Department of Neurosurgery, First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China; Department of Neurosurgery, Center of Pituitary Tumour, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Neurosurgery, Center of Pituitary Tumour, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Int Immunopharmacol. 2024 Jun 15;134:112159. doi: 10.1016/j.intimp.2024.112159. Epub 2024 Apr 30.
Medical treatment plays a critical role in pituitary neuroendocrine tumour (PitNET) treatment. Dopamine agonists and somatostatin receptor agonists are the only known drugs for effectively treating PitNET. Thus, the identification of potential therapeutic targets and drugs is urgently needed.
To discover potential drugs that can suppress PitNET growth and to further investigate the underlying mechanism involved.
High-throughput drug screening of primary cultures of 17 patient-derived PitNETs was performed to identify potential therapeutic compounds. Cell viability assays, Western blot analysis and flow cytometry were used to investigate pituitary neuroendocrine tumour cell lines and patient-derived PitNET cultures in vitro. In vivo drug efficacy was examined in a mouse xenograft model.
Seventeen primary PitNET samples were collected for high-throughput drug screening, and a class of copper ionophores that can effectively inhibit cell growth, such as zinc pyrithione, elesclomol, and disulfiram (DSF), was identified. Subsequent experiments initially validated the dose-dependent cell growth-suppressing effect of these copper ionophores on AtT20, GH3, and MMQ cells and several primary PitNET cell lines. Moreover, we confirmed that the cytotoxic effect of DSF depends on the presence of copper. Additionally, we determined that cell death occurs via cuproptosis, with events such as Fe-S cluster protein loss, dihydrolipoyl transacetylase oligomerization and heat shock protein 70 upregulation. Finally, we verified the cytotoxic effects of DSF in vivo.
The present study revealed copper ionophores as a potential class of drugs for PitNET treatment. DSF induced PitNET cell death via cuproptosis and might be a promising option for PitNET therapy.
医学治疗在垂体神经内分泌肿瘤（PitNET）的治疗中起着关键作用。多巴胺激动剂和生长抑素受体激动剂是有效治疗 PitNET 的唯一已知药物。因此，迫切需要发现潜在的治疗靶点和药物。
发现潜在的可以抑制 PitNET 生长的药物，并进一步研究其潜在的作用机制。
对 17 例患者来源的 PitNET 的原代培养物进行高通量药物筛选，以鉴定潜在的治疗化合物。细胞活力测定、Western blot 分析和流式细胞术用于研究垂体神经内分泌肿瘤细胞系和患者来源的 PitNET 培养物的体外情况。在小鼠异种移植模型中检查了药物的体内疗效。
对 17 例原发性 PitNET 样本进行了高通量药物筛选，鉴定出了一类可有效抑制细胞生长的铜离子载体，如锌吡啶硫酮、依立洛酶和双硫仑（DSF）。随后的实验初步验证了这些铜离子载体对 AtT20、GH3 和 MMQ 细胞以及几种原代 PitNET 细胞系的剂量依赖性细胞生长抑制作用。此外，我们证实了 DSF 的细胞毒性作用依赖于铜的存在。此外，我们确定细胞死亡是通过铜死亡发生的，伴随着 Fe-S 簇蛋白丢失、二氢乳清酸转乙酰酶寡聚化和热休克蛋白 70 上调等事件。最后，我们验证了 DSF 在体内的细胞毒性作用。
本研究揭示了铜离子载体作为一种治疗 PitNET 的潜在药物类别。DSF 通过铜死亡诱导 PitNET 细胞死亡，可能是 PitNET 治疗的一种有前途的选择。
