Retroviral transfer of the hENT2 nucleoside transporter cDNA confers broad-spectrum antifolate resistance in murine bone marrow cells.
Antifolate drugs such as methotrexate are commonly used in cancer chemotherapy. It may be possible to increase the antitumor activity of antifolates by the coadministration of drugs that inhibit nucleoside transport, thereby blocking the capacity of tumor cells to salvage nucleotide precursors. An important limitation of this approach is severe myelosuppression caused by many of these drug combinations. For this reason, we have developed a gene therapy strategy to protect bone marrow cells against combined treatment with antifolates and nitrobenzylmercaptopurine riboside (NBMPR), a potent inhibitor of the es nucleoside transporter. A retroviral vector (MeiIRG) was constructed that expressed the NBMPR-insensitive ei transporter, hypothesizing that transduced bone marrow cells would survive drug treatment because of the preservation of nucleoside salvage pathways. In vitro clonogenic assays confirmed that the MeiIRG vector did protect myeloid progenitors against the toxic effects of 3 different antifolates when each was combined with NBMPR. On testing this system in vivo, decreased myelosuppression was observed in mice transplanted with MeiIRG-transduced bone marrow cells and subsequently treated with trimetrexate and NBMPR-P. In these mice, significant increases were noted in absolute neutrophil count nadirs, reticulocyte indices, and the numbers of myeloid progenitors in the bone marrow. Furthermore, a survival advantage was associated with transfer of the MeiIRG vector, indicating that significant dose intensification was possible with this approach. In summary, the MeiIRG vector can decrease the toxicity associated with the combined use of antifolates and NBMPR-P and thereby may provide a strategy for simultaneously sensitizing tumor cells while protecting hematopoietic cells.
Patel
D H
DH
Division of Experimental Hematology, Department of Molecular Pharmacology, St. Jude Children's Research Hospital, Memphis, TN, USA. divyen.patel@stjude.org
Allay
J A
JA
Belt
J A
JA
Sorrentino
B P
BP
D016475:3T3 Cells; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D016923:Cell Death / Q000187:drug effects; D018076:DNA, Complementary / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics*; D004357:Drug Synergism; D033722:Equilibrative-Nucleoside Transporter 2; D005260:Female; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005971:Glutamates / Q000494:pharmacology; D006147:Guanine / Q000031:analogs & derivatives / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000187:drug effects; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D000068437:Pemetrexed; D011799:Quinazolines / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics*; D013868:Thioinosine / Q000031:analogs & derivatives / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology
D002352:Carrier Proteins; D018076:DNA, Complementary; D033722:Equilibrative-Nucleoside Transporter 2; D005493:Folic Acid Antagonists; D005971:Glutamates; D008565:Membrane Proteins; D011799:Quinazolines; C486681:Slc29a2 protein, mouse; D013876:Thiophenes; D000068437:Pemetrexed; D013868:Thioinosine; D006147:Guanine; C068874:raltitrexed; C001789:4-nitrobenzylthioinosine