Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Manuscript
  • Published:

Pharmacodynamics and Pharmacogenomics

Modulation of ara-CTP levels by fludarabine and hydroxyurea in leukemic cells

Leukemia volume 15, pages 69–73 (2001)Cite this article

Abstract

The rate of ara-cytosine triphosphate (ara-CTP) accumulation and its retention has been correlated with 1-β-D-arabinofuranosylcytosine (ara-C)-mediated toxicity and clinical outcome in childhood and adult leukemia. We tested to what extent preincubation with the ribonucleotide reductase inhibitors fludarabine (F-ara-A) and hydroxyurea (HU) enhanced ara-CTP levels in two human myeloid (HL-60, CMK) and two lymphoblastic leukemia cell lines (MOLT-4, BLIN-1) and also in blasts from 28 children with acute leukemia (AML: 14, ALL: 14). Incubation experiments carried out with cell lines showed F-ara-A and HU to be equipotent in increasing ara-CTP levels. The highest increase was observed in HL-60 cells whereas preincubation had no modulatory effect in MOLT-4 cells. Accordingly, modulation of intracellular ara-CTP levels differed between the subtypes of childhood acute leukemia: whereas in T-ALL (five) preincubation with F-ara-A and HU had no effect on intracellular ara-C metabolism, increased ara-CTP levels were seen in some cases of pre-B-ALL (seven). In myelogenous blasts (12) clinically relevant enhancement of ara-C toxification was regularly obtained with both, F-ara-A (1.9-fold) and HU (1.5-fold). In conclusion, our data suggest that combinations of ara-C and ribonucleotide reductase inhibitors are apt to increase ara-CTP levels depending on the individual cell type and its sensitivity towards ara-C modulators.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Capizzi RL, White JC, Powell BL, Perrino F . Effect of dose on the pharmacokinetic and pharmacodynamic effects of cytarabine Semin Hematol 1991 28: (Suppl. 4) 54–69

    CAS  PubMed  Google Scholar 

  2. Kufe DW, Spriggs DR . Biochemical and cellular pharmacology of cytosine arabinoside Semin Oncol 1985 12: (Suppl. 3) 34–48

    CAS  PubMed  Google Scholar 

  3. Pourquier P, Takebayashi Y, Urasaki Y, Gioffre C, Kohlhagen G, Pommier Y . Induction of topoisomerase I cleavage complexes by 1-beta-D-arabinofuranosylcytosine (ara-C) in vitro and in ara-C-treated cells Proc Natl Acad Sci USA 2000 97: 1885–1890

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Boos J, Hohenlochter B, Schulze-Westhoff P, Schiller M, Zimmermann M, Creutzig U, Ritter J, Jurgens H . Intracellular retention of cytosine arabinoside triphosphate in blast cells from children with acute myelogenous and lymphoblastic leukemia Med Pediatr Oncol 1996 26: 397–404

    Article  CAS  PubMed  Google Scholar 

  5. Kufe D, Spriggs D, Egan EM, Munroe D . Relationships among Ara-CTP pools, formation of (Ara-C)DNA, and cytotoxicity of human leukemic cells Blood 1984 64: 54–58

    CAS  PubMed  Google Scholar 

  6. Plunkett W, Iacoboni S, Keating MJ . Cellular pharmacology and optimal therapeutic concentrations of 1-beta-D-arabinofuranosylcytosine 5′-triphosphate in leukemic blasts during treatment of refractory leukemia with high-dose 1-beta-D-arabinofuranosylcytosine Scand J Haematol Suppl 1986 44: 51–59

    CAS  PubMed  Google Scholar 

  7. Zuhlsdorf M, Vormoor J, Boos J . Cytosine arabinoside in childhood leukemia Int J Pediatr Hematol/Oncol 1997 4: 565–581

    Google Scholar 

  8. Coleman CN, Stoller RG, Drake JC, Chabner BA . Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes Blood 1975 46: 791–803

    CAS  PubMed  Google Scholar 

  9. Colly LP, Peters WG, Richel D, Arentsen-Honders MW, Starrenburg CW, Willemze R . Deoxycytidine kinase and deoxycytidine deaminase values correspond closely to clinical response to cytosine arabinoside remission induction therapy in patients with acute myelogenous leukemia Semin Oncol 1987 14: (Suppl. 1) 257–261

    CAS  PubMed  Google Scholar 

  10. Kees UR, Ford J, Dawson VM, Piall E, Aherne GW . Development of resistance to 1-beta-D-arabinofuranosylcytosine after high-dose treatment in childhood lymphoblastic leukemia: analysis of resistance mechanism in established cells lines Cancer Res 1989 49: 3015–3019

    CAS  PubMed  Google Scholar 

  11. Donehower RC . An overview of the clinical experience with hydroxyurea Semin Oncol 1992 19: (Suppl. 9) 11–19

    CAS  PubMed  Google Scholar 

  12. Gandhi V, Estey E, Keating MJ, Plunkett W . Biochemical modulation of arabinosylcytosine for therapy of leukemias Leuk Lymphoma 1993 10: (Suppl) 109–114

    Article  PubMed  Google Scholar 

  13. Plunkett W, Huang P, Gandhi V . Metabolism and action of fludarabine phosphate Semin Oncol 1990 17: (Suppl. 8) 3–17

    CAS  PubMed  Google Scholar 

  14. Plunkett W, Gandhi V, Huang P, Robertson LE, Yang LY, Gregoire V, Estey E, Keating MJ . Fludarabine: pharmacokinetics, mechanisms of action, and rationales for combination therapies Semin Oncol 1993 20: (Suppl. 7) 2–12

    CAS  PubMed  Google Scholar 

  15. Walsh CT, Craig RW, Agarwal RP . Increased activation of 1-beta-D-arabinofuranosylcytosine by hydroxyurea in L1210 cells Cancer Res 1980 40: 3286–3292

    CAS  PubMed  Google Scholar 

  16. Gandhi V, Nowak B, Keating MJ, Plunkett W . Modulation of arabinosylcytosine metabolism by arabinosyl-2-fluoroadenine in lymphocytes from patients with chronic lymphocytic leukemia: implications for combination therapy Blood 1989 74: 2070–2075

    CAS  PubMed  Google Scholar 

  17. Gandhi V, Estey E, Keating MJ, Plunkett W . Fludarabine potentiates metabolism of cytarabine in patients with acute myelogenous leukemia during therapy J Clin Oncol 1993 11: 116–124

    Article  CAS  PubMed  Google Scholar 

  18. Avramis VI, Wiersma S, Krailo MD, Ramilo-Torno LV, Sharpe A, Liu-Mares W, Kowck R, Reaman GH, Sato JK . Pharmacokinetic and pharmacodynamic studies of fludarabine and cytosine arabinoside administered as loading boluses followed by continuous infusions after a phase I/II study in pediatric patients with relapsed leukemias. The Children's Cancer Group Clin Cancer Res 1998 4: 45–52

    CAS  PubMed  Google Scholar 

  19. Dinndorf PA, Avramis VI, Wiersma S, Krailo MD, Liu-Mares W, Seibel NL, Sato JK, Mosher RB, Kelleher JF, Reaman GH . Phase I/II study of idarubicin given with continuous infusion fludarabine followed by continuous infusion cytarabine in children with acute leukemia: a report from the Children's Cancer Group J Clin Oncol 1997 15: 2780–2785

    Article  CAS  PubMed  Google Scholar 

  20. Estey E, Plunkett W, Gandhi V, Rios MB, Kantarjian H, Keating MJ . Fludarabine and arabinosylcytosine therapy of refractory and relapsed acute myelogenous leukemia Leuk Lymphoma 1993 9: 343–350

    Article  CAS  PubMed  Google Scholar 

  21. Fleischhack G, Hasan C, Graf N, Mann G, Bode U . IDA-FLAG (idarubicin, fludarabine, cytarabine, G-CSF), an effective remission-induction therapy for poor-prognosis AML of childhood prior to allogeneic or autologous bone marrow transplantation: experiences of a phase II trial Br J Haematol 1998 102: 647–655

    Article  CAS  PubMed  Google Scholar 

  22. Suki S, Kantarjian H, Gandhi V, Estey E, O'Brien S, Beran M, Rios MB, Plunkett W, Keating M . Fludarabine and cytosine arabinoside in the treatment of refractory or relapsed acute lymphocytic leukemia Cancer 1993 72: 2155–2160

    Article  CAS  PubMed  Google Scholar 

  23. Visani G, Tosi P, Zinzani PL, Manfroi S, Ottaviani E, Testoni N, Clavio M, Cenacchi A, Gamberi B, Carrara P . FLAG (fludarabine + high-dose cytarabine + G-CSF): an effective and tolerable protocol for the treatment of ‘poor risk’ acute myeloid leukemias Leukemia 1994 8: 1842–1846

    CAS  PubMed  Google Scholar 

  24. Donehower RC, Karp JE, Burke PJ . Pharmacology and toxicity of high-dose cytarabine by 72-hour continuous infusion Cancer Treat Rep 1986 70: 1059–1065

    CAS  PubMed  Google Scholar 

  25. Kubota M, Takimoto T, Tanizawa A, Akiyama Y, Mikawa H . Differential modulation of 1-beta-D-arabinofuranosylcytosine metabolism by hydroxyurea in human leukemic cell lines Biochem Pharmacol 1988 37: 1745–1749

    Article  CAS  PubMed  Google Scholar 

  26. Plagemann PG, Marz R, Wohlhueter RM . Transport and metabolism of deoxycytidine and 1-beta-D-arabinofuranosylcytosine into cultured Novikoff rat hepatoma cells, relationship to phosphorylation, and regulation of triphosphate synthesis Cancer Res 1978 38: 978–989

    CAS  PubMed  Google Scholar 

  27. Rauscher F, Cadman E . Biochemical and cytokinetic modulation of L1210 and HL-60 cells by hydroxyurea and effect on 1-beta-D-arabinofuranosylcytosine metabolism and cytotoxicity Cancer Res 1983 43: 2688–2693

    CAS  PubMed  Google Scholar 

  28. Schilsky RL, Ratain MJ, Vokes EE, Vogelzang NJ, Anderson J, Peterson BA . Laboratory and clinical studies of biochemical modulation by hydroxyurea Semin Oncol 1992 19: (Suppl. 9) 84–89

    CAS  PubMed  Google Scholar 

  29. Streifel JA, Howell SB . Synergisticinteraction between 1-beta-D-arabinofuranosylcytosine, thymidine, and hydroxyurea against human B cells and leukemic blasts in vitro Proc Natl Acad Sci USA 1981 78: 5132–5136

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  30. Gallagher R, Collins S, Trujillo J, McCredie K, Ahearn M, Tsai S, Metzgar R, Aulakh G, Ting R, Ruscetti F et al. Characterization of the continuous, differentiating myeloid cell line (HL-60) from a patient with acute promyelocytic leukemia Blood 1979 54: 713–733

    CAS  PubMed  Google Scholar 

  31. Minowada J, Onuma T, Moore GE . Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes J Natl Cancer Inst 1972 49: 891–895

    CAS  PubMed  Google Scholar 

  32. Sato T, Fuse A, Eguchi M, Hayashi Y, Ryo R, Adachi M, Kishimoto Y, Teramura M, Mizoguchi H, Shima Y . Establishment of a human leukaemic cell line (CMK) with megakaryocytic characteristics from a Down's syndrome patient with acute megakaryoblastic leukaemia Br J Haematol 1989 72: 184–190

    Article  CAS  PubMed  Google Scholar 

  33. Wormann B, Anderson JM, Liberty JA, Gajl-Peczalska K, Brunning RD, Silberman TL, Arthur DC, LeBien TW . Establishment of a leukemic cell model for studying human pre-B to B cell differentiation J Immunol 1989 142: 110–117

    CAS  PubMed  Google Scholar 

  34. Gandhi V, Kemena A, Keating MJ, Plunkett W . Fludarabine infusion potentiates arabinosylcytosine metabolism in lymphocytes of patients with chronic lymphocytic leukemia Cancer Res 1992 52: 897–903

    CAS  PubMed  Google Scholar 

  35. Rossig C, Freund A, Mayer G, Hohenlochter B, Gescher A, Jurgens H, Boos J . Comparison of hydroxyurea, fludarabine and cladarabine in modulating ara-C pharmacology in the ara-C sensitive and resistant HL-60 and HL-60/Ara-C cell lines. In: Hiddemann W (ed) Acute Leukemias VII^Experimental Approaches and Novel Therapies Springer Verlag: Berlin, Heidelberg 1998 pp 645–650

    Google Scholar 

  36. Gandhi V, Plunkett W . Modulation of arabinosylnucleoside metabolism by arabinosylnucleotides in human leukemia cells Cancer Res 1988 48: 329–334

    CAS  PubMed  Google Scholar 

  37. Boos J . A simple isocratic ion-pair high-performance liquid chromatographic determination of 1-beta-D-arabinofuranosylcytosine 5′-triphosphate for intracellular drug-monitoring and in vitro incubation assays J Pharm Biomed Anal 1991 9: 47–52

    Article  CAS  PubMed  Google Scholar 

  38. Avramis VI, Wiersma S, Krailo MD, Ramilo-Torno LV, Sharpe A, Liu-Mares W, Kowck R, Reaman GH, Sato JK . Pharmacokinetic and pharmacodynamic studies of fludarabine and cytosine arabinoside administered as loading boluses followed by continuous infusions after a phase I/II study in pediatric patients with relapsed leukemias. The Children's Cancer Group Clin Cancer Res 1998 4: 45–52

    CAS  PubMed  Google Scholar 

  39. Rayappa C, McCulloch EA . A cell culture model for the treatment of acute myeloblastic leukemia with fludarabine and cytosine arabinoside Leukemia 1993 7: 992–999

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by the Federal Department of Research and Technology (No. 01EC9401). We thank Mrs G Braun-Munzinger for editing the manuscript.

Author information

Authors and Affiliations

  1. Department of Pediatric Oncology, University of Münster, Albert Schweitzer Str. 33, Münster, 48129, Germany

    M Ahlmann, C Lanvers, K Lümkemann, C Rössig, A Freund, M Baumann & J Boos

Authors
  1. M Ahlmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C Lanvers
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K Lümkemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C Rössig
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A Freund
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M Baumann
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Boos
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ahlmann, M., Lanvers, C., Lümkemann, K. et al. Modulation of ara-CTP levels by fludarabine and hydroxyurea in leukemic cells. Leukemia 15, 69–73 (2001). https://doi.org/10.1038/sj.leu.2401992

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2401992

Keywords

This article is cited by

Search

Advanced search

Quick links