High-Throughput Screening for the Identification of New Therapeutic Options for Metastatic Pheochromocytoma and Paraganglioma

Drug repurposing or repositioning is an important part of drug discovery that has been growing in the last few years for the development of therapeutic options in oncology. We applied this paradigm in a screening of a library of about 3,800 compounds (including FDA-approved drugs and pharmacologically active compounds) employing a model of metastatic pheochromocytoma, the most common tumor of the adrenal medulla in children and adults. The collection of approved drugs was screened in quantitative mode, testing the compounds in compound-titration series (dose-response curves). Analysis of the dose-response screening data facilitated the selection of 50 molecules with potential bioactivity in pheochromocytoma cells. These drugs were classified based on molecular/cellular targets and signaling pathways affected, and selected drugs were further validated in a proliferation assay and by flow cytometric cell death analysis. Using meta-analysis information from molecular targets of the top drugs identified by our screening with gene expression data from human and murine microarrays, we identified potential drugs to be used as single drugs or in combination. An example of a combination with a synergistic effect is presented. Our study exemplifies a promising model to identify potential drugs from a group of clinically approved compounds that can more rapidly be implemented into clinical trials in patients with metastatic pheochromocytoma or paraganglioma.

药物重定位（Drug repurposing）是药物研发的重要组成部分，近年来在肿瘤治疗方案开发领域的发展态势愈发显著。我们将该研究范式应用于约3800种化合物组成的化合物库筛选工作，该库涵盖美国食品药品监督管理局（FDA）批准药物及药理学活性化合物，筛选模型采用转移性嗜铬细胞瘤——儿童与成人肾上腺髓质最常见的肿瘤。我们以定量模式对其中的获批药物集合进行测试，通过化合物浓度梯度系列试验（剂量-反应曲线）评估各化合物的活性。对剂量-反应筛选数据的分析帮助我们筛选出50种对嗜铬细胞瘤细胞具有潜在生物活性的分子。上述药物依据其分子/细胞靶点及受影响的信号通路进行分类，随后我们通过增殖实验与流式细胞术细胞死亡分析，对筛选得到的药物开展进一步验证。结合本次筛选得到的顶级药物的分子靶点荟萃分析信息，与人类及小鼠微阵列基因表达数据进行整合分析后，我们确定了可单药应用或联合使用的潜在治疗药物。本文展示了一例具有协同效应的联合用药方案。本研究展示了一种极具前景的研究模型，可从已获批临床使用的化合物库中筛选潜在药物，这类药物能够更快速地推进至转移性嗜铬细胞瘤或副神经节瘤患者的临床试验阶段。