Vale-Silva Luis A, Moeckli Beat, Torelli Riccardo, Posteraro Brunella, Sanguinetti Maurizio, Sanglard Dominique
Institute of Microbiology, University of Lausanne and University Hospital Center, Lausanne, Switzerland.
Institute of Microbiology, Università Cattolica del Sacro Cuore, Rome, Italy.
mSphere. 2016 Mar 2;1(2). doi: 10.1128/mSphere.00065-15. eCollection 2016 Mar-Apr.
Candida glabrata is the second most common Candida species causing disseminated infection, after C. albicans. C. glabrata is intrinsically less susceptible to the widely used azole antifungal drugs and quickly develops secondary resistance. Resistance typically relies on drug efflux with transporters regulated by the transcription factor Pdr1. Gain-of-function (GOF) mutations in PDR1 lead to a hyperactive state and thus efflux transporter upregulation. Our laboratory has characterized a collection of C. glabrata clinical isolates in which azole resistance was found to correlate with increased virulence in vivo. Contributing phenotypes were the evasion of adhesion and phagocytosis by macrophages and an increased adhesion to epithelial cells. These phenotypes were found to be dependent on PDR1 GOF mutation and/or C. glabrata strain background. In the search for the molecular effectors, we found that PDR1 hyperactivity leads to overexpression of specific cell wall adhesins of C. glabrata. Further study revealed that EPA1 regulation, in particular, explained the increase in adherence to epithelial cells. Deleting EPA1 eliminates the increase in adherence in an in vitro model of interaction with epithelial cells. In a murine model of urinary tract infection, PDR1 hyperactivity conferred increased ability to colonize the bladder and kidneys in an EPA1-dependent way. In conclusion, this study establishes a relationship between PDR1 and the regulation of cell wall adhesins, an important virulence attribute of C. glabrata. Furthermore, our data show that PDR1 hyperactivity mediates increased adherence to host epithelial tissues both in vitro and in vivo through upregulation of the adhesin gene EPA1. IMPORTANCE Candida glabrata is an important fungal pathogen in human diseases and is also rapidly acquiring drug resistance. Drug resistance can be mediated by the transcriptional activator PDR1, and this results in the upregulation of multidrug transporters. Intriguingly, this resistance mechanism is associated in C. glabrata with increased virulence in animal models and also with increased adherence to specific host cell types. The C. glabrata adhesin gene EPA1 is a major contributor of virulence and adherence to host cells. Here, we show that EPA1 expression is controlled by PDR1 independently of subtelomeric silencing, a known EPA1 regulation mechanism. Thus, a relationship exists between PDR1, EPA1 expression, and adherence to host cells, which is critical for efficient virulence. Our results demonstrate that acquisition of drug resistance is beneficial for C. glabrata in fungus-host relationships. These findings further highlight the challenges of the therapeutic management of C. glabrata infections in human patients.
光滑念珠菌是引起播散性感染的第二常见念珠菌属物种,仅次于白色念珠菌。光滑念珠菌对广泛使用的唑类抗真菌药物天然敏感性较低,并迅速产生继发性耐药性。耐药性通常依赖于由转录因子Pdr1调控的转运蛋白介导的药物外排。PDR1中的功能获得性(GOF)突变导致高活性状态,从而使外排转运蛋白上调。我们实验室对一组光滑念珠菌临床分离株进行了特征分析,发现其中的唑类耐药性与体内毒力增加相关。相关表型包括逃避巨噬细胞的黏附和吞噬作用以及对上皮细胞黏附增加。这些表型被发现依赖于PDR1 GOF突变和/或光滑念珠菌菌株背景。在寻找分子效应器的过程中,我们发现PDR1的高活性导致光滑念珠菌特定细胞壁黏附素的过表达。进一步研究表明,特别是EPA1的调控解释了对上皮细胞黏附的增加。在与上皮细胞相互作用的体外模型中删除EPA1可消除黏附的增加。在尿路感染的小鼠模型中,PDR1的高活性以EPA1依赖的方式赋予了在膀胱和肾脏中定殖的能力增强。总之,本研究确立了PDR1与细胞壁黏附素调控之间的关系,这是光滑念珠菌的一个重要毒力属性。此外,我们的数据表明,PDR1的高活性通过上调黏附素基因EPA1在体外和体内介导了对宿主上皮组织黏附的增加。重要性 光滑念珠菌是人类疾病中的一种重要真菌病原体,并且也在迅速获得耐药性。耐药性可由转录激活因子PDR1介导,这导致多药转运蛋白上调。有趣的是,在光滑念珠菌中这种耐药机制与动物模型中毒力增加以及对特定宿主细胞类型黏附增加相关。光滑念珠菌黏附素基因EPA1是毒力和对宿主细胞黏附的主要贡献者。在这里,我们表明EPA1的表达由PDR1控制,独立于端粒旁沉默,这是一种已知的EPA1调控机制。因此,PDR1、EPA1表达和对宿主细胞黏附之间存在一种关系,这对于有效的毒力至关重要。我们的结果表明,获得耐药性对光滑念珠菌在真菌 - 宿主关系中是有益的。这些发现进一步突出了人类患者中光滑念珠菌感染治疗管理的挑战。