Host Bioenergetic Parameters Reveal Cytotoxicity of Antituberculosis Drugs Undetected Using Conventional Viability Assays.
Host Bioenergetic Parameters Reveal Cytotoxicity of Antituberculosis Drugs Undetected Using Conventional Viability Assays.
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DOI：
10.1128/aac.00932-21
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发表时间：
2021-09-17
影响因子：
4.9
通讯作者：
Steyn AJC
中科院分区：
文献类型：
--
作者：
Cumming BM;Baig Z;Addicott KW;Chen D;Steyn AJC
结核病（TB）药物开发中的高损耗率在很大程度上归因于安全性，这可能是由于使用测量细胞活力的终点测定来检测药物细胞毒性。在癌症、代谢和神经系统疾病以及抗生素的药物开发中，越来越多地使用细胞外通量（XF）分析来评估细胞毒性，该分析真实的时间测量细胞生物能量代谢。在这里，我们采用XF平台来研究目前用于TB治疗的药物对HepG 2细胞、THP-1巨噬细胞和人单核细胞衍生的巨噬细胞（hMDM）的生物能量代谢的细胞毒性。我们发现XF分析揭示了在细胞死亡之前对细胞的生物能量代谢的早期药物诱导作用，通过常规的活力测定来测量。此外，每种细胞类型对药物治疗有不同的反应，这表明在结核病药物开发中应考虑一种以上的细胞类型来检查细胞毒性。有趣的是，对结核分枝杆菌具有不同作用模式的化学上不相关的药物对细胞的生物能量参数具有相似的影响，从而阻碍了基于新化学实体的化学结构和作用模式的潜在细胞毒性的预测。药物诱导的hMDM生物能量参数效应的聚集反映在药物对hMDM中细胞因子产生的效应的聚集中，表明药物对巨噬细胞代谢和功能的效应之间存在协同效应。这些发现可以作为一个基准，以建立XF分析作为一种新的工具，以测定结核病药物开发中的细胞毒性。
High attrition rates in tuberculosis (TB) drug development have been largely attributed to safety, which is likely due to the use of endpoint assays measuring cell viability to detect drug cytotoxicity. In drug development for cancer, metabolic, and neurological disorders and for antibiotics, cytotoxicity is increasingly being assessed using extracellular flux (XF) analysis, which measures cellular bioenergetic metabolism in real time. Here, we adopt the XF platform to investigate the cytotoxicity of drugs currently used in TB treatment on the bioenergetic metabolism of HepG2 cells, THP-1 macrophages, and human monocyte-derived macrophages (hMDMs). We found that the XF analysis reveals earlier drug-induced effects on the cells’ bioenergetic metabolism prior to cell death, measured by conventional viability assays. Furthermore, each cell type has a distinct response to drug treatment, suggesting that more than one cell type should be considered to examine cytotoxicity in TB drug development. Interestingly, chemically unrelated drugs with different modes of action on Mycobacterium tuberculosis have similar effects on the bioenergetic parameters of the cells, thus discouraging the prediction of potential cytotoxicity based on chemical structure and mode of action of new chemical entities. The clustering of the drug-induced effects on the hMDM bioenergetic parameters are reflected in the clustering of the effects of the drugs on cytokine production in hMDMs, demonstrating concurrence between the effects of the drugs on the metabolism and functioning of the macrophages. These findings can be used as a benchmark to establish XF analysis as a new tool to assay cytotoxicity in TB drug development.