Generation of dual resistance to 4-hydroperoxycyclophosphamide and methotrexate by retroviral transfer of the human aldehyde dehydrogenase class 1 gene and a mutated dihydrofolate reductase gene Journal Article
| Authors: | Takebe, N.; Zhao, S. C.; Adhikari, D.; Mineishi, S.; Sadelain, M.; Hilton, J.; Colvin, M.; Banerjee, D.; Bertino, J. R. |
| Article Title: | Generation of dual resistance to 4-hydroperoxycyclophosphamide and methotrexate by retroviral transfer of the human aldehyde dehydrogenase class 1 gene and a mutated dihydrofolate reductase gene |
| Abstract: | The genetic transfer of drug resistance to hematopoietic cells is an attractive approach to overcoming myelosuppression caused by high-dose chemotherapy. Because cyclophosphamide (CTX) and methotrexate (MTX) are commonly used non-cross-resistant drugs, generation of dual drug resistance in hematopoietic cells that allows dose intensification may increase anti-tumor effects and circumvent the emergence of drug-resistant tumors. We constructed a retroviral vector containing both a human cytosolic ALDH-1 cDNA and a human doubly mutated DHFR cDNA (Phe22/Ser31; termed F/S in the description of constructs) to generate increased resistance to both CTX and MTX. Infection of NIH3T3 cells resulted in increased resistance to both 4-hydroperoxy-cyclophosphamide (4HC) (1.9 ± 0.1-fold) and MTX (73 ± 2.8-fold). Transduced human CD34+ enriched hematopoietic progenitor cells were also resistant to both 4HC and MTX by CFU-GM readout. Lethally irradiated mice transplanted with SFG-ALDH-IRES-F/S or mock-transduced bone marrow cells were treated with high-dose pulse CTX or high-dose CTX/MTX. Animals receiving marrow not transduced with ALDH-1 or mutated DHFR cDNA died from CTX or CTX/MTX toxicity, whereas mice transduced with ALDH-1 and mutated DHFR cDNA-containing marrow were able to tolerate the same doses of CTX or CTX/MTX treatment posttransplant. These data taken together indicate that ALDH-1 overexpression and mutant DHFR increased both 4HC and MTX resistance in vitro and in the in vivo mouse model. This construct may be useful for protecting patients from high-dose CTX- and MTX-induced myelosuppression. |
| Keywords: | controlled study; protein expression; gene mutation; mutation; drug tolerability; nonhuman; radiation dose; chemotherapy; methotrexate; drug megadose; polymerase chain reaction; animal cell; mouse; animals; mice; cd34 antigen; bone marrow suppression; antimetabolites, antineoplastic; serine; animal experiment; cyclophosphamide; in vivo study; antineoplastic activity; in vitro study; drug resistance; dose-response relationship, drug; drug resistance, neoplasm; time factors; gene transfer; genetic transduction; animalia; transduction, genetic; rna; blotting, western; death; irradiation; hematopoietic cell; hematopoietic stem cells; models, genetic; virus infection; hematopoietic stem cell; toxicity; dihydrofolate reductase; tetrahydrofolate dehydrogenase; bone marrow transplantation; drug pulse therapy; antigens, cd34; recombinant granulocyte macrophage colony stimulating factor; cytosol; complementary dna; retrovirus; retroviridae; phenylalanine; colony forming unit gm; 3t3 cells; aldehyde dehydrogenase; cross resistance; dna, complementary; gene construct; gene transfer techniques; recombinant stem cell factor; recombinant thrombopoietin; lethal dose; 4 hydroperoxycyclophosphamide; unidentified retrovirus; humans; article; recombinant interleukin 6 |
| Journal Title: | Molecular Therapy |
| Volume: | 3 |
| Issue: | 1 |
| ISSN: | 1525-0016 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2001-01-01 |
| Start Page: | 88 |
| End Page: | 96 |
| Language: | English |
| DOI: | 10.1006/mthe.2000.0236 |
| PUBMED: | 11162315 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 21 May 2015 -- Source: Scopus |
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