C, Husby S, Freiburghaus C, Ek S, et al. TP53 mutations identify younger mantle cell lymphoma patients that do not benefit from intensive chemoimmunotherapy: data from the Nordic MCL2 and MCL3 trial cohorts. Blood. 2017;130:1903–10. CrossRefPubMed
Grant S. Ara-C. cellular and molecular pharmacology. In: FVW G, George K, editors. Advances in Cancer research, Academic Press; 1997;72:197–233. https://doi.org/10.1016/S0065-230X(08)60703-4.
Staub M, Eriksson S. The role of deoxycytidine kinase in DNA synthesis and nucleoside analog activation. In: Deoxynucleoside analogs in cancer therapy: Springer; 2006. p. 29–52. https://doi.org/10.1007/978-1-59745-148-2_2.
Marce S, Molina-Arcas M, Villamor N, Casado F, Campo E, Pastor-Anglada M, Colomer D. Expression of human equilibrative nucleoside transporter 1 (hENT1) and its correlation with gemcitabine uptake and cytotoxicity in mantle cell lymphoma. Haematol. 2006;91:895–902.
Hubeek I, Stam R, Peters G, Broekhuizen R, Meijerink J, van Wering E, Gibson B, Creutzig U, Zwaan C, Cloos J, et al. The human equilibrative nucleoside transporter 1 mediates in vitro cytarabine sensitivity in childhood acute myeloid leukaemia. Br J Cancer. 2005;93:1388–94. CrossRefPubMedPubMedCentral
Kroep JR, Loves WJ, van der Wilt CL, Alvarez E, Talianidis I, Boven E, Braakhuis BJ, van Groeningen CJ, Pinedo HM, Peters GJ. Pretreatment deoxycytidine kinase levels predict in vivo gemcitabine sensitivity. Mol Cancer Ther. 2002;1:371–6. PubMed
Forstpointner R, Unterhalt M, Dreyling M, Bock HP, Repp R, Wandt H, Pott C, Seymour JF, Metzner B, Hanel A, et al. Maintenance therapy with rituximab leads to a significant prolongation of response duration after salvage therapy with a combination of rituximab, fludarabine, cyclophosphamide, and mitoxantrone (R-FCM) in patients with recurring and refractory follicular and mantle cell lymphomas: results of a prospective randomized study of the German low grade lymphoma study group (GLSG). Blood. 2006;108:4003–8. CrossRefPubMed
Negoro E, Yamauchi T, Urasaki Y, Nishi R, Hori H, Ueda T. Characterization of cytarabine-resistant leukemic cell lines established from five different blood cell lineages using gene expression and proteomic analyses. Int J Oncol. 2011;38:911–9. PubMed
Antonsson BE, Avramis VI, Nyce J, Holcenberg JS. Effect of 5-azacytidine and congeners on DNA methylation and expression of deoxycytidine kinase in the human lymphoid cell lines CCRF/CEM/0 and CCRF/CEM/dCk-1. Cancer Res. 1987;47:3672–8. PubMed
Candelaria M, de la Cruz-Hernandez E, Taja-Chayeb L, Perez-Cardenas E, Trejo-Becerril C, Gonzalez-Fierro A, Chavez-Blanco A, Soto-Reyes E, Dominguez G, Trujillo JE, et al. DNA methylation-independent reversion of gemcitabine resistance by hydralazine in cervical Cancer cells. PLoS One. 2012;7:e29181. CrossRefPubMedPubMedCentral
Weigert O, Weidmann E, Mueck R, Bentz M, von Schilling C, Rohrberg R, Jentsch-Ullrich K, Hiddemann W, Dreyling M. A novel regimen combining high dose cytarabine and bortezomib has activity in multiply relapsed and refractory mantle cell lymphoma – long-term results of a multicenter observation study. Leuk Lymphoma. 2009;50:716–22. CrossRefPubMed
Dreyling M. ClinicalTrials.gov: Bethesda (MD): National Library of Medicine; 2011 Sept 28. Identifier NCT01449344, Efficacy and Safety of R-HAD Alone or in Combination With Bortezomib in Patients With Relapsed or Refractory MCL ((R-HAD)). Cited in 2017 Jun 29. Available from https://clinicaltrials.gov/ct2/show/NCT01449344.
- Bortezomib prevents cytarabine resistance in MCL, which is characterized by down-regulation of dCK and up-regulation of SPIB resulting in high NF-κB activity
Venera Kuci Emruli
Christian Winther Eskelund
- BioMed Central
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