Single-cell data-driven design of armed oncolytic virus and combination therapy to activate a cooperative innate-adaptive immunity against cancer.

Oncolytic viruses have been considered promising cancer immunotherapies. However, oncovirotherapy agents impart durable responses in only a subset of cancer patients. Thus, exploring the cellular and molecular mechanisms underlying the heterogeneous responses in patients can provide guidance to develop more effective oncolytic virus therapies. Single-cell RNA sequencing (scRNA-seq) analysis of tumors responsive and non-responsive to oncovirotherapy revealed signatures of the tumor immune microenvironment associated with immune response. Thus, we designed and constructed an armed oncolytic virus OV-5A that expressed five genes with non-redundant functions. OV-5A treatment exhibits robust immune response against various tumors in multiple mouse models, peripheral blood mononuclear cell (PBMC)-patient derived xenograft (PDX) model, organoid-immune cell co-culture systems and patient tissue sections by activating a cooperative innate-adaptive immune response against tumor cells. scRNA-seq analysis of complete responder and partial responder to OV-5A treatment guided the design of combination therapy of OV-5A. This data-driven approach paves a innovative way to rationalize the design of oncolytic virus and multi-agent combination therapies.