title: Long-term protection of hematopoiesis against the cytotoxic effects of multiple doses of nitrosourea by retrovirus-mediated expression of human O6-alkylguanine-DNA-alkyltransferase.
abstract: A human O6-alkylguanine-DNA-alkyltransferase (ATase) cDNA-containing retrovirus was used to infect murine long-term primary bone marrow cultures. High levels of ATase expression were obtained, and colony-forming cells of the granulocyte-macrophage lineage from the cultures transduced with the human ATase retrovirus were three times more resistant to the alkylating agent, N-methyl-N-nitrosourea (MNU), than control cultures. Furthermore, expression of the human ATase protected long-term hematopoiesis, measured as the output of progenitor cells to the nonadherent fraction of the culture, against the cytotoxic effects of repeated exposures to MNU. These results clearly show that a human ATase cDNA-containing retrovirus can be used to infect long-term primary bone marrow cultures and that this attenuates their sensitivity to nitrosoureas.
authors.affiliation: CRC Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.
authors.forename: J
authors.initials: J
authors.lastname: Jelínek
authors.forename: L J
authors.initials: LJ
authors.lastname: Fairbairn
authors.forename: T M
authors.initials: TM
authors.lastname: Dexter
authors.forename: J A
authors.initials: JA
authors.lastname: Rafferty
authors.forename: C
authors.initials: C
authors.lastname: Stocking
authors.forename: W
authors.initials: W
authors.lastname: Ostertag
authors.forename: G P
authors.initials: GP
authors.lastname: Margison
mesh_terms: D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000633:toxicity*; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D004249:DNA Damage; D004260:DNA Repair; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D006410:Hematopoiesis / Q000187:drug effects / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006801:Humans; D008770:Methylnitrosourea / Q000633:toxicity*; D008780:Methyltransferases / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D019853:O(6)-Methylguanine-DNA Methyltransferase; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D018906:Antineoplastic Agents, Alkylating; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D008770:Methylnitrosourea; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase