Pountain Andrew W, Barrett Michael P
Wellcome Center for Integrative Parasitology, University of Glasgow, Glasgow, G12 8TA, UK.
Department of Microbiology and Molecular Genetics, McGovern Medical School, University of Texas Health Science Center at Houston, Houston, Texas, 77030, USA.
Wellcome Open Res. 2019 Nov 13;4:176. doi: 10.12688/wellcomeopenres.15452.1. eCollection 2019.
: Protozoan parasites are responsible for a range of clinical infections that represent a substantial challenge for global health. Amphotericin B (AmB) is increasingly used to treat infection, so understanding the potential for resistance to this drug is an important priority. Previously we described four independently-derived AmB-resistant lines that exhibited resistance-associated genetic lesions resulting in altered sterol content. However, substantial phenotypic variation between these lines, including differences in virulence attributes, were not fully explained by these changes. To identify alterations in cellular metabolism potentially related to phenotypic differences between wild-type and AmB-resistant lines, we extracted metabolites and performed untargeted metabolomics by liquid chromatography-mass spectrometry. We observed substantial differences in metabolite abundance between lines, arising in an apparently stochastic manner. Concerted remodeling of central carbon metabolism was not observed; however, in three lines, decreased abundance of several oligohexoses was observed. Given that the oligomannose mannogen is an important virulence factor in , this could relate to loss of virulence in these lines. Increased abundance of the reduced forms of the oxidative stress-protective thiols trypanothione and glutathione was also observed in multiple lines. This dataset will provide a useful resource for understanding the molecular basis of drug resistance in , and suggests a role for metabolic changes separate from the primary mechanism of drug resistance in determining the phenotypic profile of parasite lines subjected to experimental selection of resistance.
原生动物寄生虫会引发一系列临床感染，这对全球健康构成了重大挑战。两性霉素B（AmB）越来越多地用于治疗感染，因此了解对该药物产生耐药性的可能性是一项重要的优先事项。此前我们描述了四个独立衍生的对AmB耐药的细胞系，它们表现出与耐药相关的基因损伤，导致甾醇含量改变。然而，这些细胞系之间存在显著的表型差异，包括毒力属性的差异，这些变化并不能完全解释这些差异。为了确定可能与野生型和AmB耐药细胞系之间表型差异相关的细胞代谢变化，我们提取了代谢物，并通过液相色谱-质谱法进行了非靶向代谢组学分析。我们观察到细胞系之间代谢物丰度存在显著差异，且这种差异显然是以随机方式出现的。未观察到中心碳代谢的协同重塑；然而，在三个细胞系中，观察到几种低聚己糖的丰度降低。鉴于低聚甘露糖甘露原是[某种寄生虫]中的一种重要毒力因子，这可能与这些细胞系中毒力的丧失有关。在多个细胞系中还观察到氧化应激保护硫醇锥虫硫醇和谷胱甘肽的还原形式丰度增加。该数据集将为理解[某种寄生虫]耐药性的分子基础提供有用的资源，并表明在确定经过耐药性实验选择的寄生虫细胞系的表型特征时，代谢变化在耐药性主要机制之外发挥了作用。
