Abstract

Gastric cancer is one of the most common primary malignant tumors of the digestive system. Chemoresistance remains a major obstacle in the clinical management of gastric cancer, and targeting the metabolic reprogramming of tumor cells has emerged as a promising therapeutic strategy. N4-acetylcytidine (ac4C), an important post-transcriptional RNA modification, is catalyzed by the key acetyltransferase N -acetyltransferase 10 (NAT10). Through drug screening, we identified the phosphoinositide 3-kinase (PI3K) inhibitor GNE-493 as a potent suppressor of gastric cancer cell proliferation. GNE-493 markedly reduced glucose uptake and lactate production, indicating inhibition of aerobic glycolysis. Mechanistically, GNE-493 binds directly to NAT10, resulting in decreased ac4C acetylation within the coding sequence of hexokinase 2 (HK2), a critical glycolytic enzyme. This reduction impairs the stability and translation of HK2 transcripts, diminishes glycolytic flux, and consequently restrains gastric cancer progression. Furthermore, GNE-493 demonstrated strong efficacy in cisplatin-resistant gastric cancer cell lines, underscoring its potential to overcome conventional chemoresistance. By targeting the epitranscriptomic control of metabolic reprogramming, GNE-493 offers a novel therapeutic avenue for gastric cancer patients with highly active glucose metabolism.