Automated high-content live animal drug screening using C. elegans expressing the aggregation prone serpin α1-antitrypsin Z.
Automated high-content live animal drug screening using C. elegans expressing the aggregation prone serpin α1-antitrypsin Z.
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使用表达易于聚集的丝氨酸蛋白酶抑制剂 α1-抗胰蛋白酶 Z 的秀丽隐杆线虫进行自动化高内涵活体动物药物筛选。
DOI：
10.1371/journal.pone.0015460
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发表时间：
2010-11-12
期刊：
影响因子：
3.7
通讯作者：
Pak SC
中科院分区：
文献类型：
--
作者：
Gosai SJ;Kwak JH;Luke CJ;Long OS;King DE;Kovatch KJ;Johnston PA;Shun TY;Lazo JS;Perlmutter DH;Silverman GA;Pak SC
开发适合活体动物生物活性化合物筛选的临床前模型是发现由错误折叠和易聚集蛋白质引起的疾病的有效药理治疗的一种有吸引力的方法。然而，一般来说，活体动物药物筛选是劳动和资源密集型的，并且由于缺乏可靠的分析设计和高通量工作流程而受到阻碍。由于秀丽隐杆线虫体积小，组织透明，易于培养，因此使用秀丽隐杆线虫可以避免许多与活体动物药物筛选相关的技术障碍。此外，秀丽隐杆线虫的遗传可追溯性和对人类疾病分子和细胞基础的深入了解，应该使其成为体内药物发现活动的理想模型系统。本研究的目的是确定秀丽隐杆线虫是否可以适应类似于基于细胞的系统开发的高通量和高含量药物筛选策略。利用表达荧光标记蛋白的转基因动物，我们首先利用集成了图像采集和数据分析模块的自动荧光显微镜平台开发了一个高质量，高通量的工作流程，以定性地评估不同的生物过程，包括生长，组织发育，细胞活力和自噬。接下来，我们采用该技术进行了小分子筛选，并确定了改变α1-抗胰蛋白酶缺乏症中导致肝脏疾病的人类聚集倾向突变体细胞内积累的化合物。本研究通过在高含量成像平台上筛选模拟α1-抗胰蛋白酶缺乏症和其他复杂疾病表型的活秀丽隐杆线虫，为临床前药物发现活动的进展提供了强有力的验证。
The development of preclinical models amenable to live animal bioactive compound screening is an attractive approach to discovering effective pharmacological therapies for disorders caused by misfolded and aggregation-prone proteins. In general, however, live animal drug screening is labor and resource intensive, and has been hampered by the lack of robust assay designs and high throughput work-flows. Based on their small size, tissue transparency and ease of cultivation, the use of C. elegans should obviate many of the technical impediments associated with live animal drug screening. Moreover, their genetic tractability and accomplished record for providing insights into the molecular and cellular basis of human disease, should make C. elegans an ideal model system for in vivo drug discovery campaigns. The goal of this study was to determine whether C. elegans could be adapted to high-throughput and high-content drug screening strategies analogous to those developed for cell-based systems. Using transgenic animals expressing fluorescently-tagged proteins, we first developed a high-quality, high-throughput work-flow utilizing an automated fluorescence microscopy platform with integrated image acquisition and data analysis modules to qualitatively assess different biological processes including, growth, tissue development, cell viability and autophagy. We next adapted this technology to conduct a small molecule screen and identified compounds that altered the intracellular accumulation of the human aggregation prone mutant that causes liver disease in α1-antitrypsin deficiency. This study provides powerful validation for advancement in preclinical drug discovery campaigns by screening live C. elegans modeling α1-antitrypsin deficiency and other complex disease phenotypes on high-content imaging platforms.
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影响因子：
4.7
作者：
Artal-Sanz, Marto;de Jong, Liesbeth;Tavernarakis, Nektarios
通讯作者：
Tavernarakis, Nektarios
影响因子：
6.7
作者：
Breger J;Fuchs BB;Aperis G;Moy TI;Ausubel FM;Mylonakis E
通讯作者：
Mylonakis E
影响因子：
46.9
作者：
Dantuma, NP;Lindsten, K;Masucci, MG
通讯作者：
Masucci, MG
影响因子：
64.8
作者：
DRISCOLL, M;CHALFIE, M
通讯作者：
CHALFIE, M
DOI：
10.1073/pnas.96.1.179
发表时间：
1999-01-05
影响因子：
11.1
作者：
Faber, PW;Alter, JR;Hart, AC
通讯作者：
Hart, AC