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A three-dimensional assay for measurement of viral-induced oncolysis

Cancer Gene Therapy volume 14, pages 421–430 (2007)Cite this article

Abstract

Oncolytic viruses represent a novel cancer treatment strategy. Despite their promising preclinical data, however, corresponding clinical trials have disappointed. To aid preclinical analyses, we hypothesized that three-dimensional tumor cell clusters or spheroids might provide an assay system superior to conventional monolayer cell cultures. Spheroids show viral infection, replication and oncolytic patterns distinct from conventional monolayer assays. Therefore, viral tumor penetration and oncolysis measurements may be improved with such three-dimensional models. Also, preclinical analyses of oncolytic viruses frequently measure mitochondrial activity, but more accurate measures of oncolysis might involve quantitation of intracellular protein release. Therefore, we measured luciferase released from luciferase-expressing spheroids and found unique patterns that maintained consistency with various viruses and doses. The relative variations between viruses and doses may represent temporal differences in oncolysis dynamics. Analysis of five recombinant replicative adenoviruses with promise for clinical application showed that Ad5/3-Δ24 produced the most luciferase release 1 week after infection and achieved the earliest and highest peak luciferase release level. Ad5/3-Δ24 also effected the earliest subtotal spheroid cell death. These findings closely parallel monolayer oncolysis assays with these agents. Therefore, the luciferase-expressing tumor spheroid assay represents a promising three-dimensional model for preclinical analysis of replicative oncolytic agents.

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Acknowledgements

We thank Dr Robert S Negrin and Dr Christian Sheffold from the Division of Bone Marrow Transplantation at Stanford University Medical Center for use of the SK-OV-3luc cell line. This study was supported by NCI (R01 CA83821, P50 CA83591, P50 CA89019 and R01 CA93796), the Academy of Finland, Helsinki University Central Hospital Research Funds, University of Helsinki Internal Funds, Sohlberg Foundation, Sigrid Juselius Foundation, Finnish Cancer Society, Instrumentarium Research Fund, Emil Aaltonen Foundation, Finnish Oncology Association, Research and Science Foundation of Farmos.

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Authors and Affiliations

  1. Department of Pathology, University of Mississippi Medical Center, Jackson, MS, USA

    J T Lam

  2. Cancer Gene Therapy Group, Rational Drug Design Program, University of Helsinki, Helsinki, Finland

    A Hemminki & A Kanerva

  3. Department of Oncology, Helsinki University Central Hospital, Helsinki, Finland

    A Hemminki & A Kanerva

  4. Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Helsinki, Finland

    A Kanerva

  5. Department of Obstetrics and Gynecology, Pusan Paik Hospital, College of Medicine, Inje University, Pusan, South Korea

    K B Lee

  6. Division of Infectious Diseases, Emory University School of Medicine, Atlanta, GA, USA

    J L Blackwell

  7. Comprehensive Cancer Center Biostatistics Unit, University of Birmingham, Birmingham, AL, USA

    R Desmond

  8. Departments of Pathology, Division of Human Gene Therapy, Cell Biology, and Surgery, University of Alabama at Birmingham, Birmingham, AL, USA

    G P Siegal & D T Curiel

  9. Departments of Medicine, Surgery, and Pathology and the Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL, USA

    G P Siegal & D T Curiel

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Correspondence to D T Curiel.

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Lam, J., Hemminki, A., Kanerva, A. et al. A three-dimensional assay for measurement of viral-induced oncolysis. Cancer Gene Ther 14, 421–430 (2007). https://doi.org/10.1038/sj.cgt.7701028

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