Differential Effect of GM-CSF on the Intracellular Ara-C Metabolism in Normal Bone Marrow Mononuclear Cells and Acute Myeloid Leukemia (AML) Blasts

  • Christoph Reuter
  • Claus Rolf
  • Eberhard Schleyer
  • Michael Unterhalt
  • Bernhard Woermann
  • Thomas Buechner
  • Wolfgang Hiddemann
Conference paper
Part of the Haematology and Blood Transfusion / Hämatologie und Bluttransfusion book series (HAEMATOLOGY, volume 37)

Abstract

This study examines the effect of recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) on the intracellular metabolism of cytosine arabinoside (ara-C) in mononuclear bone marrow cells (NBMMC) from 8 healthy volunteers and leukemic blasts from 50 patients with acute myeloid leukemia (AML). Pretreatment with rh GM-CSF (100 U/ml) for 48 hrs significantly enhanced DNA synthesis: Median 3H-TdR uptake into the DNA increased from 2.15 to 3.68 pmol/105 cells in AML blasts and from 3.09 to 7.4 pmol/105 cells in NBMMC (p<0.05). 14 AML cases did not respond to GM-CSF. Median ara-C mediated inhibition of DNA synthesis was significantly higher in AML blasts as compared to NBMMC (76.5 vs 55.0% and 99.0 vs 96.0% at 0.05 and 5.0 μM ara-C, respectively, p<0.01). Similarly, intracellular ara-CTP levels were higher in AML blasts as compared to NBMMC (46.5 vs 18.7,167.8 vs 48.0 and 59.5 ng/107 cells at 1, 10, 100 μM extracellular ara-C, respectively, p< 0.01). Overall DNA polymerase and DNA polymerase α activity increased from median 80.9 to 94.1 and from 3.1 to 5.7 pmol/min × mg in AML blasts as compared to median 96.7 to 189.8 and 1.2 to 2.2 pmol/min × mg in NBMMC (p < 0.05). Median deoxycytidine and thymidine kinase activity were only slightly increased in the presence of GM-CSF. The GM-CSF induced increase of the 3H-ara-C incorporation into the DNA was slightly higher in GM-CSF responding AML blasts as compared to NBMMC (median 2.0 vs 1.7 fold at 0.06 μM ara-C, p < 0.05). The differential effect of GM-CSF on the metabolism of ara-C in normal versus leukemic cells may cause a selective increase in the antileukemic cytotoxicity of ara-C in the presence of GM-CSF.

Keywords

Leukemia Oncol Doxorubicine Thymidine Cytosine 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Hiddemann W. Cytosine arabinoside in the treatment of acute myeloid leukemia: the role and place of high-dose regimens. Ann. Hematol. 1991; 62: 119–128.PubMedCrossRefGoogle Scholar
  2. 2.
    Hiddemann W, Kiehl M, Zuehlsdorf M, Busemann C, Schleyer E, Woermann B, Buechner T. Granulocyte-macrophage colony-stimulating factor and interleukin-3 enhance the incorporation of cytosine arabinoside into the DNA of leukemic blasts and the cytotoxic effect on clonogenic cells from patients with acute myeloid leukemia. Sem. Oncol. 1992; 19: 31–37.Google Scholar
  3. 3.
    Lista P, Porcu P, Avazi GC, Pegoraro L. Interleukin-3 enhances the cytotoxic activity of 1-β D-arabino-furanosylcytosine (ara-C on acute myeloblastic leukemia (AML) cells. Br. J. Heamatol. 1988; 69: 121–123.CrossRefGoogle Scholar
  4. 4.
    Miyauchi J, Kelleher CA, Wang C, Minkin S, McCulloch EA. Growth factors influence the sensitivity of leukemic stem cells to cytosine arabinoside in culture. Blood 1989; 73: 1271–1278.Google Scholar
  5. 5.
    Cannistra SA, Groshek P, Griffin JD. Granulocyte-macrophage colony-stimulating factor enhances the cytotoxic effects of cytosine arabinoside in acute myeloblastic leukemia and in the myeloid blast crisis phase of chronic myeloid leukemia. Leukemia 1989; 3: 328–334.PubMedGoogle Scholar
  6. 6.
    Butturini A, Alessandra MA, Gale RP, Perocco P, Tur S. GM-CSF incubation prior to treatment with cytarabine or doxorubicine enhances drug activity against AML cells in vitro: A model for leukemia chemotherapy. Leukemia Res. 1990; 14: 743–749.CrossRefGoogle Scholar
  7. 7.
    Brach M, Klein H, Platzer E, Mertelsmann R, Hermann F. Effect of interleukin-3 on cytosine arabinoside-mediated cytotoxicity of leukemic myeloblasts. Exp. Hematol. 1990; 18: 748–753.PubMedGoogle Scholar
  8. 8.
    Lista P, Porcu P, Avanzi GC, Pegorarro L. Interleulin 3 enhances the cytotoxic activity of 1-β arabino-furanosyl-cytosine (ara-C) on acute myeloblastic leukaemia (AML) cells. Br. J. Haematol. 1988; 69: 121–123.CrossRefGoogle Scholar
  9. 9.
    Lista P, Rossi BM, Resegotti L, Clark SC, Pegoraro L. Different sensitivity of normal and leukemic progenitor cells to ara-C and IL-3 treatment. Br. J. Heamatol. 1990; 76: 21–26.CrossRefGoogle Scholar
  10. 10.
    Van Der Lely N, Witte T, Muus P, Raymakers R, Preijers F, Haanen C. Prolonged exposure to cytosine arabinoside in the presence of hematopoietic growth factors preferentially kills leukemic versus normal clonogenic cells. Exp. Hematol. 1991; 19: 267–272.Google Scholar
  11. 11.
    Karp JE, Burke PJ, Donehower RC. Effect of rh GM-CSF on intracellular ara-C pharmacology in vitro in acute myelocytic leukemia: comparability with drug-induced humoral stimulatory activity. Leukemia 1990; 4: 553–556.PubMedGoogle Scholar
  12. 12.
    Bhalla K, Birkhofer M, Arlin Z, Lutzky J, Graham G. Effect of recombinant GM-CSF on the metabolism of cytosine arabinoside in normal and leukemic human bone marrow cells. Leukemia 1988; 2: 810–813.PubMedGoogle Scholar
  13. 13.
    Bhalla K, Birkhofer M, Arlin Z, Grant S, Lutzky J, Safah H, Graham G. Differential effect of interleukin-3 on the metabolism of high dose cytosine arabinoside in normal versus leukemic human bone marrow cells. Exp. Hematol. 1991; 19: 669–673.PubMedGoogle Scholar
  14. 14.
    Bhalla K, Holladay C, Arlin Z, Grant S, Ibrado AM, Jasiok M. Treatment with interleukin-3 plus granulocyte-macrophage colony-stimulating factor improves the selectivity of ara-C in vitro against acute myeloid leukemia blasts. Blood 1991; 78: 2674–2679.PubMedGoogle Scholar
  15. 15.
    te Boekhorst PAW, Loewenberg B, Vlastuin M, Sonneveld P. Enhanced chemosensitivity of clonogenic blasts from patients with acute myeloid leukemia by G-CSF, IL-3 or GM-CSF stimulation. Leukemia 1993; 7: 1191–1198.Google Scholar
  16. 16.
    Yang GS, Minden MD, McCulloch EA. Influence of schedule on regulated sensitivity of AML blasts to cytosine arabinoside. Leukemia 1993; 7: 1012–1019.PubMedGoogle Scholar
  17. 17.
    Reuter C, Auf Der Landwehr U I, Auf Der Landwehr U II, Schleyer E, Zuehlsdorf M, Ameling C, Rolf C, Woermann B, Buechner T, Hiddemann W. Modulation of intracellular metabolism of cytosine arabinoside in acute myeloid leukemia by granulocyte macrophage colony stimulating factor. Leukemia 1994; 8: 217–225.PubMedGoogle Scholar
  18. 18.
    Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the prinziple of protein-dye binding. Anal. Biochem. 1976; 72: 248–254.PubMedCrossRefGoogle Scholar
  19. 19.
    Schleyer E, Ehninger G, Zuehlsdorf M, Proksch B, Hiddemann W. Detection and separation of intracellular ara-CTP by ion-pair high performance liquid chromatography: a sensitive, isocratic, highly reproducible and rapid method. J. Chromatography 1989; 497: 109–120.CrossRefGoogle Scholar
  20. 20.
    Kufe D, Spriggs D. Biochemical and cellular pharmacology of cytosine arabinoside. Sem. Oncol. 1985; 12(2): 34–48.Google Scholar
  21. 21.
    Aglietta M, De Felice L, Stacchini A, Petti MC, Bianchi ACM, Aloe Spiriti MA, Sanavio F, Apra F, Piacibello W, Stern AC, Gavosto F, Mandelli F. In vivo effect of granulocyte-macrophage colony stimulating factor on the kinetics of human acute myeloid leukemia cells. Leukemia 1991; 5: 979–984.PubMedGoogle Scholar
  22. 22.
    Bettelheim P, Valent P, Andreeff M, Tafuri A, Haimi J, Gorischek C, Muhm M, Sillaber Ch, Haas O, Vieder L, Maurer D, Schulz G, Speiser W, Geissler K, Kier P, Hinterberger W, Lechner K. Recombinant human granulocyte-macrophage colony-stimulating factor in combination with standard induction chemotherapy in de novo acute myeloid leukemia. Blood 1991; 77: 700–711.PubMedGoogle Scholar
  23. 23.
    Cannistra SA, Di Carlo J, Groshek P, Kanakura Y, Berg D, Mayer RJ, Griffen JD. Simultaneous administration of granulocyte-macrophage colony-stimulating factor and cytosine arabinoside for the treatment of relapsed acute myeloid leukemia. Leukemia 1991; 5: 230–238.PubMedGoogle Scholar
  24. 24.
    Estey E, Thall PF, Kantarjian H, O’Brien S, Koller CA, Beran M, Gutterman J, Deisseroth A, Keating M. Treatment of newly diagnosed acute myelogenous leukemia with granulocyte-macrophage colony-stimulating factor (GM-CSF) before and during continous-infusion high-dose ara-C + daunorubicin: comparison to patients treated without GM-CSF. Blood 1992; 79: 2236–2255.Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1996

Authors and Affiliations

  • Christoph Reuter
    • 1
  • Claus Rolf
    • 2
  • Eberhard Schleyer
    • 1
  • Michael Unterhalt
    • 1
  • Bernhard Woermann
    • 1
  • Thomas Buechner
    • 2
  • Wolfgang Hiddemann
    • 1
  1. 1.Department of Hematology and OncologyUniversity of GöttingenGöttingenGermany
  2. 2.Department of Internal MedicineUniversity of MünsterMünsterGermany

Personalised recommendations