Preclinical evaluation of a prostate-targeted gene-directed enzyme prodrug therapy delivered by ovine atadenovirus

• Published in: Gene therapy Vol. 11; no. 21; pp. 1559 - 1567
• Main Authors: WANG, X. Y, MARTINIELLO-WILKS, R, WEBSTER, L. K, SMITH, I. K, BOTH, G. W, RUSSELL, P. J, SHAW, J. M, HO, T, COULSTON, N, COOKE-YARBOROUGH, C, MOLLOY, P. L, CAMERON, F, MOGHADDAM, M, LOCKETT, T. J
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.11.2004
• Subjects: Adenine - analogs & derivatives; Adenine - metabolism; Androgens; Animals; Antineoplastic Agents - metabolism; Antineoplastic Agents - therapeutic use; Apoptosis; Biological and medical sciences; Biotechnology; Cell Line, Tumor; Cell Proliferation; DNA biosynthesis; DNA Replication - drug effects; Drug Evaluation, Preclinical; Enzymes; Fludarabine; Fundamental and applied biological sciences. Psychology; Gene expression; Gene therapy; Genetic Therapy - methods; Genetic Vectors - administration & dosage; Health. Pharmaceutical industry; High-performance liquid chromatography; Humans; Industrial applications and implications. Economical aspects; Infectivity; Male; Mice; Mice, Nude; Neoplasm Transplantation; ovine atadenovirus; Phosphorylase; Pnp gene; Prodrugs; Prodrugs - therapeutic use; Prostate; Prostate cancer; Prostatic Neoplasms - metabolism; Prostatic Neoplasms - therapy; Protein biosynthesis; Purine-Nucleoside Phosphorylase - genetics; Purine-Nucleoside Phosphorylase - metabolism; Reporter gene; Transcription; Transfection; Tumors; Vidarabine Phosphate - analogs & derivatives; Vidarabine Phosphate - metabolism; Vidarabine Phosphate - therapeutic use; Xenografts; Australia; Antineoplastic agent; Therapy; GDEPT; Enzyme; Prodrug; Heterograft; prostate xenografts; Fludarabine; Gene; fludarabine phosphate; PNP gene; Gene therapy; ovine atadenovirus; Urogenital system; Prostate
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3302308

Gene-directed enzyme prodrug therapy (GDEPT) based on the Escherichia coli enzyme, purine nucleoside phosphorylase (PNP), provides a novel strategy for treating slowly growing tumors like prostate cancer (CaP). PNP converts systemically administered prodrug, fludarabine phosphate, to a toxic metabolite, 2-fluoroadenine, that kills PNP-expressing and nearby cells by inhibiting DNA, RNA and protein synthesis. Reporter gene expression directed by a hybrid prostate-directed promoter and enhancer, PSMEPb, was assayed after plasmid transfection or viral transduction of prostate and non-CaP cell lines. Androgen-sensitive (AS) LNCaP-LN3 and androgen-independent (AI) PC3 human CaP xenografts in nude mice were injected intratumorally with an ovine atadenovirus vector, OAdV623, that carries the PNP gene under PSMEPb, formulated with cationic lipid for enhanced infectivity. Fludarabine phosphate was then given intraperitoneally for 5 days at 75 mg/m2/day. PNP expression was evaluated by enzymic conversion of its substrate using reverse phase HPLC. OAdV623 showed excellent in vitro transcriptional specificity for CaP cells. In vivo, expression of PNP persisted for > 6 days after OAdV623 injection and a single treatment provided 100% increase in tumor doubling time and > 50% inhibition of tumor growth for both LNCaP-LN3 and PC3 lines, with increased tumor necrosis and apoptosis and decreased tumor cell proliferation. OAdV623 significantly suppressed the growth of AS and AI human CaP xenografts in mice.