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A simple detection system for adenovirus receptor expression using a telomerase-specific replication-competent adenovirus

Gene Therapy volume 20, pages 112–118 (2013)Cite this article

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Abstract

Adenovirus serotype 5 (Ad5) is frequently used as an effective vector for induction of therapeutic transgenes in cancer gene therapy or of tumor cell lysis in oncolytic virotherapy. Ad5 can infect target cells through binding with the coxsackie and adenovirus receptor (CAR). Thus, the infectious ability of Ad5-based vectors depends on the CAR expression level in target cells. There are conventional methods to evaluate the CAR expression level in human target cells, including flow cytometry, western blotting and immunohistochemistry. Here, we show a simple system for detection and assessment of functional CAR expression in human tumor cells, using the green fluorescent protein (GFP)-expressing telomerase-specific replication-competent adenovirus OBP-401. OBP-401 infection induced detectable GFP expression in CAR-expressing tumor cells, but not in CAR-negative tumor cells, nor in CAR-positive normal fibroblasts, 24 h after infection. OBP-401-mediated GFP expression was significantly associated with CAR expression in tumor cells. OBP-401 infection detected tumor cells with low CAR expression more efficiently than conventional methods. OBP-401 also distinguished CAR-positive tumor tissues from CAR-negative tumor and normal tissues in biopsy samples. These results suggest that GFP-expressing telomerase-specific replication-competent adenovirus is a very potent diagnostic tool for assessment of functional CAR expression in tumor cells for Ad5-based antitumor therapy.

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Acknowledgements

We thank Dr Satoru Kyo (Kanazawa University) for providing the OST cells; Dr Hiroyuki Kawashima (Niigata University) for providing the NOS-10 and NMFH-1 cells; and Tomoko Sueishi for her excellent technical support. This study was supported by grants-in-Aid from the Ministry of Education, Science and Culture, Japan and grants from the Ministry of Health and Welfare, Japan.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    T Sasaki, J Hasei, S Osaki, T Kunisada, A Yoshida & T Ozaki

  2. Center for Gene and Cell Therapy, Okayama University Hospital, Okayama, Japan

    H Tazawa

  3. Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    H Tazawa, Y Hashimoto, S Yano, R Yoshida, S Kagawa, F Uno & T Fujiwara

  4. Department of Medical Materials for Musculoskeletal Reconstruction, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    T Kunisada

  5. Oncolys BioPharma, Inc., Tokyo, Japan

    Y Urata

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  1. T Sasaki
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Correspondence to T Fujiwara.

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Competing interests

Y Urata is an employee of Oncolys BioPharma, Inc., the manufacturer of OBP-401 (Telomescan). The remaining authors declare no conflict of interest.

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Sasaki, T., Tazawa, H., Hasei, J. et al. A simple detection system for adenovirus receptor expression using a telomerase-specific replication-competent adenovirus. Gene Ther 20, 112–118 (2013). https://doi.org/10.1038/gt.2011.213

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