Long-term protection of hematopoiesis against the cytotoxic effects of multiple doses of nitrosourea by retrovirus-mediated expression of human O6-alkylguanine-DNA-alkyltransferase.
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.
Jelínek
J
J
CRC Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.
Fairbairn
L J
LJ
Dexter
T M
TM
Rafferty
J A
JA
Stocking
C
C
Ostertag
W
W
Margison
G P
GP
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
D018906:Antineoplastic Agents, Alkylating; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D008770:Methylnitrosourea; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase