The transcription factor hypoxia-inducible factor 1α (HIF-1α) drives metabolic reprogramming in gliomas (GLs) under hypoxic conditions, promoting glycolysis for tumor development. Evofosfamide (EVO) releases a DNA-alkylating agent within hypoxic regions, indicating that it may serve as a hypoxia-targeted therapy. The aim of this study was to investigate the glycolytic metabolism and antitumor effects of EVO in a canine GL model. Our clinical data showed that overall survival was significantly decreased in GL dog patients with higher HIF-1α expression compared to that of those with lower HIF-1α expression, and there was a positive correlation between HIF-1α and pyruvate dehydrogenase kinase 1 (PDK1) expression, suggesting that glycolytic activity under hypoxia conditions may contribute to poor outcomes in canine GL. Our glycolysis assay tests showed that the glycolytic ATP level was higher than the mitochondrial ATP level in three types of canine GL cell lines by activating the HIF-1 signal pathway under hypoxia conditions, resulting in an overall increase in total cellular ATP production. However, treatment with EVO inhibited the glycolytic ATP level in the GL cell lines under hypoxia conditions by targeting HIF-1α-positive cells, leading to decrease in total cellular ATP production. Our in vivo tests showed that EVO significantly reduced tumor development compared to controls and temozolomide in murine GL models. A metabolic analysis demonstrated that EVO effectively suppressed glycolytic metabolism by eliminating HIF-1α-positive cells, suggesting that it may restore metabolism in canine GLs. The evidence presented here supports the favorable preclinical evaluation of EVO as a potential improvement in cancer metabolism.
转录因子缺氧诱导因子1α（HIF-1α）在缺氧条件下驱动胶质瘤（GLs）的代谢重编程，通过促进糖酵解支持肿瘤发展。依沃福酰胺（EVO）能在缺氧区域释放DNA烷化剂，表明其可作为缺氧靶向疗法。本研究旨在探究EVO在犬GL模型中的糖酵解代谢及抗肿瘤效应。临床数据显示，与低表达HIF-1α的犬GL患者相比，高表达HIF-1α患者的总生存期显著缩短，且HIF-1α与丙酮酸脱氢酶激酶1（PDK1）表达呈正相关，提示缺氧条件下的糖酵解活性可能导致犬GL预后不良。糖酵解检测表明，在缺氧条件下，三种犬GL细胞系通过激活HIF-1信号通路，其糖酵解ATP水平高于线粒体ATP水平，导致细胞总ATP产量整体上升。然而，EVO处理通过靶向HIF-1α阳性细胞，抑制了缺氧条件下GL细胞系的糖酵解ATP水平，从而降低细胞总ATP产量。体内实验显示，在小鼠GL模型中，与对照组及替莫唑胺相比，EVO能显著抑制肿瘤发展。代谢分析证实，EVO通过清除HIF-1α阳性细胞有效抑制糖酵解代谢，表明其可能恢复犬GL的代谢平衡。本研究为EVO作为改善癌症代谢的潜在药物提供了有利的临床前评估依据。
肿瘤(癌症)患者之家