Department of Medical Microbiology and Infection Control, Amsterdam UMC, Location VU Medical Center, Amsterdam, The Netherlands.
Department of Chemistry and Pharmaceutical Sciences, Amsterdam Institute of Molecular and Life Sciences (AIMMS), Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Antimicrob Agents Chemother. 2021 Mar 18;65(4). doi: 10.1128/AAC.01445-20.
Screening strategies for antituberculosis compounds using are time consuming and require biosafety level 3 (BSL3) facilities, which makes the development of high-throughput assays difficult and expensive. , a close genetic relative of , possesses several advantages as a suitable model for tuberculosis drug screening. However, despite the high genetic similarity, there are some obvious differences in susceptibility to some tuberculosis drugs between these two species, especially for the prodrugs ethionamide and isoniazid. In this study, we aimed to improve as a model for antituberculosis drug identification by heterologous expression of two common drug activators, EthA and KatG. These two activators were overexpressed in , and the strains were tested against ethionamide, isoniazid, and a library of established antimycobacterial compounds from TB Alliance to compare drug susceptibility. Both and using zebrafish larvae, these genetically modified strains showed significantly higher susceptibility against ethionamide and isoniazid, which require activation by EthA and KatG. More importantly, a strain overexpressing both and was potentially more susceptible to approximately 20% of the antituberculosis hit compounds from the TB Alliance library. Most of these compounds were activated by EthA in Four of these compounds were selected for further analysis, and three of them showed obvious EthA-dependent activity against Overall, our developed strains are valuable tools for high-throughput discovery of potential novel antituberculosis prodrugs.
使用 筛选抗结核化合物的策略既耗时又需要生物安全 3 级 (BSL3) 设施，这使得高通量检测方法的开发既困难又昂贵。 作为 结核分枝杆菌的近亲，具有作为结核药物筛选合适模型的几个优点。然而，尽管遗传相似度很高，但这两个物种对某些抗结核药物的敏感性存在一些明显差异，特别是对前药乙硫异烟胺和异烟肼。在这项研究中，我们旨在通过异源表达两种常见的药物激活剂 EthA 和 KatG 来改善 作为抗结核药物鉴定的模型。这两种激活剂在 中过表达，并对乙硫异烟胺、异烟肼和 TB 联盟的一系列已建立的抗分枝杆菌化合物库进行了测试，以比较药物敏感性。使用斑马鱼幼虫，这两种遗传修饰的 菌株对乙硫异烟胺和异烟肼的敏感性显著提高，这两种药物需要 EthA 和 KatG 的激活。更重要的是，同时过表达 和 的菌株对 TB 联盟文库中约 20%的抗结核命中化合物具有潜在的更高敏感性。这些化合物中的大多数在 中被 EthA 激活。其中四种化合物被选中进行进一步分析，其中三种对 表现出明显的 EthA 依赖性活性。总体而言，我们开发的 菌株是高通量发现潜在新型抗结核前药的有价值工具。
