Strains from the Cryptococcus gattii species complex(CGSC)have caused the Pacific Northwest cryptococcosis outbreak,the largest cluster of lifethreatening fungal infections in otherwise healthy human hosts known to da...Strains from the Cryptococcus gattii species complex(CGSC)have caused the Pacific Northwest cryptococcosis outbreak,the largest cluster of lifethreatening fungal infections in otherwise healthy human hosts known to date.In this study,we utilized a pan-phenome-based method to assess the fitness outcomes of CGSC strains under 31 stress conditions,providing a comprehensive overview of 2,821 phenotype-strain associations within this pathogenic clade.Phenotypic clustering analysis revealed a strong correlation between distinct types of stress phenotypes in a subset of CGSC strains,suggesting that shared determinants coordinate their adaptations to various stresses.Notably,a specific group of strains,including the outbreak isolates,exhibited a remarkable ability to adapt to all three of the most commonly used antifungal drugs for treating cryptococcosis(amphotericin B,5-fluorocytosine,and fluconazole).By integrating pan-genomic and pan-transcriptomic analyses,we identified previously unrecognized genes that play crucial roles in conferring multidrug resistance in an outbreak strain with high multidrug adaptation.From these genes,we identified biomarkers that enable the accurate prediction of highly multidrug-adapted CGSC strains,achieving maximum accuracy and area under the curve(AUC)of 0.79 and 0.86,respectively,using machine learning algorithms.Overall,we developed a pan-omic approach to identify cryptococcal multidrug resistance determinants and predict highly multidrug-adapted CGSC strains that may pose significant clinical concern.展开更多
Cell cycle is a fundamental process underlying growth and development in evolutionarily diverse organisms, including fungi. In human fungal pathogens, cell cycle control generally determines their life cycles, either ...Cell cycle is a fundamental process underlying growth and development in evolutionarily diverse organisms, including fungi. In human fungal pathogens, cell cycle control generally determines their life cycles, either in the environment or during infections.Thus, cell cycle components can potentially serve as important targets for the development of antifungal strategy against fungal infections. Here, in Cryptococcus neoformans, the most common cause of fatal fungal meningitis, we show that a previously uncharacterized B-type cyclin named Cbc1 is essential for both its infectious and sexual cycles. We reveal that Cbc1 coordinates various sexual differentiation and molecular processes, including meiosis. Especially, the absence of Cbc1 abolishes formation of sexual spores in C. neoformans, which are presumed infectious particles. Cbc1 is also required for the major Cryptococcus pathogenic attributes. Virulence assessment using the murine model of cryptococcosis revealed that the cbc1 mutant is avirulent.Together, our results provide an important insight into how C. neoformans employs shared cell cycle regulation to coordinate its infectious and sexual cycles, which are considered crucial for virulence evolution and the production of infectious spores.展开更多
Cryptococcus neoformans and its sister species Cryptococcus deuterogattii are important human fungal pathogens.Despite their phylogenetically close relationship,these two Cryptococcus pathogens are greatly different i...Cryptococcus neoformans and its sister species Cryptococcus deuterogattii are important human fungal pathogens.Despite their phylogenetically close relationship,these two Cryptococcus pathogens are greatly different in their clinical characteristics.However,the determinants underlying the regulatory differences of their pathogenicity remain largely unknown.Here,we show that the forkhead transcription factor Hcm1 promotes infection in C.neoformans but not in C.deuterogattii.Monitoring in vitro and in vivo fitness outcomes of multiple clinical isolates from the two pathogens indicates that Hcm1 mediates pathogenicity in C.neoformans through its key involvement in oxidative stress defense.By comparison,Hcm1 is not critical for antioxidation in C.deuterogattii.Furthermore,we identified SRX1,which encodes the antioxidant sulfiredoxin,as a conserved target of Hcm1 in two Cryptococcus pathogens.Like HCM1,SRX1 had a greater role in antioxidation in C.neoformans than in C.deuterogattii.Significantly,overexpression of SRX1 can largely rescue the defective pathogenicity caused by the absence of Hcm1 in C.neoformans.Conversely,Srx1 is dispensable for virulence in C.deuterogattii.Overall,our findings demonstrate that the difference in the contribution of the antioxidant sulfiredoxin to oxidative stress defense underlies the Hcm1-mediated regulatory differences of pathogenicity in two closely related pathogens.展开更多
基金financially supported by the National Key R&D Program of China(2021YFC2302100)the National Natural Science Foundation of China(82370005 and 82172291)+8 种基金the National Key R&D Program of China(2022YFC2303000 and 2021YFC230000)the CAS Interdisciplinary Innovation Team,the Beijing Research Center for Respiratory Infectious Diseases Project(BJRID2024-008 and BJRID2024-011)the R&D Program of Beijing Municipal Education Commission(KM202410025012)the Reform and Development Program of Beijing Institute of Respiratory Medicine(Ggyfz202328 and Ggyfz202418)the National Key R&D Program of China(2020YFA0907200)Shanghai Science and Technology Innovation Action Plan 2023“Basic Research Project”(23JC1404201)the Shanghai‘‘Belt and Road’’Joint Laboratory Project(22490750200)the National Natural Science Foundation of China(82370005)National High Level Hospital Clinical Research Funding(2022-PUMCH-C-052).
文摘Strains from the Cryptococcus gattii species complex(CGSC)have caused the Pacific Northwest cryptococcosis outbreak,the largest cluster of lifethreatening fungal infections in otherwise healthy human hosts known to date.In this study,we utilized a pan-phenome-based method to assess the fitness outcomes of CGSC strains under 31 stress conditions,providing a comprehensive overview of 2,821 phenotype-strain associations within this pathogenic clade.Phenotypic clustering analysis revealed a strong correlation between distinct types of stress phenotypes in a subset of CGSC strains,suggesting that shared determinants coordinate their adaptations to various stresses.Notably,a specific group of strains,including the outbreak isolates,exhibited a remarkable ability to adapt to all three of the most commonly used antifungal drugs for treating cryptococcosis(amphotericin B,5-fluorocytosine,and fluconazole).By integrating pan-genomic and pan-transcriptomic analyses,we identified previously unrecognized genes that play crucial roles in conferring multidrug resistance in an outbreak strain with high multidrug adaptation.From these genes,we identified biomarkers that enable the accurate prediction of highly multidrug-adapted CGSC strains,achieving maximum accuracy and area under the curve(AUC)of 0.79 and 0.86,respectively,using machine learning algorithms.Overall,we developed a pan-omic approach to identify cryptococcal multidrug resistance determinants and predict highly multidrug-adapted CGSC strains that may pose significant clinical concern.
基金financially supported by the National Science and Technology Major Project (2018ZX10101004)the Key Research Program of the Chinese Academy of Sciences (QYZDB-SSWSSMC040)the National Natural Science Foundation of China (31770163)
文摘Cell cycle is a fundamental process underlying growth and development in evolutionarily diverse organisms, including fungi. In human fungal pathogens, cell cycle control generally determines their life cycles, either in the environment or during infections.Thus, cell cycle components can potentially serve as important targets for the development of antifungal strategy against fungal infections. Here, in Cryptococcus neoformans, the most common cause of fatal fungal meningitis, we show that a previously uncharacterized B-type cyclin named Cbc1 is essential for both its infectious and sexual cycles. We reveal that Cbc1 coordinates various sexual differentiation and molecular processes, including meiosis. Especially, the absence of Cbc1 abolishes formation of sexual spores in C. neoformans, which are presumed infectious particles. Cbc1 is also required for the major Cryptococcus pathogenic attributes. Virulence assessment using the murine model of cryptococcosis revealed that the cbc1 mutant is avirulent.Together, our results provide an important insight into how C. neoformans employs shared cell cycle regulation to coordinate its infectious and sexual cycles, which are considered crucial for virulence evolution and the production of infectious spores.
基金supported by the National Key Research and Development Program of China(2021YFC2300400)[Linqi Wang],2021YFC230000[Linqi Wang],2021YFA0911300[Xiao Liu],2021YFC2100600[Xiuyun Tian]:Major Infections Diseases Such as AIDS and Viral Hepatitis Prevention and Control Technology Major Projects(2018ZX10101003[Ying Yang])and CAS Interdisciplinary Innovation Team(Linqi Wang).
文摘Cryptococcus neoformans and its sister species Cryptococcus deuterogattii are important human fungal pathogens.Despite their phylogenetically close relationship,these two Cryptococcus pathogens are greatly different in their clinical characteristics.However,the determinants underlying the regulatory differences of their pathogenicity remain largely unknown.Here,we show that the forkhead transcription factor Hcm1 promotes infection in C.neoformans but not in C.deuterogattii.Monitoring in vitro and in vivo fitness outcomes of multiple clinical isolates from the two pathogens indicates that Hcm1 mediates pathogenicity in C.neoformans through its key involvement in oxidative stress defense.By comparison,Hcm1 is not critical for antioxidation in C.deuterogattii.Furthermore,we identified SRX1,which encodes the antioxidant sulfiredoxin,as a conserved target of Hcm1 in two Cryptococcus pathogens.Like HCM1,SRX1 had a greater role in antioxidation in C.neoformans than in C.deuterogattii.Significantly,overexpression of SRX1 can largely rescue the defective pathogenicity caused by the absence of Hcm1 in C.neoformans.Conversely,Srx1 is dispensable for virulence in C.deuterogattii.Overall,our findings demonstrate that the difference in the contribution of the antioxidant sulfiredoxin to oxidative stress defense underlies the Hcm1-mediated regulatory differences of pathogenicity in two closely related pathogens.
