Abstract 6406: Radiation and STING activation limit tumor development and modulate the immune environment via distinct mechanisms
Authors: Shigemori Kay , Jiang Yanyan , Beernaert Bruno , Ryan Anderson J. , Parkes Eileen E. 
Published: April 2023

Abstract
Carcinogen-induced lung cancer in A/J mice is an established model of tumor initiation and development. The impact of radiation, immune response, and STING pathway activation on tumor development in this model is not well understood. In this study, we examined the effect of radiation and the STING agonist, DMXAA (Vadimezan), on the development of urethane-induced lung cancer. A/J mice were treated with urethane (i.p.) to induce the development of lung tumors. Three days after urethane treatment, mice received 13 Gy thorax radiation treatment (RT)/mock RT, DMXAA (20 mg/kg i.p.)/vehicle, or a combination treatment (n = 5). Mice were euthanized after 6 hours to 7 days after treatment to assess early events in the lung with RNA-sequencing and flow cytometry, or 5 months after treatment to assess tumor growth and long-term changes to the immune microenvironment with multiplex immunofluorescence. Mice treated with urethane followed by RT had fewer lung surface tumors (p &lt; 0.01) and reduced tumor area (p = 0.0732) compared to their mock RT controls. In parallel, mice that received urethane pre-treatment and RT had high numbers of tertiary lymphoid structures (TLSs; these contained T-cells, B-cells, MHCII+ APCs and CXCL13) compared to the mock RT control mice (p &lt; 0.001). Analysis of immune cell contents of the TLS revealed a notable increase in CD4, CD8 and regulatory T-cells in the RT condition (with urethane pretreatment). Important early events in the lung following RT of urethane pre-treated mice included a transient increase in type I and type II interferons, CD4 and CD8 T-cell infiltration (p &lt; 0.001) and a strong downregulation of cell cycle events. In contrast to these findings, DMXAA treatment after urethane resulted in fewer tumors (p &lt; 0.05), but larger tumors compared to the vehicle-treated group. An increase in immune cells was observed in the lung, however, without clear structures that resembled TLSs. Early events following DMXAA treatment with urethane pre-treatment included dampened levels of type I IFN and TNF-α in circulation and a decrease in T-cell infiltration into the lungs. Interestingly, RT and DMXAA combination treatment (with urethane pre-treatment) drove a synergistic anti-tumor effect, further reducing tumor numbers compared to the RT or DMXAA controls 5 months after treatment. RT and STING agonist treatment of pre-neoplastic lesions have differential impacts on tumor growth in the A/J mouse model, and work synergistically when combined. The different outcomes of RT and DMXAA on tumor growth may be driven by their distinctive mechanism of action on immune responses and capacities to form TLSs. These findings may have future implications for strategies for the early treatment of lung and other cancers, and they suggest that immune responses, including modulation of the STING pathway, may be an important aspect of early tumor development that could be targeted therapeutically.
Citation Format: Kay Shigemori, Yanyan Jiang, Bruno Beernaert, Anderson J. Ryan, Eileen E. Parkes. Radiation and STING activation limit tumor development and modulate the immune environment via distinct mechanisms [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6406.