O15.04 Metabolic reprogramming enhances stem-like and functional properties of lung cancer-derived TILs

Background Lung cancer remains the leading cause of cancer-related death, with outcomes limited by late-stage diagnosis and resistance to current therapies. Adoptive cell therapy using tumour-infiltrating lymphocytes (TILs) offers durable responses in some solid tumours, but efficacy in lung cancer is constrained by T-cell exhaustion and metabolic dysfunction. Enhancing metabolic fitness is therefore critical to improve TIL-based immunotherapy. Nicotinamide riboside (NR), a nicotinamide adenine dinucleotide (NAD +) precursor that supports mitochondrial function, may restore T-cell energy balance and reduce exhaustion. This study aimed to optimise expansion of lung cancer-derived TILs with enhanced metabolic resilience, stem-like properties, and anti-tumour functionality.Methods Following written informed consent and ethical approval, tumour tissue from lung cancer patients was processed to generate TILs. Tumour fragments (1–2 mm 2) were cultured for 15 days in interleukin-2 (IL-2)-supplemented medium with or without NR to assess effects on TIL expansion and metabolic fitness. Expanded cells were phenotyped by flow cytometry for CD3, CD4, CD8, CD39, CD69, PD-1, LAG-3, and TIM-3. Functional capacity was evaluated by cytokine secretion (IFN-γ, TNF-α, IL-2, granzyme B) following CD3/CD28/CD137 stimulation or autologous tumour co-culture. SIRT1 and C1QBP expression were quantified by quantitative PCR as markers of NR-induced mitochondrial and metabolic enhancement.Results TILs were successfully isolated and expanded from 53 lung cancer specimens, demonstrating feasibility of a robust lung TIL manufacturing workflow. Cultures with superior proliferation showed higher proportions of CD4 + T cells and an enrichment of stem-like CD39−CD69−CD8+ subsets, indicating that helper support and a less differentiated CD8+ phenotype favour expansion potential. Expanded TILs retained strong tumour reactivity, secreting high levels of IFN-γ after CD3/CD28/CD137 stimulation or autologous tumour co-culture. Elevated LAG-3 expression correlated with reduced proliferation. NR supplementation further improved TIL quality by increasing stem-like CD8+ subsets, reducing PD-1 expression, upregulating SIRT1 and C1QBP, and enhancing secretion of TNF-α, IL-2, and IFN-γ, compatible with improved metabolic and effector function.Conclusion A clinically relevant process for isolating and expanding lung cancer-derived TILs has been established, aligning with current efforts to advance cellular therapies in solid tumours. Metabolic reprogramming with NR promotes a less differentiated, metabolically resilient, and tumour-reactive TIL phenotype, supporting NR-supplemented TIL products as a promising strategy to achieve more durable and effective cell therapy for lung cancer.