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Transcriptional targeting of RGD fiber-mutant adenovirus vectors can improve the safety of suicide gene therapy for murine melanoma

Cancer Gene Therapy volume 12, pages 608–616 (2005)Cite this article

Abstract

Since RGD fiber-mutant adenovirus vector (AdRGD), which contains an αv-integrin tropism, is highly efficient in gene transduction to melanoma, the AdRGD-mediated herpes simplex virus thymidine kinase (HSVtk)/ganciclovir (GCV) system is an attractive approach for melanoma treatment. However, the intratumoral injection of AdRGD causes limited transgene expression in healthy normal tissue, due to unwanted vector spread. Herein, we describe our attempt to overcome this limitation related to the safety of HSVtk/GCV treatment by using AdRGD carrying either melanoma-specific tyrosinase (Tyr) promoter or tumor-specific telomerase reverse transcriptase (TERT) promoter instead of universal cytomegalovirus promoter. Our in vitro study revealed that Tyr promoter-regulated AdRGD exhibited high transgene expression specificity for melanoma cells, and that TERT promoter-regulated AdRGD could induce efficient gene expression in tumor cells, but was relatively quiescent in normal cells. Anti-B16BL6 melanoma effects in mice injected intratumorally with AdRGD-Tyr/HSVtk or AdRGD-TERT/HSVtk, after which GCV was injected intraperitoneally for 10 days, were comparable to those in mice injected with AdRGD-CMV/HSVtk at 10 times less vector dosage. On the other hand, AdRGD-Tyr/HSVtk and AdRGD-TERT/HSVtk did not induce severe adverse effects even when they were intravenously injected into mice at 109 plaque-forming units (PFU), whereas mice injected with AdRGD-CMV/HSVtk at 108 PFU exhibited body weight reduction and serum level increase of biochemical enzymes for hepatotoxicity. These results indicate that AdRGD combined with transcriptional regulation using Tyr or TERT promoter is a potentially useful and safe vector system for suicide gene therapy for melanoma.

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Abbreviations

Ad:

adenovirus vector

AdRGD:

RGD fiber-mutant adenovirus vector

FBS:

fetal bovine serum

GCV:

ganciclovir

GOT:

glutamic oxaloacetic transaminase

GPT:

glutamic pyruvic transaminase

HSVtk:

herpes simplex virus thymidine kinase

MOI:

multiplicity of infection

MTT, 3-(4,5-dimethylthiazol-2-yl)-2:

5-diphenyl tetrazolium bromide

PFU:

plaque-forming unit

PBS:

phosphate-buffered saline

RLU:

relative light unit

TERT:

telomerase reverse transcriptase

Tyr:

tyrosinase

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Acknowledgements

We are grateful to Dr David L Bartlett (Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD) for providing pTyrex-2. The present study was supported in part by the Sasakawa Scientific Research Grant from The Japan Science Society, and by grants from the Ministry of Health, Labour and Welfare in Japan.

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Authors and Affiliations

  1. Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Yuka Okada

  2. Department of Biopharmaceutics, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto, 607-8414, Japan

    Naoki Okada

  3. National Institute of Health Sciences, Osaka Branch, Fundamental Research Laboratories for Development of Medicine, 7-6-8 Asagi, Saito, Ibaraki, Osaka, 567-0085, Japan

    Hiroyuki Mizuguchi

  4. National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan

    Takao Hayakawa

  5. Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Shinsaku Nakagawa & Tadanori Mayumi

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  1. Yuka Okada
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Correspondence to Yuka Okada.

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Okada, Y., Okada, N., Mizuguchi, H. et al. Transcriptional targeting of RGD fiber-mutant adenovirus vectors can improve the safety of suicide gene therapy for murine melanoma. Cancer Gene Ther 12, 608–616 (2005). https://doi.org/10.1038/sj.cgt.7700824

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