Novel aspects of resistance to drugs targeted to dihydrofolate reductase and thymidylate synthase

Debabrata Banerjee, Philipp Mayer-Kuckuk, Gina Capiaux, Tulin Budak-Alpdogan, Richard Gorlick, Joseph R. Bertino

Research output: Contribution to journalArticle

188 Citations (Scopus)

Abstract

Drug resistance is often a limiting factor in successful chemotherapy. Our laboratory has been interested in studying mechanisms of resistance to drugs that are targeted to the thymidylate biosynthesis pathway especially those that target thymidylate synthase (TS) and dihydrofolate reductase (DHFR). We have used leukemia as a model system to study resistance to methotrexate (MTX) and colorectal cancer as the model system to study 5-fluorouracil (5-FU) resistance. In leukemias, we and others have shown that transport, efflux, polyglutamylation and hydrolase activities are major determinants of MTX resistance. We have further reported that some leukemic cells have an increase in DHFR gene copy number possibly contributing to the resistant phenotype. Recently, we have begun to study in detail the molecular mechanisms that govern translational regulation of DHFR in response to MTX as an additional resistance mechanism. Studies thus far involving colorectal tumors obtained from patients have focused predominantly on the predictive value of levels of TS expression and p53 mutations in determining response to 5-FU. Although the predictive value of these two measures appears to be significant, given the variety of resistance to 5-FU observed in cell lines, it is not likely that these are the only measures predictive of response or responsible for acquired resistance to this drug. The enzyme uridine-cytidine monophosphate kinase (UMPK) is an essential and rate-limiting enzyme in 5-FU activation while dihydropyrimidine dehydrogenase (DPD) is a catabolic enzyme that inactivates 5-FU. Alterations in UMPK and DPD may therefore explain failure of 5-FU response in the absence of alterations in TS or p53. Transcription factors that regulate TS may also influence drug sensitivity. We have found that mRNA levels of the E2F family of transcription factors correlates with TS message levels and are higher in lung metastases than in liver metastases of colorectal cancers. Moreover, gene copy number of the E2F-1 gene appears to be increased in a significant number of samples obtained from metastases of colorectal cancer. We have also generated mutants of both DHFR and TS that confer resistance to MTX as well as 5-FU by random as well as site-directed mutagenesis. These mutants used alone or as fusion cDNAs of the mutants have proven to be useful in transplant studies where transfer of these mutant cDNAs to bone marrow cells have been shown to confer drug resistance to recipients. The fusion cDNAs of DHFR such as the DHFR-herpes simplex virus type 1 thymidine kinase (HSVTK) are also useful for regulation of gene expression in vivo using MTX as the small molecule regulator that can be monitored by positron emission tomography (PET) scanning or by optical imaging using a fusion construct such as DHFR-EGFP.

Original languageEnglish (US)
Pages (from-to)164-173
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1587
Issue number2-3
DOIs
StatePublished - Jul 18 2002
Externally publishedYes

Fingerprint

Drug Resistance
Fluorouracil
Tetrahydrofolate Dehydrogenase
Thymidylate Synthase
Methotrexate
Colorectal Neoplasms
Uridine Kinase
Dihydrouracil Dehydrogenase (NADP)
Gene Dosage
Complementary DNA
Neoplasm Metastasis
Leukemia
Enzymes
E2F Transcription Factors
Thymidine Kinase
Optical Imaging
Human Herpesvirus 1
Gene Expression Regulation
Hydrolases
Site-Directed Mutagenesis

Keywords

  • Antifolate
  • Drug resistance
  • Fluoropyrimidine
  • In vivo imaging
  • Myeloprotection

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Biophysics

Cite this

Novel aspects of resistance to drugs targeted to dihydrofolate reductase and thymidylate synthase. / Banerjee, Debabrata; Mayer-Kuckuk, Philipp; Capiaux, Gina; Budak-Alpdogan, Tulin; Gorlick, Richard; Bertino, Joseph R.

In: Biochimica et Biophysica Acta - Molecular Basis of Disease, Vol. 1587, No. 2-3, 18.07.2002, p. 164-173.

Research output: Contribution to journalArticle

Banerjee, Debabrata ; Mayer-Kuckuk, Philipp ; Capiaux, Gina ; Budak-Alpdogan, Tulin ; Gorlick, Richard ; Bertino, Joseph R. / Novel aspects of resistance to drugs targeted to dihydrofolate reductase and thymidylate synthase. In: Biochimica et Biophysica Acta - Molecular Basis of Disease. 2002 ; Vol. 1587, No. 2-3. pp. 164-173.
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