■白色念珠菌对抗真菌药物的耐受性的出现使真菌感染的治疗复杂化。了解这种耐受性的潜在机制对于制定有效的治疗策略至关重要。
■本研究旨在阐明白色念珠菌酮康唑耐受性的遗传和分子基础,关注染色体非整倍体的作用,Hsp90和钙调磷酸酶。
■将野生型白色念珠菌菌株SC5314暴露于增加浓度的酮康唑(0.015-32μg/mL)以选择耐受性衔接子。使用圆盘扩散和斑点测定来评估耐受性。全基因组测序鉴定了衔接子的染色体变化。使用特异性抑制剂和敲除菌株研究了Hsp90和钙调磷酸酶在维持和发展酮康唑耐受性中的作用。
■衔接子表现出对酮康唑浓度高达16μg/mL的耐受性,在0.015μg/mL时,亲本菌株的抑制作用显着增加。所有耐受衔接子显示R染色体扩增,有29个适配器有三体,一个有四体。这种非整倍体是不稳定的,在无药条件下恢复整倍体并失去耐受性。Hsp90和钙调磷酸酶对于维持和发展酮康唑耐受性至关重要。这些蛋白质的抑制导致耐受性的丧失。外排基因CDR1不是耐受性发展所必需的。染色体R三体和四体诱导对其他唑类抗真菌剂的交叉耐受,包括克霉唑和咪康唑,但不是其他抗真菌药,如棘白菌素和嘧啶,例如卡泊芬净和5-氟胞嘧啶。
■白色念珠菌对酮康唑的耐受性是由染色体非整倍性介导的,特别是染色体R扩增,需要Hsp90和钙调磷酸酶。这些发现强调了治疗干预的潜在目标,以对抗抗真菌耐受和改善治疗结果。
UNASSIGNED: The emergence of tolerance to antifungal agents in Candida albicans complicates the treatment of fungal infections. Understanding the mechanisms underlying this tolerance is crucial for developing effective therapeutic strategies.
UNASSIGNED: This study aims to elucidate the genetic and molecular basis of
ketoconazole tolerance in C. albicans, focusing on the roles of chromosomal aneuploidy, Hsp90, and calcineurin.
UNASSIGNED: The wild-type C. albicans strain SC5314 was exposed to increasing concentrations of
ketoconazole (0.015-32 μg/mL) to select for tolerant adaptors. Disk diffusion and spot assays were used to assess tolerance. Whole-genome sequencing identified chromosomal changes in the adaptors. The roles of Hsp90 and calcineurin in maintaining and developing
ketoconazole tolerance were investigated using specific inhibitors and knockout strains.
UNASSIGNED: Adaptors exhibited tolerance to
ketoconazole concentrations up to 16 μg/mL, a significant increase from the parent strain\'s inhibition at 0.015 μg/mL. All tolerant adaptors showed amplification of chromosome R, with 29 adaptors having trisomy and one having tetrasomy. This aneuploidy was unstable, reverting to euploidy and losing tolerance in drug-free conditions. Both Hsp90 and calcineurin were essential for maintaining and developing
ketoconazole tolerance. Inhibition of these proteins resulted in loss of tolerance. The efflux gene CDR1 was not required for the development of tolerance. Chromosome R trisomy and tetrasomy induce cross-tolerance to other azole antifungal agents, including clotrimazole and miconazole, but not to other antifungal classes, such as echinocandins and pyrimidines, exemplified by caspofungin and 5-flucytosine.
UNASSIGNED: Ketoconazole tolerance in C. albicans is mediated by chromosomal aneuploidy, specifically chromosome R amplification, and requires Hsp90 and calcineurin. These findings highlight potential targets for therapeutic intervention to combat antifungal tolerance and improve treatment outcomes.