Abstract 162: Mapping the complex interplay between vasculogenic mimicry and immune response in lung cancer using digital spatial profiling

Abstract Background: Vasculogenic mimicry (VM) represents an alternative vascularization mechanism contributing to cancer progression and treatment resistance. Despite its clinical significance, the spatial and molecular relationships between VM networks and immune components remain poorly characterized in non-small cell lung cancer (NSCLC). Methods: We conducted a comprehensive spatial analysis of treatment-naïve, surgically resectable advanced non-small cell lung cancer (NSCLC) specimens (n=4), encompassing both EGFR-mutant and wild-type variants. High-dimensional spatial phenotyping was performed using PhenoCycler-Fusion multiplexed immunofluorescence technology, enabling precise visualization and quantification of vasculogenic mimicry (VM) architectures and immune cell infiltrates at subcellular resolution. Complementary molecular characterization of discrete tumor microenvironmental regions was achieved through GeoMx Digital Spatial Profiling platform. Spatial transcriptomics analysis was subsequently employed to validate primary observations and elucidate region-specific transcriptional signatures. Results: Through high-dimensional spatial analysis, we identified a previously undescribed vessel-like population characterized by CD31-VEGFR1+PanCK+ expression, representing vasculogenic mimicry (VM) structures in non-small cell lung cancer (NSCLC). Comprehensive spatial mapping revealed distinct topographical relationships between VM networks and immune cell populations, including CD4+ and CD8+ T lymphocytes, as well as functionally diverse macrophage subsets (CD68+, CD206+, iNOS+, and HLA-DR+). Molecular characterization through digital spatial profiling demonstrated enhanced transcriptional heterogeneity at the tumor margin, with significant enrichment of pathways involved in extracellular matrix (ECM) organization and hypoxia response (FDR < 0.01). Integration of spatial transcriptomics with an established 70-gene VM signature revealed a core set of 10 genes operating at the intersection of VM and ECM remodeling programs. This gene module, predominantly expressed at the tumor-stromal interface, represents a previously unrecognized regulatory hub coordinating VM formation through ECM modification. Conclusions: Our integrated analysis reveals a novel VM phenotype in NSCLC and identifies a core gene signature linking VM formation to ECM remodeling at the invasive front. These findings provide new insights into tumor-stromal-immune interactions and suggest potential therapeutic strategies targeting the VM-ECM-immune axis. Citation Format: Chun-Hui Lee, Chun-Hua Hung, Shu-Hsien Wang, Wei-Pang Chung, Yu-Min Yeh, Peng-Chan Lin, Wu-Chou Su. Mapping the complex interplay between vasculogenic mimicry and immune response in lung cancer using digital spatial profiling [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 162.