title: Mouse mammary tumor virus promoter-containing retroviral promoter conversion vectors for gene-directed enzyme prodrug therapy are functional in vitro and in vivo.
abstract: Gene directed-enzyme prodrug therapy (GDEPT) is an approach for sensitization of tumor cells to an enzymatically activated, otherwise nontoxic, prodrug. Cytochrome P450 2B1 (CYP2B1) metabolizes the prodrugs cyclophosphamide (CPA) and ifosfamide (IFA) to produce the cytotoxic substances phosphoramide mustard and isophosphoramide mustard as well as the byproduct acrolein. We have constructed a retroviral promoter conversion (ProCon) vector for breast cancer GDEPT. The vector allows expression of CYP2B1 from the mouse mammary tumor virus (MMTV) promoter known to be active in the mammary glands of transgenic animals. It is anticipated to be used for the generation of encapsulated viral vector producing cells which, when placed inside or close to a tumor, will act as suppliers of the therapeutic CYP2B1 protein as well as of the therapeutic vector itself. The generated vector was effectively packaged by virus producing cells and allowed the production of high levels of enzymatically active CYP2B1 in infected cells which sensitized them to killing upon treatment with both IFA and CPA. Determination of the respective IC(50) values demonstrated that the effective IFA dose was reduced by sixteen folds. Infection efficiencies in vivo were determined using a reporter gene-bearing vector in a mammary cancer cell-derived xenograft tumor mouse model.
authors.affiliation: Austrianova Biotechnology GmbH, Veterinärplatz 1, A-1210 Vienna, Austria. reinhard.klein@ccri.at
authors.forename: Reinhard
authors.initials: R
authors.lastname: Klein
authors.forename: Bärbel
authors.initials: B
authors.lastname: Ruttkowski
authors.forename: Sonja
authors.initials: S
authors.lastname: Schwab
authors.forename: Thomas
authors.initials: T
authors.lastname: Peterbauer
authors.forename: Brian
authors.initials: B
authors.lastname: Salmons
authors.forename: Walter H
authors.initials: WH
authors.lastname: Günzburg
authors.forename: Christine
authors.initials: C
authors.lastname: Hohenadl
mesh_terms: D000818:Animals; D018390:Gene Targeting / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D008325:Mammary Neoplasms, Experimental / Q000188:drug therapy* / Q000235:genetics*; D051379:Mice; D011355:Prodrugs / Q000627:therapeutic use*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics*; D016896:Treatment Outcome
chemical_list: D011355:Prodrugs