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Host- and echinocandin-induced stresses exert profound cell wall changes in Candida parapsilosis, conferring echinocandin tolerance and retaining fitness

ABSTRACT: In this study, we compared transcriptomic changes in four serial isolates of Candida parapsilosis from a single patient over three time points (90 minutes, 8 hours and 24 hours) and two growth conditions (biofilm and planktonic). The later serial isolates have increased fitness and are resistant to echinocandins compared to the initial strain. Using RNA-Seq, we identified strain-specific changes in expression of specific biological processes and cellular components including fungal-type cell wall and transmembrane transport.

ORGANISM(S): Candida parapsilosis

PROVIDER: GSE298776 | GEO | 2025/12/31

REPOSITORIES: GEO

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Host- and echinocandin-induced stresses exert profound cell wall changes in Candida parapsilosis, conferring echinocandin tolerance and retaining fitness

Project description:Host- and echinocandin-induced stresses exert profound cell wall changes in Candida parapsilosis, conferring echinocandin tolerance and retaining fitness

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Loss of sterol demethylase activity reduces both azole and echinocandin antifungal susceptibility in a clinical isolate of C. parapsilosis

Project description:The present study describes a novel mechanism of antifungal resistance affecting the susceptibility of both the azole and echinocandin antifungals in an azole-resistant isolate from a matched pair of C. parapsilosis isolates obtained from a patient with prosthetic valve endocarditis. Transcriptome analysis indicated differential expression of several genes in the resistant isolate including upregulation of ERG1, ERG2, ERG5, ERG6, ERG11, ERG24, ERG25, ERG27, DAP1 and UPC2, of the ergosterol biosynthesis pathway. Whole genome sequencing revealed a mutation in the ERG3 gene leading to a G111R amino acid substitution in the resistant isolate. Subsequent introduction of this allele in the native ERG3 locus in the susceptible isolate resulted in a fluconazole MIC of >64 mg/ml and a caspofungin MIC of 8 mg/ml. Corresponding allelic replacement of the wildtype allele for the mutant allele in the resistant isolate resulted in a drop in MIC to 1 mg/ml for both fluconazole and caspofungin. Sterol profiles indicated a loss of sterol demethylase activity as a result of this mutation. This work demonstrate that this G111R mutation is wholly responsible for the resistant phenotype in the C. parapsilosis resistant isolate and is the first report of this multidrug resistance mechanism.

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Project description:Azole resistance was induced in vitro by growth of a susceptible C. parapsilosis isolate in the presence of voriconazole. Whole genome microarrays were used to compare the transcriptional response of the voriconizole-resistant and susceptible isolates. Transcriptional profile of an in vitro derived voriconazole-resistant isolate of C. parapsilosis (BC014VRC) compared to the susceptible isolate (BC014S). Cell were grown in YPD medium in normoxia at 35 degrees. Each strain was labelled with Cy3 or Cy5. Overall, 4 independent biological replicates were compared; 2 dye swaps were performed to normalize dye effects.

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