我们最近证明，组蛋白脱乙酰酶抑制剂丙戊酸（VPA）可以重新编程顺铂诱导的三阴性乳腺癌（TNBC）细胞的代谢组，包括己糖水平的变化。因此，在这里，我们测试了 VPA 改变与顺铂敏感性相关的葡萄糖代谢的假设。分析了两种 TNBC 细胞系：MDA-MB-231（顺铂耐药细胞系）和 MDA-MB-436（顺铂敏感细胞系）。使用糖酵解压力测试试剂盒测量糖酵解和氧化代谢。通过蛋白质印迹和实时 PCR 分析研究了醛脱氢酶 (ALDH)（与耐药性相关的酶）的表达。我们还研究了双硫仑抑制 ALDH 对 MDA-MB-231 细胞活力和 TNBC 患者来源的类器官系统的影响。顺铂处理降低了 MDA-MB-436 细胞的细胞外酸化率，但没有降低 MDA-MB-231 细胞的细胞外酸化率，而添加 VPA 则增加了两种细胞系的细胞外酸化率。 VPA 进一步降低了顺铂处理的 MDA-MB-436 细胞的耗氧率，这与细胞周期的改变相关。然而，在 MDA-MB-231 细胞中，顺铂/VPA-顺铂治疗之间的细胞周期分布没有变化。在这两种细胞系中，VPA 均增加了 ALDH 异构体的表达和 ALDH1A1 的表达。然而，仅在 MDA-MB-231 细胞中，VPA 与顺铂协同作用以增强这种作用。双硫仑使细胞对 VPA-顺铂组合的细胞毒性作用敏感。此外，双硫仑-VPA-化疗组合对 TNBC 类器官最有效。我们的结果表明，ALDH 过表达可能是 TNBC 中细胞对 VPA 耐药的机制之一，并且其抑制可能会增强 VPA-化疗药物组合的治疗效果。版权所有 © 2023 Granit Mizrahi, Gugenheim, Hamad, Hamed, Tetro, Maimon,胡楚劳里、内楚什坦、尼斯曼、杜兰、萨曼、比林贝格-施瓦茨、格鲁内瓦尔德、埃亚尔和佩雷茨。

We recently demonstrated that the histone deacetylase inhibitor valproic acid (VPA) reprograms the cisplatin-induced metabolome of triple-negative breast cancer (TNBC) cells, including a shift in hexose levels. Accordingly, here, we tested the hypothesis that VPA alters glucose metabolism in correlation with cisplatin sensitivity. Two TNBC cell lines, MDA-MB-231 (a cisplatin-resistant line) and MDA-MB-436 (a cisplatin-sensitive line), were analyzed. The glycolysis and oxidative metabolism were measured using the Glycolysis Stress Test kit. The expression of aldehyde dehydrogenases (ALDHs), enzymes linked to drug resistance, was investigated by Western blot and real-time PCR analyses. We additionally studied the influence of ALDH inhibition by disulfiram on the viability of MDA-MB-231 cells and on a TNBC patient-derived organoid system. Cisplatin treatment reduced the extracellular acidification rate in MDA-MB-436 cells but not MDA-MB-231 cells, whereas VPA addition increased the extracellular acidification rate in both cell lines. VPA further reduced the oxygen consumption rate of cisplatin-treated MDA-MB-436 cells, which correlated with cell cycle alterations. However, in MDA-MB-231 cells, the cell cycle distribution did not change between cisplatin/VPA-cisplatin treatments. In both cell lines, VPA increased the expression of ALDH isoform and ALDH1A1 expression. However, only in MDA-MB-231 cells, VPA synergized with cisplatin to augment this effect. Disulfiram sensitized the cells to the cytotoxic effects of the VPA-cisplatin combination. Furthermore, the disulfiram-VPA-chemotherapy combination was most effective in TNBC organoids. Our results show that ALDH overexpression may act as one mechanism of cellular resistance to VPA in TNBC and that its inhibition may enhance the therapeutic efficacy of VPA-chemotherapeutic drug combinations.Copyright © 2023 Granit Mizrahi, Gugenheim, Hamad, Hamed, Tetro, Maimon, Khutsurauli, Nechushtan, Nisman, Duran, Samman, Birimberg-Schwartz, Grunewald, Eyal and Peretz.