Department of Microbiology and Immunology, University of Minnesota, Minneapolis, MN, USA.
Division of Infectious Diseases, Lundquist Institute for Biomedical Innovation at Harbor UCLA Medical Center, Torrance, CA, USA.
Nat Microbiol. 2024 Nov;9(11):2923-2938. doi: 10.1038/s41564-024-01795-7. Epub 2024 Sep 3.
Genomic copy number changes are associated with antifungal drug resistance and virulence across diverse fungal pathogens, but the rate and dynamics of these genomic changes in the presence of antifungal drugs are unknown. Here we optimized a dual-fluorescent reporter system in the diploid pathogen Candida albicans to quantify haplotype-specific copy number variation (CNV) and loss of heterozygosity (LOH) at the single-cell level with flow cytometry. We followed the frequency and dynamics of CNV and LOH at two distinct genomic locations in the presence and absence of antifungal drugs in vitro and in a murine model of candidiasis. Copy number changes were rapid and dynamic during adaptation to fluconazole and frequently involved competing subpopulations with distinct genotypes. This study provides quantitative evidence for the rapid speed at which diverse genotypes arise and undergo dynamic population-level fluctuations during adaptation to antifungal drugs in vitro and in vivo.
基因组拷贝数变化与不同真菌病原体的抗真菌药物耐药性和毒力有关，但在存在抗真菌药物的情况下，这些基因组变化的速度和动态尚不清楚。在这里，我们优化了二倍体病原体白色念珠菌中的双荧光报告系统，以通过流式细胞术在单细胞水平上定量检测单倍型特异性拷贝数变异（CNV）和杂合性丢失（LOH）。我们在体外和念珠菌感染的小鼠模型中，在存在和不存在抗真菌药物的情况下，跟踪两个不同基因组位置的 CNV 和 LOH 的频率和动态。在适应氟康唑的过程中，拷贝数变化迅速且动态，并且经常涉及具有不同基因型的竞争亚群。这项研究提供了定量证据，证明在体外和体内适应抗真菌药物时，不同基因型的快速出现和经历动态的群体水平波动的速度非常快。
