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

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.

• Authors: Diana J M van den Wollenberg, Iris J C Dautzenberg, Sanne K van den Hengel, Steve J Cramer, Raoul J de Groot, Rob C Hoeben
• Journal: PloS one (PLoS One) Vol. 7 Issue 10 Pg. e48064 ( 2012) ISSN: 1932-6203 [Electronic] United States
• PMID: 23110175 (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

Topics

• 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)