Genetic and genomic architecture of the evolution of resistance to antifungal drug combinations.
Genetic and genomic architecture of the evolution of resistance to antifungal drug combinations.
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DOI：
10.1371/journal.pgen.1003390
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发表时间：
2013-04
期刊：
影响因子：
4.5
通讯作者：
Cowen LE
中科院分区：
文献类型：
--
作者：
Hill JA;Ammar R;Torti D;Nislow C;Cowen LE
真菌病原体耐药性的演变损害了有限数量的抗真菌药物的疗效。药物组合已成为增强抗真菌疗效和消除耐药性的强有力策略，但对耐药性演变的影响在很大程度上尚未探索。靶向分子伴侣Hsp90或其下游效应物，蛋白磷酸酶钙调磷酸酶，消除了对最广泛使用的抗真菌剂，唑类，其抑制麦角固醇生物合成的耐药性。在这里，我们进化的模型酵母酿酒酵母和领先的人类真菌病原体白色念珠菌与唑和热休克蛋白90，格尔德霉素，或钙调神经磷酸酶，FK 506抑制剂的实验人群。为了概括其中Hsp90或钙调磷酸酶抑制剂可与唑类组合使用以使抗性病原体对治疗有反应的临床背景，进化实验以依赖于Hsp90和钙调磷酸酶的方式用对唑类有抗性的菌株开始。在最初的290个谱系中，大多数都灭绝了，但有14个进化出了对药物组合的抗性。在五个进化谱系中鉴定并验证了赋予格尔德霉素或FK506抗性的药物靶点突变。全基因组测序鉴定了编码药物外排泵转录激活因子PDR1的基因和编码麦角固醇生物合成基因MOT3的转录抑制因子的基因中的突变，所述突变将两个谱系的唑类抗性从依赖于钙调磷酸酶转变为独立于该调节剂。耐药性也是由截短钙调磷酸酶催化亚基的突变和编码鞘脂生物合成酶的LCB 1突变引起的。基因组分析显示，在四个C。白色念珠菌谱系。因此，我们确定了唑类耐药从钙调磷酸酶依赖性向独立性转变的分子决定因素，并建立了多种耐药机制，为预测和预防耐药的演变提供了基础。真菌感染是全世界死亡的主要原因，并且由于抗真菌药物数量有限而难以治疗，其有效性因耐药性的出现而受到影响。对抗耐药性的一个强有力的策略是联合治疗。抑制分子伴侣Hsp90或其下游效应物钙调磷酸酶削弱真菌应激反应并消除耐药性。在这里，我们提供了第一个分析的遗传和基因组的变化，支持抗真菌药物组合在领先的人类真菌病原体，白色念珠菌，和模型酵母，酿酒酵母的耐药性的演变。我们进化的实验人群与Hsp90或钙调神经磷酸酶的抑制剂和临床上最广泛使用的抗真菌剂，唑类，抑制麦角甾醇生物合成的组合。我们利用全基因组测序来鉴定对药物组合产生耐药性的14个谱系中的不同耐药性突变。这些包括基因突变编码的药物靶标，多药转运蛋白的转录调节因子，麦角固醇生物合成酶的转录抑制因子，和鞘脂生物合成的调节因子。我们还发现了广泛的非整倍体在几个C。白色念珠菌谱系。我们的研究揭示了多种机制，通过这些机制，对药物组合的耐药性可以演变，从而提出了对抗耐药性的新策略。
The evolution of drug resistance in fungal pathogens compromises the efficacy of the limited number of antifungal drugs. Drug combinations have emerged as a powerful strategy to enhance antifungal efficacy and abrogate drug resistance, but the impact on the evolution of drug resistance remains largely unexplored. Targeting the molecular chaperone Hsp90 or its downstream effector, the protein phosphatase calcineurin, abrogates resistance to the most widely deployed antifungals, the azoles, which inhibit ergosterol biosynthesis. Here, we evolved experimental populations of the model yeast Saccharomyces cerevisiae and the leading human fungal pathogen Candida albicans with azole and an inhibitor of Hsp90, geldanamycin, or calcineurin, FK506. To recapitulate a clinical context where Hsp90 or calcineurin inhibitors could be utilized in combination with azoles to render resistant pathogens responsive to treatment, the evolution experiment was initiated with strains that are resistant to azoles in a manner that depends on Hsp90 and calcineurin. Of the 290 lineages initiated, most went extinct, yet 14 evolved resistance to the drug combination. Drug target mutations that conferred resistance to geldanamycin or FK506 were identified and validated in five evolved lineages. Whole-genome sequencing identified mutations in a gene encoding a transcriptional activator of drug efflux pumps, PDR1, and a gene encoding a transcriptional repressor of ergosterol biosynthesis genes, MOT3, that transformed azole resistance of two lineages from dependent on calcineurin to independent of this regulator. Resistance also arose by mutation that truncated the catalytic subunit of calcineurin, and by mutation in LCB1, encoding a sphingolipid biosynthetic enzyme. Genome analysis revealed extensive aneuploidy in four of the C. albicans lineages. Thus, we identify molecular determinants of the transition of azole resistance from calcineurin dependence to independence and establish multiple mechanisms by which resistance to drug combinations evolves, providing a foundation for predicting and preventing the evolution of drug resistance. Fungal infections are a leading cause of mortality worldwide and are difficult to treat due to the limited number of antifungal drugs, whose effectiveness is compromised by the emergence of drug resistance. A powerful strategy to combat drug resistance is combination therapy. Inhibiting the molecular chaperone Hsp90 or its downstream effector calcineurin cripples fungal stress responses and abrogates drug resistance. Here we provide the first analysis of the genetic and genomic changes that underpin the evolution of resistance to antifungal drug combinations in the leading human fungal pathogen, Candida albicans, and model yeast, Saccharomyces cerevisiae. We evolved experimental populations with combinations of inhibitors of Hsp90 or calcineurin and the most widely used antifungal in the clinic, the azoles, which inhibit ergosterol biosynthesis. We harnessed whole-genome sequencing to identify diverse resistance mutations among the 14 lineages that evolved resistance to the drug combination. These included mutations in genes encoding the drug targets, a transcriptional regulator of multidrug transporters, a transcriptional repressor of ergosterol biosynthesis enzymes, and a regulator of sphingolipid biosynthesis. We also identified extensive aneuploidies in several C. albicans lineages. Our study reveals multiple mechanisms by which resistance to drug combination can evolve, suggesting new strategies to combat drug resistance.
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影响因子：
11.4
作者：
CARDENAS, ME;MUIR, RS;HEITMAN, J
通讯作者：
HEITMAN, J
影响因子：
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使用下一代 DNA 测序数据进行变异发现和基因分型的框架。
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影响因子：
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通讯作者：
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