Barker Katherine S, Santana Darian J, Zhang Qing, Peters Tracy L, Rybak Jeffrey M, Morschhäuser Joachim, Cuomo Christina A, Rogers P David
Department of Pharmacy and Pharmaceutical Sciences, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
Institute of Molecular Infection Biology, University of Würzburg, Würzburg, Germany.
Antimicrob Agents Chemother. 2025 Aug 18:e0030025. doi: 10.1128/aac.00300-25.
has emerged as a fungal pathogen of particular concern owing in part to its propensity to exhibit antifungal resistance, especially to the commonly prescribed antifungal fluconazole. A mutation in , which encodes a zinc cluster transcription factor, has been shown to confer increased resistance to fluconazole. In this work, we aimed to determine how mutations in exert this effect. mutations leading to A640V, A657V, and F862_N866del, found in fluconazole-resistant clinical isolates, were introduced into two susceptible Clade I backgrounds using CRISPR-Cas9 gene editing. These mutations conferred increased fluconazole resistance, as well as increased resistance to other triazoles as measured by broth microdilution. RNA-seq revealed that the ATP-binding cassette (ABC) transporter gene as well as the major facilitator superfamily (MFS) transporter gene were both upregulated in the presence of these mutations. Disruption of increased susceptibility in strains with mutations, whereas disruption of had little to no effect. However, disruption of both and resulted in an additional increase in susceptibility as compared with disruption of alone. mutations, leading to A640V, A657V, and F862_N866del all result in increased resistance to fluconazole and other triazole antifungals and increased expression of both and in . Together, these data suggest is the primary driver of resistance conferred by these mutations.
由于其表现出抗真菌耐药性的倾向，尤其是对常用的抗真菌药物氟康唑耐药，已成为一种特别受关注的真菌病原体。编码锌簇转录因子的基因发生突变，已被证明会导致对氟康唑的耐药性增加。在这项研究中，我们旨在确定该基因中的突变是如何产生这种作用的。利用CRISPR-Cas9基因编辑技术，将在耐氟康唑临床分离株中发现的导致A640V、A657V和F862_N866del的基因突变引入两个I类敏感背景中。这些基因突变导致氟康唑耐药性增加，同时通过肉汤微量稀释法检测发现对其他三唑类药物的耐药性也增加。RNA测序显示，在存在这些基因突变的情况下，ATP结合盒（ABC）转运蛋白基因以及主要易化子超家族（MFS）转运蛋白基因均上调。在带有这些基因突变的菌株中，破坏该基因会增加其易感性，而破坏该基因则几乎没有影响。然而，与单独破坏该基因相比，同时破坏该基因和该基因会导致易感性进一步增加。导致A640V、A657V和F862_N866del的基因突变均导致对氟康唑和其他三唑类抗真菌药物的耐药性增加，以及该基因和该基因的表达增加。总之，这些数据表明该基因是这些基因突变赋予耐药性的主要驱动因素。
