Immunotherapeutic effects of intratumorally injected Zymosan-Adenovirus conjugates encoding constant active IRF3 in a melanoma mouse model.

Abstract

M2-like tumor-associated macrophages (TAMs) play a significant role in immunosuppressive conditions in the tumor microenvironment (TME). TAM reprogramming, a dual-pronged therapy, reduces immunosuppression and induces immune favorable conditions in the TME. In this study, recombinant adenoviruses encoding active forms of interferon regulatory factor 3 (IRF3) were conjugated to zymosan particles to target phagocytic cells to create a pro-inflammatory immunomodulatory therapy. We determined TAM reprogramming by upregulation and downregulation of M1- and M2-associated genes, respectively, as well as cytokine and transcription factor expression in vitro. The overall shift to immune favorable conditions in the TME was suggested by metabolic, cytokine, and immune cell gene expression. Our data indicated that the zymosan:adenovirus (Zym:Ad) particle itself induced a shift from M2-like to M1-like TAMs, a shift in immune status of the TME, and systemic tumor immunity as determined using a double tumor melanoma mouse model and splenocyte functional assay. Notably, direct intratumoral injection of Zym:Ad IRF3 reduced tumor growth more significantly than Zym:Ad GFP, indicating additional therapeutic benefits due to incorporation of constant active IRF3.