title: 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.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Diana J M
authors.initials: DJ
authors.lastname: van den Wollenberg
authors.forename: Iris J C
authors.initials: IJ
authors.lastname: Dautzenberg
authors.forename: Sanne K
authors.initials: SK
authors.lastname: van den Hengel
authors.forename: Steve J
authors.initials: SJ
authors.lastname: Cramer
authors.forename: Raoul J
authors.initials: RJ
authors.lastname: de Groot
authors.forename: Rob C
authors.initials: RC
authors.lastname: Hoeben
mesh_terms: D000818:Animals; D016466:CHO Cells; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D005909:Glioblastoma / Q000235:genetics / Q000473:pathology / Q000821:virology; D058507:Host Specificity / Q000235:genetics; D054884:Host-Pathogen Interactions / Q000235:genetics; D006801:Humans; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D012089:Mammalian orthoreovirus 3 / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D009154:Mutation; D009369:Neoplasms / Q000235:genetics / Q000473:pathology / Q000821:virology; D050130:Oncolytic Virotherapy / Q000379:methods; D055503:Protein Multimerization; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D053586:Virus Internalization / Q000187:drug effects
chemical_list: D015815:Cell Adhesion Molecules; D015853:Cysteine Proteinase Inhibitors; C407243:F11R protein, human; D011956:Receptors, Cell Surface; D011991:Receptors, Virus; D014764:Viral Proteins; D007930:Leucine; C108192:aloxistatin