Protection of mammalian cells against chemotherapeutic agents thiotepa, 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea, and mafosfamide using the DNA base excision repair genes Fpg and α-hOgg1: Implications for protective gene therapy applications

Yi Xu, W. Kent Hansen, Thomas A. Rosenquist, David A. Williams, Melissa Limp-Foster, Mark Kelley

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Abstract

Chemotherapeutic agents used in the treatment of cancer often lead to dose-limiting bone marrow suppression and may initiate secondary leukemia. N,N′,N″-triethylenethiophosphoramide (thiotepa), a polyfunctional alkylating agent, is used in the treatment of breast, ovarian, and bladder carcinomas and is also being tested for efficacy in the treatment of central nervous system tumors. Thiotepa produces ring-opened bases such as formamidopyrimidine and 7-methyl-formamidopyrimidine, which can be recognized and repaired by the formamidopyrimidine glycosylase/AP lyase (Fpg) enzyme of Escherichia coli. Using this background information, we have created constructs using the E. coli fpg gene along with the functional equivalent human ortholog α-hOgg1. Although protection with the Fpg protein has been previously observed in Chinese hamster ovary cells, we demonstrate significant (100-fold) protection against thiotepa using the E. coli Fpg or the human α-hOgg1 cDNA in NIH3T3 cells. We have also observed a 10-fold protection by both the Fpg and α-hOgg1 transgenes against 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea (BCNU) and, to a lesser extent, mafosfamide (2-fold), an active form of the clinical agent cyclophosphamide. These latter two findings are novel and are particularly significant since the added protection was in an O6-methylguanine-DNA methyltransferase-positive background. These results support our general approach of using DNA base excision repair genes in gene therapy for cellular protection of normal cells during chemotherapy, particularly against the severe myelosuppressive effect of agents such as thiotepa, BCNU, and cyclophosphamide.

Original languageEnglish
Pages (from-to)825-831
Number of pages7
JournalJournal of Pharmacology and Experimental Therapeutics
Volume296
Issue number3
StatePublished - 2001

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DNA-(Apurinic or Apyrimidinic Site) Lyase
Thiotepa
Cytoprotection
DNA Repair
Genetic Therapy
DNA
Carmustine
Genes
Escherichia coli
Cyclophosphamide
Central Nervous System Neoplasms
Alkylating Agents
Methyltransferases
Cricetulus
Transgenes
Ovary
Leukemia
Urinary Bladder
Breast
Complementary DNA

ASJC Scopus subject areas

  • Pharmacology

Cite this

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title = "Protection of mammalian cells against chemotherapeutic agents thiotepa, 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea, and mafosfamide using the DNA base excision repair genes Fpg and α-hOgg1: Implications for protective gene therapy applications",
abstract = "Chemotherapeutic agents used in the treatment of cancer often lead to dose-limiting bone marrow suppression and may initiate secondary leukemia. N,N′,N″-triethylenethiophosphoramide (thiotepa), a polyfunctional alkylating agent, is used in the treatment of breast, ovarian, and bladder carcinomas and is also being tested for efficacy in the treatment of central nervous system tumors. Thiotepa produces ring-opened bases such as formamidopyrimidine and 7-methyl-formamidopyrimidine, which can be recognized and repaired by the formamidopyrimidine glycosylase/AP lyase (Fpg) enzyme of Escherichia coli. Using this background information, we have created constructs using the E. coli fpg gene along with the functional equivalent human ortholog α-hOgg1. Although protection with the Fpg protein has been previously observed in Chinese hamster ovary cells, we demonstrate significant (100-fold) protection against thiotepa using the E. coli Fpg or the human α-hOgg1 cDNA in NIH3T3 cells. We have also observed a 10-fold protection by both the Fpg and α-hOgg1 transgenes against 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea (BCNU) and, to a lesser extent, mafosfamide (2-fold), an active form of the clinical agent cyclophosphamide. These latter two findings are novel and are particularly significant since the added protection was in an O6-methylguanine-DNA methyltransferase-positive background. These results support our general approach of using DNA base excision repair genes in gene therapy for cellular protection of normal cells during chemotherapy, particularly against the severe myelosuppressive effect of agents such as thiotepa, BCNU, and cyclophosphamide.",
author = "Yi Xu and Hansen, {W. Kent} and Rosenquist, {Thomas A.} and Williams, {David A.} and Melissa Limp-Foster and Mark Kelley",
year = "2001",
language = "English",
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TY - JOUR

T1 - Protection of mammalian cells against chemotherapeutic agents thiotepa, 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea, and mafosfamide using the DNA base excision repair genes Fpg and α-hOgg1

T2 - Implications for protective gene therapy applications

AU - Xu, Yi

AU - Hansen, W. Kent

AU - Rosenquist, Thomas A.

AU - Williams, David A.

AU - Limp-Foster, Melissa

AU - Kelley, Mark

PY - 2001

Y1 - 2001

N2 - Chemotherapeutic agents used in the treatment of cancer often lead to dose-limiting bone marrow suppression and may initiate secondary leukemia. N,N′,N″-triethylenethiophosphoramide (thiotepa), a polyfunctional alkylating agent, is used in the treatment of breast, ovarian, and bladder carcinomas and is also being tested for efficacy in the treatment of central nervous system tumors. Thiotepa produces ring-opened bases such as formamidopyrimidine and 7-methyl-formamidopyrimidine, which can be recognized and repaired by the formamidopyrimidine glycosylase/AP lyase (Fpg) enzyme of Escherichia coli. Using this background information, we have created constructs using the E. coli fpg gene along with the functional equivalent human ortholog α-hOgg1. Although protection with the Fpg protein has been previously observed in Chinese hamster ovary cells, we demonstrate significant (100-fold) protection against thiotepa using the E. coli Fpg or the human α-hOgg1 cDNA in NIH3T3 cells. We have also observed a 10-fold protection by both the Fpg and α-hOgg1 transgenes against 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea (BCNU) and, to a lesser extent, mafosfamide (2-fold), an active form of the clinical agent cyclophosphamide. These latter two findings are novel and are particularly significant since the added protection was in an O6-methylguanine-DNA methyltransferase-positive background. These results support our general approach of using DNA base excision repair genes in gene therapy for cellular protection of normal cells during chemotherapy, particularly against the severe myelosuppressive effect of agents such as thiotepa, BCNU, and cyclophosphamide.

AB - Chemotherapeutic agents used in the treatment of cancer often lead to dose-limiting bone marrow suppression and may initiate secondary leukemia. N,N′,N″-triethylenethiophosphoramide (thiotepa), a polyfunctional alkylating agent, is used in the treatment of breast, ovarian, and bladder carcinomas and is also being tested for efficacy in the treatment of central nervous system tumors. Thiotepa produces ring-opened bases such as formamidopyrimidine and 7-methyl-formamidopyrimidine, which can be recognized and repaired by the formamidopyrimidine glycosylase/AP lyase (Fpg) enzyme of Escherichia coli. Using this background information, we have created constructs using the E. coli fpg gene along with the functional equivalent human ortholog α-hOgg1. Although protection with the Fpg protein has been previously observed in Chinese hamster ovary cells, we demonstrate significant (100-fold) protection against thiotepa using the E. coli Fpg or the human α-hOgg1 cDNA in NIH3T3 cells. We have also observed a 10-fold protection by both the Fpg and α-hOgg1 transgenes against 1,3-N,N′-bis(2-chloroethyl)-N-nitrosourea (BCNU) and, to a lesser extent, mafosfamide (2-fold), an active form of the clinical agent cyclophosphamide. These latter two findings are novel and are particularly significant since the added protection was in an O6-methylguanine-DNA methyltransferase-positive background. These results support our general approach of using DNA base excision repair genes in gene therapy for cellular protection of normal cells during chemotherapy, particularly against the severe myelosuppressive effect of agents such as thiotepa, BCNU, and cyclophosphamide.

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