A Transcriptomics Approach To Unveiling the Mechanisms of In Vitro Evolution towards Fluconazole Resistance of a Candida glabrata Clinical Isolate.
A Transcriptomics Approach To Unveiling the Mechanisms of In Vitro Evolution towards Fluconazole Resistance of a Candida glabrata Clinical Isolate.
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DOI：
10.1128/aac.00995-18
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发表时间：
2019-01
影响因子：
4.9
通讯作者：
Teixeira MC
中科院分区：
文献类型：
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作者：
Cavalheiro M;Costa C;Silva-Dias A;Miranda IM;Wang C;Pais P;Pinto SN;Mil-Homens D;Sato-Okamoto M;Takahashi-Nakaguchi A;Silva RM;Mira NP;Fialho AM;Chibana H;Rodrigues AG;Butler G;Teixeira MC
光滑念珠菌是一种新兴的真菌病原体。其患病率的增加与其迅速发展抗真菌药物耐药性的能力有关，特别是对唑类。光滑念珠菌是一种新兴的真菌病原体。其患病率的增加与其迅速发展抗真菌药物耐药性的能力有关，特别是对唑类。为了揭示唑类耐药背后的新分子机制，对一个C。通过与氟康唑长时间孵育诱导的对泊沙康唑耐药（第21天）、克霉唑耐药（第31天）和氟康唑和伏立康唑耐药（第45天）的唑类敏感性，对光滑念珠菌临床分离物（分离物044）进行了研究。发现所有进化的菌株积累的唑类药物浓度低于亲本菌株，而麦角甾醇浓度基本保持不变。然而，只有对所有唑类药物都表现出耐药性的人群被发现在C. glabrata PDR 1基因，导致编码多药耐药转运蛋白的基因上调。显示泊沙康唑/克霉唑耐药和氟康唑/伏立康唑MIC水平增加的中间菌株被发现显示出耐唑类药物的替代途径。特别是，泊沙康唑/克霉唑耐药31天后与粘附素基因的表达增加。这一发现使我们将Epa 3粘附素鉴定为唑类耐药的新决定因素。除了生物膜形成所需外，发现Epa 3表达减少唑类抗真菌药物的细胞内积累。总之，这项工作提供了一个C。glabrata人群对多唑耐药，突出了获得唑类耐药的多因素性质，并指出了唑类耐药的新参与者。
Candida glabrata is an emerging fungal pathogen. Its increased prevalence is associated with its ability to rapidly develop antifungal drug resistance, particularly to azoles. Candida glabrata is an emerging fungal pathogen. Its increased prevalence is associated with its ability to rapidly develop antifungal drug resistance, particularly to azoles. In order to unravel new molecular mechanisms behind azole resistance, a transcriptomics analysis of the evolution of a C. glabrata clinical isolate (isolate 044) from azole susceptibility to posaconazole resistance (21st day), clotrimazole resistance (31st day), and fluconazole and voriconazole resistance (45th day), induced by longstanding incubation with fluconazole, was carried out. All the evolved strains were found to accumulate lower concentrations of azole drugs than the parental strain, while the ergosterol concentration remained mostly constant. However, only the population displaying resistance to all azoles was found to have a gain-of-function mutation in the C. glabrata PDR1 gene, leading to the upregulation of genes encoding multidrug resistance transporters. Intermediate strains, exhibiting posaconazole/clotrimazole resistance and increased fluconazole/voriconazole MIC levels, were found to display alternative ways to resist azole drugs. Particularly, posaconazole/clotrimazole resistance after 31 days was correlated with increased expression of adhesin genes. This finding led us to identify the Epa3 adhesin as a new determinant of azole resistance. Besides being required for biofilm formation, Epa3 expression was found to decrease the intracellular accumulation of azole antifungal drugs. Altogether, this work provides a glimpse of the transcriptomics evolution of a C. glabrata population toward multiazole resistance, highlighting the multifactorial nature of the acquisition of azole resistance and pointing out a new player in azole resistance.