Abstract
Candida species are one of the predominant causes of fungal infections and show drug-resistant infections in immune-compromised individuals. Simultaneous targeting of existing antifungal drugs with heat shock protein 90 (Hsp90) inhibitors may be an approach that increases the efficacy of antifungal drugs. Also, since most of the patients at risk for invasive fungal infections use these anticancer or immunosuppressive drugs, synergistic interaction in combination treatment can reduce the dose of antifungal drugs and create an alternative for the toxicity problem. In this study, in vitro efficacy of commonly used antifungals (amphotericin B, caspofungin, itraconazole, voriconazole, and fluconazole) in combination with four heat shock protein inhibitors geldanamycin, 17-allylamino-17-demethoxygeldanamycin, radicicol, and novobiocin against 30 clinical Candida isolates (C. albicans n = 13, C. krusei n = 7, and C. glabrata n = 10) were evaluated by time kill and checkerboard methods. The significant synergistic interaction determined especially in the combinations of geldanamycin with antifungal drugs suggests that substances with inhibitory effects on Hsp90 increase the effectiveness of antifungals or reduce the antifungal resistance. Although Hsp90 inhibitors alone did not have any significant antifungal activity, they did not show adverse interactions in combination with antifungals, and at some concentrations, they increased the effectiveness of the antifungals. These in vitro results have been found promising for the development of new therapeutic approaches in the treatment of invasive fungal infections. However, detailed studies are needed.
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Abu-Romman S (2016) Genotypic response to heat stress in durum wheat and the expression of small HSP genes. Rend Fis Acc Lincei 27(2):261–267. https://doi.org/10.1007/s12210-015-0471-9
Campitelli M, Zeineddine N, Samaha G, Maslak S (2017) Combination antifungal therapy: a review of current data. J Clin Med Res 9(6):451–456. https://doi.org/10.14740/jocmr2992w
Carradori S, Ammazzalorso A, De Filippis B, Şahin AF, Akdemir A, Orekhova A, Simonetti G (2022) Azole-based compounds that are active against candida biofilm: in vitro, in vivo and in silico studies. Antibiotics 11(10):1375. https://doi.org/10.3390/antibiotics11101375
Castanheira M, Deshpande LM, Davis AP, Carvalhaes CG, Pfaller MA (2022) Azole resistance in Candida glabrata clinical isolates from global surveillance is associated with efflux overexpression. J Glob Antimicrob Resist. https://doi.org/10.1016/j.jgar.2022.05.004
Chen SC, Lewis RE, Kontoyiannis DP (2011) Direct effects of non-antifungal agents used in cancer chemotherapy and organ transplantation on the development and virulence of Candida and Aspergillus species. Virulence 2(4):280–295. https://doi.org/10.4161/viru.2.4.16764
Clinical Laboratory Standards Institute (CLSI) (2008) Reference method for broth dilution antifungal susceptibility testing of yeasts. In: Approved Standard-Third Edition, CLSI document M27-A3. Wayne, PA: Clinical and Laboratory Standards Institute
Clinical Laboratory Standards Institute (CLSI) (2012) Reference method for broth microdilution antifungal susceptibility testing of yeasts. In: Fourth Informational Supplement, M27-S4. USA: Wayne
Cowen LE (2008) The evolution of fungal drug resistance: modulating the trajectory from genotype to phenotype. Nat Rev Microbiol 6:187–198
Cowen LE, Singh SD, Köhler JR, Collins C, Zaas AK, Schell WA, Aziz H, Mylonakis E, Perfect JR, Whitesell L, Lindquist S (2009) Harnessing Hsp90 function as a powerful, broadly effective therapeutic strategy for fungal infectious disease. Proc Natl Acad Sci USA 106:2818–2823. https://doi.org/10.1073/pnas.0813394106
Cruz MC, Del Poeta M, Wang P, Wenger R, Zenke G, Quesniaux VF, Movva NR, Perfect JR, Cardenas ME, Heitman J (2000) Immunosuppressive and nonimmunosuppressive cyclosporine analogs are toxic to the opportunistic fungal pathogen Cryptococcus neoformans via cyclophilin-dependent inhibition of calcineurin. Antimicrob Agents Chemother 44(1):143–149. https://doi.org/10.1128/AAC.44.1.143-149.2000
Derf A, Verekar SA, Jain SK, Deshmukh SK, Bharate SB, Chaudhuri B (2019) Radicicol rescues yeast cell death triggered by expression of human α-synuclein and its A53T mutant, but not by human βA4 peptide and proapoptotic protein bax. Bioorg Chem 85:152–158. https://doi.org/10.1016/j.bioorg.2018.12.033
Donnelly A, Blagg BS (2008) Novobiocin and additional inhibitors of the Hsp90 C-terminal nucleotide-binding pocket. Curr Med Chem 15(26):2702–2717. https://doi.org/10.2174/092986708786242895
El-Darier SM, El-Ahwany A, Elkenany ET, Abdeldaim AA (2018) An in vitro study on antimicrobial and anticancer potentiality of thyme and clove oils. Rend Fis Acc Lincei 29(1):131–139. https://doi.org/10.1007/s12210-018-0672-0
Gao L, Sun Y, He C, Li M, Zeng T (2018) In vitro interactions between 17-AAG and azoles against Exophiala dermatitidis. Mycoses 61(11):853–856. https://doi.org/10.1111/myc.12824
Hosseini Bafghi M, Safdari H, Nazari R, Darroudi M, Sabouri Z, Zargar M, Zarrinfar H (2021) Evaluation and comparison of the effects of biosynthesized selenium and silver nanoparticles using plant extracts with antifungal drugs on the growth of Aspergillus and Candida species. Rend Fis Acc Lincei 32(4):791–803. https://doi.org/10.1007/s12210-021-01021-0
Infectious Diseases Society of America (IDSA) (2011) Combating antimicrobial resistance: policy recommendations to save lives. Clin Infect Dis 52(suppl_5):S397-428. https://doi.org/10.1093/cid/cir153
Jacob TR, Peres NT, Martins MP, Lang EA, Sanches PR, Rossi A, Martinez-Rossi NM (2015) Heat shock protein 90 (Hsp90) as a molecular target for the development of novel drugs against the dermatophyte Trichophyton rubrum. Front Microbiol 6:1241. https://doi.org/10.3389/fmicb.2015.01241
Johnson MD, MacDougall C, Ostrosky-Zeichner L, Perfect JR, Rex JH (2004) Combination antifungal therapy. Antimicrob Agents Chemother 48(3):693–715. https://doi.org/10.1128/AAC.48.3.693-715.2004
Kaneko Y, Ohno H, Imamura Y, Kohno S, Miyazaki Y (2009) The effects of an Hsp90 inhibitor on the paradoxical effect. Jpn J Infect Dis 62:392–393
Kashefi E, Seyedi SJ, Zarrinfar H, Fata A, Mehrad-Majd H, Najafzadeh MJ (2021) Molecular identification of Candida species in bronchoalveolar lavage specimens of hospitalized children with pulmonary disorders. J Babol Univ Med Sci 23(1):331–336. https://doi.org/10.22088/jbums.23.1.331
Kaya SŞ, Kiraz N, Bariş A, Turan D, Öz Y, Dağ İ, Aygün G (2021) Effects of calcineurin inhibitors, cyclosporine A and tacrolimus (FK506), on the activity of antifungal drugs against Candida spp. J Med Microbiol 70(4):001354. https://doi.org/10.1099/jmm.0.001354
Kiraz N, Dag I, Yamac M, Kiremitci A, Kasifoglu N, Oz Y (2010) Synergistic activities of three triazoles with caspofungin against Candida glabrata isolates determined by time–kill, Etest, and disk diffusion methods. Antimicrob Agents Chemother 54:2244–2247. https://doi.org/10.1128/AAC.01527-09
Lamping E, Ranchod A, Nakamura K, Tyndall JD, Niimi K, Holmes AR, Cannon RD (2009) Abc1p is a multidrug efflux transporter that tips the balance in favor of innate azole resistance in Candida krusei. Antimicrob Agents Chemother 53(2):354–369
Leach MD, Budge S, Walker L, Munro C, Cowen LE, Brown AJ (2012) Hsp90 orchestrates transcriptional regulation by Hsf1 and cell wall remodelling by MAPK signalling during thermal adaptation in a pathogenic yeast. PLoS Pathog 8(12):e1003069. https://doi.org/10.1371/journal.ppat.1003069
Mahmoudi S, Rezaie S, Ghazvini RD, Hashemi SJ, Badali H, Foroumadi A, Diba K, Chowdhary A, Meis JF, Khodavaisy S (2019) In Vitro interaction of geldanamycin with triazoles and echinocandins against common and emerging Candida species. Mycopathologia 184(5):607–613. https://doi.org/10.1007/s11046-019-00370-7(0123456789
Mavor AL, Thewes S, Hube B (2005) Systemic fungal infections caused by Candida species: epidemiology, infection process and virulence attributes. Curr Drug Targets 6(8):863–874. https://doi.org/10.2174/138945005774912735
Medina N, Soto-Debrán JC, Seidel D, Akyar I, Badali H, Barac A, MixInYeast Study Group from EFISG (2020) MixInYeast: a multicenter study on mixed yeast infections. J Fungi 7(1):13. https://doi.org/10.3390/jof701001
Minooeianhaghighi MH, Sehatpour M, Zarrinfar H, Sen T (2020) Recurrent vulvovaginal candidiasis: the causative agents, clinical signs and susceptibility to fluconazole in Gonabad city, northeast Iran. Curr Women’s Health Rev 16(1):46–51. https://doi.org/10.2174/1573404815666191104142813
Mout R, Xu ZD, Wolf AK, Davisson VJ, Jarori GK (2012). Anti-malarial activity of geldanamycin derivatives in mice infected with Plasmodium yoelii. Malar J 11(1): 1–10. http://www.malariajournal.com/content/11/1/54
Neckers L, Blagg B, Haystead T, Trepel JB, Whitesell L, Picard D (2018) Methods to validate Hsp90 inhibitor specificity, to identify off-target effects, and to rethink approaches for further clinical development. Cell Stress Chaperones 23(4):467–482. https://doi.org/10.1007/s12192-018-0877-2
Oz Y, Dag I, Kiraz N (2012) Broth microdilution and Time-Kill testing of caspofungin, voriconazole, amphotericin B and their combinations against clinical isolates of Candida krusei. Mycopathologia 173(1):27–34. https://doi.org/10.1007/s11046-011-9459-x
Pfaller MA, Messer SA, Boyken L, Hollis RJ, Rice C, Tendolkar S, Diekema DJ (2004) In vitro activities of voriconazole, posaconazole, and fluconazole against 4,169 clinical isolates of Candida spp. and Cryptococcus neoformans collected during 2001 and 2002 in the ARTEMIS global antifungal surveillance program. Diagn Microbiol Infect Dis 48(3):201–205. https://doi.org/10.1016/j.diagmicrobio.2003.09.008
Piper PW, Millson SH (2012) Spotlight on the microbes that produce heat shock protein 90-targeting antibiotics. Open Biol 2(12):120138. https://doi.org/10.1098/rsob.120138
Robbins N, Uppuluri P, Nett J, Rajendran R, Ramage G, Lopez-Ribot JL, Andes D, Cowen LE (2011) Hsp90 governs dispersion and drug resistance of fungal biofilms. PLoS Pathog 7(9):e1002257. https://doi.org/10.1371/journal.ppat.1002257
Scorzoni L, de Paula e Silva AC, Marcos CM, Assato PA, de Melo WC, de Oliveira HC, Costa-Orlandi CB, Mendes-Giannini MJ, Fusco-Almeida AM (2017) Antifungal therapy: new advances in the understanding and treatment of mycosis. Front Microbiol 8:36. https://doi.org/10.3389/fmicb.2017.00036
Shevtsov M, Multhoff G, Mikhaylova E, Shibata A, Guzhova I, Margulis B (2019) Combination of anti-cancer drugs with molecular chaperone inhibitors. Int J Mol Sci 20(21):5284. https://doi.org/10.3390/ijms20215284
Singh SD, Robbins N, Zaas AK, Schell WA, Perfect JR, Cowen LE (2009) Hsp90 governs echinocandin resistance in the pathogenic yeast Candida albicans via calcineurin. PLoS Pathog 5(7):e1000532. https://doi.org/10.1371/journal.ppat.1000532
Sobel JD (2007) Vulvovaginal candidosis. The Lancet 369(9577):1961–1971. https://doi.org/10.1016/S0140-6736(07)60917-9
Thati B, Noble A, Rowan R, Creaven BS, Walsh M, McCann M, Egan D, Kavanagh K (2007) Mechanism of action of coumarin and silver (I)–coumarin complexes against the pathogenic yeast Candida albicans. Toxicol in Vitro 21(5):801–808. https://doi.org/10.1016/j.tiv.2007.01.022
Tome M, Zupan J, Tomičić Z, Matos T, Raspor P (2018) Synergistic and antagonistic effects of immunomodulatory drugs on the action of antifungals against Candida glabrata and Saccharomyces cerevisiae. PeerJ 6:e4999. https://doi.org/10.7717/peerj.4999
Wirk B (2011) Heat shock protein inhibitors for the treatment of fungal infections. Recent Pat Anti-Infect Drug Discov 6(1):38–44
Zarrinfar H, Kaboli S, Dolatabadi S, Mohammadi R (2016) Rapid detection of Candida species in bronchoalveolar lavage fluid from patients with pulmonary symptoms. Braz J Microbiol 47:172–176. https://doi.org/10.1016/j.bjm.2015.02.001
Zarrinfar H, Kord Z, Fata A (2021) High incidence of azole resistance among Candida albicans and C. glabrata isolates in Northeastern Iran. Curr Med Mycol 7(3):18
Zhang J, Liu W, Tan J, Sun Y, Wan Z, Li R (2013) Antifungal activity of geldanamycin alone or in combination with fluconazole against Candida species. Mycopathologia 175(3–4):273–279. https://doi.org/10.1007/s11046-012-9612-1
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This work was supported by a grant from TUBİTAK (Project No. 112S503).
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Nuri Kiraz conceived the study. Sümeyye Şen Kaya and Yasemin Oz carried out microbiological studies. Nuri Kiraz, Ilknur Dag, and Yasemin Oz provided critical readings of the manuscript at several stages. All authors read and approved the final manuscript.
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Kiraz, N., Şen Kaya, S., Öz, Y. et al. Evaluation of the efficacy of heat shock protein inhibitors and antifungal drug combinations against Candida spp.. Rend. Fis. Acc. Lincei 34, 179–188 (2023). https://doi.org/10.1007/s12210-022-01118-0
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DOI: https://doi.org/10.1007/s12210-022-01118-0