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
| Autor: | Naoko Takebe, John Hilton, Shi Cheng Zhao, Michael Colvin, Shin Mineishi, Debasis Adhikari, Michel Sadelain, Debabrata Banerjee, Joseph R. Bertino |
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| Rok vydání: | 2001 |
| Předmět: |
Antimetabolites
Antineoplastic DNA Complementary Time Factors Blotting Western CD34 Antigens CD34 Drug resistance Biology Polymerase Chain Reaction Viral vector Mice Transduction Genetic In vivo Drug Discovery Genetics medicine Animals Humans heterocyclic compounds Cyclophosphamide Molecular Biology Bone Marrow Transplantation Pharmacology Dose-Response Relationship Drug Models Genetic Genetic transfer Gene Transfer Techniques 3T3 Cells Aldehyde Dehydrogenase Hematopoietic Stem Cells Virology Molecular biology Tetrahydrofolate Dehydrogenase Haematopoiesis Methotrexate Retroviridae medicine.anatomical_structure Drug Resistance Neoplasm Mutation RNA Molecular Medicine Bone marrow medicine.drug |
| Zdroj: | Molecular Therapy. 3:88-96 |
| ISSN: | 1525-0016 |
| DOI: | 10.1006/mthe.2000.0236 |
| Popis: | 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-hydroperoxycyclophosphamide (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. |
| Databáze: | OpenAIRE |
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