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Combination gene therapy for glioblastoma involving herpes simplex virus vector-mediated codelivery of mutant IκBα and HSV thymidine kinase

Cancer Gene Therapy volume 12, pages 487–496 (2005)Cite this article

Abstract

To improve the effectiveness of herpes simplex virus (HSV) thymidine kinase/ganciclovir (HSV-tk/GCV) suicide gene therapy, the replication-defective HSV vector TOIκB expressing both HSV-TK and a mutant form of the NF-κB inhibitor IκBα (IκBαM) was developed. TOIκB was constructed by recombining the IκBαM gene into the UL41 locus of a replication-defective lacZ expression vector, TOZ.1. Expression of IκBαM was confirmed by Western blotting, and the ability of the mutant protein to inhibit NF-κB nuclear translocation was examined by electrophoretic mobility shift assay. In human glioblastoma U-87MG cells, the p50/p50 dimer of NF-κB was already translocated to the nucleus without receptor-dependent signaling by TNF-α. Following infection with TOIκB, nuclear translocation of NF-κB in U-87MG cells was significantly inhibited and caspase-3 activity increased compared with TOZ.1-infected cells. The cytotoxicity of TOIκB for U-87MG cells was investigated by colorimetric MTT assay. At an MOI of 3, TOIκB infection killed 85% of the cells compared to 20% killed by TOZ.1 infection. In the presence of GCV, these numbers increased to 95–100% for TOIκB and 80–85% for TOZ.1. TOIκB neurotoxicity measured on cultured murine neurons was relatively low and similar to that of TOZ.1. The survival of nude mice implanted into the brain with U-87MG tumor cells was markedly prolonged by intratumoral TOIκB injection and GCV administration. Survival of TOIκB+GCV group was significantly longer (P<.02, Wilcoxon test) than for the control groups (TOZ.1 or TOIκB only, PBS or PBS+GCV). These results suggest that IκBαM expression may be a safe enhancement of replication-defective HSV-based suicide gene therapy in vitro and in vivo.

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Acknowledgements

This work was supported by NIH Grant GM34534 to JCG, by GenVec, Inc. (Rockville, MD), and by a Grant-in-Aid for Scientific Research to SM (No. 12671357) from the Ministry of Education, Science and Culture, Ministry of Health and Human Welfare, Japan.

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Authors and Affiliations

  1. Department of Neurosurgery, Osaka Graduate University Medical School, 2-2 Yamada-oka, Suita city, Osaka, 565, Japan

    Shusuke Moriuchi, Motohiko Maruno, Shuichi Izumoto & Toshiki Yoshimine

  2. Department of Neurosurgery, National Hospital Organization Osaka National Hospital, 2-1-14 Hoenzaka, Chuou-ku, Osaka, Osaka, 540-0006, Japan

    Shusuke Moriuchi

  3. Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, E1240 Biomedical Science Tower, Pittsburgh, Pennsylvania, 15261, USA

    Joseph C Glorioso, Darren Wolfe, Shaohua Huang & Justus B Cohen

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  1. Shusuke Moriuchi
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Correspondence to Shusuke Moriuchi.

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Moriuchi, S., Glorioso, J., Maruno, M. et al. Combination gene therapy for glioblastoma involving herpes simplex virus vector-mediated codelivery of mutant IκBα and HSV thymidine kinase. Cancer Gene Ther 12, 487–496 (2005). https://doi.org/10.1038/sj.cgt.7700816

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