Dual inhibition of glycolysis and glutaminolysis by targeting FOXO3 for hepatocellular carcinoma treatment.

Metabolic reprogramming is a hallmark of tumorigenesis and progression in hepatocellular carcinoma (HCC) and has emerged as a promising therapeutic strategy. Forkhead box O3 (FOXO3), a critical nuclear transcription factor, is dysregulated in multiple cancers; however, its precise role in HCC progression remains unclear. In this study, we demonstrate that enhanced glycolysis and glutaminolysis are pivotal metabolic features of HCC, with tumor cells heavily relying on both pathways for survival and proliferation. We identify FOXO3 as a tumor suppressor in HCC that inhibits key metabolic enzymes and metabolites involved in these pathways. This inhibitory effect is mediated through suppression of yes-associated protein (YAP). Mechanistically, FOXO3 directly binds to the GTGAACAT motif (-1824 to -1817) within the YAP promoter, leading to transcription repression of YAP and subsequent disruption of YAP-driven metabolic programs. Pharmacological activation of FOXO3 using specific inducers markedly reduced YAP expression, resulting in inhibition of glycolysis, glutaminolysis, and proliferation in HCC cells. In vivo, activation of the FOXO3/YAP axis effectively suppressed HCC progression through the coordinated inhibition of glycolysis and glutaminolysis. Moreover, FOXO3 inducers significantly impaired the growth and viability of patient-derived HCC organoid models. Hence, these findings identify FOXO3 as a key regulator of metabolic reprogramming in HCC and establish the FOXO3/YAP axis as a promising therapeutic target, suggesting potential strategies for metabolic-based interventions in HCC treatment.