Phenotypic and transcriptional profiling in Entamoeba histolytica reveal costs to fitness and adaptive responses associated with metronidazole resistance
Phenotypic and transcriptional profiling in Entamoeba histolytica reveal costs to fitness and adaptive responses associated with metronidazole resistance
复制标题
DOI：
10.3389/fmib.2015.00154
复制
发表时间：
2015-05-05
影响因子：
5.2
通讯作者：
Nozaki, Tomoyoshi
中科院分区：
文献类型：
--
作者：
Penuliar, Gil M.;Nakada-Tsukui, Kumiko;Nozaki, Tomoyoshi
抗微生物化疗在对抗由溶组织内阿米巴引起的传染病中至关重要。在可用于治疗阿米巴病的药物中，甲硝唑（MTZ）被认为是首选药物。最近，体外研究已经描述了MTZ耐药性和相关的潜在机制。然而，与抗性相关的适应性和适应性反应的成本尚未调查。在这项研究中，我们产生了HM-1衍生的菌株耐12 μ M MTZ（MTZR）。我们研究了它的表型和转录谱，以确定与MTZ抗性相关的后果和mRNA水平变化。我们的研究结果表明，细胞大小和粒度增加，细胞分裂，粘附，吞噬作用，细胞致病性和葡萄糖消耗率下降。转录组分析显示MTZR中有142个差异表达基因。与其他MTZ耐药寄生虫相比，MTZR没有下调丙酮酸：铁氧还蛋白氧化还原酶，但表现出一种假设蛋白（HP 1）和几种铁硫黄素蛋白的基因表达增加，以及富含亮氨酸蛋白的基因下调。Fisher精确检验显示MTZR中有24个显著富集的GO术语，MTZR中调节基因与无MTZ培养的MTZR和有MTZ培养的HM-1中调节基因的3向比较显示88个基因对MTZR特异。总体而言，我们的研究结果表明，MTZ抗性与特定的转录变化和降低寄生虫毒力有关。
Antimicrobial chemotherapy is critical in the fight against infectious diseases caused by Entamoeba histolytica. Among the drugs available for the treatment of amebiasis, metronidazole (MTZ) is considered the drug of choice. Recently, in vitro studies have described MTZ resistance and the potential mechanisms involved. Costs to fitness and adaptive responses associated with resistance, however, have not been investigated. In this study we generated an HM-1 derived strain resistant to 12 mu M MTZ (MTZR). We examined its phenotypic and transcriptional profile to determine the consequences and mRNA level changes associated with MTZ resistance. Our results indicated increased cell size and granularity, and decreased rates in cell division, adhesion, phagocytosis, cytopathogenicity, and glucose consumption. Transcriptome analysis revealed 142 differentially expressed genes in MTZR. In contrast to other MTZ resistant parasites, MTZR did not down-regulate pyruvate:ferredoxin oxidoreductase, but showed increased expression of genes for a hypothetical protein (HP1) and several iron-sulfur flavoproteins, and downregulation of genes for leucine-rich proteins. Fisher's exact test showed 24 significantly enriched GO terms in MTZR, and a 3-way comparison of modulated genes in MTZR against those of MTZR cultured without MTZ and HM-1 cultured with MTZ, showed that 88 genes were specific to MTZR. Overall, our findings suggested that MTZ resistance is associated with specific transcriptional changes and decreased parasite virulence.