Identification of Oncolytic Avian Reovirus Receptors in B16-F10 Cells and the Signaling-Mediated Pathways Involved in...

Area of Science:

• Virology
• Oncology
• Cell Biology

Background:

• Avian reovirus (ARV) is a poultry pathogen with oncolytic potential.
• ARV selectively infects and kills cancer cells, sparing healthy cells.
• The specific receptors for ARV entry into cancer cells are not well understood.

Purpose of the Study:

• To identify the receptors mediating ARV entry into cancer cells.
• To elucidate the molecular mechanisms of ARV-mediated oncolysis.
• To provide insights for enhancing ARV as an oncolytic virus therapy.

Main Methods:

• Viral overlay protein binding assay (VOPBA), SDS-PAGE, and LC-MS/MS to identify ARV-binding proteins.
• Plaque-forming assays (PFAs) to assess viral replication.
• Co-immunoprecipitation (Co-IP) and proximity ligation assay (PLA) to confirm protein interactions.
• shRNA knockdown and antibody blocking assays to evaluate receptor function.
• Western blot analysis to investigate signaling pathway activation.

Main Results:

• Plg-RKT was identified as a key receptor for ARV σC binding to B16-F10 melanoma cells.
• ARV binding to Plg-RKT activates Src and p38 MAPK signaling pathways.
• Activation of these pathways promotes caveolin-1 phosphorylation and caveolae-mediated endocytosis.
• Inhibition of Plg-RKT expression significantly reduced ARV infection.

Conclusions:

• Plg-RKT is a crucial receptor for ARV entry into melanoma cells.
• The Src-p38 MAPK signaling pathway is essential for ARV internalization.
• Understanding these mechanisms can improve the selectivity and therapeutic efficacy of ARV oncolytic virus therapy.