肾上腺皮质癌（ACC）是一种侵袭性内分泌恶性肿瘤，治疗选择有限。使用米托坦这一基础疗法治疗晚期 ACC 仍然具有挑战性，因此强调了在治疗前预测米托坦反应并寻求其他有效治疗策略的重要性。我们的目的是通过使用患者来源的 ACC 细胞进行体外测定来确定米托坦的疗效对17个PDC进行了体外米托坦敏感性试验，对8个PDC进行了40种化合物的高通量筛选。使用外显子组和转录组测序对 9 个样本的遗传特征进行了评估。PDC 对米托坦治疗表现出不同的敏感性。应答者 (n=8) 和无应答者 (n=9) 的中位细胞活力抑制率分别为 48.4% (IQR: 39.3-59.3%) 和 -1.2% (IQR: -26.4-22.1%)。应答者的中位 IC50 和 AUC 显着较低（IC50：53.4 µM vs 74.7 µM，P<0.0001；AUC：158.0 vs 213.5，P<0.0001）。基因组分析显示 CTNNB1 体细胞改变仅在有反应者中发现 (3/5)，而 ZNRF3 改变仅在无反应者中发现 (3/4)。转录组分析发现，响应肿瘤中与脂质代谢相关的通路上调，而 CYP27A1 和 ABCA1 表达与体外米托坦敏感性呈正相关。此外，药理学分析发现，包括双硫仑、氯硝柳胺和硼替佐米在内的化合物对 PDC 表现出功效。ACC PDC 可用于测试药物反应、药物再利用和指导个性化治疗。我们的结果表明对米托坦的反应可能与脂质代谢的依赖性有关。 CYP27A1 和 ABCA1 表达可能是米托坦反应的预测标志物，双硫仑、氯硝柳胺和硼替佐米可能是潜在的治疗方法，两者都值得进一步研究。版权所有 © 2024 张，吴，苏，刘，江，江，吴，陈，李，郑、孙、彭、韩、宁、叶、王。

Adrenocortical carcinoma (ACC) is an aggressive endocrine malignancy with limited therapeutic options. Treating advanced ACC with mitotane, the cornerstone therapy, remains challenging, thus underscoring the significance to predict mitotane response prior to treatment and seek other effective therapeutic strategies.We aimed to determine the efficacy of mitotane via an in vitro assay using patient-derived ACC cells (PDCs), identify molecular biomarkers associated with mitotane response and preliminarily explore potential agents for ACC.In vitro mitotane sensitivity testing was performed in 17 PDCs and high-throughput screening against 40 compounds was conducted in 8 PDCs. Genetic features were evaluated in 9 samples using exomic and transcriptomic sequencing.PDCs exhibited variable sensitivity to mitotane treatment. The median cell viability inhibition rate was 48.4% (IQR: 39.3-59.3%) and -1.2% (IQR: -26.4-22.1%) in responders (n=8) and non-responders (n=9), respectively. Median IC50 and AUC were remarkably lower in responders (IC50: 53.4 µM vs 74.7 µM, P<0.0001; AUC: 158.0 vs 213.5, P<0.0001). Genomic analysis revealed CTNNB1 somatic alterations were only found in responders (3/5) while ZNRF3 alterations only in non-responders (3/4). Transcriptomic profiling found pathways associated with lipid metabolism were upregulated in responder tumors whilst CYP27A1 and ABCA1 expression were positively correlated to in vitro mitotane sensitivity. Furthermore, pharmacologic analysis identified that compounds including disulfiram, niclosamide and bortezomib exhibited efficacy against PDCs.ACC PDCs could be useful for testing drug response, drug repurposing and guiding personalized therapies. Our results suggested response to mitotane might be associated with the dependency on lipid metabolism. CYP27A1 and ABCA1 expression could be predictive markers for mitotane response, and disulfiram, niclosamide and bortezomib could be potential therapeutics, both warranting further investigation.Copyright © 2024 Zhang, Wu, Su, Liu, Jiang, Jiang, Wu, Chen, Li, Zheng, Sun, Peng, Han, Ning, Ye and Wang.