1887

Abstract

Biofilms are the natural growth state for most microorganisms. biofilms are composed of multiple cell types (round budding yeast-form cells, oval pseudohyphal cells, and elongated hyphal cells) encased in an extracellular matrix. biofilms are notorious for resistance to antimicrobial treatments, a property that might be determined by complex mechanisms. Exogenous farnesol exerts a certain antifungal activity against with medical implications. Different from other microbes, biofilms can tolerate exogenous farnesol at high concentration with some cells still surviving and even maintaining proliferation, but the mechanism is unclear.

The study hypothesizes that resists farnesol by activating the PKC signalling pathway.

The study aims to discuss the molecular mechanism of resistance to farnesol.

The ROS levels, the genes and proteins of the PKC pathway were compared between the farnesol-tolerant and non-tolerant groups using ROS levels assay, -RT PCR and Western blot, respectively. Further, the mutant strains (Δ/Δ and Δ/Δ) were constructed, then the survival rates and ROS levels of biofilms exposed to farnesol were compared between mutant and wild strains. The morphological changes were observed using TEM.

The survival rates of biofilms decreased rapidly under the lower concentration of farnesol (<0.05), and kept stable (>0.05) as the concentration rose up to 200 µM. The gene expression of the PKC pathway increased, while ROS levels remained stable and even decreased in the farnesol-tolerant biofilms, compared with those in the farnesol-nontolerant biofilms after farnesol treatment (<0.05); and were significantly upregulated by during the development of biofilm tolerance to farnesol. The cell wall and cytoplasm of Δ/Δ and Δ/Δ were damaged, and the ROS level increased (<0.05); meanwhile, the survival rate of biofilms decreased compared with that of wild-type strain under the same farnesol concentrations (<0.05). ROS inhibitors reversed these changes in Δ/Δ and Δ/Δ when the mutant strains exposed to farnesol.

biofilms can tolerate high concentrations of farnesol by activating and of the PKC pathway and stabilizing ROS levels. The and are two key genes regulated by in the process of biofilm tolerance to farnesol.

Funding
This study was supported by the:
  • The Foundation of the Priority Academic Program Development of Jiangsu Higher Education Institutions (Award PAPD, 2018-87)
    • Principle Award Recipient: XinWei
  • National Natural Sciences Foundation of China (Award 81970945)
    • Principle Award Recipient: XinWei
  • National Natural Sciences Foundation of China (Award 81371156)
    • Principle Award Recipient: XinWei
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001476
2022-01-12
2022-01-17
Loading full text...

Full text loading...

References

  1. Kullberg BJ, Vasquez J, Mootsikapun P, Nucci M, Paiva J-A et al. Efficacy of anidulafungin in 539 patients with invasive candidiasis: a patient-level pooled analysis of six clinical trials. J Antimicrob Chemother 2017; 72:2368–2377 [View Article] [PubMed]
    [Google Scholar]
  2. Wall G, Montelongo-Jauregui D, Vidal Bonifacio B, Lopez-Ribot JL, Uppuluri P. Candida albicans biofilm growth and dispersal: contributions to pathogenesis. Curr Opin Microbiol 2019; 52:1–6 [View Article] [PubMed]
    [Google Scholar]
  3. Taff HT, Mitchell KF, Edward JA, Andes DR. Mechanisms of Candida biofilm drug resistance. Future Microbiol 2013; 8:1325–1337 [View Article] [PubMed]
    [Google Scholar]
  4. Yu L-H, Wei X, Ma M, Chen X-J, Xu S-B. Possible inhibitory molecular mechanism of farnesol on the development of fluconazole resistance in Candida albicans biofilm. Antimicrob Agents Chemother 2012; 56:770–775 [View Article] [PubMed]
    [Google Scholar]
  5. Egbe NE, Dornelles TO, Paget CM, Castelli LM, Ashe MP. Farnesol inhibits translation to limit growth and filamentation in C. albicans and S. cerevisiae. Microb Cell 2017; 4:294–304 [View Article] [PubMed]
    [Google Scholar]
  6. Hasim S, Vaughn EN, Donohoe D, Gordon DM, Pfiffner S et al. Influence of phosphatidylserine and phosphatidylethanolamine on farnesol tolerance in Candida albicans. Yeast 2018; 35:343–351 [View Article] [PubMed]
    [Google Scholar]
  7. Polke M, Leonhardt I, Kurzai O, Jacobsen ID. Farnesol signalling in Candida albicans - more than just communication. Crit Rev Microbiol 2018; 44:230–243 [View Article] [PubMed]
    [Google Scholar]
  8. Xia J, Qian F, Xu W, Zhang Z, Wei X. In vitro inhibitory effects of farnesol and interactions between farnesol and antifungals against biofilms of Candida albicans resistant strains. Biofouling 2017; 33:283–293 [View Article] [PubMed]
    [Google Scholar]
  9. Gulati M, Nobile CJ. Candida albicans biofilms: development, regulation, and molecular mechanisms. Microbes Infect 2016; 18:310–321 [View Article] [PubMed]
    [Google Scholar]
  10. Höfs S, Mogavero S, Hube B. Interaction of Candida albicans with host cells: virulence factors, host defense, escape strategies, and the microbiota. J Microbiol 2016; 54:149–169 [View Article] [PubMed]
    [Google Scholar]
  11. Liu N-N, Uppuluri P, Broggi A, Besold A, Ryman K et al. Intersection of phosphate transport, oxidative stress and TOR signalling in Candida albicans virulence. PLoS Pathog 2018; 14:e1007076 [View Article] [PubMed]
    [Google Scholar]
  12. da Silva Dantas A, Day A, Ikeh M, Kos I, Achan B et al. Oxidative stress responses in the human fungal pathogen, Candida albicans. Biomolecules 2015; 5:142–165 [View Article] [PubMed]
    [Google Scholar]
  13. Yu Q, Zhang B, Li J, Zhang B, Wang H et al. Endoplasmic reticulum-derived reactive oxygen species (ROS) is involved in toxicity of cell wall stress to Candida albicans. Free Radic Biol Med 2016; 99:572–583 [View Article] [PubMed]
    [Google Scholar]
  14. LaFayette SL, Collins C, Zaas AK, Schell WA, Betancourt-Quiroz M et al. PKC signaling regulates drug resistance of the fungal pathogen Candida albicans via circuitry comprised of Mkc1, calcineurin, and Hsp90. PLoS Pathog 2010; 6:e1001069 [View Article] [PubMed]
    [Google Scholar]
  15. Navarro-García F, Eisman B, Fiuza SM, Nombela C, Pla J. The MAP kinase Mkc1p is activated under different stress conditions in Candida albicans. Microbiology (Reading) 2005; 151:2737–2749 [View Article] [PubMed]
    [Google Scholar]
  16. Nett JE, Sanchez H, Cain MT, Ross KM, Andes DR. Interface of Candida albicans biofilm matrix-associated drug resistance and cell wall integrity regulation. Eukaryot Cell 2011; 10:1660–1669 [View Article] [PubMed]
    [Google Scholar]
  17. Machida K, Tanaka T, Fujita K, Taniguchi M. Farnesol-induced generation of reactive oxygen species via indirect inhibition of the mitochondrial electron transport chain in the yeast Saccharomyces cerevisiae. J Bacteriol 1998; 180:4460–4465 [View Article] [PubMed]
    [Google Scholar]
  18. Fairn GD, MacDonald K, McMaster CR. A chemogenomic screen in Saccharomyces cerevisiae uncovers a primary role for the mitochondria in farnesol toxicity and its regulation by the Pkc1 pathway. J Biol Chem 2007; 282:4868–4874 [View Article] [PubMed]
    [Google Scholar]
  19. Heinisch JJ, Rodicio R. Protein kinase C in fungi—more than just cell wall integrity. FEMS Microbiol Rev 2017; 42:fux051 [View Article]
    [Google Scholar]
  20. Ramage G, Vande Walle K, Wickes BL, López-Ribot JL. Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. Antimicrob Agents Chemother 2001; 45:2475–2479 [View Article] [PubMed]
    [Google Scholar]
  21. Pierce CG, Uppuluri P, Tummala S, Lopez-Ribot JL. A 96 well microtiter plate-based method for monitoring formation and antifungal susceptibility testing of Candida albicans biofilms. JoVE 201044 [View Article]
    [Google Scholar]
  22. Kuhn DM, Balkis M, Chandra J, Mukherjee PK, Ghannoum MA. Uses and limitations of the XTT assay in studies of Candida growth and metabolism. J Clin Microbiol 2003; 41:506–508 [View Article] [PubMed]
    [Google Scholar]
  23. Pumeesat P, Muangkaew W, Ampawong S, Luplertlop N. Candida albicans biofilm development under increased temperature. New Microbiol 2017; 40:279–283 [PubMed]
    [Google Scholar]
  24. Taskova RM, Zorn H, Krings U, Bouws H, Berger RG. A comparison of cell wall disruption techniques for the isolation of intracellular metabolites from Pleurotus and Lepista sp. Z Naturforsch C J Biosci 2006; 61:347–350 [View Article] [PubMed]
    [Google Scholar]
  25. Janeczko M. Emodin reduces the activity of (1,3)-β-D-glucan Synthase from Candida albicans and does not interact with Caspofungin. Pol J Microbiol 2018; 67:463–470 [View Article] [PubMed]
    [Google Scholar]
  26. Liu H-T, Li W-M, Xu G, Li X-Y, Bai X-F et al. Chitosan oligosaccharides attenuate hydrogen peroxide-induced stress injury in human umbilical vein endothelial cells. Pharmacol Res 2009; 59:167–175 [View Article] [PubMed]
    [Google Scholar]
  27. Lee W, Lee DG. Reactive oxygen species modulate itraconazole-induced apoptosis via mitochondrial disruption in Candida albicans. Free Radic Res 2018; 52:39–50 [View Article] [PubMed]
    [Google Scholar]
  28. Zeng X, Ye G, Tang W, Ouyang T, Tian L et al. Fungicidal efficiency of electrolyzed oxidizing water on Candida albicans and its biochemical mechanism. J Biosci Bioeng 2011; 112:86–91 [View Article] [PubMed]
    [Google Scholar]
  29. Chen S, Xia J, Li C, Zuo L, Wei X. The possible molecular mechanisms of farnesol on the antifungal resistance of C. albicans biofilms: the regulation of CYR1 and PDE2. BMC Microbiol 2018; 18:203. [View Article] [PubMed]
    [Google Scholar]
  30. Fuchs BB, Mylonakis E. Our paths might cross: the role of the fungal cell wall integrity pathway in stress response and cross talk with other stress response pathways. Eukaryot Cell 2009; 8:1616–1625 [View Article] [PubMed]
    [Google Scholar]
  31. Zhao X, Mehrabi R, Xu J-R. Mitogen-activated protein kinase pathways and fungal pathogenesis. Eukaryot Cell 2007; 6:1701–1714 [View Article] [PubMed]
    [Google Scholar]
  32. Delattin N, Cammue BPA, Thevissen K. Reactive oxygen species-inducing antifungal agents and their activity against fungal biofilms. Future Med Chem 2014; 6:77–90 [View Article] [PubMed]
    [Google Scholar]
  33. Cáp M, Váchová L, Palková Z. Reactive oxygen species in the signaling and adaptation of multicellular microbial communities. Oxid Med Cell Longev 2012; 2012:976753. [View Article] [PubMed]
    [Google Scholar]
  34. Frost DJ, Brandt KD, Cugier D, Goldman R. A whole-cell Candida albicans assay for the detection of inhibitors towards fungal cell wall synthesis and assembly. J Antibiot 1995; 48:306–310 [View Article]
    [Google Scholar]
  35. Román E, Alonso-Monge R, Gong Q, Li D, Calderone R et al. The Cek1 MAPK is a short-lived protein regulated by quorum sensing in the fungal pathogen Candida albicans. FEMS Yeast Res 2009; 9:942–955 [View Article] [PubMed]
    [Google Scholar]
  36. Noble SM, Johnson AD. Strains and strategies for large-scale gene deletion studies of the diploid human fungal pathogen Candida albicans. Eukaryot Cell 2005; 4:298–309 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001476
Loading
/content/journal/jmm/10.1099/jmm.0.001476
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error