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A novel dual function retrovirus expressing multidrug resistance 1 and O6-alkylguanine-DNA-alkyltransferase for engineering resistance of haemopoietic progenitor cells to multiple chemotherapeutic agents

Gene Therapy volume 6, pages 1489–1493 (1999)Cite this article

Abstract

Following transduction with a retrovirus (SF1MIH) expressing both the multidrug resistance 1 (MDR1) and O6-alkylguanine-DNA-alkyltransferase (ATase) proteins, human erythroleukaemic progenitor (K562) cells were isolated which were resistant to killing by the MDR1 substrate, colchicine. In colony-forming survival assays, K562-SF1MIH cells exhibited resistance to colchicine and doxorubicin, as well as to the O6-alkylating agents N-Methyl-N-nitrosourea (MNU) and temozolomide. Furthermore, the resistance to doxorubicin was abolished by preincubation with the MDR1 inhibitor verapamil while resistance to MNU was ablated by the specific ATase inactivator, O6-benzylguanine (O6-beG) confirming that resistance to doxorubicin and MNU was conferred by MDR1 and ATase, respectively. When K562-SF1MIH were exposed to combinations of colchicine and MNU or doxorubicin and temozolomide, simultaneous resistance to these agents was observed. Thus, transduction of K562 with SF1MIH conferred dual resistance to these cells. These data offer the prospect of designing vectors that will confer resistance to entire regimens of chemotherapy rather than just to individual components of such drug cocktails, thereby substantially increasing the efficacy of therapy. Furthermore, the use of such dual expression constructs is likely to be highly informative for the design of effective in vivo selection protocols, an issue likely to make a major impact in a clinical context in gene therapy in the near future.

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Acknowledgements

JJ and RC were supported by grant 3557–3 from the Internal Grant Agency, Ministry of Health, Czech Republic. JAR, LJF, GPM and TMD were supported by grants from the Medical Research Council and the Cancer Research Campaign. MH was supported by the German José-Carreras-Leukemia-Fond, CB by the Deutsche Krebshilfe (10–1063-BaI). Our thanks to Gill Foster for provision of technical assistance with aspects of immunocytochemistry and to Eva Mikulova and Eva Podzimkova for cell culture work.

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Authors and Affiliations

  1. Institute of Haematology and Blood Transfusion, Praha, Czech Republic

    J Jelinek & R Cmejla

  2. Sections of Genome Damage and Repair, Paterson Institute for Cancer Research, Manchester, UK

    J A Rafferty & G P Margison

  3. Sections of Haemopoietic Cell and Gene Therapeutics, Paterson Institute for Cancer Research, Manchester, UK

    D Chinnasamy, T M Dexter & L J Fairbairn

  4. Sections of Academic Unit of Paediatric Oncology, Christie Hospital (NHS) Trust, Manchester, UK

    L S Lashford

  5. Heinrich-Pette-Institute for Experimental Virology and Immunology, Hamburg, Germany

    M Hildinger, W Ostertag & C Baum

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  1. J Jelinek
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Jelinek, J., Rafferty, J., Cmejla, R. et al. A novel dual function retrovirus expressing multidrug resistance 1 and O6-alkylguanine-DNA-alkyltransferase for engineering resistance of haemopoietic progenitor cells to multiple chemotherapeutic agents. Gene Ther 6, 1489–1493 (1999). https://doi.org/10.1038/sj.gt.3300962

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