重编程葡萄糖代谢，也被称为Warburg效应（有氧糖酵解），是肿瘤的一个标志。肿瘤糖酵解的增加不仅有利于癌细胞的快速增殖，还会重新调整免疫微环境以促进肿瘤进展。转录因子ONECUT3在肝脏和胰腺的发育中发挥关键作用，然而，关于其肿瘤促进作用，特别是代谢重编程方面的了解有限。我们采用免疫组化和免疫印迹等方法，确定了PDAC中ONECUT3的表达模式和其临床相关性。我们使用基因沉默和过表达策略，确定了ONECUT3的体外和体内功能。我们采用染色质免疫沉淀、荧光素酶报告实验和基因集富集分析等方法，揭示了其分子机制。糖酵解代谢与PDAC中T细胞浸润呈负相关。我们发现ONECUT3是PDAC糖酵解和CD8+ T细胞浸润的关键调节因子。ONECUT3的基因沉默抑制了细胞增殖，促进了细胞凋亡，并通过葡萄糖摄取、乳酸产生和细胞外酸化率的证据减少了糖酵解代谢。过表达研究观察到了与之相反的效果。ONECUT3通过转录调控PDK1来增强有氧糖酵解。靶向ONECUT3有效抑制了PDAC的肿瘤生长，增加了CD8+ T细胞浸润，并发挥了对抗PD-1疗法的增效作用。PDK1的药物抑制也与抗PD-1疗法具有协同效应。在临床环境中，ONECUT3与PDAC中的PDK1表达和T细胞浸润密切相关，并起到独立的预后因子作用。我们的研究揭示了ONECUT3在PDAC糖酵解中的前所未有的调控作用，并提供了增加糖酵解与免疫抑制微环境相关的体内证据。此外，靶向ONECUT3-PDK1轴可能成为PDAC治疗的一个有希望的治疗方法。© 2023. Springer Nature Switzerland AG.

Reprogramming glucose metabolism, also known as the Warburg effect (aerobic glycolysis), is a hallmark of cancers. Increased tumor glycolysis not only favors rapid cancer cell proliferation but reprograms the immune microenvironment to enable tumor progression. The transcriptional factor ONECUT3 plays key roles in the development of the liver and pancreas, however, limited is known about its oncogenic roles, particularly metabolic reprogramming.Immunohistochemistry and Western blotting are applied to determine the expression pattern of ONECUT3 and its clinical relevance in pancreatic ductal adenocarcinoma (PDAC). Knockdown and overexpression strategies are employed to determine the in vitro and in vivo functions of ONECUT3. Chromatin immunoprecipitation, luciferase reporter assay, and gene set enrichment analysis are used to decipher the molecular mechanisms.The glycolytic metabolism is inversely associated with T-cell infiltration in PDAC. ONECUT3 is identified as a key regulator for PDAC glycolysis and CD8+ T-cell infiltration. Genetic silencing of ONECUT3 inhibits cell proliferation, promotes cell apoptosis, and reduces glycolytic metabolism as evidenced by glucose uptake, lactate production, and extracellular acidification rate. Opposite effects of ONECUT3 are observed in overexpression studies. ONECUT3 enhances aerobic glycolysis via transcriptional regulation of PDK1. Targeting ONECUT3 effectively suppresses tumor growth, increases CD8+ T-cell infiltration, and potentiates anti-PD-1 therapy in PDAC. Pharmacological inhibition of PDK1 also shows a synergistic effect with anti-PD-1 therapy. In clinical setting, ONECUT3 is closely associated with PDK1 expression and T-cell infiltration in PDAC and acts as an independent prognostic factor.Our study reveals a previous unprecedented regulatory role of ONECUT3 in PDAC glycolysis and provides in vivo evidence that increased glycolysis is linked to an immunosuppressive microenvironment. Moreover, targeting ONECUT3-PDK1 axis may serve as a promising therapeutic approach for the treatment of PDAC.© 2023. Springer Nature Switzerland AG.