Pan-azole-resistant Meyerozyma guilliermondii clonal isolates harbouring a double F126L and L505F mutation in Erg11.
ERG11
Meyerozyma guilliermondii
antifungal resistance
azoles
Journal
Mycoses
ISSN: 1439-0507
Titre abrégé: Mycoses
Pays: Germany
ID NLM: 8805008
Informations de publication
Date de publication:
Mar 2024
Mar 2024
Historique:
revised:
29
01
2024
received:
16
11
2023
accepted:
02
02
2024
medline:
2
3
2024
pubmed:
2
3
2024
entrez:
1
3
2024
Statut:
ppublish
Résumé
Meyerozyma guilliermondii is a yeast species responsible for invasive fungal infections. It has high minimum inhibitory concentrations (MICs) to echinocandins, the first-line treatment of candidemia. In this context, azole antifungal agents are frequently used. However, in recent years, a number of azole-resistant strains have been described. Their mechanisms of resistance are currently poorly studied. The aim of this study was consequently to understand the mechanisms of azole resistance in several clinical isolates of M. guilliermondii. Ten isolates of M. guilliermondii and the ATCC 6260 reference strain were studied. MICs of azoles were determined first. Whole genome sequencing of the isolates was then carried out and the mutations identified in ERG11 were expressed in a CTG clade yeast model (C. lusitaniae). RNA expression of ERG11, MDR1 and CDR1 was evaluated by quantitative PCR. A phylogenic analysis was developed and performed on M. guilliermondii isolates. Lastly, in vitro experiments on fitness cost and virulence were carried out. Of the ten isolates tested, three showed pan-azole resistance. A combination of F126L and L505F mutations in Erg11 was highlighted in these three isolates. Interestingly, a combination of these two mutations was necessary to confer azole resistance. An overexpression of the Cdr1 efflux pump was also evidenced in one strain. Moreover, the three pan-azole-resistant isolates were shown to be genetically related and not associated with a fitness cost or a lower virulence, suggesting a possible clonal transmission. In conclusion, this study identified an original combination of ERG11 mutations responsible for pan-azole-resistance in M. guilliermondii. Moreover, we proposed a new MLST analysis for M. guilliermondii that identified possible clonal transmission of pan-azole-resistant strains. Future studies are needed to investigate the distribution of this clone in hospital environment and should lead to the reconsideration of the treatment for this species.
Sections du résumé
BACKGROUND
BACKGROUND
Meyerozyma guilliermondii is a yeast species responsible for invasive fungal infections. It has high minimum inhibitory concentrations (MICs) to echinocandins, the first-line treatment of candidemia. In this context, azole antifungal agents are frequently used. However, in recent years, a number of azole-resistant strains have been described. Their mechanisms of resistance are currently poorly studied.
OBJECTIVE
OBJECTIVE
The aim of this study was consequently to understand the mechanisms of azole resistance in several clinical isolates of M. guilliermondii.
METHODS
METHODS
Ten isolates of M. guilliermondii and the ATCC 6260 reference strain were studied. MICs of azoles were determined first. Whole genome sequencing of the isolates was then carried out and the mutations identified in ERG11 were expressed in a CTG clade yeast model (C. lusitaniae). RNA expression of ERG11, MDR1 and CDR1 was evaluated by quantitative PCR. A phylogenic analysis was developed and performed on M. guilliermondii isolates. Lastly, in vitro experiments on fitness cost and virulence were carried out.
RESULTS
RESULTS
Of the ten isolates tested, three showed pan-azole resistance. A combination of F126L and L505F mutations in Erg11 was highlighted in these three isolates. Interestingly, a combination of these two mutations was necessary to confer azole resistance. An overexpression of the Cdr1 efflux pump was also evidenced in one strain. Moreover, the three pan-azole-resistant isolates were shown to be genetically related and not associated with a fitness cost or a lower virulence, suggesting a possible clonal transmission.
CONCLUSION
CONCLUSIONS
In conclusion, this study identified an original combination of ERG11 mutations responsible for pan-azole-resistance in M. guilliermondii. Moreover, we proposed a new MLST analysis for M. guilliermondii that identified possible clonal transmission of pan-azole-resistant strains. Future studies are needed to investigate the distribution of this clone in hospital environment and should lead to the reconsideration of the treatment for this species.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e13704Informations de copyright
© 2024 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.
Références
McCarty TP, White CM, Pappas PG. Candidemia and invasive candidiasis. Infect Dis Clin N Am. 2021;35(2):389-413.
Savini V, Catavitello C, Onofrillo D, et al. What do we know about Candida guilliermondii? A voyage throughout past and current literature about this emerging yeast. Mycoses. 2011;54(5):434-441.
Marcos-Zambrano LJ, Puig-Asensio M, Pérez-García F, et al. Candida guilliermondii complex is characterized by high antifungal resistance but low mortality in 22 cases of candidemia. Antimicrob Agents Chemother. 2017;61(7):e00099-17.
Girmenia C, Pizzarelli G, Cristini F, et al. Candida guilliermondii fungemia in patients with hematologic malignancies. J Clin Microbiol. 2006;44(7):2458-2464.
Pfaller MA, Diekema DJ, Mendez M, et al. Candida guilliermondii, an opportunistic fungal pathogen with decreased susceptibility to fluconazole: geographic and temporal trends from the ARTEMIS DISK antifungal surveillance program. J Clin Microbiol. 2006;44(10):3551-3556.
Cheng JW, Yu SY, Xiao M, et al. Identification and antifungal susceptibility profile of Candida guilliermondii and Candida fermentati from a multicenter study in China. J Clin Microbiol. 2016;54(8):2187-2189.
Krcmery V, Grausova S, Mraz M, Pichnova E, Jurga L. Candida guilliermondii fungemia in cancer patients: report of three cases. J Infect Chemother. 1999;5(1):58-59.
Pfaller MA, Boyken L, Hollis RJ, et al. Wild-type MIC distributions and epidemiological cutoff values for the echinocandins and Candida spp. J Clin Microbiol. 2010;48(1):52-56.
Pfaller MA, Boyken L, Hollis RJ, et al. In vitro susceptibility of invasive isolates of Candida spp. to anidulafungin, caspofungin, and micafungin: six years of global surveillance. J Clin Microbiol. 2008;46(1):150-156.
Dudiuk C, Macedo D, Leonardelli F, et al. Molecular confirmation of the relationship between Candida guilliermondii Fks1p naturally occurring amino acid substitutions and its intrinsic reduced echinocandin susceptibility. Antimicrob Agents Chemother. 2017;61(5):e02644-16.
Barchiesi F, Spreghini E, Tomassetti S, et al. Effects of caspofungin against Candida guilliermondii and Candida parapsilosis. Antimicrob Agents Chemother. 2006;50(8):2719-2727.
Kabbara N, Lacroix C, Peffault de Latour R, et al. Guilliermondii blood stream infections in allogeneic hematopoietic stem cell transplant recipients receiving long-term caspofungin therapy. Haematologica. 2008;93(4):639-640.
Nishida R, Eriguchi Y, Miyake N, et al. Breakthrough candidemia with hematological disease: results from a single-center retrospective study in Japan, 2009-2020. Med Mycol. 2023;61(6):myad056.
Ghasemi R, Lotfali E, Rezaei K, et al. Meyerozyma guilliermondii species complex: review of current epidemiology, antifungal resistance, and mechanisms. Braz J Microbiol. 2022;53(4):1761-1779.
Hamill RJ. Amphotericin B formulations: a comparative review of efficacy and toxicity. Drugs. 2013;73(9):919-934.
Desnos-Ollivier M, Bretagne S, Boullié A, Gautier C, Dromer F, Lortholary O. Isavuconazole MIC distribution of 29 yeast species responsible for invasive infections (2015-2017). Clin Microbiol Infect. 2019;25(5):634.e1-634.e4.
Tietz HJ, Czaika V, Sterry W. Case report. Osteomyelitis caused by high resistant Candida guilliermondii. Mycoses. 1999;42(9-10):577-580.
Savini V, Catavitello C, Di Marzio I, et al. Pan-azole-resistant Candida guilliermondii from a leukemia patient's silent funguria. Mycopathologia. 2010;169(6):457-459.
Cebeci Güler N, Tosun I, Aydin F. The identification of Meyerozyma guilliermondii from blood cultures and surveillance samples in a university hospital in Northeast Turkey: a ten-year survey. J Mycol Médicale. 2017;27(4):506-513.
Cheng JW, Liao K, Kudinha T, et al. Molecular epidemiology and azole resistance mechanism study of Candida guilliermondii from a Chinese surveillance system. Sci Rep. 2017;7(1):907.
Pristov KE, Ghannoum MA. Resistance of Candida to azoles and echinocandins worldwide. Clin Microbiol Infect. 2019;25(7):792-798.
Arendrup MC, Meletiadis J, Mouton JW, Lagrou K, Hamal P, Guinea J. EUCAST DEFINITIVE DOCUMENT E.DEF 7.3.2. 2020.
Wick RR. Filtlong [Internet]. GitHub. 2017 [cited 2022 Feb 7]. Available from: https://github.com/rrwick/Filtlong/
Kolmogorov M, Yuan J, Lin Y, Pevzner PA. Assembly of long, error-prone reads using repeat graphs. Nat Biotechnol. 2019;37(5):540-546.
Candida Genome Database. [cited 2023 Feb 1]. Available from: http://www.candidagenome.org/
Accoceberry I, Rougeron A, Biteau N, Chevrel P, Fitton-Ouhabi V, Noël T. A CTG clade Candida yeast genetically engineered for the genotype-phenotype characterization of azole antifungal resistance in human-pathogenic yeasts. Antimicrob Agents Chemother. 2018;62(1):e01483-17.
El-Kirat-Chatel S, Dementhon K, Noël T. A two-step cloning-free PCR-based method for the deletion of genes in the opportunistic pathogenic yeast Candida lusitaniae. Yeast. 2011;28(4):321-330.
Bougnoux ME, Tavanti A, Bouchier C, et al. Collaborative consensus for optimized multilocus sequence typing of Candida albicans. J Clin Microbiol. 2003;41(11):5265-5266.
Tavanti A, Davidson AD, Johnson EM, et al. Multilocus sequence typing for differentiation of strains of Candida tropicalis. J Clin Microbiol. 2005;43(11):5593-5600.
Curtis A, Binder U, Kavanagh K. Galleria mellonella larvae as a model for investigating fungal-host interactions. Front Fungal Biol. 2022;3:893494.
MIC EUCAST. [cited 2023 Feb 1]. Available from: https://mic.eucast.org/search/?search%5Bmethod%5D=mic&search%5Bantibiotic%5D=-1&search%5Bspecies%5D=131&search%5Bdisk_content%5D=-1&search%5Blimit%5D=50
Espinel-Ingroff A, Turnidge J. The role of epidemiological cutoff values (ECVs/ECOFFs) in antifungal susceptibility testing and interpretation for uncommon yeasts and moulds. Rev Iberoam Micol. 2016;33(2):63-75.
Salsé M, Gangneux JP, Cassaing S, et al. Multicentre study to determine the Etest epidemiological cut-off values of antifungal drugs in Candida spp. and aspergillus fumigatus species complex. Clin Microbiol Infect. 2019;25(12):1546-1552.
Espinel-Ingroff A, Sasso M, Turnidge J, et al. Etest ECVs/ECOFFs for detection of resistance in prevalent and three nonprevalent Candida spp. to triazoles and amphotericin B and aspergillus spp. to Caspofungin: further assessment of modal variability. Antimicrob Agents Chemother. 2021;65(11):e0109321.
Desnos-Ollivier M, Lortholary O, Bretagne S, Dromer F. Azole susceptibility profiles of more than 9,000 clinical yeast isolates belonging to 40 common and rare species. Antimicrob Agents Chemother. 2021;65(6):e02615-20.
Morio F, Loge C, Besse B, Hennequin C, Le Pape P. Screening for amino acid substitutions in the Candida albicans Erg11 protein of azole-susceptible and azole-resistant clinical isolates: new substitutions and a review of the literature. Diagn Microbiol Infect Dis. 2010;66(4):373-384.
Favre B, Didmon M, Ryder NS. Multiple amino acid substitutions in lanosterol 14alpha-demethylase contribute to azole resistance in Candida albicans. Microbiology (Reading). 1999;145(Pt 10):2715-2725.
Perea S, López-Ribot JL, Kirkpatrick WR, et al. Prevalence of molecular mechanisms of resistance to azole antifungal agents in Candida albicans strains displaying high-level fluconazole resistance isolated from human immunodeficiency virus-infected patients. Antimicrob Agents Chemother. 2001;45(10):2676-2684.
Chow NA, Muñoz JF, Gade L, et al. Tracing the evolutionary history and global expansion of Candida auris using population genomic analyses. MBio. 2020;11(2):e03364-19.
Rybak JM, Sharma C, Doorley LA, Barker KS, Palmer GE, Rogers PD. Delineation of the direct contribution of Candida auris ERG11 mutations to clinical triazole resistance. Microbiol Spectr. 2021;9(3):e0158521.
Williamson B, Wilk A, Guerrero KD, et al. Impact of Erg11 amino acid substitutions identified in Candida auris clade III isolates on triazole drug susceptibility. Antimicrob Agents Chemother. 2022;66(1):e0162421.
Naicker SD, Maphanga TG, Chow NA, et al. Clade distribution of Candida auris in South Africa using whole genome sequencing of clinical and environmental isolates. Emerg Microbes Infect. 2021;10(1):1300-1308.
Maphanga TG, Naicker SD, Kwenda S, et al. In vitro antifungal resistance of Candida auris isolates from bloodstream infections, South Africa. Antimicrob Agents Chemother. 2021;65(9):e0051721.
Keniya MV, Sabherwal M, Wilson RK, et al. Crystal structures of full-length lanosterol 14α-demethylases of prominent fungal pathogens Candida albicans and Candida glabrata provide tools for antifungal discovery. Antimicrob Agents Chemother. 2018;62(11):e01134-18.
Prakash SMU, Nazeer Y, Jayanthi S, Kabir MA. Computational insights into fluconazole resistance by the suspected mutations in lanosterol 14α-demethylase (Erg11p) of Candida albicans. Mol Biol Res Commun. 2020;9(4):155-167.
Debnath S, Addya S. Structural basis for heterogeneous phenotype of ERG11 dependent azole resistance in C.Albicans clinical isolates. Springerplus. 2014;3:660.
Borgeat V, Brandalise D, Grenouillet F, Sanglard D. Participation of the ABC transporter CDR1 in azole resistance of Candida lusitaniae. J Fungi (Basel). 2021;7(9):760.
Li J, Coste AT, Liechti M, Bachmann D, Sanglard D, Lamoth F. Novel ERG11 and TAC1b mutations associated with azole resistance in Candida auris. Antimicrob Agents Chemother. 2023;65(5):e02663-20.
Mayr EM, Ramírez-Zavala B, Krüger I, Morschhäuser J. A zinc cluster transcription factor contributes to the intrinsic fluconazole resistance of Candida auris. mSphere. 2020;5(2):e00279-20.
Liu Z, Myers LC. Mediator tail module is required for Tac1-activated CDR1 expression and azole resistance in Candida albicans. Antimicrob Agents Chemother. 2017;61(11):e01342-17.
Wang D, An N, Yang Y, Yang X, Fan Y, Feng J. Candida tropicalis distribution and drug resistance is correlated with ERG11 and UPC2 expression. Antimicrob Resist Infect Control. 2021;10(1):54.
Presente S, Bonnal C, Normand AC, et al. Hospital clonal outbreak of fluconazole-resistant Candida parapsilosis harboring the Y132F ERG11p substitution in a French intensive care unit. Antimicrob Agents Chemother. 2023;67(3):e0113022.
Díaz-García J, Gómez A, Alcalá L, et al. Evidence of fluconazole-resistant Candida parapsilosis genotypes spreading across hospitals located in Madrid, Spain and harboring the Y132F ERG11p substitution. Antimicrob Agents Chemother. 2022;66(8):e0071022.
Alcoceba E, Gómez A, Lara-Esbrí P, et al. Fluconazole-resistant Candida parapsilosis clonally related genotypes: first report proving the presence of endemic isolates harbouring the Y132F ERG11 gene substitution in Spain. Clin Microbiol Infect. 2022;28(8):1113-1119.