Retroviral Transfer of a Bacterial Alkyltransferase Gene into Murine Bone Marrow Protects Against Chloroethylnitrosourea Cytotoxicity

Linda C. Harris, Upendra K. Marathi, Carol C. Edwards, Peter J Houghton, Deo Kumar Srivastava, Elio F. Vanin, Brian P. Sorrentino, Thomas P. Brent

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The chloroethylnitrosoureas (CENUs) are important antineoplastic drugs for which clinical utility has been restricted by the development of severe delayed myelosuppression in most patients. To investigate the potential of DNA repair proteins to reduce bone marrow sensitivity to the CENUs, we transferred the Escherichia coli ada gene, which encodes a Mr 39, 000 O6-alkylguanine-DNA alkyltransferase (ATase), into murine bone marrow cells by the use of a high-titer ecotropic retrovirus. The ada-encoded ATase is resistant to O6-benzylguanine (O6-BG), a potent inhibitor of the mammalian ATases, thus affording the bone marrow an additional level of protection against CENUs. In methylcellulose cultures, ada-infected hematopoietic progenitor cells were twice as resistant as uninfected cells to the toxic effects of l, 3-bis(2-chloroethyl)-l-nitrosourea (BCNU) following treatment with O6-BG. Although showing no obvious protective effects against leukopenia, overexpression of the bacterial ATase activity reduced the severity of anemia and thrombocytopenia in mice treated with O6-BG and BCNU. These effects, which were maximal at a BCNU dose of 12.5 mg/kg, were associated with improved survival when BCNU was given at this dose. At lower BCNU doses cytotoxicity was limited in both transduced and control mice, and at higher doses the protective effect was saturated due to cytotoxicity. These results suggest that ada gene therapy may be a feasible approach to amelioration of delayed myelosuppression following O6-BG plus CENU combination chemotherapy.

Original languageEnglish (US)
Pages (from-to)1359-1368
Number of pages10
JournalClinical Cancer Research
Volume1
Issue number11
StatePublished - Nov 1 1995
Externally publishedYes

Fingerprint

Alkyl and Aryl Transferases
Bacterial Genes
Carmustine
Bone Marrow
Methylcellulose
Poisons
Leukopenia
Retroviridae
Hematopoietic Stem Cells
Combination Drug Therapy
DNA Repair
Bone Marrow Cells
Thrombocytopenia
Genetic Therapy
Antineoplastic Agents
Anemia
Escherichia coli
Survival
O(6)-benzylguanine
Genes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Harris, L. C., Marathi, U. K., Edwards, C. C., Houghton, P. J., Srivastava, D. K., Vanin, E. F., ... Brent, T. P. (1995). Retroviral Transfer of a Bacterial Alkyltransferase Gene into Murine Bone Marrow Protects Against Chloroethylnitrosourea Cytotoxicity. Clinical Cancer Research, 1(11), 1359-1368.

Retroviral Transfer of a Bacterial Alkyltransferase Gene into Murine Bone Marrow Protects Against Chloroethylnitrosourea Cytotoxicity. / Harris, Linda C.; Marathi, Upendra K.; Edwards, Carol C.; Houghton, Peter J; Srivastava, Deo Kumar; Vanin, Elio F.; Sorrentino, Brian P.; Brent, Thomas P.

In: Clinical Cancer Research, Vol. 1, No. 11, 01.11.1995, p. 1359-1368.

Research output: Contribution to journalArticle

Harris, LC, Marathi, UK, Edwards, CC, Houghton, PJ, Srivastava, DK, Vanin, EF, Sorrentino, BP & Brent, TP 1995, 'Retroviral Transfer of a Bacterial Alkyltransferase Gene into Murine Bone Marrow Protects Against Chloroethylnitrosourea Cytotoxicity', Clinical Cancer Research, vol. 1, no. 11, pp. 1359-1368.
Harris, Linda C. ; Marathi, Upendra K. ; Edwards, Carol C. ; Houghton, Peter J ; Srivastava, Deo Kumar ; Vanin, Elio F. ; Sorrentino, Brian P. ; Brent, Thomas P. / Retroviral Transfer of a Bacterial Alkyltransferase Gene into Murine Bone Marrow Protects Against Chloroethylnitrosourea Cytotoxicity. In: Clinical Cancer Research. 1995 ; Vol. 1, No. 11. pp. 1359-1368.
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abstract = "The chloroethylnitrosoureas (CENUs) are important antineoplastic drugs for which clinical utility has been restricted by the development of severe delayed myelosuppression in most patients. To investigate the potential of DNA repair proteins to reduce bone marrow sensitivity to the CENUs, we transferred the Escherichia coli ada gene, which encodes a Mr 39, 000 O6-alkylguanine-DNA alkyltransferase (ATase), into murine bone marrow cells by the use of a high-titer ecotropic retrovirus. The ada-encoded ATase is resistant to O6-benzylguanine (O6-BG), a potent inhibitor of the mammalian ATases, thus affording the bone marrow an additional level of protection against CENUs. In methylcellulose cultures, ada-infected hematopoietic progenitor cells were twice as resistant as uninfected cells to the toxic effects of l, 3-bis(2-chloroethyl)-l-nitrosourea (BCNU) following treatment with O6-BG. Although showing no obvious protective effects against leukopenia, overexpression of the bacterial ATase activity reduced the severity of anemia and thrombocytopenia in mice treated with O6-BG and BCNU. These effects, which were maximal at a BCNU dose of 12.5 mg/kg, were associated with improved survival when BCNU was given at this dose. At lower BCNU doses cytotoxicity was limited in both transduced and control mice, and at higher doses the protective effect was saturated due to cytotoxicity. These results suggest that ada gene therapy may be a feasible approach to amelioration of delayed myelosuppression following O6-BG plus CENU combination chemotherapy.",
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