Reduced Folate Carrier Mutations Are Not the Mechanism Underlying Methotrexate Resistance in Childhood Acute Lymphoblastic Leukemia

Yotam Kaufman, Stavit Drori, Peter D. Cole, Barton A. Kamen, Jenny Sirota, Ilan Ifergan, Myriam Weyl Ben Arush, Ronit Elhasid, Dvora Sahar, Gert Jan L Kaspers, Gerrit Jansen, Larry H. Matherly, Gideon Rechavi, Amos Toren, Yehuda G. Assaraf

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

BACKGROUND. Although the majority of children with acute lymphoblastic leukemia (ALL) are cured with combination chemotherapy containing methotrexate (MTX), drug resistance contributes to treatment failure for a substantial fraction of patients. The primary transporter for folates and MTX is the reduced folate carrier (RFC). Impaired drug transport is a documented mechanism of MTX resistance in patients with ALL; however, to the authors' knowledge it is not known whether inactivating RFC mutations are a contributing factor. METHODS. The authors devised a genomic polymerase chain reaction-single strand conformational polymorphism assay followed by sequencing and screened the entire RFC coding region for sequence alterations in DNA from 246 leukemia specimens from patients with diverse ethnic variation, 24 at the time of recurrence and the rest at the time of diagnosis. This cohort was comprised of 203 β-precursor ALL specimens (82.5%), 32 T-lineage ALL specimens (13%), and 11 acute myeloblastic leukemia specimens (4.5%). RESULTS. Of 246 DNA samples, only 3 diagnosis β-precursor ALL specimens (1.2%) were found to harbor alterations in the RFC gene, including heterozygous single nucleotide changes resulting in D56H and D522N substitutions in the first extra-cellular loop and the C-terminus of this transporter, respectively. The third sample had a sequence alteration in exon 3 that could not be identified because of the lack of availability of DNA. CONCLUSIONS. Whereas inactivating RFC mutations are a frequent mechanism of MTX resistance in human leukemia cell lines and in patients with osteosarcoma, they are not common and do not appear to play any significant role in intrinsic or acquired resistance to MTX in childhood leukemia. This is the first study of RFC mutations in multiple pediatric leukemia specimens.

Original languageEnglish (US)
Pages (from-to)773-782
Number of pages10
JournalCancer
Volume100
Issue number4
DOIs
StatePublished - Feb 15 2004
Externally publishedYes

Fingerprint

Reduced Folate Carrier Protein
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Methotrexate
Mutation
Leukemia
Folic Acid Transporters
DNA
Osteosarcoma
Combination Drug Therapy
Treatment Failure
Drug Resistance
Acute Myeloid Leukemia
Exons
Nucleotides
Pediatrics
Recurrence
Cell Line
Polymerase Chain Reaction
Pharmaceutical Preparations

Keywords

  • Acute lymphoblastic leukemia (ALL)
  • Chemotherapy
  • Drug resistance
  • Drug transport
  • Methotrexate (MTX)
  • Mutations
  • Reduced folate carrier (RFC)

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Kaufman, Y., Drori, S., Cole, P. D., Kamen, B. A., Sirota, J., Ifergan, I., ... Assaraf, Y. G. (2004). Reduced Folate Carrier Mutations Are Not the Mechanism Underlying Methotrexate Resistance in Childhood Acute Lymphoblastic Leukemia. Cancer, 100(4), 773-782. https://doi.org/10.1002/cncr.20018

Reduced Folate Carrier Mutations Are Not the Mechanism Underlying Methotrexate Resistance in Childhood Acute Lymphoblastic Leukemia. / Kaufman, Yotam; Drori, Stavit; Cole, Peter D.; Kamen, Barton A.; Sirota, Jenny; Ifergan, Ilan; Arush, Myriam Weyl Ben; Elhasid, Ronit; Sahar, Dvora; Kaspers, Gert Jan L; Jansen, Gerrit; Matherly, Larry H.; Rechavi, Gideon; Toren, Amos; Assaraf, Yehuda G.

In: Cancer, Vol. 100, No. 4, 15.02.2004, p. 773-782.

Research output: Contribution to journalArticle

Kaufman, Y, Drori, S, Cole, PD, Kamen, BA, Sirota, J, Ifergan, I, Arush, MWB, Elhasid, R, Sahar, D, Kaspers, GJL, Jansen, G, Matherly, LH, Rechavi, G, Toren, A & Assaraf, YG 2004, 'Reduced Folate Carrier Mutations Are Not the Mechanism Underlying Methotrexate Resistance in Childhood Acute Lymphoblastic Leukemia', Cancer, vol. 100, no. 4, pp. 773-782. https://doi.org/10.1002/cncr.20018
Kaufman, Yotam ; Drori, Stavit ; Cole, Peter D. ; Kamen, Barton A. ; Sirota, Jenny ; Ifergan, Ilan ; Arush, Myriam Weyl Ben ; Elhasid, Ronit ; Sahar, Dvora ; Kaspers, Gert Jan L ; Jansen, Gerrit ; Matherly, Larry H. ; Rechavi, Gideon ; Toren, Amos ; Assaraf, Yehuda G. / Reduced Folate Carrier Mutations Are Not the Mechanism Underlying Methotrexate Resistance in Childhood Acute Lymphoblastic Leukemia. In: Cancer. 2004 ; Vol. 100, No. 4. pp. 773-782.
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abstract = "BACKGROUND. Although the majority of children with acute lymphoblastic leukemia (ALL) are cured with combination chemotherapy containing methotrexate (MTX), drug resistance contributes to treatment failure for a substantial fraction of patients. The primary transporter for folates and MTX is the reduced folate carrier (RFC). Impaired drug transport is a documented mechanism of MTX resistance in patients with ALL; however, to the authors' knowledge it is not known whether inactivating RFC mutations are a contributing factor. METHODS. The authors devised a genomic polymerase chain reaction-single strand conformational polymorphism assay followed by sequencing and screened the entire RFC coding region for sequence alterations in DNA from 246 leukemia specimens from patients with diverse ethnic variation, 24 at the time of recurrence and the rest at the time of diagnosis. This cohort was comprised of 203 β-precursor ALL specimens (82.5{\%}), 32 T-lineage ALL specimens (13{\%}), and 11 acute myeloblastic leukemia specimens (4.5{\%}). RESULTS. Of 246 DNA samples, only 3 diagnosis β-precursor ALL specimens (1.2{\%}) were found to harbor alterations in the RFC gene, including heterozygous single nucleotide changes resulting in D56H and D522N substitutions in the first extra-cellular loop and the C-terminus of this transporter, respectively. The third sample had a sequence alteration in exon 3 that could not be identified because of the lack of availability of DNA. CONCLUSIONS. Whereas inactivating RFC mutations are a frequent mechanism of MTX resistance in human leukemia cell lines and in patients with osteosarcoma, they are not common and do not appear to play any significant role in intrinsic or acquired resistance to MTX in childhood leukemia. This is the first study of RFC mutations in multiple pediatric leukemia specimens.",
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author = "Yotam Kaufman and Stavit Drori and Cole, {Peter D.} and Kamen, {Barton A.} and Jenny Sirota and Ilan Ifergan and Arush, {Myriam Weyl Ben} and Ronit Elhasid and Dvora Sahar and Kaspers, {Gert Jan L} and Gerrit Jansen and Matherly, {Larry H.} and Gideon Rechavi and Amos Toren and Assaraf, {Yehuda G.}",
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T1 - Reduced Folate Carrier Mutations Are Not the Mechanism Underlying Methotrexate Resistance in Childhood Acute Lymphoblastic Leukemia

AU - Kaufman, Yotam

AU - Drori, Stavit

AU - Cole, Peter D.

AU - Kamen, Barton A.

AU - Sirota, Jenny

AU - Ifergan, Ilan

AU - Arush, Myriam Weyl Ben

AU - Elhasid, Ronit

AU - Sahar, Dvora

AU - Kaspers, Gert Jan L

AU - Jansen, Gerrit

AU - Matherly, Larry H.

AU - Rechavi, Gideon

AU - Toren, Amos

AU - Assaraf, Yehuda G.

PY - 2004/2/15

Y1 - 2004/2/15

N2 - BACKGROUND. Although the majority of children with acute lymphoblastic leukemia (ALL) are cured with combination chemotherapy containing methotrexate (MTX), drug resistance contributes to treatment failure for a substantial fraction of patients. The primary transporter for folates and MTX is the reduced folate carrier (RFC). Impaired drug transport is a documented mechanism of MTX resistance in patients with ALL; however, to the authors' knowledge it is not known whether inactivating RFC mutations are a contributing factor. METHODS. The authors devised a genomic polymerase chain reaction-single strand conformational polymorphism assay followed by sequencing and screened the entire RFC coding region for sequence alterations in DNA from 246 leukemia specimens from patients with diverse ethnic variation, 24 at the time of recurrence and the rest at the time of diagnosis. This cohort was comprised of 203 β-precursor ALL specimens (82.5%), 32 T-lineage ALL specimens (13%), and 11 acute myeloblastic leukemia specimens (4.5%). RESULTS. Of 246 DNA samples, only 3 diagnosis β-precursor ALL specimens (1.2%) were found to harbor alterations in the RFC gene, including heterozygous single nucleotide changes resulting in D56H and D522N substitutions in the first extra-cellular loop and the C-terminus of this transporter, respectively. The third sample had a sequence alteration in exon 3 that could not be identified because of the lack of availability of DNA. CONCLUSIONS. Whereas inactivating RFC mutations are a frequent mechanism of MTX resistance in human leukemia cell lines and in patients with osteosarcoma, they are not common and do not appear to play any significant role in intrinsic or acquired resistance to MTX in childhood leukemia. This is the first study of RFC mutations in multiple pediatric leukemia specimens.

AB - BACKGROUND. Although the majority of children with acute lymphoblastic leukemia (ALL) are cured with combination chemotherapy containing methotrexate (MTX), drug resistance contributes to treatment failure for a substantial fraction of patients. The primary transporter for folates and MTX is the reduced folate carrier (RFC). Impaired drug transport is a documented mechanism of MTX resistance in patients with ALL; however, to the authors' knowledge it is not known whether inactivating RFC mutations are a contributing factor. METHODS. The authors devised a genomic polymerase chain reaction-single strand conformational polymorphism assay followed by sequencing and screened the entire RFC coding region for sequence alterations in DNA from 246 leukemia specimens from patients with diverse ethnic variation, 24 at the time of recurrence and the rest at the time of diagnosis. This cohort was comprised of 203 β-precursor ALL specimens (82.5%), 32 T-lineage ALL specimens (13%), and 11 acute myeloblastic leukemia specimens (4.5%). RESULTS. Of 246 DNA samples, only 3 diagnosis β-precursor ALL specimens (1.2%) were found to harbor alterations in the RFC gene, including heterozygous single nucleotide changes resulting in D56H and D522N substitutions in the first extra-cellular loop and the C-terminus of this transporter, respectively. The third sample had a sequence alteration in exon 3 that could not be identified because of the lack of availability of DNA. CONCLUSIONS. Whereas inactivating RFC mutations are a frequent mechanism of MTX resistance in human leukemia cell lines and in patients with osteosarcoma, they are not common and do not appear to play any significant role in intrinsic or acquired resistance to MTX in childhood leukemia. This is the first study of RFC mutations in multiple pediatric leukemia specimens.

KW - Acute lymphoblastic leukemia (ALL)

KW - Chemotherapy

KW - Drug resistance

KW - Drug transport

KW - Methotrexate (MTX)

KW - Mutations

KW - Reduced folate carrier (RFC)

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