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Improved replication efficiency of echovirus 5 after transfection of colon cancer cells using an authentic 5’ RNA genome end methodology

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Summary

Oncolytic virotherapy is a promising novel form of cancer treatment, but the therapeutic efficiency needs improvement. A potential strategy to enhance the therapeutic effect of oncolytic viruses is to use infectious nucleic acid as therapeutic agent to initiate an oncolytic infection, without administrating infectious viral particles. Here we demonstrate improved viral replication activation efficiency when transfecting cells with 5’ end authentic in vitro transcribed enterovirus RNA as compared to genomic RNA with additional non-genomic 5’ nucleotides generated by conventional cloning methods. We used echovirus 5 (E5) as an oncolytoc model virus due to its ability to replicate in and completely destroy five out of six colon cancer cell lines and kill artificial colon cancer tumors (HT29 spheroids), as shown here. An E5 infectious cDNA clone including a hammerhead ribozyme sequence was used to generate in vitro transcripts with native 5’ genome ends. In HT29 cells, activation of virus replication is approximately 20-fold more efficient for virus genome transcripts with native 5’ genome ends compared to E5 transcripts generated from a standard cDNA clone. This replication advantage remains when viral progeny release starts by cellular lysis 22 h post transfection. Hence, a native 5’ genomic end improves infection activation efficacy of infectious nucleic acid, potentially enhancing its therapeutic effect when used for cancer treatment. The clone design with a hammerhead ribozyme is likely to be applicable to a variety of oncolytic positive sense RNA viruses for the purpose of improving the efficacy of oncolytic virotherapy.

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Acknowledgements

We thank Christina Gustafson-Svärd and Hans-Christoph Selinka for providing cells. This study was supported by grants from the National Graduate School of Pharmaceutical Sciences (FLäK) and the Faculty of Health and Life Science, Linnaeus University.

Conflict of interests

The authors declare that they have no conflict of interests.

Author information

Author notes

  1. S. Israelsson

    Present address: Centre for Ecology and Evolution in Microbial model Systems - EEMiS, Linnaeus University, Kalmar, Sweden

  2. J-O. Ekström

    Present address: Department of Molecular Biology, Umeå University, 901 87, Umeå, Sweden

  3. N. Jonsson

    Present address: Centre for Biomaterials Chemistry, Linnaeus University, Kalmar, Sweden

Authors and Affiliations

  1. Department of Chemistry and Biomedical Sciences, Linnaeus University, 391 82, Kalmar, Sweden

    S. Israelsson, A. Sävneby, J-O. Ekström, N. Jonsson, K. Edman & A. M. Lindberg

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  1. S. Israelsson
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Correspondence to A. M. Lindberg.

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Israelsson, S., Sävneby, A., Ekström, JO. et al. Improved replication efficiency of echovirus 5 after transfection of colon cancer cells using an authentic 5’ RNA genome end methodology. Invest New Drugs 32, 1063–1070 (2014). https://doi.org/10.1007/s10637-014-0136-z

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  • DOI: https://doi.org/10.1007/s10637-014-0136-z

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