UHRF1-regulated aerobic glycolysis mediates TOX lactylation to induce CD8⁺ T cell exhaustion in laryngeal cancer

Laryngeal squamous cell carcinoma (LSCC) is a malignancy characterized by metabolic reprogramming and immune evasion, where the immunosuppressive tumor microenvironment contributes to poor therapeutic response. The role of Ubiquitin-like with PHD and RING finger domain 1 (UHRF1), an epigenetic regulator, in regulating metabolic-immune crosstalk in LSCC remains unclear. This study aimed to explore whether UHRF1 drives aerobic glycolysis and lactate-mediated TOX lactylation to induce CD8⁺ T cell exhaustion. UHRF1 expression and glycolytic activity were assessed in LSCC cell lines and tissues. Glycolysis was inhibited via UHRF1 knockdown or 2-deoxyglucose (2-DG), and lactate levels were modulated with sodium lactate (NaL) or oxamate (OX). CD8⁺ T cell function was evaluated in co-culture systems using flow cytometry and ELISA. TOX lactylation was detected by Western blot and Co-IP assays. UHRF1 was overexpressed in LSCC and correlated with upregulation of glycolytic enzymes (HK2, PKM2, LDHA) and increased lactate secretion ( P < 0.01). UHRF1 knockdown or 2-DG treatment reduced tumor proliferation, migration, and glycolytic output, and restored CD8⁺ T-cell effector function by reducing exhaustion markers (PD-1, TIM3, CTLA-4, LAG3; all P < 0.01). UHRF1-driven lactate production induced TOX lactylation, which suppressed T-bet and enhanced Eomes expression. OX reversed this effect, while NaL restored TOX lactylation in UHRF1-deficient cells. In conclusion, UHRF1-driven lactate promotes TOX lactylation and contributes to CD8 + T-cell exhaustion. Combinatorial targeting of UHRF1 and glycolysis may help reverse this immunosuppressive circuit and provide a potential strategy for improving LSCC immunotherapy.