title: Characterization of a full-length infectious cDNA clone and a GFP reporter derivative of the oncolytic picornavirus SVV-001.
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.
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD 21287, USA.
authors.forename: John T
authors.initials: JT
authors.lastname: Poirier
authors.affiliation: Neotropix, Inc., 351 Phoenixville Pike, Malvern, PA 19355, USA.
authors.forename: P Seshidhar
authors.initials: PS
authors.lastname: Reddy
authors.affiliation: Neotropix, Inc., 351 Phoenixville Pike, Malvern, PA 19355, USA.
authors.forename: Neeraja
authors.initials: N
authors.lastname: Idamakanti
authors.affiliation: Neotropix, Inc., 351 Phoenixville Pike, Malvern, PA 19355, USA.
authors.forename: Shawn S
authors.initials: SS
authors.lastname: Li
authors.affiliation: Neotropix, Inc., 351 Phoenixville Pike, Malvern, PA 19355, USA.
authors.forename: Kristine L
authors.initials: KL
authors.lastname: Stump
authors.affiliation: Neotropix, Inc., 351 Phoenixville Pike, Malvern, PA 19355, USA.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Burroughs
authors.affiliation: Neotropix, Inc., 351 Phoenixville Pike, Malvern, PA 19355, USA.
authors.forename: Paul L
authors.initials: PL
authors.lastname: Hallenbeck
authors.affiliation: The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD 21287, USA.
authors.forename: Charles M
authors.initials: CM
authors.lastname: Rudin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018288:Carcinoma, Small Cell / Q000628:therapy / Q000821:virology; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D003588:Cytopathogenic Effect, Viral / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000628:therapy / Q000821:virology; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D050130:Oncolytic Virotherapy; D050504:Oncolytic Viruses / Q000235:genetics*; D010849:Picornaviridae / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D004279:DNA, Viral; D049452:Green Fluorescent Proteins