Journal of Veterinary Science

The Korean Society of Veterinary Science (대한수의학회)
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Transcriptome sequencing revealed the inhibitory mechanism of ketoconazole on clinical Microsporum canis

  • Wang, Mingyang (Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University) ;
  • Zhao, Yan (Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University) ;
  • Cao, Lingfang (Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University) ;
  • Luo, Silong (Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University) ;
  • Ni, Binyan (Qingdao Vetlab Biotechnology Co., Ltd.) ;
  • Zhang, Yi (Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University) ;
  • Chen, Zeliang (Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University)
  • Received : 2020.09.04
  • Accepted : 2020.11.05
  • Published : 2021.01.31
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Abstract

Background: Microsporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans. Objectives: In clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood. The antifungal mechanism of KTZ needs to be studied in detail. Methods: In this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis. The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto. Results: At half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated (p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ. Conclusions: The results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.

Keywords

Acknowledgement

This research was funded by the Education Department of Liaoning Province (LSNYB201612), National Key Research and Development Program of China (2017YFD0500901).

References

  1. Binder B, Lackner HK, Poessl BD, Propst E, Weger W, Smolle J, et al. Prevalence of tinea capitis in Southeastern Austria between 1985 and 2008: up-to-date picture of the current situation. Mycoses. 2011;54(3):243-247. https://doi.org/10.1111/j.1439-0507.2009.01804.x
  2. Bond R, Morris DO, Guillot J, Bensignor EJ, Robson D, Mason KV, et al. Biology, diagnosis and treatment of Malassezia dermatitis in dogs and cats: clinical consensus guidelines of the world association for veterinary dermatology. Vet Dermatol. 2020;31(1):75. https://doi.org/10.1111/vde.12834
  3. Yin B, Xiao Y, Ran Y, Kang D, Dai Y, Lama J. Microsporum canis infection in three familial cases with tinea capitis and tinea corporis. Mycopathologia. 2013;176(3-4):259-265. https://doi.org/10.1007/s11046-013-9685-5
  4. Anemuller W, Baumgartner S, Brasch J. Atypical Microsporum canis variant in an immunosuppressed child. J Dtsch Dermatol Ges. 2008;6(6):473-475. https://doi.org/10.1111/j.1610-0387.2007.06458.x
  5. Ginter-Hanselmayer G, Weger W, Ilkit M, Smolle J. Epidemiology of tinea capitis in Europe: current state and changing patterns. Mycoses. 2007;50 Suppl 2:6-13. https://doi.org/10.1111/j.1439-0507.2007.01424.x
  6. Verma SB. Emergence of recalcitrant dermatophytosis in India. Lancet Infect Dis. 2018;18(7):718-719. https://doi.org/10.1016/S1473-3099(18)30338-4
  7. Aneke CI, Otranto D, Cafarchia C. Therapy and antifungal susceptibility profile of Microsporum canis. J Fungi (Basel). 2018;4(3):107. https://doi.org/10.3390/jof4030107
  8. Ghannoum MA, Rice LB. Antifungal agents: mode of action, mechanisms of resistance, and correlation of these mechanisms with bacterial resistance. Clin Microbiol Rev. 1999;12(4):501-517. https://doi.org/10.1128/cmr.12.4.501
  9. Vanden Bossche H. Biochemical targets for antifungal azole derivatives: hypothesis on the mode of action. Curr Top Med Mycol. 1985;1:313-351. https://doi.org/10.1007/978-1-4613-9547-8_12
  10. Borgers M, Van den Bossche H, De Brabander M. The mechanism of action of the new antimycotic ketoconazole. Am J Med. 1983;74(1B):2-8.
  11. Wang Z, Gerstein M, Snyder M. RNA-Seq: a revolutionary tool for transcriptomics. Nat Rev Genet. 2009;10(1):57-63. https://doi.org/10.1038/nrg2484
  12. Lockhart DJ, Winzeler EA. Genomics, gene expression and DNA arrays. Nature. 2000;405:827-836. https://doi.org/10.1038/35015701
  13. Costa V, Angelini C, De Feis I, Ciccodicola A. Uncovering the complexity of transcriptomes with RNA-Seq. J Biomed Biotechnol. 2010;2010:853916.
  14. Sheehan DJ, Hitchcock CA, Sibley CM. Current and emerging azole antifungal agents. Clin Microbiol Rev. 1999;12(1):40-79. https://doi.org/10.1128/cmr.12.1.40
  15. Sanati H, Belanger P, Fratti R, Ghannoum M. A new triazole, voriconazole (UK-109,496), blocks sterol biosynthesis in Candida albicans and Candida krusei. Antimicrob Agents Chemother. 1997;41(11):1249-6.
  16. Ghannoum MA, Spellberg BJ, Ibrahim AS, Ritchie JA, Currie B, Spitzer ED, et al. Sterol composition of Cryptococcus neoformans in the presence and absence of fluconazole. Antimicrob Agents Chemother. 1994;38(9):2029-2033. https://doi.org/10.1128/AAC.38.9.2029
  17. Vanden Bossche H, Marichal P, Le Jeune L, Coene MC, Gorrens J, Cools W. Effects of itraconazole on cytochrome P-450-dependent sterol 14 alpha-demethylation and reduction of 3-ketosteroids in Cryptococcus neoformans. Antimicrob Agents Chemother. 1993;37(10):2101-2105. https://doi.org/10.1128/AAC.37.10.2101
  18. Xiao CW, Ji QA, Wei Q, Liu Y, Pan LJ, Bao GL. Digital gene expression analysis of Microsporum canis exposed to berberine chloride. PLoS One. 2015;10(4):e0124265. https://doi.org/10.1371/journal.pone.0124265
  19. Galani K, Nissan TA, Petfalski E, Tollervey D, Hurt E. Rea1, a dynein-related nuclear AAA-ATPase, is involved in late rRNA processing and nuclear export of 60 S subunits. J Biol Chem. 2004;279(53):55411-55418. https://doi.org/10.1074/jbc.M406876200
  20. Lepesheva GI, Waterman MR. Sterol 14alpha-demethylase cytochrome P450 (CYP51), a P450 in all biological kingdoms. Biochim Biophys Acta. 2007;1770(3):407-477.
  21. Lepesheva GI, Friggeri L, Waterman MR. CYP51 as drug targets for fungi and protozoan parasites: past, present and future. Parasitology. 2018;145(14):1820-1836. https://doi.org/10.1017/s0031182018000562
  22. Zhang J, Li L, Lv Q, Yan L, Wang Y, Jiang Y. The fungal CYP51s: their functions, structures, related drug resistance, and inhibitors. Front Microbiol. 2019;10:691. https://doi.org/10.3389/fmicb.2019.00691
  23. Nowosielski M, Hoffmann M, Wyrwicz LS, Stepniak P, Plewczynski DM, Lazniewski M, et al. Detailed mechanism of squalene epoxidase inhibition by terbinafine. J Chem Inf Model. 2011;51(2):455-462. https://doi.org/10.1021/ci100403b
  24. Zare B, Sepehrizadeh Z, Faramarzi MA, Soltany-Rezaee-Rad M, Rezaie S, Shahverdi AR. Antifungal activity of biogenic tellurium nanoparticles against Candida albicans and its effects on squalene monooxygenase gene expression. Biotechnol Appl Biochem. 2014;61(4):395-400. https://doi.org/10.1002/bab.1180
  25. Konecna A, Toth Hervay N, Valachovic M, Gbelska Y. ERG6 gene deletion modifies Kluyveromyces lactis susceptibility to various growth inhibitors. Yeast. 2016;33(12):621-632. https://doi.org/10.1002/yea.3212
  26. Morio F, Pagniez F, Lacroix C, Miegeville M, Le Pape P. Amino acid substitutions in the Candida albicans sterol Δ5,6-desaturase (Erg3p) confer azole resistance: characterization of two novel mutants with impaired virulence. J Antimicrob Chemother. 2012;67(9):2131-2138. https://doi.org/10.1093/jac/dks186
  27. Xiao CW, Ji QA, Wei Q, Liu Y, Bao GL. Antifungal activity of berberine hydrochloride and palmatine hydrochloride against Microsporum canis -induced dermatitis in rabbits and underlying mechanism. BMC Complement Altern Med. 2015;15(4):177. https://doi.org/10.1186/s12906-015-0680-x
  28. Ruiz-Herrera J, San-Blas G. Chitin synthesis as target for antifungal drugs. Curr Drug Targets Infect Disord. 2003;3(1):77-91. https://doi.org/10.2174/1568005033342064
  29. Yang J, Zhang KQ. Chitin synthesis and degradation in fungi: biology and enzymes. Adv Exp Med Biol. 2019;1142:153-167. https://doi.org/10.1007/978-981-13-7318-3_8
  30. Shen Q, Zheng X, McNutt MA, Guang L, Sun Y, Wang J, et al. NAT10, a nucleolar protein, localizes to the midbody and regulates cytokinesis and acetylation of microtubules. Exp Cell Res. 2009;315(10):1653-1667. https://doi.org/10.1016/j.yexcr.2009.03.007
  31. Neyer S, Kunz M, Geiss C, Hantsche M, Hodirnau VV, Seybert A, et al. Structure of RNA polymerase I transcribing ribosomal DNA genes. Nature. 2016;540(7634):607-610. https://doi.org/10.1038/nature20561
  32. Vannini A. A structural perspective on RNA polymerase I and RNA polymerase III transcription machineries. Biochim Biophys Acta. 2013;1829(3-4):258-264. https://doi.org/10.1016/j.bbagrm.2012.09.009
  33. Hopper AK, Nostramo RT. tRNA Processing and subcellular trafficking proteins multitask in pathways for other RNAs. Front Genet. 2019;10:96. https://doi.org/10.3389/fgene.2019.00096
  34. Keene JD. Ribonucleoprotein infrastructure regulating the flow of genetic information between the genome and the proteome. Proc Natl Acad Sci U S A. 2001;98(13):7018-7024. https://doi.org/10.1073/pnas.111145598