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

J. T. Lam, A. Hemminki, A. Kanerva, K. B. Lee, J. L. Blackwell, R. Desmond, G. P. Siegal, D. T. Curiel

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

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.

Original languageEnglish
Pages (from-to)421-430
Number of pages10
JournalCancer gene therapy
Volume14
Issue number4
DOIs
StatePublished - Apr 2007

Keywords

  • Adenovirus
  • Oncolytic virus
  • Ovarian cancer
  • Replication-competent viruses
  • Spheroid

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