多形核巨细胞的代谢重编程决定了胶质母细胞瘤从阿霉素诱导的应激中恢复。

BACKGROUND: Multi-drug resistance of poly(morpho)nuclear giant cells (PGCs) determines their cytoprotective and generative potential in cancer ecosystems. However, mechanisms underlying the involvement of PGCs in glioblastoma multiforme (GBM) adaptation to chemotherapeutic regimes remain largely obscure. In particular, metabolic reprogramming of PGCs has not yet been considered in terms of GBM recovery from doxorubicin (DOX)-induced stress. METHODS: Long-term proteomic and metabolic cell profiling was applied to trace the phenotypic dynamics of GBM populations subjected to pulse DOX treatment in vitro, with a particular focus on PGC formation and its metabolic background. The links between metabolic reprogramming, drug resistance and drug retention capacity of PGCs were assessed, along with their significance for GBM recovery from DOX-induced stress. RESULTS: Pulse DOX treatment triggered the transient formation of PGCs, followed by the appearance of small expanding cell (SEC) clusters. Development of PGCs was accompanied by the mobilization of their metabolic proteome, transient induction of oxidative phosphorylation (OXPHOS), and differential intracellular accumulation of NADH, NADPH, and ATP. The metabolic background of PGC formation was confirmed by the attenuation of GBM recovery from DOX-induced stress following the chemical inhibition of GSK-3β, OXPHOS, and the pentose phosphate pathway. Concurrently, the mobilization of reactive oxygen species (ROS) scavenging systems and fine-tuning of NADPH-dependent ROS production systems in PGCs was observed. These processes were accompanied by perinuclear mobilization of ABCB1 and ABCG2 transporters and DOX retention in the perinuclear PGC compartments. CONCLUSIONS: These data demonstrate the cooperative pattern of GBM recovery from DOX-induced stress and the crucial role of metabolic reprogramming of PGCs in this process. Metabolic reprogramming enhances the efficiency of self-defense systems and increases the DOX retention capacity of PGCs, potentially reducing DOX bioavailability in the proximity of SECs. Consequently, the modulation of PGC metabolism is highlighted as a potential target for intervention in glioblastoma treatment.

背景：多药耐药多形核巨细胞（PGC）决定了它们在癌症生态系统中的细胞保护和生成潜力。然而，PGC 参与胶质母细胞瘤多形性（GBM）适应化疗方案的机制在很大程度上仍不清楚。特别是，PGC 的代谢重编程在 GBM 从阿霉素（DOX）诱导的应激中恢复方面尚未得到考虑。

方法：长期蛋白质组学和代谢细胞分析用于追踪体外接受脉冲 DOX 处理的 GBM 群体的表型动态，特别关注 PGC 的形成及其代谢背景。评估了代谢重编程与 PGC 耐药性和药物保留能力之间的联系，以及它们对 GBM 从 DOX 诱导的应激中恢复的意义。

结果：脉冲 DOX 处理触发了 PGC 的短暂形成，随后出现了小扩展细胞（SEC）簇。PGC 的形成伴随着其代谢蛋白质组的动员、氧化磷酸化（OXPHOS）的短暂诱导以及 NADH、NADPH 和 ATP 的细胞内差异积累。PGC 形成的代谢背景通过化学抑制 GSK-3β、OXPHOS 和戊糖磷酸途径来减弱 GBM 从 DOX 诱导的应激中恢复得到证实。同时，观察到 PGC 中活性氧（ROS）清除系统的动员和 NADPH 依赖性 ROS 产生系统的微调。这些过程伴随着 ABCB1 和 ABCG2 转运体的核周动员和 DOX 在核周 PGC 隔室中的保留。

结论：这些数据表明了 GBM 从 DOX 诱导的应激中恢复的协同模式，以及 PGC 代谢重编程在这一过程中的关键作用。代谢重编程提高了自我防御系统的效率，并增加了 PGC 的 DOX 保留能力，从而降低了 SEC 附近 DOX 的生物利用度。因此，调节 PGC 代谢被强调为干预胶质母细胞瘤治疗的潜在靶点。