Li Huanjuan, Li Yanchun, Yu Yanhua, Ren Xueying, Yang Chen, Jin Weidong, Li Keyi, Zhou Yi, Wu Cuiyun, Shen Yuhuan, Hu Wanye, Liu Yingchao, Yu Lingyan, Tong Xiangmin, Du Jing, Wang Ying
Laboratory Medicine Center, Department of Clinical Laboratory, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, Zhejiang, 310014, China.
Department of Clinical Laboratory, Affiliated Hangzhou First People's Hospital, School of Medicine, Westlake University, Hangzhou, Zhejiang, 310006, China.
Free Radic Biol Med. 2024 Sep;222:130-148. doi: 10.1016/j.freeradbiomed.2024.06.006. Epub 2024 Jun 11.
The clinical application of the therapeutic approach in myelodysplastic syndromes (MDS) remains an insurmountable challenge for the high propensity for progressing to acute myeloid leukemia and predominantly affecting elderly individuals. Thus, the discovery of molecular mechanisms underlying the regulatory network of different programmed cell death holds great promise for the identification of therapeutic targets and provides insights into new therapeutic avenues. Herein, we found that disulfiram/copper (DSF/Cu) significantly repressed the cell viability, increased reactive oxygen species (ROS) accumulation, destroyed mitochondrial morphology, and altered oxygen consumption rate. Further studies verified that DSF/Cu induces cuproptosis, as evidenced by the depletion of glutathione (GSH), aggregation of lipoylated DLAT, and induced loss of Fe-S cluster-containing proteins, which could be rescued by tetrathiomolybdate and knockdown of ferredoxin 1 (FDX1). Additionally, GSH contributed to the tolerance of DSF/Cu-mediated cuproptosis, while pharmacological chelation of GSH triggered ROS accumulation and sensitized cell death. The xCT-GSH-GPX4 axis is the ideal downstream component of ferroptosis that exerts a powerful protective mechanism. Notably, classical xCT inhibitors were capable of leading to the catastrophic accumulation of ROS and exerting synergistic cell death, while xCT overexpression restored these phenomena. Simvastatin, an inhibitor of HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) reductase, has beneficial effects in repurposing for inhibiting GPX4. Similarly, the combination treatment of DSF/Cu and simvastatin dramatically decreased the expression of GPX4 and Fe-S proteins, ultimately accelerating cell death. Moreover, we identified that the combination treatment of DSF/Cu and simvastatin also had a synergistic antitumor effect in the MDS mouse model, with the reduced GPX4, increased COX-2 and accumulated lipid peroxides. Overall, our study provided insight into developing a novel synergistic strategy to sensitize MDS therapy by targeting ferroptosis and cuproptosis.
对于骨髓增生异常综合征（MDS）而言，这种治疗方法的临床应用仍然是一项难以克服的挑战，因为该疾病进展为急性髓系白血病的倾向很高，且主要影响老年人。因此，发现不同程序性细胞死亡调控网络背后的分子机制，对于确定治疗靶点具有巨大潜力，并为新的治疗途径提供了思路。在此，我们发现双硫仑/铜（DSF/Cu）显著抑制细胞活力，增加活性氧（ROS）积累，破坏线粒体形态，并改变氧消耗率。进一步研究证实，DSF/Cu诱导铜死亡，表现为谷胱甘肽（GSH）耗竭、脂酰化DLAT聚集以及含Fe-S簇蛋白的诱导性丢失，而四硫钼酸盐和铁氧化还原蛋白1（FDX1）的敲低可以挽救这些现象。此外，GSH有助于细胞对DSF/Cu介导的铜死亡产生耐受性，而GSH的药理学螯合会引发ROS积累并使细胞死亡敏感化。xCT-GSH-GPX4轴是铁死亡的理想下游成分，发挥着强大的保护机制。值得注意的是，经典的xCT抑制剂能够导致ROS的灾难性积累并发挥协同细胞死亡作用，而xCT的过表达可恢复这些现象。辛伐他汀是HMG-CoA（3-羟基-3-甲基戊二酰辅酶A）还原酶的抑制剂，在重新用于抑制GPX4方面具有有益作用。同样，DSF/Cu与辛伐他汀的联合治疗显著降低了GPX4和Fe-S蛋白的表达，并最终加速细胞死亡。此外，我们发现DSF/Cu与辛伐他汀的联合治疗在MDS小鼠模型中也具有协同抗肿瘤作用，表现为GPX4降低、COX-2增加以及脂质过氧化物积累。总体而言，我们的研究为通过靶向铁死亡和铜死亡来开发一种使MDS治疗敏感化的新型协同策略提供了思路。
