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.

Highlights

  • In the United Kingdom, over 40,000 new cases of prostate cancer are diagnosed every year and 11,000 patients die as a direct result of this disease (Source: Office for National Statistics)

  • In order to determine the capacity of BV to differentially transduce malignant vs. non-malignant prostate targets, we evaluated the susceptibility of a panel of established human prostate cell lines to BV transduction

  • Malignant prostate cell lines (LNCaP and PC-3) demonstrated rapid and efficient uptake of BV-[CMV-EGFPCAT], requiring only short incubation times (,45 or,90 min, respectively) to reach 50% of the levels recorded at 48 h (Figure 1B)

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Summary

Introduction

In the United Kingdom, over 40,000 new cases of prostate cancer are diagnosed every year and 11,000 patients die as a direct result of this disease (Source: Office for National Statistics). While the bulk of most prostate tumours comprises cells of epithelial origin [3], multiple cell types have the capacity to contribute towards malignancy, and must be eradicated in order to achieve an effective clinical outcome. The stage of prostate disease and the responsiveness of the tumour to androgens are two key factors that direct treatment strategies. Androgen ablation therapy, removes the supply of androgens that many prostate tumours rely on for growth and survival; the recurrence of more aggressive castration-resistant prostate cancer (CRPC) is a common outcome. Since CRPC is resistant to radiotherapy and chemotherapy, novel strategies are essential to provide improved outcomes for prostate cancer patients. Gene therapy is one potential solution, and would ideally be positioned to target both primary and secondary prostate tumours following systemic administration

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