Stimulator of Interferon Genes Agonist Synergistically Amplifies Programmed Cell Death Protein-1 Blockade and Radiation-Induced Systemic Antitumor Responses via Tumor Microenvironment Enrichment.

Abstract

The effectiveness of immune checkpoint inhibitors in solid tumors is limited and heavily dependent on the tumor microenvironment (TME). Radiation therapy (RT) reshapes the TME, promoting T cell infiltration. We explored the combined antitumor effects of the stimulator of interferon genes (STING) agonist with low-dose RT and immunotherapy. Tumor cell lines (PRM-SCLC, MC38, and LL2) were treated with the STING agonist diABZI (0.001-10 µM) to assess cytotoxicity. The mRNA expression levels of chemokines and cytokines in tumor cells were quantitatively analyzed in conjunction with RT to assess immune activation. Flow cytometry assessed bone marrow-derived dendritic cell and macrophage maturation. Subcutaneous tumor-bearing mouse models (PRM-SCLC, MC38, LL2) were used to monitor tumor volume, body weight, and survival. Tumor samples were collected for flow cytometry, immunofluorescence, immunohistochemistry, and transcriptome sequencing. Bilateral tumor models assessed the abscopal effect, with tumor and tumor-draining lymph node samples collected. The STING agonist diABZI did not directly inhibit tumor cell proliferation at tested concentrations. However, when combined with RT, diABZI significantly upregulated chemokines and IFN-β mRNA levels in tumor cells, while mitigating the RT-induced rise in TGF-β levels. In vitro, bone marrow-derived dendritic cells and macrophages treated with STING agonist + RT showed increased maturation. In tumor-bearing mice, the STING agonist enhanced the efficacy of RT, chemotherapy, and immunotherapy. Adding STING agonist to low-dose RT + αPD-1 activated tumor-infiltrating CD45⁺, CD8⁺, CD4⁺ T cells, natural killer cells, and dendritic cells, and promoted M1 macrophage polarization. Transcriptome analysis showed enhanced antigen presentation and T cell activation. In bilateral tumor models, triple therapy reduced both primary and distant tumor volumes, with increased T cell infiltration and a higher presence of TCF1⁺ PD-1⁺ T_SL cells in tumor-draining lymph nodes. STING agonist boosts immune activation and cell recruitment in the TME, enhancing immunotherapy response. It also amplifies the abscopal effect of RT, promoting systemic antitumor immunity with clinical translational potential.