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. 1994 Nov;94(5):1996–2001. doi: 10.1172/JCI117552

Blast cell methotrexate-polyglutamate accumulation in vivo differs by lineage, ploidy, and methotrexate dose in acute lymphoblastic leukemia.

T W Synold 1, M V Relling 1, J M Boyett 1, G K Rivera 1, J T Sandlund 1, H Mahmoud 1, W M Crist 1, C H Pui 1, W E Evans 1
PMCID: PMC294625  PMID: 7525652

Abstract

High-dose methotrexate (HDMTX) is a component of most treatment protocols for childhood acute lymphoblastic leukemia (ALL), yet recent studies of receptor-mediated transport and saturable polyglutamylation have questioned its rationale. To investigate this in vivo, methotrexate and its active polyglutamated metabolites (MTX-PG) were measured in bone marrow blasts obtained from 101 children randomized to single-agent therapy with either HDMTX (1 g/m2 per 24 h i.v., n = 47) or low-dose MTX (LDMTX, 30 mg/m2 by mouth every 6 h x 6, n = 54), before remission induction therapy. Blast concentrations of total MTX-PGs (median 460 vs 1380 pmol/10(9) cells) and of long-chain MTX-glu4-6 were both significantly higher after HDMTX (P < 0.001). With either treatment, MTX-PGs were significantly higher in B-lineage blasts than in T-lineage blasts (LDMTX P = 0.001, HDMTX P = 0.03). In a multiple regression analysis of B-lineage ALL, blast MTX-PG was significantly related to MTX dose (or plasma MTX concentration), lymphoblast ploidy (hyperdiploid > nonhyperdiploid), and percentage S-phase. This is the first evidence that HDMTX achieves higher MTX-PG concentrations in ALL blasts in vivo, establishing a rationale for HDMTX in the treatment of childhood ALL, especially T-lineage or nonhyperdiploid B-lineage ALL, disease characteristics associated with a poor prognosis on conventional therapy.

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Selected References

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  1. Allegra C. J., Chabner B. A., Drake J. C., Lutz R., Rodbard D., Jolivet J. Enhanced inhibition of thymidylate synthase by methotrexate polyglutamates. J Biol Chem. 1985 Aug 15;260(17):9720–9726. [PubMed] [Google Scholar]
  2. Allegra C. J., Drake J. C., Jolivet J., Chabner B. A. Inhibition of phosphoribosylaminoimidazolecarboxamide transformylase by methotrexate and dihydrofolic acid polyglutamates. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4881–4885. doi: 10.1073/pnas.82.15.4881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson R. G., Kamen B. A., Rothberg K. G., Lacey S. W. Potocytosis: sequestration and transport of small molecules by caveolae. Science. 1992 Jan 24;255(5043):410–411. doi: 10.1126/science.1310359. [DOI] [PubMed] [Google Scholar]
  4. Balinska M., Nimec Z., Galivan J. Characteristics of methotrexate polyglutamate formation in cultured hepatic cells. Arch Biochem Biophys. 1982 Jul;216(2):466–476. doi: 10.1016/0003-9861(82)90235-1. [DOI] [PubMed] [Google Scholar]
  5. Barredo J. C., Synold T. W., Laver J., Relling M. V., Pui C. H., Priest D. G., Evans W. E. Differences in constitutive and post-methotrexate folylpolyglutamate synthetase activity in B-lineage and T-lineage leukemia. Blood. 1994 Jul 15;84(2):564–569. [PubMed] [Google Scholar]
  6. Bertino J. R. Karnofsky memorial lecture. Ode to methotrexate. J Clin Oncol. 1993 Jan;11(1):5–14. doi: 10.1200/JCO.1993.11.1.5. [DOI] [PubMed] [Google Scholar]
  7. Camitta B., Leventhal B., Lauer S., Shuster J. J., Adair S., Casper J., Civin C., Graham M., Mahoney D., Munoz L. Intermediate-dose intravenous methotrexate and mercaptopurine therapy for non-T, non-B acute lymphocytic leukemia of childhood: a Pediatric Oncology Group study. J Clin Oncol. 1989 Oct;7(10):1539–1544. doi: 10.1200/JCO.1989.7.10.1539. [DOI] [PubMed] [Google Scholar]
  8. Chabner B. A., Allegra C. J., Curt G. A., Clendeninn N. J., Baram J., Koizumi S., Drake J. C., Jolivet J. Polyglutamation of methotrexate. Is methotrexate a prodrug? J Clin Invest. 1985 Sep;76(3):907–912. doi: 10.1172/JCI112088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cowan K. H., Jolivet J. A methotrexate-resistant human breast cancer cell line with multiple defects, including diminished formation of methotrexate polyglutamates. J Biol Chem. 1984 Sep 10;259(17):10793–10800. [PubMed] [Google Scholar]
  10. D'Argenio D. Z., Schumitzky A. A program package for simulation and parameter estimation in pharmacokinetic systems. Comput Programs Biomed. 1979 Mar;9(2):115–134. doi: 10.1016/0010-468x(79)90025-4. [DOI] [PubMed] [Google Scholar]
  11. Evans W. E., Crom W. R., Abromowitch M., Dodge R., Look A. T., Bowman W. P., George S. L., Pui C. H. Clinical pharmacodynamics of high-dose methotrexate in acute lymphocytic leukemia. Identification of a relation between concentration and effect. N Engl J Med. 1986 Feb 20;314(8):471–477. doi: 10.1056/NEJM198602203140803. [DOI] [PubMed] [Google Scholar]
  12. Evans W. E., Schell M. J., Pui C. H. MTX clearance is more important for intermediate-risk ALL. J Clin Oncol. 1990 Jun;8(6):1115–1116. doi: 10.1200/JCO.1990.8.6.1115. [DOI] [PubMed] [Google Scholar]
  13. Fabre G., Matherly L. H., Favre R., Catalin J., Cano J. P. In vitro formation of polyglutamyl derivatives of methotrexate and 7-hydroxymethotrexate in human lymphoblastic leukemia cells. Cancer Res. 1983 Oct;43(10):4648–4652. [PubMed] [Google Scholar]
  14. Fleming T. R., Harrington D. P., O'Brien P. C. Designs for group sequential tests. Control Clin Trials. 1984 Dec;5(4):348–361. doi: 10.1016/s0197-2456(84)80014-8. [DOI] [PubMed] [Google Scholar]
  15. Fry D. W., Yalowich J. C., Goldman I. D. Rapid formation of poly-gamma-glutamyl derivatives of methotrexate and their association with dihydrofolate reductase as assessed by high pressure liquid chromatography in the Ehrlich ascites tumor cell in vitro. J Biol Chem. 1982 Feb 25;257(4):1890–1896. [PubMed] [Google Scholar]
  16. Galivan J. Evidence for the cytotoxic activity of polyglutamate derivatives of methotrexate. Mol Pharmacol. 1980 Jan;17(1):105–110. [PubMed] [Google Scholar]
  17. Galivan J., Nimec Z., Balinska M. Regulation of methotrexate polyglutamate accumulation in vitro: effects of cellular folate content. Biochem Pharmacol. 1983 Nov 1;32(21):3244–3247. doi: 10.1016/0006-2952(83)90211-3. [DOI] [PubMed] [Google Scholar]
  18. Göker E., Lin J. T., Trippett T., Elisseyeff Y., Tong W. P., Niedzwiecki D., Tan C., Steinherz P., Schweitzer B. I., Bertino J. R. Decreased polyglutamylation of methotrexate in acute lymphoblastic leukemia blasts in adults compared to children with this disease. Leukemia. 1993 Jul;7(7):1000–1004. [PubMed] [Google Scholar]
  19. Jacobs S. A., Adamson R. H., Chabner B. A., Derr C. J., Johns D. C. Stoichiometric inhibition of mammalian dihydrofolate reductase by the gamma-glutamyl metabolite of methotrexiate, 4-amino-4-deoxy-N-10-methylpteroylglutamyl-gamma-glutamate. Biochem Biophys Res Commun. 1975 Apr 7;63(3):692–698. doi: 10.1016/s0006-291x(75)80439-6. [DOI] [PubMed] [Google Scholar]
  20. Jolivet J., Chabner B. A. Intracellular pharmacokinetics of methotrexate polyglutamates in human breast cancer cells. Selective retention and less dissociable binding of 4-NH2-10-CH3-pteroylglutamate4 and 4-NH2-10-CH3-pteroylglutamate5 to dihydrofolate reductase. J Clin Invest. 1983 Sep;72(3):773–778. doi: 10.1172/JCI111048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kamen B. A., Capdevila A. Receptor-mediated folate accumulation is regulated by the cellular folate content. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5983–5987. doi: 10.1073/pnas.83.16.5983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kamen B. A., Winick N. J. High dose methotrexate therapy: insecure rationale? Biochem Pharmacol. 1988 Jul 15;37(14):2713–2715. doi: 10.1016/0006-2952(88)90032-9. [DOI] [PubMed] [Google Scholar]
  23. Kamen B. A., Winick N. Analysis of methotrexate polyglutamate derivatives in vivo. Methods Enzymol. 1986;122:339–346. doi: 10.1016/0076-6879(86)22191-6. [DOI] [PubMed] [Google Scholar]
  24. Keefe D. A., Capizzi R. L., Rudnick S. A. Methotrexate cytotoxicity for L5178Y/Asn- lymphoblasts: relationship of dose and duration of exposure to tumor cell viability. Cancer Res. 1982 May;42(5):1641–1645. [PubMed] [Google Scholar]
  25. Look A. T., Melvin S. L., Williams D. L., Brodeur G. M., Dahl G. V., Kalwinsky D. K., Murphy S. B., Mauer A. M. Aneuploidy and percentage of S-phase cells determined by flow cytometry correlate with cell phenotype in childhood acute leukemia. Blood. 1982 Oct;60(4):959–967. [PubMed] [Google Scholar]
  26. McCloskey D. E., McGuire J. J., Russell C. A., Rowan B. G., Bertino J. R., Pizzorno G., Mini E. Decreased folylpolyglutamate synthetase activity as a mechanism of methotrexate resistance in CCRF-CEM human leukemia sublines. J Biol Chem. 1991 Apr 5;266(10):6181–6187. [PubMed] [Google Scholar]
  27. McGuire J. J., Hsieh P., Coward J. K., Bertino J. R. Enzymatic synthesis of folylpolyglutamates. Characterization of the reaction and its products. J Biol Chem. 1980 Jun 25;255(12):5776–5788. [PubMed] [Google Scholar]
  28. Myers C. E., Lippman M. E., Elliot H. M., Chabner B. A. Competitive protein binding assay for methotrexate. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3683–3686. doi: 10.1073/pnas.72.9.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Niemeyer C. M., Gelber R. D., Tarbell N. J., Donnelly M., Clavell L. A., Blattner S. R., Donahue K., Cohen H. J., Sallan S. E. Low-dose versus high-dose methotrexate during remission induction in childhood acute lymphoblastic leukemia (Protocol 81-01 update). Blood. 1991 Nov 15;78(10):2514–2519. [PubMed] [Google Scholar]
  30. Pizzorno G., Mini E., Coronnello M., McGuire J. J., Moroson B. A., Cashmore A. R., Dreyer R. N., Lin J. T., Mazzei T., Periti P. Impaired polyglutamylation of methotrexate as a cause of resistance in CCRF-CEM cells after short-term, high-dose treatment with this drug. Cancer Res. 1988 Apr 15;48(8):2149–2155. [PubMed] [Google Scholar]
  31. Pui C. H., Behm F. G., Crist W. M. Clinical and biologic relevance of immunologic marker studies in childhood acute lymphoblastic leukemia. Blood. 1993 Jul 15;82(2):343–362. [PubMed] [Google Scholar]
  32. Pui C. H., Crist W. M. Biology and treatment of acute lymphoblastic leukemia. J Pediatr. 1994 Apr;124(4):491–503. doi: 10.1016/s0022-3476(05)83125-7. [DOI] [PubMed] [Google Scholar]
  33. Pui C. H., Crist W. M., Look A. T. Biology and clinical significance of cytogenetic abnormalities in childhood acute lymphoblastic leukemia. Blood. 1990 Oct 15;76(8):1449–1463. [PubMed] [Google Scholar]
  34. Rivera G. K., Raimondi S. C., Hancock M. L., Behm F. G., Pui C. H., Abromowitch M., Mirro J., Jr, Ochs J. S., Look A. T., Williams D. L. Improved outcome in childhood acute lymphoblastic leukaemia with reinforced early treatment and rotational combination chemotherapy. Lancet. 1991 Jan 12;337(8733):61–66. doi: 10.1016/0140-6736(91)90733-6. [DOI] [PubMed] [Google Scholar]
  35. Rosenblatt D. S., Whitehead V. M., Vera N., Pottier A., Dupont M., Vuchich M. J. Prolonged inhibition of DNA synthesis associated with the accumulation of methotrexate polyglutamates by cultured human cells. Mol Pharmacol. 1978 Nov;14(6):1143–1147. [PubMed] [Google Scholar]
  36. Taylor R. T., Hanna M. L. Folate-dependent enzymes in cultured Chinese hamster cells: folypolyglutamate synthetase and its absence in mutants auxotrophic for glycine + adenosine + thymidine. Arch Biochem Biophys. 1977 May;181(1):331–334. doi: 10.1016/0003-9861(77)90512-4. [DOI] [PubMed] [Google Scholar]
  37. Teresi M. E., Crom W. R., Choi K. E., Mirro J., Evans W. E. Methotrexate bioavailability after oral and intramuscular administration in children. J Pediatr. 1987 May;110(5):788–792. doi: 10.1016/s0022-3476(87)80025-2. [DOI] [PubMed] [Google Scholar]
  38. Whitehead V. M., Rosenblatt D. S., Vuchich M. J., Shuster J. J., Witte A., Beaulieu D. Accumulation of methotrexate and methotrexate polyglutamates in lymphoblasts at diagnosis of childhood acute lymphoblastic leukemia: a pilot prognostic factor analysis. Blood. 1990 Jul 1;76(1):44–49. [PubMed] [Google Scholar]
  39. Whitehead V. M. Synthesis of methotrexate polyglutamates in L1210 murine leukemia cells. Cancer Res. 1977 Feb;37(2):408–412. [PubMed] [Google Scholar]
  40. Whitehead V. M., Vuchich M. J., Lauer S. J., Mahoney D., Carroll A. J., Shuster J. J., Esseltine D. W., Payment C., Look A. T., Akabutu J. Accumulation of high levels of methotrexate polyglutamates in lymphoblasts from children with hyperdiploid (greater than 50 chromosomes) B-lineage acute lymphoblastic leukemia: a Pediatric Oncology Group study. Blood. 1992 Sep 1;80(5):1316–1323. [PubMed] [Google Scholar]
  41. Williams D. L., Harris A., Williams K. J., Brosius M. J., Lemonds W. A direct bone marrow chromosome technique for acute lymphoblastic leukemia. Cancer Genet Cytogenet. 1984 Nov;13(3):239–257. doi: 10.1016/0165-4608(84)90046-3. [DOI] [PubMed] [Google Scholar]

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