Oncolytic virus-antibody combinations enhance immune-mediated killing of osteosarcoma

Abstract

A major barrier to effective immunotherapy in osteosarcoma (OS) is the highly immunosuppressive tumour microenvironment (TME), which limits immune recognition and elimination of tumour cells. We evaluated a panel of oncolytic herpes simplex viruses (oHSVs) for their direct oncolytic activity, immune-modulatory properties and capacity to counteract OS-associated immunosuppression using immunologically relevant in vitro models. We demonstrate that established OS cell lines, primary cell cultures and dissociated OS cells from freshly resected tumour samples are susceptible to direct oncolysis by three oHSVs; HSV1716, HSV1716-GMCSF, and HSV47{Delta}, although susceptibility levels varied. Treatment of peripheral blood mononuclear cells from healthy donors and OS patients with oHSVs enhanced natural killer (NK) cell activation and promoted immune-mediated killing of OS cell lines and primary OS cell cultures. Among the three viruses, HSV1716-GMCSF exhibited the strongest immune-stimulatory effects and was uniquely capable of reducing the abundance of CD163+CD206+ immunosuppressive TAMs; use of this oHSV was therefore prioritised. We developed a multicellular spheroid model of OS, incorporating OS cells, mesenchymal stem cells and TAMs, which exhibits resistance to immune-mediated killing, better reflecting the immunosuppressive TME in patients. In this model, pairing HSV1716-GMCSF treatment with either anti-GD2 or anti-EGFR monoclonal antibodies (mAbs), selected according to OS tumour antigen expression, significantly increased immune-mediated tumour cell killing. These findings suggest that personalised combination strategies pairing oHSVs with appropriate mAbs provide a promising therapeutic approach for OS by integrating direct oncolysis, remodelling of the immunosuppressive TME and enhanced immune-mediated tumour destruction.