Experimental Resistance to Drug Combinations in Leishmania donovani: Metabolic and Phenotypic Adaptations
Experimental Resistance to Drug Combinations in Leishmania donovani: Metabolic and Phenotypic Adaptations
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DOI：
10.1128/aac.04231-14
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发表时间：
2015-04-01
影响因子：
4.9
通讯作者：
Gamarro, Francisco
中科院分区：
文献类型：
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作者：
Berg, Maya;Garcia-Hernandez, Raquel;Gamarro, Francisco
与病媒控制一起，化疗是控制内脏利什曼病（VL）的重要工具，但其疗效因一线药物耐药性增加和治疗失败而受到损害。为了延迟耐药性的出现，在治疗内脏利什曼病（VL）的临床试验中系统地测试了现有抗利什曼病药物的药物组合的使用。在体外，杜氏利什曼原虫前鞭毛体能够在药物压力10周后对不同抗利什曼原虫药物的几种组合产生实验性耐药性。使用非靶向液相色谱-质谱（LC-MS）代谢组学方法，我们确定了实验上对药物组合耐药的品系及其各自的单一耐药品系的代谢变化。这突出了集体代谢变化（在所有联合治疗耐药[CTR]系中发现）和特定代谢变化（在某些CTR系中发现）。我们证明，单耐药和CTR寄生虫细胞系显示出不同的代谢适应，这些适应都集中在实验验证的相同防御机制上：保护免受药物诱导的和外部氧化应激以及膜流动性的变化。膜流动性的变化伴随着药物吸收的变化，只有在对药物与锑剂的组合具有抗性的品系中，并且令人惊讶的是，这些品系中的药物积累更高。总之，这些结果突出了在不同抗性品系中保护免受氧化应激的重要性和核心作用。最终，这些表型变化可能会干扰目前用于治疗VL的所有药物的作用模式，并应在药物开发中加以考虑。
Together with vector control, chemotherapy is an essential tool for the control of visceral leishmaniasis (VL), but its efficacy is jeopardized by growing resistance and treatment failure against first-line drugs. To delay the emergence of resistance, the use of drug combinations of existing antileishmanial agents has been tested systematically in clinical trials for the treatment of visceral leishmaniasis (VL). In vitro, Leishmania donovani promastigotes are able to develop experimental resistance to several combinations of different antileishmanial drugs after 10 weeks of drug pressure. Using an untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics approach, we identified metabolic changes in lines that were experimentally resistant to drug combinations and their respective single-resistant lines. This highlighted both collective metabolic changes (found in all combination therapy-resistant [CTR] lines) and specific ones (found in certain CTR lines). We demonstrated that single-resistant and CTR parasite cell lines show distinct metabolic adaptations, which all converge on the same defensive mechanisms that were experimentally validated: protection against drug-induced and external oxidative stress and changes in membrane fluidity. The membrane fluidity changes were accompanied by changes in drug uptake only in the lines that were resistant against drug combinations with antimonials, and surprisingly, drug accumulation was higher in these lines. Together, these results highlight the importance and the central role of protection against oxidative stress in the different resistant lines. Ultimately, these phenotypic changes might interfere with the mode of action of all drugs that are currently used for the treatment of VL and should be taken into account in drug development.