Viral delivery of P450 reductase recapitulates the ability of constitutive overexpression of reductase enzymes to potentiate the activity of mitomycin C in human breast cancer xenografts

Rachel L Cowen, Adam V Patterson, Brian A Telfer, Rachel E Airley, Steve Hobbs, Roger M Phillips, Mohammed Jaffar, Ian J Stratford, Kaye J Williams

Research output: Contribution to journalArticle

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Abstract

Indolequinones such as mitomycin C (MMC) require enzymatic bioreduction to yield cytotoxic moieties. An attractive approach to overcome the potential variability in reductive bioactivation between tumors is to exploit specific enzyme-bioreductive drug combinations in an enzyme-directed gene therapy (GDEPT) approach. To this end, human breast cancer cell lines (T47D, MDA468, and MDA231) that overexpress either DT-diaphorase (DTD) or NADPH:cytochrome P450 reductase (P450R) have been developed. Cytotoxicity of MMC was evaluated in the panel of cell lines following aerobic or anoxic exposure in vitro. DTD and/or P450R overexpression sensitized cells to MMC in air with no further increase in the cytotoxicity of MMC under anoxia. The most profound effect was seen in the MDA468 cells, where a 27-fold increase in potency was observed for MMC in the DTD-overexpressing cell line. The MMC sensitization achieved through DTD and P450R overexpression in MDA468 cells was maintained in vivo. Xenografts established from the clonal lines exhibited significant tumor control following MMC treatment (treated/control [T/C] 17% and 51% for DTD and P450R xenografts, respectively) that was not seen in wild-type tumors (T/C 102%). Delivery of a clinically relevant adenoviral vector encoding P450R to MDA468 wild-type tumors yielded comparable P450R activity to that seen in the P450R clonal xenografts and resulted in greater MMC sensitization (T/C 46%). The model systems developed will facilitate the identification of novel indolequinone agents that are targeted toward a specific enzyme for bioactivation and are consequently of potential use in a GDEPT approach.

Original languageEnglish
Pages (from-to)901-909
Number of pages9
JournalMolecular Cancer Therapeutics
Volume2
Issue number9
Publication statusPublished - Sep 2003
Externally publishedYes

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Mitomycin
Heterografts
NAD(P)H Dehydrogenase (Quinone)
Oxidoreductases
Breast Neoplasms
Enzymes
Indolequinones
Cell Line
Neoplasms
NADPH-Ferrihemoprotein Reductase
Drug Combinations
Genetic Therapy
Air

Cite this

Cowen, Rachel L ; Patterson, Adam V ; Telfer, Brian A ; Airley, Rachel E ; Hobbs, Steve ; Phillips, Roger M ; Jaffar, Mohammed ; Stratford, Ian J ; Williams, Kaye J. / Viral delivery of P450 reductase recapitulates the ability of constitutive overexpression of reductase enzymes to potentiate the activity of mitomycin C in human breast cancer xenografts. In: Molecular Cancer Therapeutics. 2003 ; Vol. 2, No. 9. pp. 901-909.
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abstract = "Indolequinones such as mitomycin C (MMC) require enzymatic bioreduction to yield cytotoxic moieties. An attractive approach to overcome the potential variability in reductive bioactivation between tumors is to exploit specific enzyme-bioreductive drug combinations in an enzyme-directed gene therapy (GDEPT) approach. To this end, human breast cancer cell lines (T47D, MDA468, and MDA231) that overexpress either DT-diaphorase (DTD) or NADPH:cytochrome P450 reductase (P450R) have been developed. Cytotoxicity of MMC was evaluated in the panel of cell lines following aerobic or anoxic exposure in vitro. DTD and/or P450R overexpression sensitized cells to MMC in air with no further increase in the cytotoxicity of MMC under anoxia. The most profound effect was seen in the MDA468 cells, where a 27-fold increase in potency was observed for MMC in the DTD-overexpressing cell line. The MMC sensitization achieved through DTD and P450R overexpression in MDA468 cells was maintained in vivo. Xenografts established from the clonal lines exhibited significant tumor control following MMC treatment (treated/control [T/C] 17{\%} and 51{\%} for DTD and P450R xenografts, respectively) that was not seen in wild-type tumors (T/C 102{\%}). Delivery of a clinically relevant adenoviral vector encoding P450R to MDA468 wild-type tumors yielded comparable P450R activity to that seen in the P450R clonal xenografts and resulted in greater MMC sensitization (T/C 46{\%}). The model systems developed will facilitate the identification of novel indolequinone agents that are targeted toward a specific enzyme for bioactivation and are consequently of potential use in a GDEPT approach.",
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Viral delivery of P450 reductase recapitulates the ability of constitutive overexpression of reductase enzymes to potentiate the activity of mitomycin C in human breast cancer xenografts. / Cowen, Rachel L; Patterson, Adam V; Telfer, Brian A; Airley, Rachel E; Hobbs, Steve; Phillips, Roger M; Jaffar, Mohammed; Stratford, Ian J; Williams, Kaye J.

In: Molecular Cancer Therapeutics, Vol. 2, No. 9, 09.2003, p. 901-909.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Viral delivery of P450 reductase recapitulates the ability of constitutive overexpression of reductase enzymes to potentiate the activity of mitomycin C in human breast cancer xenografts

AU - Cowen, Rachel L

AU - Patterson, Adam V

AU - Telfer, Brian A

AU - Airley, Rachel E

AU - Hobbs, Steve

AU - Phillips, Roger M

AU - Jaffar, Mohammed

AU - Stratford, Ian J

AU - Williams, Kaye J

PY - 2003/9

Y1 - 2003/9

N2 - Indolequinones such as mitomycin C (MMC) require enzymatic bioreduction to yield cytotoxic moieties. An attractive approach to overcome the potential variability in reductive bioactivation between tumors is to exploit specific enzyme-bioreductive drug combinations in an enzyme-directed gene therapy (GDEPT) approach. To this end, human breast cancer cell lines (T47D, MDA468, and MDA231) that overexpress either DT-diaphorase (DTD) or NADPH:cytochrome P450 reductase (P450R) have been developed. Cytotoxicity of MMC was evaluated in the panel of cell lines following aerobic or anoxic exposure in vitro. DTD and/or P450R overexpression sensitized cells to MMC in air with no further increase in the cytotoxicity of MMC under anoxia. The most profound effect was seen in the MDA468 cells, where a 27-fold increase in potency was observed for MMC in the DTD-overexpressing cell line. The MMC sensitization achieved through DTD and P450R overexpression in MDA468 cells was maintained in vivo. Xenografts established from the clonal lines exhibited significant tumor control following MMC treatment (treated/control [T/C] 17% and 51% for DTD and P450R xenografts, respectively) that was not seen in wild-type tumors (T/C 102%). Delivery of a clinically relevant adenoviral vector encoding P450R to MDA468 wild-type tumors yielded comparable P450R activity to that seen in the P450R clonal xenografts and resulted in greater MMC sensitization (T/C 46%). The model systems developed will facilitate the identification of novel indolequinone agents that are targeted toward a specific enzyme for bioactivation and are consequently of potential use in a GDEPT approach.

AB - Indolequinones such as mitomycin C (MMC) require enzymatic bioreduction to yield cytotoxic moieties. An attractive approach to overcome the potential variability in reductive bioactivation between tumors is to exploit specific enzyme-bioreductive drug combinations in an enzyme-directed gene therapy (GDEPT) approach. To this end, human breast cancer cell lines (T47D, MDA468, and MDA231) that overexpress either DT-diaphorase (DTD) or NADPH:cytochrome P450 reductase (P450R) have been developed. Cytotoxicity of MMC was evaluated in the panel of cell lines following aerobic or anoxic exposure in vitro. DTD and/or P450R overexpression sensitized cells to MMC in air with no further increase in the cytotoxicity of MMC under anoxia. The most profound effect was seen in the MDA468 cells, where a 27-fold increase in potency was observed for MMC in the DTD-overexpressing cell line. The MMC sensitization achieved through DTD and P450R overexpression in MDA468 cells was maintained in vivo. Xenografts established from the clonal lines exhibited significant tumor control following MMC treatment (treated/control [T/C] 17% and 51% for DTD and P450R xenografts, respectively) that was not seen in wild-type tumors (T/C 102%). Delivery of a clinically relevant adenoviral vector encoding P450R to MDA468 wild-type tumors yielded comparable P450R activity to that seen in the P450R clonal xenografts and resulted in greater MMC sensitization (T/C 46%). The model systems developed will facilitate the identification of novel indolequinone agents that are targeted toward a specific enzyme for bioactivation and are consequently of potential use in a GDEPT approach.

KW - Adenoviridae/genetics

KW - Animals

KW - Antibiotics, Antineoplastic/therapeutic use

KW - DNA, Neoplasm/biosynthesis

KW - Drug Delivery Systems

KW - Drug Resistance, Neoplasm

KW - Drug Screening Assays, Antitumor

KW - Drug Synergism

KW - Female

KW - Genetic Vectors

KW - Humans

KW - Immunoenzyme Techniques

KW - Mammary Neoplasms, Experimental/drug therapy

KW - Mice

KW - Mice, Nude

KW - Mitomycin/therapeutic use

KW - NAD(P)H Dehydrogenase (Quinone)/genetics

KW - NADPH-Ferrihemoprotein Reductase/genetics

KW - Oxygen/metabolism

KW - Transplantation, Heterologous

KW - Tumor Cells, Cultured/transplantation

M3 - Article

VL - 2

SP - 901

EP - 909

JO - Molecular Cancer Therapeutics

JF - Molecular Cancer Therapeutics

SN - 1535-7163

IS - 9

ER -