Targeted NAT10 Degradation by PROTAC NP1192 Suppresses Hypoxia-Adaptive Glycolysis and Reinvigorates CD8⁺ Effector...

Tumor resistance to immune checkpoint blockade (ICB) therapy is frequently driven by adaptive metabolic reprogramming in the hypoxic tumor microenvironment (TME). The key N4-acetylcytidine (ac4C) RNA modification mediator N-acetyltransferase 10 (NAT10) emerges as a promising therapeutic target, despite the lack of potent targeting agents. Here, we engineered NP1192, a PROTAC degrader targeting NAT10. NP1192 achieved nearly 70% NAT10 degradation and a 26.8% lower IC₅₀ than canonical NAT10 inhibitor Remodelin in cervical cancer cells, outperforming Remodelin in antitumor effect in vivo, in vitro, and across three tumor organoids. It abrogated ac4C modifications on HIF1A mRNA and translation, reducing hypoxic lactate production and depleted ATP, and suppressed HIF-1α-mediated PD-L1 upregulation. In xenograft models, NP1192 combined with anti-PD-L1 inhibited subcutaneous xenograft growth and reduced tumor-core lactate gradients by > 80%. Furthermore, scRNA-seq and in vitro coculture experiments identified expansion of IFN-γ⁺ effector CD8⁺ T cells (T_eff) and decline in exhausted CD8⁺ T cells (T_ex). NP1192 in combination with anti-PD-L1 enhanced proliferation and effector function of CD8⁺ T_eff cells, thereby reversing resistance to anti-PD-L1 blockade therapy and synergizing with immunotherapy. These findings establish PROTAC-mediated NAT10 degradation as a dual metabolic-immune strategy to enhance checkpoint blockade efficacy.