Shir Jui-Chia, Chen Pin-Yu, Kuo Chuan-Hao, Hsieh Chiao-Hui, Chang Hsin-Yi, Lee Hong-Chih, Huang Chen-Hao, Hsu Chun-Hua, Hsu Wen-Ming, Huang Hsuan-Cheng, Juan Hsueh-Fen
Institute of Molecular and Cellular Biology, National Taiwan University, Taipei, Taiwan.
Department of Life Science, National Taiwan University, Taipei, Taiwan.
Mol Cell Proteomics. 2025 Jun 11;24(7):101014. doi: 10.1016/j.mcpro.2025.101014.
Neuroblastoma is the most common heterogeneous solid tumor in children, and current treatment options remain limited, especially for high-risk patients. Previous studies have identified dihydroorotate dehydrogenase (DHODH), a key enzyme in pyrimidine synthesis, as a potential therapeutic target in cancer. However, none of the existing FDA-approved DHODH inhibitors have shown effective inhibition of neuroblastoma cell growth. To address this challenge, we employed virtual screening to discover potential DHODH-targeting drugs, identifying Regorafenib as a promising candidate. Regorafenib significantly inhibited neuroblastoma growth in both neuroblastoma cells and patient-derived organoids. To unravel the underlying molecular mechanisms, we conducted Tandem Mass Tag (TMT)-based quantitative proteomics using LC-MS/MS. Our proteomic profiling revealed substantial regulation of lipid metabolism proteins, specifically those in the mevalonate pathway, correlating with ferroptosis induction. Further analysis showed that DHODH inhibition led to a reduction in total cholesterol, cholesterol esters, disrupted lipid droplet formation, and significantly decreased the expression of Squalene Epoxidase (SQLE), a key enzyme in lipid metabolism. Notably, we also observed an increase in nuclear SQLE expression following DHODH inhibition. In summary, our study highlights DHODH blockade as a novel approach to induce ferroptosis through lipid metabolism reprogramming, underscoring DHODH as a viable therapeutic target for neuroblastoma treatment. These insights open new avenues for metabolism-based interventions in aggressive pediatric cancers.
神经母细胞瘤是儿童中最常见的异质性实体瘤，目前的治疗选择仍然有限，尤其是对于高危患者。先前的研究已将嘧啶合成中的关键酶二氢乳清酸脱氢酶（DHODH）确定为癌症的潜在治疗靶点。然而，现有的美国食品药品监督管理局（FDA）批准的DHODH抑制剂均未显示出对神经母细胞瘤细胞生长的有效抑制作用。为应对这一挑战，我们采用虚拟筛选来发现潜在的靶向DHODH的药物，确定瑞戈非尼为有前景的候选药物。瑞戈非尼在神经母细胞瘤细胞和患者来源的类器官中均显著抑制神经母细胞瘤的生长。为了阐明潜在的分子机制，我们使用液相色谱-串联质谱（LC-MS/MS）进行了基于串联质量标签（TMT）的定量蛋白质组学研究。我们的蛋白质组分析揭示了脂质代谢蛋白的大量调控，特别是甲羟戊酸途径中的那些蛋白，这与铁死亡诱导相关。进一步分析表明，DHODH抑制导致总胆固醇、胆固醇酯减少，脂滴形成受到破坏，脂质代谢关键酶角鲨烯环氧酶（SQLE）的表达显著降低。值得注意的是，我们还观察到DHODH抑制后细胞核中SQLE表达增加。总之，我们的研究强调了阻断DHODH作为通过脂质代谢重编程诱导铁死亡的新方法，突出了DHODH作为神经母细胞瘤治疗的可行靶点。这些见解为侵袭性儿科癌症基于代谢的干预开辟了新途径。
