Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001

J Gen Virol. 2012 Dec;93(Pt 12):2606-2613. doi: 10.1099/vir.0.046011-0. Epub 2012 Sep 12.

Abstract

Seneca Valley virus (SVV-001) is an oncolytic picornavirus with selective tropism for a subset of human cancers with neuroendocrine differentiation. To characterize further the specificity of SVV-001 and its patterns and kinetics of intratumoral spread, bacterial plasmids encoding a cDNA clone of the full-length wild-type virus and a derivative virus expressing GFP were generated. The full-length cDNA of the SVV-001 RNA genome was cloned into a bacterial plasmid under the control of the T7 core promoter sequence to create an infectious cDNA clone, pNTX-09. A GFP reporter virus cDNA clone, pNTX-11, was then generated by cloning a fusion protein of GFP and the 2A protein from foot-and-mouth disease virus immediately following the native SVV-001 2A sequence. Recombinant GFP-expressing reporter virus, SVV-GFP, was rescued from cells transfected with in vitro RNA transcripts from pNTX-11 and propagated in cell culture. The proliferation kinetics of SVV-001 and SVV-GFP were indistinguishable. The SVV-GFP reporter virus was used to determine that a subpopulation of permissive cells is present in small-cell lung cancer cell lines previously thought to lack permissivity to SVV-001. Finally, it was shown that SVV-GFP administered to tumour-bearing animals homes in to and infects tumours whilst having no detectable tropism for normal mouse tissues at 1×10(11) viral particles kg(-1), a dose equivalent to that administered in ongoing clinical trials. These infectious clones will be of substantial value in further characterizing the biology of this virus and as a backbone for the generation of additional oncolytic derivatives.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Carcinoma, Small Cell / therapy
  • Carcinoma, Small Cell / virology
  • Cell Line, Tumor
  • Cloning, Molecular
  • Cytopathogenic Effect, Viral / genetics
  • DNA, Complementary / genetics
  • DNA, Viral / genetics
  • Female
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Humans
  • Lung Neoplasms / therapy
  • Lung Neoplasms / virology
  • Mice
  • Mice, Nude
  • Molecular Sequence Data
  • Oncolytic Virotherapy
  • Oncolytic Viruses / genetics*
  • Picornaviridae / genetics*

Substances

  • DNA, Complementary
  • DNA, Viral
  • Green Fluorescent Proteins