Establishment of biological and pharmacokinetic assays of telomerase-specific replication-selective adenovirus

Yuuri Hashimoto, Yuichi Watanabe, Yoshiko Shirakiya, Futoshi Uno, Shunsuke Kagawa, Hitoshi Kawamura, Katsuyuki Nagai, Noriaki Tanaka, Horomi Kumon, Yasuo Urata, Toshiyoshi Fujiwara

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38 Citations (Scopus)

Abstract

The use of replication-selective tumor-specific viruses represents a novel approach for the treatment of neoplastic disease. We constructed an attenuated adenovirus, telomerase-specific replication-selective adenovirus (TRAD), in which the human telomerase reverse transcriptase promoter element drives the expression of the E1A and E1B genes linked with an internal ribosome entry site (IRES). Forty-eight hours after TRAD infection at a multiplicity of infection of 1.0, the cell viability of H1299 human lung cancer cells was consistently less than 50% and therefore this procedure could be used as a potency assay to assess the biological activity of TRAD. We also established a quantitative real-time polymerase chain reaction (PCR) analysis with consensus primers for either the adenovirus E1A or IRES sequence. The linear ranges of quantitation with E1A and IRES primers were 103 -108 and 102-108 plaque-forming units/ mL in the plasma, respectively. The PCR analysis demonstrated that the levels of E1A in normal tissues were more than 103 lower than in the tumors of A549 human lung tumor xenografts in nu/nμ mice at 28 days after intratumoral injection. Our results suggest that the cell-killing assay against H1299 cells and real-time PCR can be used to assess the biological activity and biodistribution of TRAD in clinical trials.

Original languageEnglish
Pages (from-to)385-390
Number of pages6
JournalCancer Science
Volume99
Issue number2
DOIs
Publication statusPublished - Feb 2008

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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    Hashimoto, Y., Watanabe, Y., Shirakiya, Y., Uno, F., Kagawa, S., Kawamura, H., Nagai, K., Tanaka, N., Kumon, H., Urata, Y., & Fujiwara, T. (2008). Establishment of biological and pharmacokinetic assays of telomerase-specific replication-selective adenovirus. Cancer Science, 99(2), 385-390. https://doi.org/10.1111/j.1349-7006.2007.00665.x