The antimycotic 5-fluorocytosine is a virulence inhibitor of uropathogenic Escherichia coli and eradicates biofilm-embedded bacteria synergizing with β-lactams

TL;DR

5-Fluorocytosine, an antifungal drug, inhibits virulence and biofilm formation in uropathogenic E. coli and works synergistically with β-lactam antibiotics.

Contribution

5-FC's antivirulence and antibiofilm effects in UPEC are temperature- and pH-independent, and it synergizes with β-lactams against biofilms.

Findings

5-FC reduced curli fiber gene expression and inhibited virulence factors in UPEC strains.

5-FC impaired UPEC adherence to bladder cells and improved host cell survival.

Combining 5-FC with β-lactams significantly reduced viability of biofilm-embedded bacteria.

Abstract

Biofilm can enhance antibiotic tolerance in bacteria, making treatment of biofilm-associated infections in clinical settings a significant challenge. 5-Fluorocytosine (5-FC), an FDA-approved drug mostly used as an antifungal, can hinder biofilm formation and production of virulence factors in Gram-negative bacteria. In this study, we tested 5-FC on nine uropathogenic Escherichia coli (UPEC) strains plus a fecal isolate. Our data indicated that 5-FC reduced curli fiber gene expression and inhibited virulence factors in UPEC strains. Unlike what was observed in other microorganisms, 5-FC antivirulence and antibiofilm properties were unaffected by either growth temperature or the medium pH, which might prove critical in urinary tract infection (UTI) treatment. Additionally, 5-FC impaired the expression of various UPEC virulence factors, including secreted toxins and type I and P fimbriae, thus leading to decreased UPEC adherence to bladder epithelial cells and improved survival of host cells. Finally, we found that a combination of 5-FC with β-lactams, but not other classes of antibiotics, significantly lowered the viability of bacteria in preformed biofilms. Despite a small set of pathogenic E. coli strains and an in vitro infection model, our findings strongly suggest that 5-FC might be a possible candidate as an antivirulence agent, particularly in a synergistic approach with β-lactam antibiotics.

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