Abstract
Tumor heterogeneity and the dynamic evolution of tumor immune microenvironment (TIME) contribute to therapeutic resistance and poor clinical prognosis. To elucidate this mechanism, we first established a murine tumor evolution model (TEM) and systematically identified evolutionary core genes demonstrating progressive alterations during evolution. Subsequently, we developed a single-cell TEM through integrative analysis of hepatocellular carcinoma (HCC) clinical specimens (n=10) with external cohorts (n=11), enabling dynamic characterization of tumor-immune interactions during evolution, while addressing ethical challenges associated with obtaining tumor tissues from multiple stages in a single patient. Through TEMs analyses, we identified a contrasting glucose metabolism pattern between malignant cells and CD8(+) T cells during tumor evolution. Mechanistically, glucose metabolic dominance triggers NSUN2 upregulation in tumor cells, where this functional RNA methyltransferase stabilizes key glycolytic transcripts (GLUT1, HK2, PFKM) through mRNA methylation. The NSUN2-mediated GLUT1 stabilization enhances the competitive advantage of tumor cells in glucose acquisition, creating a positive feedback loop that accelerates malignancy and exacerbates CD8(+) T cell dysfunction. Building on these insights, we designed a dual-targeting strategy combining GLUT1/NSUN2 axis inhibitor WZB117 with PD-L1 blockade, which synergistically suppressed tumor evolution and reversed immunosuppression in preclinical models, suggesting a novel synergistic therapeutic strategy for treatment-resistant HCC.