Abstract
Bendamustine is used in the treatment of chronic lymphocytic leukemia (CLL). Routes for bendamustine entry into target cells are unknown. This study aimed at identifying transporter proteins implicated in bendamustine uptake. Our results showed that hOCT1 is a bendamustine transporter, as bendamustine could cis-inhibit the uptake of a canonical hOCT1 substrate, with a Ki in the micromolar range, consistent with the EC50 values of the cytotoxicity triggered by this drug in HEK293 cells expressing hOCT1. hOCT1 polymorphic variants determining impaired bendamustine-transporter interaction, consistently reduced bendamustine cytotoxicity in HEK293 cells stably expressing them. Exome genotyping of the SLC22A1 gene, encoding hOCT1, was undertaken in a cohort of 241 CLL patients. Ex vivo cytotoxicity to bendamustine was measured in a subset of cases and shown to correlate with SLC22A1 polymorphic variants. In conclusion, hOCT1 is a suitable bendamustine transporter, thereby contributing to its cytotoxic effect depending upon the hOCT1 genetic variants expressed.
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References
Chang JE, Kahl BS . Bendamustine for treatment of chronic lymphocytic leukemia. Expert Opin Pharmacother 2012; 13: 1495–1505.
Hoy SM . Bendamustine: a review of its use in the management of chronic lymphocytic leukaemia, rituximab-refractory indolent non-Hodgkin’s lymphoma and multiple myeloma. Drugs 2012; 72: 1929–1950.
Cheson BD, Wendtner CM, Pieper A, Dreyling M, Friedberg J, Hoelzer D et al. Optimal use of bendamustine in chronic lymphocytic leukemia, non-Hodgkin lymphomas, and multiple myeloma: treatment recommendations from an international consensus panel. Clin Lymphoma Myeloma Leuk 2010; 10: 21–27.
Gaidano G, Foa R, Dalla-Favera R . Molecular pathogenesis of chronic lymphocytic leukemia. J Clin Invest 2012; 122: 3432–3438.
Damle RN, Wasil T, Fais F, Ghiotto F, Valetto A, Allen SL et al. Ig V gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood 1999; 94: 1840–1847.
Crespo M, Bosch F, Villamor N, Bellosillo B, Colomer D, Rozman M et al. ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia. N Engl J Med 2003; 348: 1764–1775.
Rossi D, Rasi S, Spina V, Bruscaggin A, Monti S, Ciardullo C et al. Integrated mutational and cytogenetic analysis identifies new prognostic subgroups in chronic lymphocytic leukemia. Blood 2013; 121: 1403–1412.
Quesada V, Conde L, Villamor N, Ordóñez GR, Jares P, Bassaganyas L et al. Exome sequencing identifies recurrent mutations of the splicing factor SF3B1 gene in chronic lymphocytic leukemia. Nat Genet 2011; 44: 47–52.
Puente XS, Pinyol M, Quesada V, Conde L, Ordóñez GR, Villamor N et al. Whole-genome sequencing identifies recurrent mutations in chronic lymphocytic leukaemia. Nature 2011; 475: 101–105.
Hallek M . Signaling the end of chronic lymphocytic leukemia: new frontline treatment strategies. Blood 2013; 122: 3723–3734.
Traynor K . Treanda approved for chronic lymphocytic leukemia. Am J Health Syst Pharm 2008; 65: 793.
Knauf WU, Lissichkov T, Aldaoud A, Liberati A, Loscertales J, Herbrecht R et al. Phase III randomized study of bendamustine compared with chlorambucil in previously untreated patients with chronic lymphocytic leukemia. J Clin Oncol 2009; 27: 4378–4384.
www.ema.europa.eu/ E.M.A.w. (2012). Accessed February 11.
Bergmann MA, Goebeler ME, Herold M, Emmerich B, Wilhelm M, Ruelfs C et al. Efficacy of bendamustine in patients with relapsed or refractory chronic lymphocytic leukemia: results of a phase I/II study of the German CLL Study Group. Haematologica 2005; 90: 1357–1364.
Cortelezzi A, Sciumè M, Liberati AM, Vincenti D, Cuneo A, Reda G et al. Bendamustine in combination with ofatumumab in relapsed or refractory chronic lymphocytic leukemia: a GIMEMA Multicenter Phase II Trial. Leukemia 2013; 28: 642–648.
Hill BT, Caimi P, Kindwall-Keller T, Habecker B, Kalaycio M . Bendamustine plus alemtuzumab is safe and feasible treatment for fludarabine refractory chronic lymphocytic leukaemia (CLL). Br J Haematol 2014; 164: 297–299.
Fischer K, Cramer P, Busch R, Böttcher S, Bahlo J, Schubert J et al. Bendamustine in combination with rituximab for previously untreated patients with chronic lymphocytic leukemia: a multicenter phase II trial of the German Chronic Lymphocytic Leukemia Study Group. J Clin Oncol 2012; 30: 3209–3216.
Fischer K, Cramer P, Busch R, Stilgenbauer S, Bahlo J, Schweighofer CD et al. Bendamustine combined with rituximab in patients with relapsed and/or refractory chronic lymphocytic leukemia: a multicenter phase II trial of the German Chronic Lymphocytic Leukemia Study Group. J Clin Oncol 2011; 29: 3559–3566.
Molina-Arcas M, Bellosillo B, Casado FJ, Montserrat E, Gil J, Colomer D et al. Fludarabine uptake mechanisms in B-cell chronic lymphocytic leukemia. Blood 2003; 101: 2328–2334.
Molina-Arcas M, Marcé S, Villamor N, Huber-Ruano I, Casado FJ, Bellosillo B et al. Equilibrative nucleoside transporter-2 (hENT2) protein expression correlates with ex vivo sensitivity to fludarabine in chronic lymphocytic leukemia (CLL) cells. Leukemia 2005; 19: 64–68.
Marce S, Molina-Arcas M, Villamor N, Casado FJ, Campo E, Pastor-Anglada M et al. Expression of human equilibrative nucleoside transporter 1 (hENT1) and its correlation with gemcitabine uptake and cytotoxicity in mantle cell lymphoma. Haematologica 2006; 91: 895–902.
Minuesa G, Purcet S, Erkizia I, Molina-Arcas M, Bofill M, Izquierdo-Useros N et al. Expression and functionality of anti-human immunodeficiency virus and anticancer drug uptake transporters in immune cells. J Pharmacol Exp Ther 2008; 324: 558–567.
Gupta S, Wulf G, Henjakovic M, Koepsell H, Burckhardt G, Hagos Y . Human organic cation transporter 1 is expressed in lymphoma cells and increases susceptibility to irinotecan and paclitaxel. J Pharmacol Exp Ther 2012; 341: 16–23.
Mata JF, García-Manteiga JM, Lostao MP, Fernández-Veledo S, Guillén-Gómez E, Larrayoz IM et al. Role of the human concentrative nucleoside transporter (hCNT1) in the cytotoxic action of 5[Prime]-deoxy-5-fluorouridine, an active intermediate metabolite of capecitabine, a novel oral anticancer drug. Mol Pharmacol 2001; 59: 1542–1548.
Errasti-Murugarren E, Pastor-Anglada M, Casado FJ . Role of CNT3 in the transepithelial flux of nucleosides and nucleoside-derived drugs. J Physiol 2007; 582: 1249–1260.
Gorboulev V, Ulzheimer JC, Akhoundova A, Ulzheimer-Teuber I, Karbach U, Quester S et al. Cloning and characterization of two human polyspecific organic cation transporters. DNA Cell Biol 1997; 16: 871–881.
Nies AT, Koepsell H, Winter S, Burk O, Klein K, Kerb R et al. Expression of organic cation transporters OCT1 (SLC22A1) and OCT3 (SLC22A3) is affected by genetic factors and cholestasis in human liver. Hepatology 2009; 50: 1227–1240.
del Santo B, Valdes R, Mata J, Felipe A, Casado FJ, Pastor-Anglada M . Differential expression and regulation of nucleoside transport systems in rat liver parenchymal and hepatoma cells. Hepatology 1998; 28: 1504–1511.
Hudson TJ, Anderson W, Artez A, Barker AD, Bell C, Bernabé RR et al. International network of cancer genome projects. Nature 2010; 464: 993–998.
Minuesa G, Huber-Ruano I, Pastor-Anglada M, Koepsell H, Clotet B, Martinez-Picado J . Drug uptake transporters in antiretroviral therapy. Pharmacol Ther 2011; 132: 268–279.
Minuesa G, Volk C, Molina-Arcas M, Gorboulev V, Erkizia I, Arndt P et al. Transport of lamivudine [(-)-beta-L-2',3'-dideoxy-3'-thiacytidine] and high-affinity interaction of nucleoside reverse transcriptase inhibitors with human organic cation transporters 1, 2, and 3. J Pharmacol Exp Ther 2009; 329: 252–261.
Errasti-Murugarren E, Pastor-Anglada M . Drug transporter pharmacogenetics in nucleoside-based therapies. Pharmacogenomics 2010; 11: 809–841.
Shu Y, Leabman MK, Feng B, Mangravite LM, Huang CC, Stryke D et al. Evolutionary conservation predicts function of variants of the human organic cation transporter, OCT1. Proc Natl Acad Sci USA 2003; 100: 5902–5907.
Shu Y, Sheardown SA, Brown C, Owen RP, Zhang S, Castro RA et al. Effect of genetic variation in the organic cation transporter 1 (OCT1) on metformin action. J Clin Invest 2007; 117: 1422–1431.
Kerb R, Brinkmann U, Chatskaia N, Gorbunov D, Gorboulev V, Mornhinweg E et al. Identification of genetic variations of the human organic cation transporter hOCT1 and their functional consequences. Pharmacogenetics 2002; 12: 591–595.
Dubbelman AC, Rosing H, Darwish M, D'Andrea D, Bond M, Hellriegel E et al. Pharmacokinetics and excretion of 14C-bendamustine in patients with relapsed or refractory malignancy. Drugs R D 2013; 13: 17–28.
Martinez-Becerra P, Vaquero J, Romero MR, Lozano E, Anadon C, Macias RI et al. No correlation between the expression of FXR and genes involved in multidrug resistance phenotype of primary liver tumors. Mol Pharm 2012; 9: 1693–1704.
Herraez E, Lozano E, Macias RI, Vaquero J, Bujanda L, Banales JM et al. Expression of SLC22A1 variants may affect the response of hepatocellular carcinoma and cholangiocarcinoma to sorafenib. Hepatology 2013; 58: 1065–1073.
Schaeffeler E, Hellerbrand C, Nies AT, Winter S, Kruck S, Hofmann U et al. DNA methylation is associated with downregulation of the organic cation transporter OCT1 (SLC22A1) in human hepatocellular carcinoma. Genome Med 2011; 3: 82.
Hayer M, Bonisch H, Bruss M . Molecular cloning, functional characterization and genomic organization of four alternatively spliced isoforms of the human organic cation transporter 1 (hOCT1/SLC22A1). Ann Hum Genet 1999; 63: 473–482.
Wu M, Akinleye A, Zhu X . Novel agents for chronic lymphocytic leukemia. J Hematol Oncol 2013; 6: 36.
Byrd JC, Furman RR, Coutre SE, Flinn IW, Burger JA, Blum KA et al. Targeting BTK with ibrutinib in relapsed chronic lymphocytic leukemia. N Engl J Med 2013; 369: 32–42.
Brown JR, Barrientos JC, Barr PM, Flinn I, Burger JA, Salman Z et al. Ibrutinib in combination with bendamustine and rituximab is active and tolerable in patients with relapsed/refractory CLL/SLL: final results of a phase 1b study. 55th ASH Annual Meeting and Exposition. New Orleans, LA.
Acknowledgements
We thank Ingrid Iglesias, Jocabed Roldan, Laura Jiménez and Sandra Cabezas for their technical support. We also thank all individuals with CLL who have participated in this study from the CLL Spanish Consortium. The UB laboratory is a member of the Oncology Program of the National Biomedical Research Institute of Liver and Gastrointestinal Diseases (CIBER ehd). CIBER ehd is an initiative of Instituto de Salud Carlos III (Spain). Part of this work was carried out at the Esther Koplowitz Center, Barcelona. This work has been funded by Mundipharma Research Ltd. This study was also partially supported by research funding from Ministerio de Ciencia e Innovación (SAF2011-23660 to MP-A. and SAF 12/31242 to DC), Redes Temáticas de Investigación Cooperativa de Cáncer from the Instituto de Salud Carlos III (ISCIII), Spanish Ministry of Economy and Competitiveness & European Regional Development Fund (ERDF) ‘Una manera de hacer Europa’ RD12/0036/0004, RD12/0036/0036; RD12/0036/0067 and Generalitat de Catalunya 2009SGR967; 2014SGR346 (to DC) and 2009SGR624 (to MP-A). CAN, AM and EL are recipients of predoctoral fellowships FPI from Ministerio de Ciencia e Innovación.
Author Contributions
Cristina Arimany Nardi: wrote manuscript, designed research, performed research, analyzed data. Arnau Montraveta: wrote manuscript, performed research, analyzed data. Eriong Lee-Vergés: wrote manuscript, performed research, analyzed data. Xose S. Puente: wrote manuscript, contributed new reagents/analytical tools. Hermann Koepsell: reviewed manuscript, contributed new reagents/analytical tools. Elías Campo: reviewed manuscript. Dolors Colomer: wrote manuscript, designed research, analyzed data. Marçal Pastor-Anglada: wrote manuscript, designed research, analyzed data.
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This study was partially funded by Mundipharma.
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Arimany-Nardi, C., Montraveta, A., Lee-Vergés, E. et al. Human organic cation transporter 1 (hOCT1) as a mediator of bendamustine uptake and cytotoxicity in chronic lymphocytic leukemia (CLL) cells. Pharmacogenomics J 15, 363–371 (2015). https://doi.org/10.1038/tpj.2014.77
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DOI: https://doi.org/10.1038/tpj.2014.77
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