Targeted NAT10 Degradation by PROTAC NP1192 Suppresses Hypoxia-Adaptive Glycolysis and Reinvigorates CD8 ⁺ Effector T-Cell Function for Synergistic Cancer Immunotherapy

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 <i>in vivo</i>, <i>in vitro</i>, and across three tumor organoids. It abrogated ac4C modifications on <i>HIF1A</i> 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 <i>in vitro</i> 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.