Ottaviano Emerenziana, Dei Cas Michele, Ancona Silvia, Triva Francesca, Casati Sara, Sisto Francesca, Borghi Elisa
Department of Health Sciences, Università degli Studi di Milano, Milan, Lombardy, Italy.
Department of Biomedical, Surgical and Dental Sciences, Università degli Studi di Milano, Milan, Lombardy, Italy.
Microbiol Spectr. 2025 May 6;13(5):e0298724. doi: 10.1128/spectrum.02987-24. Epub 2025 Mar 20.
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 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.
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.
菌丝形成和生物膜形成是与组织侵袭和抗真菌耐受性相关的关键毒力因子。毛果芸香碱盐酸盐（PHCl），一种毒蕈碱受体激动剂，可抑制生物膜成熟，但其机制尚不清楚。我们通过分析经处理的SC5314生物膜中的鞘脂和脂质组成以及蛋白质组学来探究PHCl的作用。PHCl显著降低了生物膜中的极性脂质和麦角固醇水平，同时诱导了植物鞘氨醇和葡糖神经酰胺的积累。PHCl还诱导了活性氧的产生和早期凋亡。蛋白质组学分析表明，PHCl处理使生物膜中与代谢、细胞壁重塑和DNA修复相关的蛋白质下调至与浮游细胞中观察到的水平相当。与麦角固醇减少一致，发现Erg2减少。总体而言，PHCl破坏了生物膜完整性所必需的关键途径，降低了其稳定性并促进了表面脱离，突出了其作为一种通用抗真菌化合物的潜力。
菌丝形成和生物膜形成是促进真菌持续存在和耐药性的关键毒力因子。哺乳动物细胞常见的真核性质对新型活性无毒化合物的开发构成了重大限制。了解PHCl对酵母-菌丝转变、生物膜粘附和成熟的抑制活性的潜在机制可为在缺乏药物投资的领域中进行有效的药物重新利用铺平道路。
