胶质母细胞瘤（GBM）是一种原发性恶性脑肿瘤，即使采用放疗和化疗等标准治疗，预后也很差。在这项研究中，我们探讨了多巴胺受体 D2/3 (DRD2/3) 拮抗剂奋乃静 (PER) 与 GBM 标准治疗药物替莫唑胺 (TMZ) 协同组合在患者来源的人 GBM 中的抗癌作用。通过测量细胞活力、ATP、干性、侵袭性和凋亡来评估 PER 和 TMZ 组合在 GBM TS 中的生物学效应。使用蛋白质印迹和 RNA 测序分析蛋白质和 mRNA 表达的变化。使用小鼠原位异种移植模型对 PER 和 TMZ 的联合给药进行了体内评估。Severance 数据集显示，GBM 患者肿瘤组织中的 DRD2 和 DRD3 表达高于无肿瘤皮质中的表达。在源自患者的人 GBM TS 中，通过 CRISPR/Cas9 敲除 DRD2/3，抑制细胞生长和 ATP 产生。与单一治疗组相比，PER 和 TMZ 联合治疗对细胞活力和 ATP 测定产生了更好的效果。流式细胞术、蛋白质印迹和 RNA 测序证实联合治疗后 GBM TS 细胞凋亡增加。此外，PER 和 TMZ 的组合下调了与干性和侵袭性相关的蛋白质和 mRNA 的表达。体内评估表明，PER 和 TMZ 组合可延长小鼠原位异种移植模型的存活期。PER 和 TMZ 的协同组合有潜力成为 GBM 的新型联合治疗策略。© 作者 2024。牛津大学出版代表神经肿瘤学会新闻。

Glioblastoma (GBM), a primary malignant brain tumor, has a poor prognosis, even with standard treatments such as radiotherapy and chemotherapy. In this study, we explored the anticancer effects of the synergistic combination of perphenazine (PER), a dopamine receptor D2/3 (DRD2/3) antagonist, and temozolomide (TMZ), a standard treatment for GBM, in patient-derived human GBM tumorspheres (TSs).The biological effects of the combination of PER and TMZ in GBM TSs were assessed by measuring cell viability, ATP, stemness, invasiveness, and apoptosis. Changes in protein and mRNA expression were analyzed using western blotting and RNA sequencing. Co-administration of PER and TMZ was evaluated in vivo using a mouse orthotopic xenograft model.The Severance dataset showed that DRD2 and DRD3 expression was higher in tumor tissues than in the tumor-free cortex of patients with GBM. DRD2/3 knockout by CRISPR/Cas9 in patient-derived human GBM TSs inhibited cell growth and ATP production. The combined treatment with PER and TMZ resulted in superior effects on cell viability and ATP assays compared to those in single treatment groups. Flow cytometry, western blotting, and RNA sequencing confirmed elevated apoptosis in GBM TSs following combination treatment. Additionally, the combination of PER and TMZ downregulated the expression of protein and mRNA associated with stemness and invasiveness. In vivo evaluation showed that combining PER and TMZ extended the survival period of the mouse orthotopic xenograft model.The synergistic combination of PER and TMZ has potential as a novel combination treatment strategy for GBM.© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Neuro-Oncology.