Abstract 6788: Spatial single-cell analysis reveals stage-dependent mechanisms of anti-IL-1β therapy in lung adenocarcinoma

Abstract Introduction: Lung adenocarcinoma (LUAD) is a leading cause of cancer mortality, with IL-1β-driven inflammation playing a central role in tumor progression. Although IL-1β blockade shows promising antitumor effects, its efficacy varies across tumor stages, and the mechanisms behind this stage-dependent response remain unclear. Here, we investigate how tumor-immune interactions and microenvironmental remodeling shape differential responses to anti-IL-1β therapy during LUAD progression. Methods: We performed spatial single-cell analysis of the immune microenvironment using Visium HD on human (10 adjacent normal, 17 AAH, 12 IAC) and mouse samples (21 adjacent normal, 57 hyperplasia, 13 IAC). Integrating these data with stage-matched scRNA-seq, we mapped IL-1β-IL1R1-mediated cell-cell interactions across LUAD stages. Long-term anti-IL-1β treatment was evaluated in two mouse models—C57BL/6 Kras^G12D (early: 4-14 weeks; late: 20-30 weeks) and 129S4 urethane-induced (early: 4-14 weeks; late: 30-40 weeks)—with efficacy and mechanisms assessed via scRNA-seq and Xenium 5K. Short-term treatments (0, 6, 30 weeks) in C57BL/6 Kras^G12D were performed to further explore mechanisms. Results: In human tissues, IL-1β-IL1R1 interactions (“IL-1βnets”) occurred primarily between myeloid cells and fibroblasts, epithelial, and endothelial cells, a pattern conserved in mouse tissues. Long-term anti-IL-1β treatment was effective only at the early stage. Early-stage scRNA-seq showed enhanced antitumor immunity and a marked reduction in interactions between IL1R1+ fibroblasts and IL-1β+ myeloid cells (monocytes, macrophages, cDCs, neutrophils), whereas late-stage tissues showed minimal changes. Spatial analysis with Xenium 5K confirmed reduced IL1R1+ cell fractions within IL-1βnets at the early stage, with IL-1βnet number and size largely unchanged. Short-term treatment at the early stage reduced interactions between IL1R1+ epithelial cells and neutrophils and attenuated NF-κB signatures in fibroblasts and epithelial cells; late-stage treatment had no significant effects. Conclusion: Spatial single-cell profiling revealed the IL-1β-IL1R1 interaction landscape across LUAD progression in humans and mice. Anti-IL-1β therapy was effective in early-stage LUAD, disrupting key interactions between myeloid and fibroblast/epithelial cells, but was largely ineffective at later stages. These findings suggest that fibroblast and epithelial remodeling are critical determinants of stage-dependent anti-IL-1β efficacy. Citation Format: Bo Zhu, Muhammad Aminu, Yanhua Tian, Shao-Wei Lu, Ou Shi, Wei Lu, Hong Chen, Hui Li, Zhubo Wei, Monique B. Nilsson, Luisa Maren Solis Soto, Futreal P. Andrew, Don L. Gibbons, John V. Heymach, Chao Cheng, Jia Wu, Jianjun Zhang. Spatial single-cell analysis reveals stage-dependent mechanisms of anti-IL-1β therapy in lung adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6788.