Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance
ABSTRACT Resistance to fluconazole is one of clinical characteristics most frequently challenging the treatment of invasive Candida auris infections, and is observed among >90% of all characterized clinical isolates. In this work, the native C. auris ERG11 allele in a previously characterized flu...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2021-12-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/Spectrum.01585-21 |
| _version_ | 1818973831963869184 |
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| author | Jeffrey M. Rybak Cheshta Sharma Laura A. Doorley Katherine S. Barker Glen E. Palmer P. David Rogers |
| author_facet | Jeffrey M. Rybak Cheshta Sharma Laura A. Doorley Katherine S. Barker Glen E. Palmer P. David Rogers |
| author_sort | Jeffrey M. Rybak |
| collection | DOAJ |
| description | ABSTRACT Resistance to fluconazole is one of clinical characteristics most frequently challenging the treatment of invasive Candida auris infections, and is observed among >90% of all characterized clinical isolates. In this work, the native C. auris ERG11 allele in a previously characterized fluconazole-susceptible clinical isolate was replaced with the ERG11 alleles from three highly fluconazole-resistant clinical isolates (MIC ≥256 mg/L), encoding the amino acid substitutions VF125AL, Y132F, and K143R, using Cas9-ribonucleoprotein (RNP) mediated transformation system. Reciprocally, the ERG11WT allele from the same fluconazole-susceptible clinical isolate, lacking any resistance-associated mutation, was introduced into a previously characterized fluconazole-resistant clinical isolate, replacing the native ERG11K143R allele, using the same methods. The resulting collection of strains was subjected to comprehensive triazole susceptibility testing, and the direct impact each of these clinically-derived ERG11 mutations on triazole MIC was determined. Introduction of each of the three mutant ERG11 alleles was observed to increase fluconazole and voriconazole MIC by 8- to 16-fold. The MIC for the other clinically available triazoles were not significantly impacted by any ERG11 mutation. In the fluconazole-resistant clinical isolate background, correction of the K143R encoding mutation led to a similar 16-fold decrease in fluconazole MIC, and 8-fold decrease in voriconazole MIC, while the MIC of other triazoles were minimally changed. Taken together, these findings demonstrate that mutations in C. auris ERG11 significantly contribute to fluconazole and voriconazole resistance, but alone cannot explain the substantially elevated MIC observed among clinical isolates of C. auris. IMPORTANCE Candida auris is an emerging multidrug-resistant and health care-associated pathogen of urgent clinical concern. The triazoles are the most widely prescribed antifungal agents worldwide and are commonly utilized for the treatment of invasive Candida infections. Greater than 90% of all C. auris clinical isolates are observed to be resistant to fluconazole, and nearly all fluconazole-resistant isolates of C. auris are found to have one of three mutations (encoding VF125AL, Y132F, or K143R) in the gene encoding the target of the triazoles, ERG11. However, the direct contribution of these mutations in ERG11 to fluconazole resistance and the impact these mutations may have the susceptibility of the other triazoles remains unknown. The present study seeks to address this knowledge gap and potentially inform the future application the triazole antifungals for the treatment of infections caused by C. auris. |
| first_indexed | 2024-12-20T15:30:25Z |
| format | Article |
| id | doaj.art-11fc8a65bf524a0eb9afa9a462436f19 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-12-20T15:30:25Z |
| publishDate | 2021-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-11fc8a65bf524a0eb9afa9a462436f192022-12-21T19:35:37ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972021-12-019310.1128/Spectrum.01585-21Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole ResistanceJeffrey M. Rybak0Cheshta Sharma1Laura A. Doorley2Katherine S. Barker3Glen E. Palmer4P. David Rogers5Department of Pharmacy and Pharmaceutical Sciences, St. Jude Children’s Research Hospital, Memphis, Tennessee, USADepartment of Clinical Pharmacy and Translational Science, University of Tennessee College of Pharmacy, Memphis, Tennessee, USADepartment of Pharmacy and Pharmaceutical Sciences, St. Jude Children’s Research Hospital, Memphis, Tennessee, USADepartment of Pharmacy and Pharmaceutical Sciences, St. Jude Children’s Research Hospital, Memphis, Tennessee, USADepartment of Clinical Pharmacy and Translational Science, University of Tennessee College of Pharmacy, Memphis, Tennessee, USADepartment of Pharmacy and Pharmaceutical Sciences, St. Jude Children’s Research Hospital, Memphis, Tennessee, USAABSTRACT Resistance to fluconazole is one of clinical characteristics most frequently challenging the treatment of invasive Candida auris infections, and is observed among >90% of all characterized clinical isolates. In this work, the native C. auris ERG11 allele in a previously characterized fluconazole-susceptible clinical isolate was replaced with the ERG11 alleles from three highly fluconazole-resistant clinical isolates (MIC ≥256 mg/L), encoding the amino acid substitutions VF125AL, Y132F, and K143R, using Cas9-ribonucleoprotein (RNP) mediated transformation system. Reciprocally, the ERG11WT allele from the same fluconazole-susceptible clinical isolate, lacking any resistance-associated mutation, was introduced into a previously characterized fluconazole-resistant clinical isolate, replacing the native ERG11K143R allele, using the same methods. The resulting collection of strains was subjected to comprehensive triazole susceptibility testing, and the direct impact each of these clinically-derived ERG11 mutations on triazole MIC was determined. Introduction of each of the three mutant ERG11 alleles was observed to increase fluconazole and voriconazole MIC by 8- to 16-fold. The MIC for the other clinically available triazoles were not significantly impacted by any ERG11 mutation. In the fluconazole-resistant clinical isolate background, correction of the K143R encoding mutation led to a similar 16-fold decrease in fluconazole MIC, and 8-fold decrease in voriconazole MIC, while the MIC of other triazoles were minimally changed. Taken together, these findings demonstrate that mutations in C. auris ERG11 significantly contribute to fluconazole and voriconazole resistance, but alone cannot explain the substantially elevated MIC observed among clinical isolates of C. auris. IMPORTANCE Candida auris is an emerging multidrug-resistant and health care-associated pathogen of urgent clinical concern. The triazoles are the most widely prescribed antifungal agents worldwide and are commonly utilized for the treatment of invasive Candida infections. Greater than 90% of all C. auris clinical isolates are observed to be resistant to fluconazole, and nearly all fluconazole-resistant isolates of C. auris are found to have one of three mutations (encoding VF125AL, Y132F, or K143R) in the gene encoding the target of the triazoles, ERG11. However, the direct contribution of these mutations in ERG11 to fluconazole resistance and the impact these mutations may have the susceptibility of the other triazoles remains unknown. The present study seeks to address this knowledge gap and potentially inform the future application the triazole antifungals for the treatment of infections caused by C. auris.https://journals.asm.org/doi/10.1128/Spectrum.01585-21CandidatriazoleresistanceERG11CRISPR |
| spellingShingle | Jeffrey M. Rybak Cheshta Sharma Laura A. Doorley Katherine S. Barker Glen E. Palmer P. David Rogers Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance Microbiology Spectrum Candida triazole resistance ERG11 CRISPR |
| title | Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance |
| title_full | Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance |
| title_fullStr | Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance |
| title_full_unstemmed | Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance |
| title_short | Delineation of the Direct Contribution of Candida auris ERG11 Mutations to Clinical Triazole Resistance |
| title_sort | delineation of the direct contribution of candida auris erg11 mutations to clinical triazole resistance |
| topic | Candida triazole resistance ERG11 CRISPR |
| url | https://journals.asm.org/doi/10.1128/Spectrum.01585-21 |
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