Isolation of reovirus T3D mutants capable of infecting human tumor cells independent of junction adhesion molecule-A.

Mammalian Reovirus is a double-stranded RNA virus with a distinctive preference to replicate in and lyse transformed cells. On that account, Reovirus type 3 Dearing (T3D) is clinically evaluated as oncolytic agent. The therapeutic efficacy of this approach depends in part on the accessibility of the reovirus receptor Junction Adhesion Molecule-A (JAM-A) on the target cells. Here, we describe the isolation and characterization of reovirus T3D mutants that can infect human tumor cells independent of JAM-A. The JAM-A-independent (jin) mutants were isolated on human U118MG glioblastoma cells, which do not express JAM-A. All jin mutants harbour mutations in the S1 segments close to the region that encodes the sialic acid-binding pocket in the shaft of the spike protein. In addition, two of the jin mutants encode spike proteins with a Q336R substitution in their head domain. The jin mutants can productively infect a wide range of cell lines that resist wt reovirus T3D infection, including chicken LMH cells, hamster CHO cells, murine endothelioma cells, human U2OS and STA-ET2.1 cells, but not primary human fibroblasts. The jin-mutants rely on the presence of sialic-acid residues on the cell surface for productive infection, as is evident from wheat germ agglutinin (WGA) inhibition experiments, and from the jin-reovirus resistance of CHO-Lec2 cells, which have a deficiency of sialic-acids on their glycoproteins. The jin mutants may be useful as oncolytic agents for use in tumors in which JAM-A is absent or inaccessible.
AuthorsDiana J M van den Wollenberg, Iris J C Dautzenberg, Sanne K van den Hengel, Steve J Cramer, Raoul J de Groot, Rob C Hoeben
JournalPloS one (PLoS One) Vol. 7 Issue 10 Pg. e48064 ( 2012) ISSN: 1932-6203 [Electronic] United States
PMID23110175 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Cell Adhesion Molecules
  • Cysteine Proteinase Inhibitors
  • F11R protein, human
  • Receptors, Cell Surface
  • Receptors, Virus
  • Viral Proteins
  • Leucine
  • aloxistatin
  • Animals
  • CHO Cells
  • Cell Adhesion Molecules (genetics, metabolism)
  • Cell Line
  • Cell Line, Tumor
  • Cricetinae
  • Cricetulus
  • Cysteine Proteinase Inhibitors (pharmacology)
  • Glioblastoma (genetics, pathology, virology)
  • Host Specificity (genetics)
  • Host-Pathogen Interactions (genetics)
  • Humans
  • Leucine (analogs & derivatives, pharmacology)
  • Mammalian orthoreovirus 3 (genetics, metabolism, physiology)
  • Molecular Sequence Data
  • Mutation
  • Neoplasms (genetics, pathology, virology)
  • Oncolytic Virotherapy (methods)
  • Protein Multimerization
  • Receptors, Cell Surface (genetics, metabolism)
  • Receptors, Virus (genetics, metabolism)
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Viral Proteins (chemistry, genetics, metabolism)
  • Virus Internalization (drug effects)

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