Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content

ABSTRACT Candida albicans filamentation and biofilm formation are key virulence factors tied to tissue invasion and antifungal tolerance. Pilocarpine hydrochloride (PHCl), a muscarinic receptor agonist, inhibits biofilm maturation, although its mechanism remains unclear. We explored PHCl effects by...

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Main Authors: Emerenziana Ottaviano, Michele Dei Cas, Silvia Ancona, Francesca Triva, Sara Casati, Francesca Sisto, Elisa Borghi
Format: Article
Language:English
Published: American Society for Microbiology 2025-05-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.02987-24
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author Emerenziana Ottaviano
Michele Dei Cas
Silvia Ancona
Francesca Triva
Sara Casati
Francesca Sisto
Elisa Borghi
author_facet Emerenziana Ottaviano
Michele Dei Cas
Silvia Ancona
Francesca Triva
Sara Casati
Francesca Sisto
Elisa Borghi
author_sort Emerenziana Ottaviano
collection DOAJ
description ABSTRACT Candida albicans filamentation and biofilm formation are key virulence factors tied to tissue invasion and antifungal tolerance. Pilocarpine hydrochloride (PHCl), a muscarinic receptor agonist, inhibits biofilm maturation, although its mechanism remains unclear. We explored PHCl effects by analyzing sphingolipid and lipid composition and proteomics in treated C. albicans SC5314 biofilms. PHCl significantly decreased polar lipid and ergosterol levels in biofilms while inducing phytoceramide and glucosylceramide accumulation. PHCl also induced reactive oxygen species and early apoptosis. Proteomic analysis revealed that PHCl treatment downregulated proteins associated with metabolism, cell wall remodeling, and DNA repair in biofilms to levels comparable to those observed in planktonic cells. Consistent with ergosterol reduction, Erg2 was found to be reduced. Overall, PHCl disrupts key pathways essential for biofilm integrity, decreasing its stability and promoting surface detachment, underscoring its potential as a versatile antifungal compound.IMPORTANCECandida albicans filamentation and biofilm formation represent crucial virulence factors promoting fungus persistence and drug tolerance. The common eukaryotic nature of mammalian cells poses significant limitations to the development of new active nontoxic compounds. Understanding the mechanism underlying PHCl inhibitory activity on yeast–hypha transition, biofilm adhesion, and maturation can pave the way to efficient drug repurposing in a field where pharmaceutical investment is lacking.
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spelling doaj-art-bfdd2c3433b147eabb2055b19b4377792025-08-20T03:52:15ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-05-0113510.1128/spectrum.02987-24Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein contentEmerenziana Ottaviano0Michele Dei Cas1Silvia Ancona2Francesca Triva3Sara Casati4Francesca Sisto5Elisa Borghi6Department of Health Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyDepartment of Health Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyDepartment of Health Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyDepartment of Health Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyDepartment of Biomedical, Surgical and Dental Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyDepartment of Biomedical, Surgical and Dental Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyDepartment of Health Sciences, Università degli Studi di Milano, Milan, Lombardy, ItalyABSTRACT Candida albicans filamentation and biofilm formation are key virulence factors tied to tissue invasion and antifungal tolerance. Pilocarpine hydrochloride (PHCl), a muscarinic receptor agonist, inhibits biofilm maturation, although its mechanism remains unclear. We explored PHCl effects by analyzing sphingolipid and lipid composition and proteomics in treated C. albicans SC5314 biofilms. PHCl significantly decreased polar lipid and ergosterol levels in biofilms while inducing phytoceramide and glucosylceramide accumulation. PHCl also induced reactive oxygen species and early apoptosis. Proteomic analysis revealed that PHCl treatment downregulated proteins associated with metabolism, cell wall remodeling, and DNA repair in biofilms to levels comparable to those observed in planktonic cells. Consistent with ergosterol reduction, Erg2 was found to be reduced. Overall, PHCl disrupts key pathways essential for biofilm integrity, decreasing its stability and promoting surface detachment, underscoring its potential as a versatile antifungal compound.IMPORTANCECandida albicans filamentation and biofilm formation represent crucial virulence factors promoting fungus persistence and drug tolerance. The common eukaryotic nature of mammalian cells poses significant limitations to the development of new active nontoxic compounds. Understanding the mechanism underlying PHCl inhibitory activity on yeast–hypha transition, biofilm adhesion, and maturation can pave the way to efficient drug repurposing in a field where pharmaceutical investment is lacking.https://journals.asm.org/doi/10.1128/spectrum.02987-24Candida albicansbiofilmsdrug repurposing
spellingShingle Emerenziana Ottaviano
Michele Dei Cas
Silvia Ancona
Francesca Triva
Sara Casati
Francesca Sisto
Elisa Borghi
Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content
Microbiology Spectrum
Candida albicans
biofilms
drug repurposing
title Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content
title_full Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content
title_fullStr Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content
title_full_unstemmed Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content
title_short Pilocarpine inhibits Candida albicans SC5314 biofilm maturation by altering lipid, sphingolipid, and protein content
title_sort pilocarpine inhibits candida albicans sc5314 biofilm maturation by altering lipid sphingolipid and protein content
topic Candida albicans
biofilms
drug repurposing
url https://journals.asm.org/doi/10.1128/spectrum.02987-24
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