Ye Rongxu, Zhang Wanghao, Zhang Huayang, Qu Shanqiang, Xu Junyi, Xu Rongyang, Zhu Ye, Huang Guanglong, Zhang Xi-An, Yi Guo-Zhong
Department of Neurosurgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.
The Laboratory for Precision Neurosurgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People's Republic of China.
CNS Neurosci Ther. 2025 Aug;31(8):e70546. doi: 10.1111/cns.70546.
The clinical efficacy of temozolomide (TMZ) in glioblastoma (GBM) patients is often limited by the development of resistance. To date, no clinically validated therapeutic strategies exist to restore sensitivity to TMZ treatment. In this study, we investigated the potential of FR054, a hexosamine biosynthesis pathway (HBP) inhibitor, to sensitize GBM cells to TMZ and elucidated its underlying molecular mechanism.
TMZ-resistant U87-MG and A172 cell lines were generated through stepwise exposure to increasing concentrations of TMZ. Proteomics and bioinformatics analyses revealed HBP activation in these resistant cells. The effects of FR054 alone or in combination with TMZ were assessed using cell line models, GBM organoid models, and intracranial xenograft models. Transcriptomic analysis and validation experiments were further conducted to explore the molecular mechanisms involved.
Long-term exposure to TMZ induced resistance in U87-MG and A172 GBM cells, which was associated with the activation of the HBP pathway. PGM3, a key enzyme in the HBP, was found to correlate with poor prognosis in GBM patients. The combination of FR054, a specific PGM3 inhibitor, with TMZ exhibited synergistic inhibitory effects in vitro and superior inhibitory efficacy in GBM organoid models. In vivo, this combination significantly suppressed tumor progression and prolonged survival in orthotopic xenograft mice with minimal side effects. Mechanistically, FR054 enhanced TMZ sensitivity by inhibiting protein O-GlcNAcylation and promoting ferroptosis via the upregulation of HMOX1 and downregulation of GPX4.
Our findings demonstrate that targeting the HBP pathway with FR054 can overcome TMZ resistance in GBM by reducing O-GlcNAc modification and inducing ferroptosis. This novel approach enhances the efficacy of TMZ, offering a promising therapeutic strategy for GBM patients with limited treatment options.
替莫唑胺（TMZ）在胶质母细胞瘤（GBM）患者中的临床疗效常常受到耐药性发展的限制。迄今为止，尚无经临床验证的治疗策略可恢复对TMZ治疗的敏感性。在本研究中，我们研究了己糖胺生物合成途径（HBP）抑制剂FR054使GBM细胞对TMZ敏感的潜力，并阐明了其潜在的分子机制。
通过逐步暴露于浓度递增的TMZ产生TMZ耐药的U87-MG和A172细胞系。蛋白质组学和生物信息学分析揭示了这些耐药细胞中HBP的激活。使用细胞系模型、GBM类器官模型和颅内异种移植模型评估了FR054单独或与TMZ联合使用的效果。进一步进行转录组分析和验证实验以探索其中涉及的分子机制。
长期暴露于TMZ会诱导U87-MG和A172 GBM细胞产生耐药性，这与HBP途径的激活有关。发现HBP中的关键酶PGM3与GBM患者的不良预后相关。特异性PGM3抑制剂FR054与TMZ联合使用在体外表现出协同抑制作用，在GBM类器官模型中具有优异的抑制效果。在体内，这种联合用药显著抑制了原位异种移植小鼠的肿瘤进展并延长了生存期，且副作用最小。从机制上讲，FR054通过抑制蛋白质O-连接的N-乙酰葡糖胺化并上调HMOX1和下调GPX4来促进铁死亡，从而增强TMZ敏感性。
我们的研究结果表明，用FR054靶向HBP途径可通过减少O-GlcNAc修饰和诱导铁死亡来克服GBM中的TMZ耐药性这种新方法提高了TMZ的疗效，为治疗选择有限的GBM患者提供了一种有前景的治疗策略。
