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In vivo RNA sequencing identifies the lncRNA ReLoV critical for morphological transition and virulence in Candida albicans II

ABSTRACT: Candida albicans is the most prevalent fungal pathogen of humans responsible for nosocomial infections with high mortality rates. Despite its clinical importance, mechanisms governing C. albicans pathogenicity are incompletely understood. To identify long noncoding RNAs (lncRNAs) implicated in the pathogenicity of C. albicans, we performed in vivo Strand-specific RNA sequencing (RNA-seq) in a murine model of systemic infection. We found that hundreds of lncRNAs were expressed by C. albicans during invasive infection, among which 170 were unannotated so far. We further showed that a considerable number of lncRNAs were differentially expressed in fungal cells that invade into murine kidneys relative to uninfected (UI) cells. Differential expression of lncRNAs was validated by qRT-PCR assay. We identified ReLoV, an uncharacterized lncRNA, whose expression was significantly induced upon in vivo infection. The relov Δ/Δ mutant failed to develop elongated filaments in murine kidneys and displayed an attenuated virulence in a systemic infection model. The expression of ReLoV was also increased in response to multiple filament-inducing stimuli, including serum, neutral pH (Lee’s) and rapamycin. ReLoV was critical for hyphal maintenance, but is dispensable to initiate hypha formation. The lack of ReLoV reduced the transcript of UME6, a hypha-specific transcriptional activator that controls the level and duration of hypha-specific transcription, during hyphal elongation but not initiation. We suggest that misregulation of UME6 level in the relov Δ/Δ mutant causes its defects in hyphal development during invasive infection, as well as under in vitro hypha-inducing conditions. Collectively, these data uncover the lncRNA ReLoV as an additional layer of genetic regulation of morphogenesis and pathogenesis in C. albicans.

ORGANISM(S): Candida albicans

PROVIDER: GSE327269 | GEO | 2026/04/12

REPOSITORIES: GEO

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Project description:Candida albicans and Candida dubliniensis are closely related species displaying differences in virulence and genome content, therefore providing potential opportunities to identify novel C. albicans virulence genes. C. albicans gene arrays were used for comparative analysis of global gene expression in the two species in reconstituted human oral epithelium (RHE). C. albicans (SC5314) showed upregulation of hypha-specific and virulence genes within 30 min postinoculation, coinciding with rapid induction of filamentation and increased RHE damage. C. dubliniensis (CD36) showed no detectable upregulation of hypha-specific genes, grew as yeast, and caused limited RHE damage. Several genes absent or highly divergent in C. dubliniensis were upregulated in C. albicans. One such gene, SFL2 (orf19.3969), encoding a putative heat shock factor, was deleted in C. albicans. ÎÎsfl2 cells failed to filament under a range of hypha-inducing conditions and exhibited greatly reduced RHE damage, reversed by reintroduction of SFL2 into the ÎÎsfl2 strain. Moreover, SFL2 overexpression in C. albicans triggered hyphal morphogenesis. Although SFL2 deletion had no apparent effect on host survival in the murine model of systemic infection, ÎÎsfl2 strain-infected kidney tissues contained only yeast cells. These results suggest a role for SFL2 in morphogenesis and an indirect role in C. albicans pathogenesis in epithelial tissues. Global gene expression in Candida albicans SC5314 or Candida dubliniensis CD36 on reconstituted human oral epithelium (RHE) 90 min postinoculation. Gene expression on RHE was compared to gene expression on polycarbonate filter (PCF; used as RHE support matrix) 90 min postinoculation. Performed in two biological replicates with reciprocal dye swaps for each biological replicate. Gene expression of Candida cells on RHE was normalized to gene expression in reference control (PCF); log2 ratios were calculated by dividing spot intensity of experimental by that of the reference control.

Project description:Candida albicans and Candida dubliniensis are closely related species displaying differences in virulence and genome content, therefore providing potential opportunities to identify novel C. albicans virulence genes. C. albicans gene arrays were used for comparative analysis of global gene expression in the two species in reconstituted human oral epithelium (RHE). C. albicans (SC5314) showed upregulation of hypha-specific and virulence genes within 30 min postinoculation, coinciding with rapid induction of filamentation and increased RHE damage. C. dubliniensis (CD36) showed no detectable upregulation of hypha-specific genes, grew as yeast, and caused limited RHE damage. Several genes absent or highly divergent in C. dubliniensis were upregulated in C. albicans. One such gene, SFL2 (orf19.3969), encoding a putative heat shock factor, was deleted in C. albicans. ÎÎsfl2 cells failed to filament under a range of hypha-inducing conditions and exhibited greatly reduced RHE damage, reversed by reintroduction of SFL2 into the ÎÎsfl2 strain. Moreover, SFL2 overexpression in C. albicans triggered hyphal morphogenesis. Although SFL2 deletion had no apparent effect on host survival in the murine model of systemic infection, ÎÎsfl2 strain-infected kidney tissues contained only yeast cells. These results suggest a role for SFL2 in morphogenesis and an indirect role in C. albicans pathogenesis in epithelial tissues. Global gene expression in Candida albicans SC5314 or Candida dubliniensis CD36 on reconstituted human oral epithelium (RHE) 30 min postinoculation. Gene expression on RHE was compared to gene expression in cells used as inoculum (0 min). Performed in three biological replicates with dye swaps across the biological replicates. Gene expression of Candida cells on RHE was normalized to gene expression in reference control (0 min); log2 ratios were calculated by dividing spot intensity of experimental by that of the reference control.

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In vivo RNA sequencing identifies the lncRNA ReLoV critical for morphological transition and virulence in Candida albicans II

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