Department of Biological Sciences, Vanderbilt University, Nashville, Tennessee, USA.
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
Microbiol Spectr. 2021 Sep 3;9(1):e0001021. doi: 10.1128/Spectrum.00010-21. Epub 2021 Jun 9.
The ongoing global pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for coronavirus disease 2019 (COVID-19), first described in Wuhan, China. A subset of COVID-19 patients has been reported to have acquired secondary infections by microbial pathogens, such as opportunistic fungal pathogens from the genus Aspergillus. To gain insight into COVID-19-associated pulmonary aspergillosis (CAPA), we analyzed the genomes and characterized the phenotypic profiles of four CAPA isolates of Aspergillus fumigatus obtained from patients treated in the area of North Rhine-Westphalia, Germany. By examining the mutational spectrum of single nucleotide polymorphisms, insertion-deletion polymorphisms, and copy number variants among 206 genes known to modulate A. fumigatus virulence, we found that CAPA isolate genomes do not exhibit significant differences from the genome of the Af293 reference strain. By examining a number of factors, including virulence in an invertebrate moth model, growth in the presence of osmotic, cell wall, and oxidative stressors, secondary metabolite biosynthesis, and the MIC of antifungal drugs, we found that CAPA isolates were generally, but not always, similar to A. fumigatus reference strains Af293 and CEA17. Notably, CAPA isolate D had more putative loss-of-function mutations in genes known to increase virulence when deleted. Moreover, CAPA isolate D was significantly more virulent than the other three CAPA isolates and the A. fumigatus reference strains Af293 and CEA17, but similarly virulent to two other clinical strains of A. fumigatus. These findings expand our understanding of the genomic and phenotypic characteristics of isolates that cause CAPA. The global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of coronavirus disease 2019 (COVID-19), has already killed millions of people. COVID-19 patient outcome can be further complicated by secondary infections, such as COVID-19-associated pulmonary aspergillosis (CAPA). CAPA is caused by Aspergillus fungal pathogens, but there is little information about the genomic and phenotypic characteristics of CAPA isolates. We conducted genome sequencing and extensive phenotyping of four CAPA isolates of Aspergillus fumigatus from Germany. We found that CAPA isolates were often, but not always, similar to other reference strains of A. fumigatus across 206 genetic determinants of infection-relevant phenotypes, including virulence. For example, CAPA isolate D was more virulent than other CAPA isolates and reference strains in an invertebrate model of fungal disease, but similarly virulent to two other clinical strains. These results expand our understanding of COVID-19-associated pulmonary aspergillosis.
由严重急性呼吸系统综合症冠状病毒 2（SARS-CoV-2）引起的持续全球大流行是导致 2019 年冠状病毒病（COVID-19）的原因，该病毒最初在中国武汉被描述。据报道，有一部分 COVID-19 患者发生了微生物病原体的二次感染，例如来自曲霉菌属的机会性真菌病原体。为了深入了解 COVID-19 相关的肺曲霉病（CAPA），我们分析了从德国北莱茵-威斯特法伦地区治疗的患者中获得的四个烟曲霉 CAPA 分离株的基因组，并对其表型特征进行了表征。通过检查已知调节烟曲霉毒力的 206 个基因中的单核苷酸多态性、插入缺失多态性和拷贝数变异的突变谱，我们发现 CAPA 分离株的基因组与 Af293 参考菌株的基因组没有显著差异。通过检查包括在无脊椎鳞翅目昆虫模型中的毒力、在渗透压、细胞壁和氧化应激物存在下的生长、次生代谢产物生物合成以及抗真菌药物的 MIC 等多种因素，我们发现 CAPA 分离株通常与 Af293 和 CEA17 等烟曲霉参考菌株相似，但并不总是如此。值得注意的是，在已知缺失时会增加毒力的基因中，CAPA 分离株 D 具有更多潜在的功能丧失突变。此外，CAPA 分离株 D 比其他三个 CAPA 分离株和烟曲霉参考菌株 Af293 和 CEA17 更具毒力，但与两个其他临床烟曲霉分离株的毒力相似。这些发现扩展了我们对引起 CAPA 的分离株的基因组和表型特征的理解。由严重急性呼吸系统综合症冠状病毒 2（SARS-CoV-2）引起的全球大流行，是导致 2019 年冠状病毒病（COVID-19）的病因，已经导致数百万人死亡。COVID-19 患者的预后可能会因二次感染而变得更加复杂，例如 COVID-19 相关的肺曲霉病（CAPA）。CAPA 是由曲霉菌属真菌病原体引起的，但关于 CAPA 分离株的基因组和表型特征的信息很少。我们对来自德国的四个烟曲霉 CAPA 分离株进行了基因组测序和广泛的表型分析。我们发现，在与感染相关表型的 206 个遗传决定因素中，CAPA 分离株通常与其他烟曲霉参考菌株相似，包括毒力。例如，在真菌疾病的无脊椎模型中，CAPA 分离株 D 比其他 CAPA 分离株和参考菌株更具毒力，但与两个其他临床烟曲霉分离株的毒力相似。这些结果扩展了我们对 COVID-19 相关肺曲霉病的理解。
