The impact of Candida albicans on the transcriptome of Enterococcus faecalis when grown in a biofilm
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ABSTRACT:
ORGANISM(S): Enterococcus faecalis OG1RF
PROVIDER: GSE290415 | GEO | 2025/03/03
REPOSITORIES: GEO
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Project description:Catheter-associated urinary tract infections (CAUTI) account for 40% of all nosocomial infections and can lead to significant life-threatening complications such as bacteremia. Microbial biofilms play an important role in the development and pathogenesis of CAUTI, and these biofilms are often polymicrobial. Proteus mirabilis and Enterococcus faecalis are two of the most common causes of CAUTI, and they often persistently co-colonize the catheterized urinary tract. We previously demonstrated that co-culture of E. faecalis with P. mirabilis increased biofilm biomass, antimicrobial resistance, and disease severity. In this study, we uncover the metabolic interplay that drives biofilm enhancement and examine the contribution of this polymicrobial interaction to CAUTI severity. Through compositional and proteomic biofilm analyses, we determined that the increase in biofilm biomass stems from an increase in the protein fraction of the polymicrobial biofilm matrix. We further observed an enrichment in proteins associated with ornithine and arginine metabolism in polymicrobial biofilms compared to single-species biofilms. By testing mutants of E. faecalis and P. mirabilis, we found that L-ornithine secreted by the E. faecalis ArcD antiporter promotes L-arginine biosynthesis in P. mirabilis via ArgF, which ultimately fuels production of proteins that facilitate contact-dependent interactions to enhance biofilm biomass. We further demonstrate that ArcD and ArgF are not important for urinary tract colonization by either species when alone, but ornithine/arginine interplay is critical for the increased disease severity that occurs during coinfection. This study provides deeper insight into the polymicrobial interactions occurring during CAUTI and highlights how these interactions can have significant impacts on pathogenesis and bacterial persistence.
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Project description:The fungal species Candida albicans and the bacterium Enterococcus faecalis are both common members of the gut microbiome, with co-occurrence of these two species widely reported in dysbiotic gut environments. While prior research suggests these species interact in the gut, the underlying mechanisms of these interactions remain unclear. To explore these interactions further, we utilized dual RNA-sequencing to transcriptionally profile both C. albicans and E. faecalis during co-culture and mono-culture under two conditions: (1) an in vitro condition designed to mimic certain aspects of the gut environment and (2) the germ-free mouse gut. Gene expression analysis revealed that both species strongly upregulate citrate-related genes during co-culture: C. albicans upregulates CIT1 (citrate synthase) which produces citrate, while E. faecalis upregulates its cit operon, responsible for citrate metabolism. In vitro co-cultures revealed that citrate cross-feeding supports increased growth of E. faecalis, which depends both on C. albicans' expression of CIT1 and E. faecalis' expression of the cit operon. A main byproduct of citrate metabolism in E. faecalis is formate, a short chain fatty acid (SCFA) toxic to fungi. Our RNA profiling revealed that C. albicans strongly upregulates three formate dehydrogenases (FDHs) during co-culture, enabling formate detoxification and conferring a growth advantage to C. albicans when grown in E. faecalis conditioned medium. These findings reveal a metabolically driven interaction between C. albicans and E. faecalis in the gut, where cross-feeding of citrate and detoxification of formate facilitates the growth of both species when they are cultured together. Using a simplified fungal-bacterial co-culture system, our studies begin to reveal the mechanistic complexities of metabolic sharing between a eukaryote and a bacterium.
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Project description:Transcriptional profiling of C. albicans in biofilms after 90 min of adherence or 8 h, 24 h, or 48 h of development; compared to C. albicans in suspension cultures grown to log at 30 deg or 37 deg or grown to stationary phase at 30 deg or collected from the unadhered cells in the biofilm assay. At least 2 biological replicates for each condition. All experiments are multiple condition: C. albicans in particular growth condition vs. mixed reference of all conditions in that experiment.
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Project description:Enterococcus faecalis is a major nosocomial pathogen frequently isolated from non- healing wound infections. The major factor responsible for the virulence and establishment of infection is its ability to form robust biofilm. This renders the recalcitrant nature of Enterococci towards the current treatment strategies. In the present study, the quantitative proteomic approach is carried out to elucidate the protein expression levels in E. faecalis at planktonic and biofilm stages. This will help to identify biofilm associated pathways in E. faecalis which inturn can be considered as novel targets for biofilm inhibition.
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Project description:Enterococcus faecalis is often co-isolated with Pseudomonas aeruginosa in mixed-species biofilm-associated infections of wounds and the urinary tract. As a defence strategy, the host innately restricts iron availability at infection sites. Despite their co-prevalence, the polymicrobial interactions of these two pathogens in low iron conditions, such as those found in the host, remains unexplored. Here we show that E. faecalis inhibits P. aeruginosa growth within macrocolony biofilms when iron is restricted. E. faecalis lactate dehydrogenase (ldh1) gives rise to L-lactate production during fermentative growth. We find that E. faecalis ldh1 mutant fails to inhibit P. aeruginosa growth. Additionally, we demonstrate that ldh1 expression is induced when iron is restricted, resulting in increased lactic acid exported and consequently, a reduction in pH. Together, our results suggest that E. faecalis synergistically impact P. aeruginosa growth negatively by decreasing environmental pH and L-lactate-mediated iron chelation. Overall, this study highlights that the microenvironment in which the infection occurs is important for understanding its pathophysiology.
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