title: Evaluating baculovirus as a vector for human prostate cancer gene therapy.
abstract: Gene therapy represents an attractive strategy for the non-invasive treatment of prostate cancer, where current clinical interventions show limited efficacy. Here, we evaluate the use of the insect virus, baculovirus (BV), as a novel vector for human prostate cancer gene therapy. Since prostate tumours represent a heterogeneous environment, a therapeutic approach that achieves long-term regression must be capable of targeting multiple transformed cell populations. Furthermore, discrimination in the targeting of malignant compared to non-malignant cells would have value in minimising side effects. We employed a number of prostate cancer models to analyse the potential for BV to achieve these goals. In vitro, both traditional prostate cell lines as well as primary epithelial or stromal cells derived from patient prostate biopsies, in two- or three-dimensional cultures, were used. We also evaluated BV in vivo in murine prostate cancer xenograft models. BV was capable of preferentially transducing invasive malignant prostate cancer cell lines compared to early stage cancers and non-malignant samples, a restriction that was not a function of nuclear import. Of more clinical relevance, primary patient-derived prostate cancer cells were also efficiently transduced by BV, with robust rates observed in epithelial cells of basal phenotype, which expressed BV-encoded transgenes faster than epithelial cells of a more differentiated, luminal phenotype. Maximum transduction capacity was observed in stromal cells. BV was able to penetrate through three-dimensional structures, including in vitro spheroids and in vivo orthotopic xenografts. BV vectors containing a nitroreductase transgene in a gene-directed enzyme pro-drug therapy approach were capable of efficiently killing malignant prostate targets following administration of the pro-drug, CB1954. Thus, BV is capable of transducing a large proportion of prostate cell types within a heterogeneous 3-D prostate tumour, can facilitate cell death using a pro-drug approach, and shows promise as a vector for the treatment of prostate cancer.
authors.affiliation: Yorkshire Cancer Research Unit, Department of Biology, University of York, Heslington, York, United Kingdom.
authors.forename: Stephanie L
authors.initials: SL
authors.lastname: Swift
authors.forename: Guillermo C
authors.initials: GC
authors.lastname: Rivera
authors.forename: Vincent
authors.initials: V
authors.lastname: Dussupt
authors.forename: Regina M
authors.initials: RM
authors.lastname: Leadley
authors.forename: Lucy C
authors.initials: LC
authors.lastname: Hudson
authors.forename: Corrina
authors.initials: C
authors.lastname: Ma de Ridder
authors.forename: Robert
authors.initials: R
authors.lastname: Kraaij
authors.forename: Julie E
authors.initials: JE
authors.lastname: Burns
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Maitland
authors.forename: Lindsay J
authors.initials: LJ
authors.lastname: Georgopoulos
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal; D011471:Prostatic Neoplasms / Q000628:therapy*; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays