Transcriptome profiling of tumor-infiltrating lymphocyte-mediated cytotoxicity against patient-derived lung cancer organoids.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer mortality, and therapies utilizing tumor-infiltrating lymphocytes (TILs) show significant promise. However, molecular signatures defining a productive TIL-mediated response remain poorly characterized. Here, we establish a patient-derived organoid and autologous TIL co-culture platform to show that expanded TILs mediate potent, specific cytotoxicity against NSCLC organoids. This functional response is associated with a crucial shift in T-cell states from proliferative towards effector memory phenotypes and involves activating key signaling networks, including the TNF and IL-17 pathways. Furthermore, T-cell receptor (TCR) analysis confirms the expansion process selectively enriches tumor-associated clonotypes, resulting in a more focused repertoire. This work delineates the transcriptional and clonal signatures of an effective anti-tumor immune response, providing a robust framework to guide next-generation personalized TIL therapies.