METHODS: Cell lines and primary cells from acute myeloid leukaemia (AML) patients were used and the effect of the CNB derivative WIN-55 was evaluated in vitro, ex vivo and in vivo.
RESULTS: We demonstrate a potent antileukemic effect of WIN-55 which is abolished with CB antagonists. WIN-treated mice, xenografted with AML cells, had better survival as compared to vehicle or cytarabine. DNA damage-related genes were affected upon exposure to WIN. Co-incubation with the PARP inhibitor Olaparib prevented WIN-induced cell death, suggesting PARP-mediated apoptosis which was further confirmed with the translocation of AIF to the nucleus observed in WIN-treated cells. Nicotinamide prevented WIN-related apoptosis, indicating NAD+ depletion. Finally, WIN altered glycolytic enzymes levels as well as the activity of G6PDH. These effects are reversed through PARP1 inhibition.
CONCLUSIONS: WIN-55 exerts an antileukemic effect through Parthanatos, leading to translocation of AIF to the nucleus and depletion of NAD+, which are reversed through PARP1 inhibition. It also induces metabolic disruptions. These effects are not observed in normal HSC.
方法:使用来自急性髓系白血病(AML)患者的细胞系和原代细胞,并在体外评估CNB衍生物WIN-55的作用,离体和体内。
结果:我们证明了WIN-55的有效抗白血病作用,该作用被CB拮抗剂消除。胜利治疗的小鼠,与AML细胞异种移植,与媒介物或阿糖胞苷相比,存活率更好。DNA损伤相关基因在暴露于WIN时受到影响。与PARP抑制剂Olaparib共同孵育可防止WIN诱导的细胞死亡,提示PARP介导的细胞凋亡,在WIN处理的细胞中观察到AIF易位到细胞核进一步证实。烟酰胺可预防WIN相关细胞凋亡,表示NAD+耗尽。最后,WIN改变了糖酵解酶水平以及G6PDH的活性。这些作用通过PARP1抑制而逆转。
结论:WIN-55通过Parthanatos发挥抗白血病作用,导致AIF易位到细胞核和NAD+耗尽,通过PARP1抑制逆转。它还诱导代谢破坏。在正常HSC中未观察到这些作用。