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

PLoS One. 2012;7(10):e48064. doi: 10.1371/journal.pone.0048064. Epub 2012 Oct 24.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CHO Cells
  • Cell Adhesion Molecules / genetics*
  • Cell Adhesion Molecules / metabolism
  • Cell Line
  • Cell Line, Tumor
  • Cricetinae
  • Cricetulus
  • Cysteine Proteinase Inhibitors / pharmacology
  • Glioblastoma / genetics
  • Glioblastoma / pathology
  • Glioblastoma / virology
  • Host Specificity / genetics
  • Host-Pathogen Interactions / genetics
  • Humans
  • Leucine / analogs & derivatives
  • Leucine / pharmacology
  • Mammalian orthoreovirus 3 / genetics*
  • Mammalian orthoreovirus 3 / metabolism
  • Mammalian orthoreovirus 3 / physiology
  • Molecular Sequence Data
  • Mutation*
  • Neoplasms / genetics
  • Neoplasms / pathology
  • Neoplasms / virology
  • Oncolytic Virotherapy / methods
  • Protein Multimerization
  • Receptors, Cell Surface / genetics*
  • Receptors, Cell Surface / metabolism
  • Receptors, Virus / genetics
  • Receptors, Virus / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / metabolism
  • Virus Internalization / drug effects

Substances

  • Cell Adhesion Molecules
  • Cysteine Proteinase Inhibitors
  • F11R protein, human
  • Receptors, Cell Surface
  • Receptors, Virus
  • Viral Proteins
  • Leucine
  • aloxistatin

Associated data

  • GENBANK/GU991659
  • GENBANK/GU991660
  • GENBANK/GU991661
  • GENBANK/GU991662
  • GENBANK/GU991663
  • GENBANK/GU991664
  • GENBANK/GU991665
  • GENBANK/GU991666
  • GENBANK/GU991667
  • GENBANK/GU991668
  • GENBANK/GU991669
  • GENBANK/GU991670
  • GENBANK/GU991671
  • GENBANK/GU991672
  • GENBANK/GU991673
  • GENBANK/GU991674
  • GENBANK/GU991675
  • GENBANK/GU991676
  • GENBANK/GU991677
  • GENBANK/GU991678

Grant support

This work was supported by the European Union through the 6th Framework Program GIANT (Contract No. 512087). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.