A High-Throughput Screen for Tuberculosis Progression
A High-Throughput Screen for Tuberculosis Progression
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DOI：
10.1371/journal.pone.0016779
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发表时间：
2011-02-16
期刊：
影响因子：
3.7
通讯作者：
Spaink, Herman P.
中科院分区：
文献类型：
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作者：
Carvalho, Ralph;de Sonneville, Jan;Spaink, Herman P.
世界三分之一的人口感染结核分枝杆菌，多重耐药菌株正在迅速进化。用于研究结核病进展的全有机体高通量筛选系统的明显缺乏正在迅速成为结核病研究的瓶颈。我们使用斑马鱼海洋分枝杆菌感染模型成功开发了这样一个系统，该模型是一种具有生物医学意义的、具有良好特征的结核病进展模型，模仿了人类结核病病理学的特征。重要的是，我们证明了我们的系统直接研究结核分枝杆菌的适用性，首次表明人类病原体可以在这种脊椎动物模型中传播，从而导致与海洋分枝杆菌感染时观察到的相似的早期疾病症状。我们的系统能够通过早期胚胎的机器人卵黄注射和晚期幼虫的视觉流筛查来筛查疾病进展。我们还表明，该系统可以可靠地重现标准尾静脉注射方法，吞吐量为每小时 2,000 个胚胎。我们还证明了使用高通量水平的反向遗传学研究导致疾病进展的信号转导的可能性。重要的是，我们使用参考化合物来验证我们的系统测试预防结核病进展的分子，使其非常适合在体内研究新型抗结核化合物。
One-third of the world population is infected with Mycobacterium tuberculosis and multi-drug resistant strains are rapidly evolving. The noticeable absence of a whole organism high-throughput screening system for studying the progression of tuberculosis is fast becoming the bottleneck in tuberculosis research. We successfully developed such a system using the zebrafish Mycobacterium marinum infection model, which is a well-characterized model for tuberculosis progression with biomedical significance, mimicking hallmarks of human tuberculosis pathology. Importantly, we demonstrate the suitability of our system to directly study M. tuberculosis, showing for the first time that the human pathogen can propagate in this vertebrate model, resulting in similar early disease symptoms to those observed upon M. marinum infection. Our system is capable of screening for disease progression via robotic yolk injection of early embryos and visual flow screening of late-stage larvae. We also show that this system can reliably recapitulate the standard caudal vein injection method with a throughput level of 2,000 embryos per hour. We additionally demonstrate the possibility of studying signal transduction leading to disease progression using reverse genetics at high-throughput levels. Importantly, we use reference compounds to validate our system in the testing of molecules that prevent tuberculosis progression, making it highly suited for investigating novel anti-tuberculosis compounds in vivo.