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, the molecular signatures that define a productive TIL-mediated response against tumors remain poorly characterized. Here, we establish a patient-derived organoid and autologous TIL co-culture platform to dissect this interaction at high resolution. We show that expanded TILs mediate potent and 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 the activation of key signaling networks including the Tumor Necrosis Factor (TNF) and Interleukin-17 (IL-17) pathways. Furthermore, analysis of the T-cell receptor (TCR) repertoire confirms that 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 the development of next-generation personalized TIL therapies.