Abstract 5299: BZW2: a novel therapeutic target for liver metastasis in lung cancer

Abstract Introduction: Liver metastasis (LM) occurs in approximately 13% of patients with lung cancer at the time of diagnosis and is associated with poor prognosis. Currently, there are no effective preventive or therapeutic strategies specifically targeting LM. Investigating the LM microenvironment can not only elucidate the mechanisms underlying its development but also provide opportunities to identify novel therapeutic targets. Methods: An LM-prone mouse model was established by repeatedly injecting and harvesting Lewis Lung Carcinoma (LLC) cells from LM lesions. The spatial transcriptomics (Visium, 10X Genomics) was applied to explore the microenvironment and intratumoral heterogeneity of LM. Validation was conducted using single-cell RNA sequencing and spatial proteomics (PhenoCycler System, Akoya Bioscience) on human and murine LM samples, respectively. Bioinformatics analyses, including Gene Set Enrichment Analysis (GSEA), were applied to infer gene function and associated behaviors. Gene set variation analysis (GSVA) was performed to evaluate lung cancer patient survival in relation to the expression of the BZW2-associated gene set. Functional validation of BZW2 was conducted in vitro by assessing cell migration using transwell and wound healing assays, and by examining epithelial-to-mesenchymal transition (EMT) markers via immunoblotting. Results: Bzw2 is the significantly upregulated differential expressed gene in LM, particularly at the invasive front, based on the murine spatial transcriptomic data. The finding was further validated in murine LM samples using spatial proteomics as well. In human single-cell RNA sequencing data, BZW2 exhibited high expression in LM subclusters characterized by aggressive phenotypes, including enhanced proliferation, hypoxia, mesenchymal traits, invasiveness, and cancer stemness. GSVA analysis demonstrated that elevated expression of the BZW2-associated gene set was associated with poorer patient survival and enrichment of EMT-related pathways using the TCGA LUAD dataset. Furthermore, in vitro functional assays showed that BZW2 inhibition suppressed cell migration and reduced the expression of EMT markers, as confirmed by immunoblotting. Conclusion: By integrating findings from spatial transcriptomics, proteomics, and single-cell RNA sequencing, this study elucidates the role of BZW2 in promoting the aggressiveness of LM in lung cancer through its influence on cell migration and metastasis. These results suggest that BZW2 could serve as a promising therapeutic target worthy of further advanced research for LM in lung cancer. Citation Format: Kuan-Li Wu, Ying-Ming Tsai, Ya-Ling Hsu. BZW2: a novel therapeutic target for liver metastasis in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 5299.