Long-Term Protection of Hematopoiesis Against the Cytotoxic Effects of Multiple Doses of Nitrosourea by Retrovirus-Mediated Expression of Human O6-Alkylguanine-DNA-Alkyltransferase

• Published in: Blood Vol. 87; no. 5; pp. 1957 - 1961
• Main Authors: Jelinek, Jaroslav, Fairbairn, Leslie J., Dexter, T.Michael, Rafferty, Joseph A., Stocking, Carol, Ostertag, Wolfram, Margison, Geoffrey P.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 01-03-1996; The Americain Society of Hematology
• Subjects: Animals; Antineoplastic Agents, Alkylating - toxicity; Biological and medical sciences; Bone Marrow Cells; Cells, Cultured; DNA Damage; DNA Repair; DNA, Complementary - genetics; Drug toxicity and drugs side effects treatment; Genetic Vectors; Hematopoiesis - drug effects; Hematopoiesis - physiology; Hematopoietic Stem Cells - drug effects; Humans; Medical sciences; Methylnitrosourea - toxicity; Methyltransferases - biosynthesis; Methyltransferases - genetics; Methyltransferases - physiology; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; O-Methylguanine-DNA Methyltransferase; Pharmacology. Drug treatments; Recombinant Fusion Proteins - metabolism; Retroviridae - genetics; Toxicity: blood; Transfection; Tumor Cells, Cultured; Antineoplastic agent; Human; Cell culture; Enzyme; Nitrosoureas; Toxicity; Transferases; Rodentia; Retroviridae; In vitro; Genetic transfer; Virus; Prevention; Methylated-DNA-protein-cysteine S-methyltransferase; Vertebrata; Alkylating agent; Mammalia; Methyltransferases; Mouse; Hematopoiesis; Animal; Bone marrow; Gene therapy
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V87.5.1957.1957

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-nirtrosourea (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.