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Mechanisms of Methotrexate Resistance in Acute Leukemia

Decreased Transport and Polyglutamylation
  • Richard Gorlick
  • Peter Cole
  • Debabrata Banerjee
  • Giuseppe Longo
  • Wei Wei Li
  • Daniel Hochhauser
  • Joseph R. Bertino
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 457)

Abstract

Drug resistance limits the effectiveness of methotrexate (MTX) for the treatment of acute leukemia. An increased understanding of the pathways involved in folate metabolism has allowed investigations of the mechanisms of resistance observed in leukemic blasts obtained from patients. Acute lymphocytic leukemia (ALL) was studied for mechanisms of acquired MTX resistance. MTX transport in 27 patients with untreated ALL and 31 patients with relapsed ALL was measured using a previously described competitive displacement assay. Only 13% of the untreated patients were considered to have impaired MTX transport whereas over 70% of the relapsed patients had evidence of impaired MTX transport. Northern analyses and quantitative RT-PCR for the reduced folate carrier (RFC) were performed on the RNA available from the leukemic blasts of 24 patients in whom MTX transport had been measured. Six of 9 samples with impaired MTX transport had decreased RFC expression (one had no detectable RFC expression), while three had no decrease in RFC expression. Acute myelocytic leukemia (AML) was studied to determine the basis of the decreased MTX polyglutamylation. Enzyme kinetics of the enzyme folylpolyglutamate synthetase (FPGS) were studied, demonstrating FPGS in the myeloid cell lines and patient samples had a higher Km for MTX as a substrate than lymphoid cells. Measuring gamma-glutamyl hydrolase enzyme activity allowed a more accurate prediction of steady state levels of MTX polyglutamates. A knowledge of the mechanisms of MTX resistance that occur in leukemic blasts obtained from patients may allow the development of therapeutic strategies to circumvent resistance.

Keywords

Folylpolyglutamate synthetase gamma-glutamyl hydrolase methotrexate reduced folate carrier acute lymphocytic leukemia acute myelocytic leukemia 

Abbreviations

DHFR

dihydrofolate reductase

RFC

reduced folate carrier

FPGS

folylpolyglutamate synthetase

GGH

gamma-glutamyl hydrolase

MTX

methotrexate

ALL

acute lymphocytic leukemia

AML

acute myelocytic leukemia

TMTX

trimetrexate

GAR

glycinamide ribonucleotide

AICAR

aminoimidazole carboxamide

LV

leucovorin

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Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • Richard Gorlick
    • 1
  • Peter Cole
    • 1
  • Debabrata Banerjee
    • 2
  • Giuseppe Longo
    • 2
  • Wei Wei Li
    • 2
  • Daniel Hochhauser
    • 2
  • Joseph R. Bertino
    • 2
  1. 1.Department of PediatricsMemorial Sloan-Kettering Cancer CenterNew YorkUSA
  2. 2.Program of Molecular Pharmacology and TherapeuticsMemorial Sloan-Kettering Cancer CenterNew YorkUSA

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