Delbaje Endrews, de Castro Patrícia Alves, Calise Dante G, Mengyao Niu, Horta Maria Augusta Crivelente, Akiyama Daniel Yuri, Pontes João Guilherme, Fill Taícia, Kniemeyer Olaf, Krüger Thomas, Brakhage Axel A, Wong Koon Ho, Keller Nancy P, Goldman Gustavo H
School of Pharmaceutical Sciences of Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto 14040-903, Brazil.
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA.
J Fungi (Basel). 2024 Oct 30;10(11):749. doi: 10.3390/jof10110749.
can cause invasive pulmonary aspergillosis (IPA). Fungicidal azoles and fungistatic caspofungin (CAS) are the first- and second-line therapies, respectively, used to treat IPA. Treatment of with CAS or micafungin induces the production of the oxylipin 5,8-diHODE by the fungal oxygenase PpoA. For this article, we investigated the influence of ppo genes, which encode the fatty acid oxygenases responsible for oxylipin biosynthesis, on CAS tolerance. The influence of PpoA and PpoC on CAS tolerance is mediated by MpkA phosphorylation and protein kinase A (PKA) activity. RNAseq transcriptional profiling and the label-free quantitative proteomics of the ppoA and ppoC mutants showed that differentially expressed genes and proteins are related to secondary metabolites and carbohydrate metabolism. We also characterized two clinical isolates, CM7555 and IFM61407, which decrease and increase susceptibility to CAS, respectively. CM7555 does not exhibit increased oxylipin production in the presence of CAS but oxylipin induction upon CAS exposure is increased in IFM61407, suggesting that oxylipins are not the only mechanism involved in CAS tolerance in these isolates. Upon CAS exposure, CM7555 has higher MpkA phosphorylation and PKA activity than IFM61407. Our results reveal the different aspects and genetic determinants involved in CAS tolerance.
可导致侵袭性肺曲霉病（IPA）。杀真菌唑类和抑菌性卡泊芬净（CAS）分别是用于治疗IPA的一线和二线疗法。用CAS或米卡芬净治疗会诱导真菌加氧酶PpoA产生氧化脂质5,8 - 二羟基十八碳二烯酸（5,8 - diHODE）。在本文中，我们研究了编码负责氧化脂质生物合成的脂肪酸加氧酶的ppo基因对CAS耐受性的影响。PpoA和PpoC对CAS耐受性的影响是由MpkA磷酸化和蛋白激酶A（PKA）活性介导的。ppoA和ppoC突变体的RNAseq转录谱分析和无标记定量蛋白质组学表明，差异表达的基因和蛋白质与次级代谢产物和碳水化合物代谢有关。我们还对两株临床分离株CM7555和IFM61407进行了表征，它们对CAS的敏感性分别降低和增加。CM7555在存在CAS的情况下不会表现出氧化脂质产量增加，但IFM61407在接触CAS后氧化脂质的诱导增加，这表明氧化脂质不是这些分离株中CAS耐受性的唯一机制。在接触CAS后,CM7555的MpkA磷酸化和PKA活性高于IFM61407。我们的结果揭示了CAS耐受性涉及的不同方面和遗传决定因素。
