Multimodal and spatial single-cell profiling reveals immune-suppressed niches in tumor-adjacent lymph nodes of early-stage NSCLC patients 4608

Abstract Description Objective Lymph node (LN) metastasis is a critical prognostic factor for lung cancer, but the influence of the LN immune microenvironment (IM) on metastasis remains unclear. We hypothesize that changes in the LN IM predict LN metastasis and indicate tumor progression in premetastatic early-stage non-small cell lung cancer (ES-NSCLC). Methods Multi-modal single-cell sequencing was used to profile the transcriptome and proteome of 88,203 cells from LN and tumor biopsies of 10 ES-NSCLC patients. Imaging Mass Cytometry (IMC) spatially profiled 1,024,996 cells from FFPE slides of the same biopsies using 39 antibodies. Results Subtypes of exhausted CD8+ T cells were identified in N1-stationed LNs of ES-NSCLC patients and were enriched in corresponding patient tumors. These LNs were also enriched with mature regulatory dendritic cells (mregDCs), plasmacytoid dendritic cells (pDCs), plasma cells, B cell subsets, and regulatory T cells. IMC revealed spatial interactions among these cells, forming immune niches in the LNs of patients with more advanced disease. Conclusion The observed immune dysregulation may suggest the formation of a “pre-metastatic immune niche” in specific LNs, increasing their susceptibility to tumor metastasis. This research has the potential to identify LN biomarkers, leading to earlier and more effective treatments for ES-NSCLC patients. Future work will address the limitation of a small patient cohort by enrolling additional patients for validation. Funding Sources Supported by NCI HTAN 1U2CCA233238-01; Elsa Pardee Foundation; J&J-BU Pilot Grant; CTSI Pilot Grant Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)