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Applied Microbiology and Biotechnology

, Volume 102, Issue 12, pp 5255–5264 | Cite as

Lovastatin synergizes with itraconazole against planktonic cells and biofilms of Candida albicans through the regulation on ergosterol biosynthesis pathway

  • Yujie Zhou
  • Hong Yang
  • Xuedong Zhou
  • Hongke Luo
  • Fan Tang
  • Jin Yang
  • Gil Alterovitz
  • Lei Cheng
  • Biao Ren
Applied microbial and cell physiology

Abstract

The increase of fungal infectious diseases and lack of safe and efficacious antifungal drugs result in the urgent need of new therapeutic strategies. Here, we repurposed the lovastatin (LOV) as a synergistic antifungal potentiator to itraconazole (ITZ) against Candida albicans planktonic cells and biofilms in vitro for the first time. Mutants from ergosterol biosynthesis pathway were employed and key gene expression profiles of ergosterol pathway were also measured. LOV single treatment was unable to inhibit C. albicans strains except the ERG3 and ERG11 double mutant. LOV and ITZ combination was capable of inhibiting the C. albicans planktonic cells and biofilms synergistically including the ITZ resistant mutants. The synergistic antifungal ability was stronger in either ERG11 or ERG3 dysfunctional mutants compared to wild type. The combination lost the synergistic activities in the ERG11 and ERG3 double mutant, while it was sensitive to LOV single treatment. The expression of HMG1, encoding HMG-CoA the target of LOV, was significantly upregulated in ERG11 and ERG3 double mutant strain by the treatment of the combination at 1.5 and 3 h. The combination also significantly increased the HMG1 expression in mutants from ergosterol pathway compared with wild type. The ERG11 and ERG3 gene expressions were upregulated by ITZ and its combination with LOV, but seemingly not by LOV single treatment after 1.5 and 3 h. The combination of LOV and ITZ on C. albicans planktonic cells and biofilms highlights its potential clinical practice especially against the azole drug-resistant mutants.

Keywords

Fungal infection Drug resistance Biofilm Ergosterol biosynthesis Statin 

Notes

Acknowledgements

We greatly thank Prof. Dominique Sanglard for providing the C. albicans mutants. This work was supported by the National Natural Science Foundation of China (grant nos. 81600858, 81372889, and 81430011), National Key Research and Development Program of China (2016YFC1102700), and Youth Grant of the Science and Technology Department of Sichuan Province, China (2017JQ0028).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_8959_MOESM1_ESM.pdf (207 kb)
ESM 1 (PDF 207 kb)

References

  1. Aftab BT, Dobromilskaya I, Liu JO, Rudin CM (2011) Itraconazole inhibits angiogenesis and tumor growth in non–small cell lung cancer. Cancer Res 71(21):6764–6772CrossRefPubMedPubMedCentralGoogle Scholar
  2. Akins RA (2005) An update on antifungal targets and mechanisms of resistance in Candida albicans. Med Mycol 43(4):285–318CrossRefPubMedGoogle Scholar
  3. Anderson JB (2005) Evolution of antifungal-drug resistance: mechanisms and pathogen fitness. Rev Microbiol 3(7):547–556Google Scholar
  4. Cannon RD, Lamping E, Holmes AR, Niimi K, Baret PV, Keniya MV, Tanabe K, Niimi M, Goffeau A, Monk BC (2009) Efflux-mediated antifungal drug resistance. Clin Microbiol Rev 22(2):291–321.  https://doi.org/10.1128/CMR.00051-08 CrossRefPubMedPubMedCentralGoogle Scholar
  5. Carole AS, Gerald RD (1994) Infections in bone marrow transplant recipients. Clin Infect Dis 18(3):273–281CrossRefGoogle Scholar
  6. Chen X, Ren B, Chen M, Liu M-X, Ren W, Wang Q-X, Zhang L-X, Yan G-Y (2014) ASDCD: antifungal synergistic drug combination database. PLoS One 9(1):e86499CrossRefPubMedPubMedCentralGoogle Scholar
  7. Chen X, Ren B, Chen M, Wang Q, Zhang L, Yan G (2016) NLLSS: predicting synergistic drug combinations based on semi-supervised learning. PLoS Comput Biol 12(7):e1004975CrossRefPubMedPubMedCentralGoogle Scholar
  8. Coste A, Selmecki A, Forche A, Diogo D, Bougnoux ME, d'Enfert C, Berman J, Sanglard D (2007) Genotypic evolution of azole resistance mechanisms in sequential Candida albicans isolates. Eukaryot Cell 6(10):1889–1904.  https://doi.org/10.1128/ec.00151-07 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Coste A, Turner V, Ischer F, Morschhauser J, Forche A, Selmecki A, Berman J, Bille J, Sanglard D (2006) A mutation in Tac1p, a transcription factor regulating CDR1 and CDR2, is coupled with loss of heterozygosity at chromosome 5 to mediate antifungal resistance in Candida albicans. Genetics 172(4):2139–2156.  https://doi.org/10.1534/genetics.105.054767 CrossRefPubMedPubMedCentralGoogle Scholar
  10. Crowley JH, Parks LW (1999) Dual physiological effects of antifungal sterol biosynthetic inhibitors on enzyme targets and on transcriptional regulation. Pest Manag Sci 55(4):393–397CrossRefGoogle Scholar
  11. Cuervo G, Garcia-Vidal C, Nucci M, Puchades F, Fernández-Ruiz M, Mykietiuk A, Manzur A, Gudiol C, Pemán J, Viasus D (2013) Effect of statin use on outcomes of adults with candidemia. PLoS One 8(10):e77317CrossRefPubMedPubMedCentralGoogle Scholar
  12. Enoch DA, Ludlam HA, Brown NM (2006) Invasive fungal infections: a review of epidemiology and management options. J Med Microbiol 55(7):809–818.  https://doi.org/10.1099/jmm.0.46548-0 CrossRefPubMedGoogle Scholar
  13. Fonzi WA, Irwin MY (1993) Isogenic strain construction and gene mapping in Candida albicans. Genetics 134(3):717–728PubMedPubMedCentralGoogle Scholar
  14. Istvan ES, Deisenhofer J (2001) Structural mechanism for statin inhibition of HMG-CoA reductase. Science 292(5519):1160–1164CrossRefPubMedGoogle Scholar
  15. Liu Z, Myers LC (2017) Mediator tail module is required for Tac1-activated CDR1 expression and azole resistance in Candida albicans. Antimicrob Agents Chemother 61(11):e01342–e01317.  https://doi.org/10.1128/aac.01342-17 PubMedPubMedCentralCrossRefGoogle Scholar
  16. Loeffler J, Stevens DA (2003) Antifungal drug resistance. Clin Infect Dis 36(Suppl 1):S31–S41.  https://doi.org/10.1086/344658 CrossRefPubMedGoogle Scholar
  17. Lorenz R, Parks L (1990) Effects of lovastatin (mevinolin) on sterol levels and on activity of azoles in Saccharomyces cerevisiae. Antimicrob Agents Chemother 34(9):1660–1665CrossRefPubMedPubMedCentralGoogle Scholar
  18. Macreadie IG, Johnson G, Schlosser T, Macreadie PI (2006) Growth inhibition of Candida species and Aspergillus fumigatus by statins. FEMS Microbiol Lett 262(1):9–13CrossRefPubMedGoogle Scholar
  19. Martin MV (2000) The use of fluconazole and itraconazole in the treatment of Candida albicans infections: a review. J Antimicrob Chemother 45(4):555CrossRefPubMedGoogle Scholar
  20. Mathé L, Van Dijck P (2013) Recent insights into Candida albicans biofilm resistance mechanisms. Curr Genet 59(4):251–264CrossRefPubMedPubMedCentralGoogle Scholar
  21. Menezes EA, AAd VJ, Silva CLF, Plutarco FX, Cunha MCS, Cunha FA (2012) In vitro synergism of simvastatin and fluconazole against Candida species. Rev Inst Med Trop São Paulo 54(4):197–199CrossRefPubMedGoogle Scholar
  22. Morschhauser J (2010) Regulation of multidrug resistance in pathogenic fungi. Fungal Genet Biol 47(2):94–106.  https://doi.org/10.1016/j.fgb.2009.08.002 CrossRefPubMedGoogle Scholar
  23. Nash JD, Burgess DS, Talbert RL (2002) Effect of fluvastatin and pravastatin, HMG-CoA reductase inhibitors, on fluconazole activity against Candida albicans. J Med Microbiol 51(2):105–109CrossRefPubMedGoogle Scholar
  24. Nobile CJ, Johnson AD (2015) Candida albicans biofilms and human disease. Annu Rev Microbiol 69:71–92CrossRefPubMedPubMedCentralGoogle Scholar
  25. Nyilasi I, Kocsubé S, Krizsán K, Galgóczy L, Pesti M, Papp T, Vágvölgyi C (2010) In vitro synergistic interactions of the effects of various statins and azoles against some clinically important fungi. FEMS Microbiol Lett 307(2):175–184CrossRefPubMedGoogle Scholar
  26. Odds FC (2003) Synergy, antagonism, and what the chequerboard puts between them. J Antimicrob Chemother 52(1):1–1CrossRefPubMedGoogle Scholar
  27. Odds FC, Brown AJP, Gow NAR (2003) Antifungal agents: mechanisms of action. Trends Microbiol 11(6):272–279.  https://doi.org/10.1016/S0966-842X(03)00117-3 CrossRefPubMedGoogle Scholar
  28. Perchellet JP, Perchellet EM, Crow KR, Buszek KR, Brown N, Ellappan S, Gao G, Luo D, Minatoya M, Lushington GH (2009) Novel synthetic inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme a (HMG-CoA) reductase activity that inhibit tumor cell proliferation and are structurally unrelated to existing statins. Int J Mol Med 24(5):633–643CrossRefPubMedPubMedCentralGoogle Scholar
  29. Pfaller MA (2012) Antifungal drug resistance: mechanisms, epidemiology, and consequences for treatment. Am J Med 125(1 Suppl):S3–S13.  https://doi.org/10.1016/j.amjmed.2011.11.001 CrossRefPubMedGoogle Scholar
  30. Qiao J, Kontoyiannis DP, Wan Z, Li R, Liu W (2007) Antifungal activity of statins against Aspergillus species. Med Mycol 45(7):589–593CrossRefPubMedGoogle Scholar
  31. Sangeorzan JA, Bradley SF, He X, Zarins LT, Ridenour GL, Tiballi RN, Kauffman CA (1994) Epidemiology of oral candidiasis in HIV-infected patients: colonization, infection, treatment, and emergence of fluconazole resistance. Am J Med 97(4):339–346.  https://doi.org/10.1016/0002-9343(94)90300-X CrossRefPubMedGoogle Scholar
  32. Sanglard D, Coste A, Ferrari S (2009) Antifungal drug resistance mechanisms in fungal pathogens from the perspective of transcriptional gene regulation. FEMS Yeast Res 9(7):1029–1050.  https://doi.org/10.1111/j.1567-1364.2009.00578.x CrossRefPubMedGoogle Scholar
  33. Sanglard D, Ischer F, Parkinson T, Falconer D, Bille J (2003) Candida albicans mutations in the ergosterol biosynthetic pathway and resistance to several antifungal agents. Antimicrob Agents Chemother 47(8):2404–2412CrossRefPubMedPubMedCentralGoogle Scholar
  34. Shapiro RS, Robbins N, Cowen LE (2011) Regulatory circuitry governing fungal development, drug resistance, and disease. Microbiol Mol Bio Rev 75(2):213–267.  https://doi.org/10.1128/mmbr.00045-10 CrossRefGoogle Scholar
  35. Song JL, Lyons CN, Holleman S, Oliver BG, White TC (2003) Antifungal activity of fluconazole in combination with lovastatin and their effects on gene expression in the ergosterol and prenylation pathways in Candida albicans. Med Mycol 41(5):417–425CrossRefPubMedGoogle Scholar
  36. Sun J, Qi C, Lafleur MD, Qi Q-g (2009) Fluconazole susceptibility and genotypic heterogeneity of oral Candida albicans colonies from the patients with cancer receiving chemotherapy in China. Int J Oral Sci 1(3):156–162CrossRefPubMedPubMedCentralGoogle Scholar
  37. Xie JL, O'Meara TR, Polvi EJ, Robbins N, Cowen LE (2017) Staurosporine induces filamentation in the human fungal pathogen Candida albicans via signaling through Cyr1 and protein kinase A. mSphere 2(2):e00056.  https://doi.org/10.1128/mSphere.00056-17 CrossRefPubMedPubMedCentralGoogle Scholar
  38. Xu Y, Chen L, Li C (2008) Susceptibility of clinical isolates of Candida species to fluconazole and detection of Candida albicans ERG11 mutations. J Antimicrob Chemother 61(4):798–804.  https://doi.org/10.1093/jac/dkn015 CrossRefPubMedGoogle Scholar
  39. Zhang L, Yan K, Zhang Y, Huang R, Bian J, Zheng C, Sun H, Chen Z, Sun N, An R (2007) High-throughput synergy screening identifies microbial metabolites as combination agents for the treatment of fungal infections. Proc Nati Acade Sci U S A 104(11):4606–4611CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Oral Diseases and National Clinical Research Center for Oral Diseases, West China Hospital of StomatologySichuan UniversityChengduChina
  2. 2.Department of Operative Dentistry and Endodontics, West China Hospital of StomatologySichuan UniversityChengduChina
  3. 3.Department of Orthodontics, West China Hospital of StomatologySichuan UniversityChengduChina
  4. 4.Harvard Medical SchoolBoston Children’s HospitalBostonUSA

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