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In vivo depletion of O6-alkylguanine-DNA-alkyltransferase in lymphocytes and melanoma of patients treated with CB 10-277, a new DTIC analogue

  • Original Articles
  • O6-Alkylguanine-DNA-Alkyltransferase, Lymphocytes, Melanoma, CB10-277, DTIC Analogue
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Summary

there is increasing evidence to suggest that alkylation of guanine residues in DNA at the O6 position is the critical cytotoxic event following treatment with dacarbazine (DTIC) and related drugs and that endogenous O6-alkylguanine-DNA alkyltransferase (ATase) gene expression may be a major factor in resistance to such agents. 1-p-Carboxyl-3,3-dimethylphenyltriazene (CB 10-277) was recently selected for clinical evaluation as a DTIC analogue with improved solubility, stability and (possibly) metabolic activation. Serial ATase levels were measured in peripheral blood lymphocytes of nine patients and in biopsied melanoma samples of two patients undergoing treatment with 24-h continuous infusion of CB 10-277 (12 g/m2). Wide individual variations in pre-treatment levels as well as in the post-treatment depletion of lymphocyte ATase were seen. Progressive depletion of lymphocyte ATase was seen during continuous infusion of CB 10-277 in all patients. Complete suppression of lymphocyte ATase activity occurred in two patients whose pre-treatment ATase levels were low. Immediately following completion of the CB 10-277 infusion, the median ATase activity was 17% of pre-treatment levels (range, 0–67%). At 24 h after the end of the infusion, no recovery of lymphocyte ATase activity was observed in six patients, but significant recovery to 50%, 100% and 102% of pre-treatment activity occurred in the other three. In three patients who returned for subsequent cycles of chemotherapy at 4 weeks after the first dose, pre-treatment ATase levels showed a 3-to 4-fold increase relative to the original pre-treatment values. A significant correlation was found between the extent of ATase depletion and the initial lymphocyte ATase levels (r=0.725,P<0.05). Haematological toxicity developed in two patients and was associated with low pre-treatment ATase activity. Depletion of tumour ATase activity was noted in these patients, with residual activity amounting to 8% and 11% of pre-treatment levels, respectively, in the biopsied melanoma tissues. These results indicate extensive metabolism of CB 10-277 to a methylating agent capable of mediating alkylation of DNA and subsequent depletion of lymphocyte and tumor ATase levels and further indicate that the effects on lymphocytes may reflect effects on the target tumour.

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Abbreviations

ATase:

O6-alkylguanine-DNA alkyltransferase

CB 10-277:

1-p-carboxyl-3,3-dimethylphenyltriazene

DTIC:

dacarbazine, 5-(3,3-dimethyl-1-triazeno)imidazole-4-carboxamide

MTIC:

5-(3-methyl-1-triazeno)imidazole-4-carboxamide

References

  1. Aamdal S, Lee SM, Radford JA, Thatcher N, Calabresi F, Israels SP, Kerger J, Stamatakis L, Kleeberg UR, Brocker E, Gerard B (1991) Sequential administration of dacarbazine (DTIC) and fotemustine in disseminated melanoma. Proc Am Assoc Cancer Res 32: 191

    Google Scholar 

  2. Balch CM, Houghton A, Peters L (1989) Cutaneous melanoma. In: De Vita VT, Hellman S, Rosenberg SA (eds) Cancer: principles and practice of oncology. J. B. Lippincott, Philadelphia, p 1522

    Google Scholar 

  3. Bignami M, Terlizzese M, Zijno A, Calcagnile A, Frosima G, Abbondandolo A, Dogliotti E (1987) Cytotoxicity, mutations and SCEs induced by methylating agents are reduced in CHO cells expressing an active mammalian O6-methylguanine-DNA methyltransferase gene. Carcinogenesis 8: 1417

    Google Scholar 

  4. Bodell WJ (1990) Molecular dosimetry for sister-chromatid exchange induction and cytotoxicity by monofunctional and bifunctional alkylating agents. Mutat Res 233: 203

    Google Scholar 

  5. Bøyum A (1968) Isolation of mononuclear cells and granulocytes from human blood. Scand J Clin Lab Invest 21: 77

    Google Scholar 

  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem 72: 248

    Google Scholar 

  7. Brennand J, Margison GP (1986) Reduction of the toxicity and mutagenicity of alkylating agents in mammalian cells harboring theEscherichia coli alkyltransferase gene. Proc Natl Acad Sci USA 83: 6292

    Google Scholar 

  8. Catapano CV, Broggini M, Erba E, Ponti M, Mariani L, Citti L, D'Incalci M (1987) In vitro and in vivo methazolostone-induced DNA damage and repair in L1210 leukemia sensitive and resistant to chloroethylnitrosoureas. Cancer Res 47: 4884

    Google Scholar 

  9. Clark PC (1976) The evolution of therapy for malignant melanoma at the University of Texas M. D. Anderson Hospital and Tumour Institute 1950 to 1975. Pigm Cell 2: 365

    Google Scholar 

  10. Colombo T, D'Incalci M (1984) Comparison of the tumour activity of DTIC and 1-p-(3,3-dimethyl-1-triazeno) benzoic acid potassium salt on murine transplantable tumors and their hematological toxicity. Cancer Chemother Pharmacol 13: 139

    Google Scholar 

  11. D'Incalci M, Citti L, Taverna P, Catapano CV (1988) Importance of DNA repair enzyme O6-alkyltransferase (AT) in cancer chemotherapy. Cancer Treat Rev 15: 279

    Google Scholar 

  12. Ding R, Ghosh K, Eastman A, Bresnick E (1985) DNA-mediated transfer and expression of a human DNA repair gene that demethylates O6-methylguanine. Mol Cell Biol 5: 3293

    Google Scholar 

  13. Dolan ME, Corsico CD, Pegg AE (1985) Exposure of HeLa cells to O6-alkylguanines increases sensitivity to the cytotoxic effects of alkylating agents. Biochem Biophys Res Commun 132: 178

    Google Scholar 

  14. Dolan ME, Norbeck L, Clyde C, Hora NK, Erickson LC, Pegg AE (1989) Expression of mammalian O6-alkylguanine-DNA alkyltransferase in a cell line sensitive to alkylating agents. Carcinogenesis 10: 1613

    Google Scholar 

  15. Dolan ME, Mitchell RB, Mummert C, Moschel RC, Pegg AE (1991) Effect of O6-benzylguanine analogues on sensitivity of human tumor cells to the cytotoxic effects of alkylating agents. Cancer Res 51: 3367

    Google Scholar 

  16. Foster BJ, Newell DR, Lunn JM, Jones M, Calvert AH (1990) Correlation of dacarbazine and CB 10-277 activity against human melanoma xenografts with O6-alkyltransferase. Proc Am Assoc Cancer Res 31: 401

    Google Scholar 

  17. Gerson SL, Miller K, Berger NA (1985) O6-alkylguanine-DNA alkyltransferase activity in myeloid cells. J Clin Invest 76: 2106

    Google Scholar 

  18. Gerson SL, Trey JE, Miller K (1988) Potentiation of nitrosourea cytotoxicity in human leukemic cells by inactivation of O6-alkylguanine-DNA alkyltransferase. Cancer Res 48: 1521

    Google Scholar 

  19. Gibson NW, Hartley JA, Barnes D, Erickson LC (1986) Combined effects of streptozotocin and mitozolomide against four human cell lines of the Mer+ phenotype. Cancer Res 46: 4995

    Google Scholar 

  20. Gibson NW, Hartley J, La-France RJ, Vaughan K (1986) Differential cytotoxicity and DNA-damaging effects produced in human cells of the Mer+ and Mer− phenotypes by a series of alkyltriazenylimidazoles. Carcinogenesis 7: 259

    Google Scholar 

  21. Hayward IP, Parsons PG (1984) Comparison of virus reactivation, DNA base damage, and cell cycle effects in autologous melanoma cells resistant to methylating agents. Cancer Res 44: 55

    Google Scholar 

  22. Jelinek J, Kleibl K, Dexter TM, Margison GP (1988) Transfection of murine multi-potent haemopoietic stem cells with anE. coli DNA alkyltransferase gene confers resistance to the toxic effects of alkylating agents. Carcinogenesis 9: 81

    Google Scholar 

  23. Kataoka H, Hall J, Karran P (1986) Complementation of sensitivity to alkylating agents inEscherichia coli and Chinese hamster cells by expression of a cloned bacterial repair gene. EMBO J 5: 3195

    Google Scholar 

  24. Lee SM, Thatcher N, Margison GP (1991) O6-Alkylguanine-DNA alkyltransferase depletion and regeneration in human peripheral lymphocytes following dacarbazine and fotemustine. Cancer Res 51: 619

    Google Scholar 

  25. Lee SM, Rafferty JA, Elder RH, Fan C-Y, Bromley M, Harris M, Thatcher N, Potter PM, Altermatt HJ, Perinat-Frey T, Cerny T, O'Connor PJ, Margison GP (1992) Immunohistological examination of the inter- and intracellular distribution of O6-alkylguanine-DNA alkyltransferase in human liver and melanoma. Br J Cancer 66, 355

    Google Scholar 

  26. Lee SM, Thatcher N, Margison GP (1992) Dosage and cycle effects of dacarbazine (DTIC) and fotemustine on O6-alkylguanine-DNA alkyltransferase in human peripheral lymphocytes. Br J Cancer (in press)

  27. Lunn JM, Harris AL (1988) Cytotoxicity of 5-(3-methyl-1-triazeno)imidazole-4-carboxamide (MTIC) on Mer+, Mer+ Rem−, and Mer− cell lines: differential potentiation by 3-acetamidobenzamide. Br J Cancer 57: 54

    Google Scholar 

  28. Maynard K, Parsons PG, Cerny T, Margison GP (1989) Relationships among cell survival, O6-alkylguanine-DNA alkyltransferase activity, and reactivation of methylated adenovirus 5 and herpes simplex virus type 1 in human melanoma cell lines. Cancer Res 49: 4813

    Google Scholar 

  29. Meer L, Janzer RC, Kleihues P, Kolar GF (1986) In vivo metabolism and reaction with DNA of the cytostatic agent, 5-(3,3-dimethyl-1-triazeno)imidazole-4-carboxamide (DTIC). Biochem Pharmacol 35: 3243

    Google Scholar 

  30. Newell DR, Foster B, Carmicheal J, Harris AL, Jenns K, Gumbrell L, Calvert H (1990) Clinical studies with thep-carboxyl dimethyl phenyl triazene CB 10-277. In: Giraldi T, Connors TA, Cartei G (eds) Triazenes. Chemical, biological, and clinical aspects. Plenum, New York, p 119

    Google Scholar 

  31. Parson PG, Smellie SG, Morrison LE, Hayward IP (1982) Properties of human melanoma cells resistant to 5-(3′-3′-dimethyl-1-triazeno)imidazole-4-carboxamide and other methylating agents. Cancer Res 42: 1454

    Google Scholar 

  32. Pegg AE (1990) Mammalian O6-alkylguanine-DNA alkyltransferase regulation and importance in response to alkylating carcinogenic and therapeutic agents. Cancer Res 50: 6119

    Google Scholar 

  33. Pieper RO, Costello JF, Kroes RA, Futscher BW, Maranthi U, Erickson LC (1991) Direct correlation between methylation status and expression of the human O6-methylguanine DNA methyltransferase gene. Cancer Commun 3: 241

    Google Scholar 

  34. Rutty CJ, Newell DR, Vincent RB, Abel G, Goddard PM, Harland SJ, Calvert AH (1983) The species dependent pharmacokinetics of DTIC. Br J Cancer 48: 140

    Google Scholar 

  35. Rutty CJ, Graham MA, Abel G, Judson IR, Goddard PM (1986) Preclinical evaluation of 1-p-carboxy-3,3-dimethyl phenyltriazene (CB 10-277). An alternative to DTIC. Br J Cancer 54: 194

    Google Scholar 

  36. Samson L, Derfler B, Waldstein EA (1986) Suppression of human alkylation-repair defects byEscherichia coli DNA-repair genes. Proc Natl Acad Sci USA 83: 5607

    Google Scholar 

  37. Yarosh DB, Hurst-Calderone S, Babich MA, Day RS III (1986) Inactivation of O6-methylguanine-DNA methyltransferase and sensitization of human tumour cells to killing by chloroethylnitrosourea by O6-methylguanine as a free base. Cancer Res 46: 1663

    Google Scholar 

  38. Zlotogorski C, Erickson LC (1983) Pretreatment of normal human fibroblasts and human colon carcinoma cells with MNNG allows chloroethylnitrosourea to produce DNA interstrand cross-links not observed in cells treated with chloroethylnitrosourea alone. Carcinogenesis 4: 759

    Google Scholar 

  39. Zlotogorski C, Erickson LC (1984) Pretreatment of human colon tumour cells with DNA methylating agents inhibits their ability to repair chloroethyl monoadducts. Carcinogenesis 5: 83

    Google Scholar 

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Authors and Affiliations

  1. CRC Department of Medical Oncology, Christie Hospital NHS Trust, M20 9BX, Manchester, UK

    Siow Ming Lee, Nick Thatcher & Derek Crowther

  2. CRC Department of Carcinogenesis, Paterson Institute for Cancer Research, M20 9BX, Manchester, UK

    Siow Ming Lee & Geoffrey P Margison

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  1. Siow Ming Lee
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  2. Nick Thatcher
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  3. Derek Crowther
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  4. Geoffrey P Margison
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Lee, S.M., Thatcher, N., Crowther, D. et al. In vivo depletion of O6-alkylguanine-DNA-alkyltransferase in lymphocytes and melanoma of patients treated with CB 10-277, a new DTIC analogue. Cancer Chemother. Pharmacol. 31, 240–246 (1992). https://doi.org/10.1007/BF00685554

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  • DOI: https://doi.org/10.1007/BF00685554

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