Microprinted tumor spheroids enable anti-cancer drug screening.
Microprinted tumor spheroids enable anti-cancer drug screening.
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微印刷肿瘤球体可用于抗癌药物筛选。
DOI：
10.1109/embc.2016.7591647
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Tavana,Hossein
中科院分区：
文献类型：
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作者：
Thakuri,PradipS;Ham,StephanieL;Tavana,Hossein
球体是无血管肿瘤的生物学相关模型，也是发现抗癌药物的独特工具。尽管在研究实验室中使用了几十年，但球状体并没有常规地用于药物发现，主要是由于大规模生产均匀尺寸的球状体的困难以及处理、药物治疗和分析它们所涉及的密集劳动。我们使用一种新技术克服了这一障碍，在标准384孔板中机器人微打印球状体。将含有癌细胞的水性液滴分配到第二不混溶水相的浴中。液滴保持细胞紧密靠近以聚集成单个球状体。使用U-87 MG脑癌细胞，我们表明，这种方法产生的球体的明确定义的大小与其平均直径的偏差为± 10%。我们证明了机器人的可行性，高通量化合物筛选肿瘤球体使用的25个标准的化疗药物和分子抑制剂对U-87 MG球体的集合。每种药物的使用浓度范围很广。使用PrestoBlue测定法测量药物处理的球状体中癌细胞的活力。形态学变化被用作分析药物作用的次要指标。我们确定了几种化合物，有效地抑制球体的生长。为了生成药物有效性的评分系统，我们使用半最大抑制浓度（IC 50）、最大抑制（Emax）和剂量-反应曲线下面积（AUC）来呈现考虑药物的效力和功效的多参数方法。我们的机器人技术提供了一个低成本和方便的平台，用于在昂贵和繁琐的体内测试之前针对真实的肿瘤模型筛选大量化合物，从而大大提高测试通量和效率，并降低成本。
Spheroids present a biologically relevant model of avascular tumors and a unique tool for discovery of anti-cancer drugs. Despite being used in research laboratories for several decades, spheroids are not routinely used for drug discovery primarily due to the difficulty of mass-producing uniformly-sized spheroids and intense labor involved in handling, drug treatment, and analyzing them. We overcome this barrier using a novel technology to robotically microprint spheroids in standard 384-well plates. An aqueous drop containing cancer cells is dispensed into a bath of a second, immiscible aqueous phase. The drop maintains cells in close proximity to aggregate into a single spheroid. Using U-87 MG brain cancer cells, we show that this approach produces spheroids of well-defined size with ∼10% deviation from their mean diameter. We demonstrate the feasibility of robotic, high throughput compound screening against tumor spheroids using a collection of 25 standard chemotherapeutics and molecular inhibitors against U-87 MG spheroids. Each drug is used in a wide range of concentrations. Viability of cancer cells in drug-treated spheroids is measured using a PrestoBlue assay. Morphological changes are used as a secondary measure for analysis of drug effect. We identify several compounds that effectively inhibit growth of spheroids. To generate a scoring system for effectiveness of drugs, we use half-maximum inhibitory concentration (IC50), maximum inhibition (Emax), and area under the dose-response curve (AUC) to present a multi-parametric approach that takes into account both potency and efficacy of drugs. Our robotic technology offers a low cost and convenient platform for screening large collections of chemical compounds against realistic tumor models prior to expensive and tedious in vivo tests, dramatically improving testing throughput and efficiency, and reducing costs.