Natural Killer Cell-Mediated Immunosurveillance Drives Liver Cancer Evolution Through Cancer Stemness Enhancement and Lipid Metabolism Reprogramming

Abstract Tumor cells acquire survival advantages and evade therapeutic-induced cell death through continuous evolution. The liver has a unique immune microenvironment, with a higher proportion of natural killer (NK) cells compared to other organs. While NK cells are closely associated with liver precancerous conditions and liver cancer, their role in liver cancer evolution remains unclear. This study aims to comprehensively reveal core regulatory factors and key molecular events in liver cancer evolution, and develop novel therapeutic strategies. In this study, Immune-humanized spatiotemporal models were established to simulate liver cancer evolution. Multi-omics approaches, including scRNA-seq, snRNA-seq, spatial transcriptome and CRISPR/Cas9 screening, were subsequently used to establish the functional relevance of NK cells and liver cancer evolution. We demonstrated that early immunosurveillance induced by NK cells is the pioneering power to trigger transition in tumor cell state and impedes subsequent adaptive immunosurveillance. Mechanistically, NK cells induced the reprogramming of lipid metabolism, especially cholesterol accumulation in tumor cells, and tumor stemness enhancement, which are demonstrated to be crucial for liver cancer evolution. Furthermore, combining anti-LAG-3 and LXR activation halts tumor evolution, enhancing the efficacy and durability of immune checkpoint inhibitors against advanced liver cancer. In conclusion, NK cell-mediated early immunosurveillance drives liver cancer evolution. Immunometabolic therapy targeting these evolved tumor cells presents a promising strategy for advanced liver cancer.