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A strategy for genetic modification of the spike-encoding segment of human reovirus T3D for reovirus targeting

Gene Therapy volume 15, pages 1567–1578 (2008)Cite this article

Abstract

Human Orthoreovirus Type 3 Dearing is not pathogenic to humans and has been evaluated clinically as an oncolytic agent. Its transduction efficiency and the tumor cell selectivity may be enhanced by incorporating ligands for alternative receptors. However, the genetic modification of reoviruses has been difficult, and genetic targeting of reoviruses has not been reported so far. Here we describe a technique for generating genetically targeted reoviruses. The propagation of wild-type reoviruses on cells expressing a modified σ1-encoding segment embedded in a conventional RNA polymerase II transcript leads to substitution of the wild-type genome segment by the modified version. This technique was used for generating reoviruses that are genetically targeted to an artificial receptor expressed on U118MG cells. These cells lack the junction adhesion molecule-1 and therefore resist infection by wild-type reoviruses. The targeted reoviruses were engineered to carry the ligand for this receptor at the C terminus of the σ1 spike protein. This demonstrates that the C terminus of the σ1 protein is a suitable locale for the insertion of oligopeptide ligands and that targeting of reoviruses is feasible. The genetically targeted viruses can be propagated using the modified U118MG cells as helper cells. This technique may be applicable for the improvement of human reoviruses as oncolytic agents.

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Acknowledgements

We thank Martijn Rabelink and Cynthia Sitaram for their expert technical assistance. We gratefully acknowledge Drs Lee Fradkin, Hans Tanke, Danijella Koppers-Lalic and Twan de Vries for their helpful discussions and critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands

    D J M van den Wollenberg, S K van den Hengel, I J C Dautzenberg, S J Cramer & R C Hoeben

  2. Department of Neurology, Erasmus Medical Center, Rotterdam, The Netherlands

    S K van den Hengel

  3. Department of Surgery, University Medical Center Utrecht, Utrecht, The Netherlands

    O Kranenburg

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  1. D J M van den Wollenberg
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Correspondence to R C Hoeben.

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van den Wollenberg, D., van den Hengel, S., Dautzenberg, I. et al. A strategy for genetic modification of the spike-encoding segment of human reovirus T3D for reovirus targeting. Gene Ther 15, 1567–1578 (2008). https://doi.org/10.1038/gt.2008.118

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