title: Enhancing hemopoietic drug resistance: a rationale for reconsidering the clinical use of mitozolomide.
abstract: Retroviral gene transfer was used to achieve expression in mouse bone marrow of a mutant form of the DNA repair protein O6-alkylguanine-DNA alkyltransferase (hATPA/GA), which exhibits resistance to inactivation by O6-benzylguanine (O6-beG). After reconstitution of mice with transduced bone marrow, approximately 50% of the bipotent granulocyte-macrophage colony-forming cell (GM-CFC) and multipotent spleen colony-forming unit (CFU-S) hemopoietic populations showed expression of the transgene; this expression was associated with resistance to either mitozolomide or to a combination of O6-beG and mitozolomide, relative to mock-transduced controls. Thus, at a dose of mitozolomide in vivo that allowed only 70% and 62% survival of mock-transduced GM-CFC and CFU-S, respectively, the hATPA/GA CFC were totally resistant to the same dose of mitozolomide (P < .05 and .001, respectively). In the presence of O6-beG, the toxicity of mitozolomide was greatly potentiated. Only 24% and 18%, respectively, of mock-transduced GM-CFC and CFU-S survived combination treatment, whereas 45% (P < .05) and 37% (P < .01) of GM-CFC and CFU-S, respectively, from hATPA/GA mice survived the same combination of doses. Furthermore, as a result of transgene expression, the number of micronucleated polychromatic erythrocytes induced by mitozolomide was significantly reduced (P < .05) by 40% relative to mock-transduced controls, indicating the potential of this approach to reduce the frequency of mutation associated with chemotherapy exposure. The protection against the toxic and clastogenic effects of mitozolomide in both primitive and more mature hemopoietic cells suggests that the severe myelosuppression that halted further clinical investigation of this drug could be substantially ameliorated by the exogenous expression of O6-alkylguanine-DNA alkyltransferase. Therefore, these data raise the prospect for the reinvestigation of mitozolomide and other proscribed drugs in the context of genetically protected hemopoiesis.
authors.affiliation: Cancer Research Campaign Sections of Hemopoietic Cell and Gene Therapeutics, Paterson Institute for Cancer Research, Manchester, United Kingdom.
authors.forename: L J
authors.initials: LJ
authors.lastname: Fairbairn
authors.forename: N
authors.initials: N
authors.lastname: Chinnasamy
authors.forename: L S
authors.initials: LS
authors.lastname: Lashford
authors.forename: D
authors.initials: D
authors.lastname: Chinnasamy
authors.forename: J A
authors.initials: JA
authors.lastname: Rafferty
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D016026:Bone Marrow Transplantation; D003114:Colony-Forming Units Assay; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D004357:Drug Synergism; D018014:Gene Transfer Techniques; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D009153:Mutagens / Q000494:pharmacology; D009588:Nitrogen Mustard Compounds / Q000494:pharmacology*; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D009153:Mutagens; D009588:Nitrogen Mustard Compounds; C040369:mitozolomide; D019853:O(6)-Methylguanine-DNA Methyltransferase