Targeted expression of Escherichia coli purine nucleoside phosphorylase and Fludara® for prostate cancer therapy

• Published in: The journal of gene medicine Vol. 13; no. 12; pp. 680 - 691
• Main Authors: Xie, Xinhua, Guo, Jiaoli, Kong, Yanan, Xie, Gene Xianquan, Li, Laishen, Lv, Ning, Xiao, Xiangsheng, Tang, Jun, Wang, Xi, Liu, Peng, Yang, Mingtian, Xie, Zeming, Wei, Weidong, Xie, Xiaoming
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.12.2011; Wiley Periodicals Inc
• Subjects: Animals; Arrestins - genetics; beta-Arrestins; Cell Death - drug effects; Cell Death - genetics; Cell Line, Tumor; E. coli PNP; Escherichia coli; fludarabine; Gene Expression Regulation, Neoplastic - drug effects; Gene Expression Regulation, Neoplastic - genetics; Gene therapy; Genes, Transgenic, Suicide - genetics; Genetic Therapy; Genetic Vectors; Green Fluorescent Proteins - metabolism; Humans; Male; Metribolone - administration & dosage; Mice; probasin promoter; Promoter Regions, Genetic; prostate cancer; Prostatic Neoplasms - genetics; Prostatic Neoplasms - pathology; Prostatic Neoplasms - therapy; Purine-Nucleoside Phosphorylase - genetics; Purine-Nucleoside Phosphorylase - therapeutic use; Rats; transcriptional targeting; Vidarabine Phosphate - analogs & derivatives; Vidarabine Phosphate - therapeutic use
• ISSN: 1099-498X; 1521-2254
• DOI: 10.1002/jgm.1620

Background Previous studies have shown that Herpes Simplex Virus thymidine kinase (HSV‐tk)/ganciclovir (GCV) comprised the most commonly used suicide gene therapy for prostate cancer, with modest results being obtained. However, novel suicide genes, such as Escherichia coli purine nucleoside phosphorylase (PNP), have been utilized to demonstrate more potent tumor killing and an enhanced bystander effect on local, non‐expressing cells compared to HSV‐tk. Methods PNP/fludarabine (Fludara®; fludarabine phosphate; Berlex Labs, Richmond, CA, USA) was deliveried by prostate‐specific, rat probasin‐based promoter, ARR2PB. After infection of various cell lines with ADV.ARR2PB‐PNP and administration of androgen analog, R1881, expression of PNP mRNA was detected; in vivo, the antitumor effect of the ARR2PB‐PNP/Fludara system was monitored and analyzed, as well as animal survival. Results After in vitro infection with ADV.ARR2PB‐PNP (multiplicity of infection = 10), LNCaP cells were more sensitive to a lower concentration Fludara (LD50, approximately 0.1 µg/ml) in the presence of R1881. Furthermore, robust bystander effects after R1881/Fludara treatment were observed in LNCaP cells after infection with bicistronic vector ADV.ARR2PB/PNP‐IRES‐EGFP in contrast to a much weaker effect in cells treated with ADV.CMV‐HSV‐tk/GCV. In vivo, tumor size in the ADV.ARR2PB‐PNP/Fludara treatment group was dramatically smaller than in the control groups, and the mice treated with our system had a significantly prolonged survival, with three of eight mice surviving up to the 160‐day termination point, as well as no systemic toxicity. Conclusions The ARR2PB‐PNP/Fludara system induced massive tumor cell death and a prolonged life span without systemic cytotoxicity; therefore, it might be a more attractive strategy for suicide gene therapy of prostate cancer. Copyright © 2011 John Wiley & Sons, Ltd.