Abstract
Pseudomonas aeruginosa is a ubiquitous pathogen that is the leading cause of chronic infections. Bacterial biofilm formation facilitates CF development and restricts the anti-bacterial potential of many current antibiotics. The capacity of P. aeruginosa to form biofilms and resist antibiotics is closely correlated with quorum sensing (QS). Disrupting QS by QS inhibitors is a promising strategy for treating chronic infections. Here, we evaluated the effect of hordenine, a recently characterized QS inhibitor, on the susceptibility of aminoglycoside antibiotics against P. aeruginosa biofilms. Hordenine significantly enhanced the susceptibility of aminoglycoside antibiotics tobramycin, gentamycin, and amikacin against P. aeruginosa PAO1 biofilm formation. Combinations of hordenine and aminoglycoside antibiotics showed potent efficiency in disrupting the preformed biofilms of P. aeruginosa. Microscopic observations showed flat, scattered, and unstructured biofilm architecture after treatment with hordenine. Mechanistic study further revealed that hordenine treatment led to the downregulation of genes involved in QS and biofilm formation. Thus, our results suggest that hordenine has the potential to function as an antibiotic accelerant in treating P. aeruginosa infections.
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Allesen-Holm M, Barken KB, Yang L, Klausen M, Webb JS, Kjelleberg S, Molin S, Givskov M, Tolker-Nielsen T (2006) A characterization of DNA release in Pseudomonas aeruginosa cultures and biofilms. Mol Microbiol 59(4):1114–1128. https://doi.org/10.1111/j.1365-2958.2005.05008.x
Babii O, Afonin S, Berditsch M, Reibetaer S, Mykhailiuk PK, Kubyshkin VS, Steinbrecher T, Ulrich AS, Komarov IV (2014) Controlling biological activity with light: diarylethene-containing cyclic peptidomimetics. Angew Chem Int Ed Engl 53(13):3392–3395. https://doi.org/10.1002/anie.201310019
Beury-Cirou A, Tannieres M, Minard C, Soulere L, Rasamiravaka T, Dodd RH, Queneau Y, Dessaux Y, Guillou C, Vandeputte OM, Faure D (2013) At a supra-physiological concentration, human sexual hormones act as quorum-sensing inhibitors. PLoS One 8(12):e83564. https://doi.org/10.1371/journal.pone.0083564
Bjarnsholt T, Jensen PO, Burmolle M, Hentzer M, Haagensen JAJ, Hougen HP, Calum H, Madsen KG, Moser C, Molin S, Hoiby N, Givskov M (2005) Pseudomonas aeruginosa tolerance to tobramycin, hydrogen peroxide and polymorphonuclear leukocytes is quorum-sensing dependent. Microbiol-Sgm 151:373–383. https://doi.org/10.1099/mic.0.27463-0
Brackman G, Cos P, Maes L, Nelis HJ, Coenye T (2011) Quorum sensing inhibitors increase the susceptibility of bacterial biofilms to antibiotics in vitro and in vivo. Antimicrob Agents Chemother 55(6):2655–2661. https://doi.org/10.1128/AAC.00045-11
Colvin KM, Gordon VD, Murakami K, Borlee BR, Wozniak DJ, Wong GCL, Parsek MR (2011) The pel polysaccharide can serve a structural and protective role in the biofilm matrix of Pseudomonas aeruginosa. PLoS Pathog 7(1):e1001264. https://doi.org/10.1371/journal.ppat.1001264
Corral-Lugo A, Daddaoua A, Ortega A, Espinosa-Urgel M, Krell T (2016) Rosmarinic acid is a homoserine lactone mimic produced by plants that activates a bacterial quorum-sensing regulator. Sci Signal 9(409):ra1. https://doi.org/10.1126/scisignal.aaa8271
Davies D (2003) Understanding biofilm resistance to antibacterial agents. Nat Rev Drug Discov 2(2):114–122. https://doi.org/10.1038/nrd1008
Davies DG, Parsek MR, Pearson JP, Iglewski BH, Costerton JW, Greenberg EP (1998) The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280(5361):295–298
de la Fuente-Nunez C, Reffuveille F, Fairfull-Smith KE, Hancock REW (2013) Effect of nitroxides on swarming motility and biofilm formation, multicellular behaviors in Pseudomonas aeruginosa. Antimicrob Agents Chemother 57(10):4877–4881. https://doi.org/10.1128/Aac.01381-13
Doring G, Flume P, Heijerman H, Elborn JS, Consensus Study G (2012) Treatment of lung infection in patients with cystic fibrosis: current and future strategies. J Cyst Fibros 11(6):461–479. https://doi.org/10.1016/j.jcf.2012.10.004
Dumas JL, van Delden C, Perron K, Kohler T (2006) Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR. FEMS Microbiol Lett 254(2):217–225. https://doi.org/10.1111/j.1574-6968.2005.00008.x
Fernandez L, Gooderham WJ, Bains M, McPhee JB, Wiegand I, Hancock REW (2010) Adaptive resistance to the “last hope” antibiotics polymyxin B and colistin in Pseudomonas aeruginosa is mediated by the novel two-component regulatory system ParR-ParS. Antimicrob Agents Chemother 54(8):3372–3382. https://doi.org/10.1128/Aac.00242-10
Flemming HC, Wingender J (2010) The biofilm matrix. Nat Rev Microbiol 8:623–633. https://doi.org/10.1038/nrmicro2415
Habash MB, Park AJ, Vis EC, Harris RJ, Khursigara CM (2014) Synergy of silver nanoparticles and aztreonam against Pseudomonas aeruginosa PAO1 biofilms. Antimicrob Agents Chemother 58(10):5818–5830. https://doi.org/10.1128/AAC.03170-14
Hall S, McDermott C, Anoopkumar-Dukie S, McFarland AJ, Forbes A, Perkins AV, Davey AK, Chess-Williams R, Kiefel MJ, Arora D, Grant GD (2016) Cellular effects of pyocyanin, a secreted virulence factor of Pseudomonas aeruginosa. Toxins (Basel) 8(8):236. https://doi.org/10.3390/toxins8080236
Hentzer M, Riedel K, Rasmussen TB, Heydorn A, Andersen JB, Parsek MR, Rice SA, Eberl L, Molin S, Hoiby N, Kjelleberg S, Givskov M (2002) Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound. Microbiol-Sgm 148(Pt 1):87–102. https://doi.org/10.1099/00221287-148-1-87
Li Y, Huang J, Li L, Liu L (2017) Synergistic activity of berberine with azithromycin against Pseudomonas aeruginosa isolated from patients with cystic fibrosis of lung in vitro and in vivo. Cell Physiol Biochem 42(4):1657–1669. https://doi.org/10.1159/000479411
Linde L, Boelz S, Nissim-Rafinia M, Oren YS, Wilschanski M, Yaacov Y, Virgilis D, Neu-Yilik G, Kulozik AE, Kerem E, Kerem B (2007) Nonsense-mediated mRNA decay affects nonsense transcript levels and governs response of cystic fibrosis patients to gentamicin. J Clin Invest 117(3):683–692. https://doi.org/10.1172/JCI28523
Loo CY, Rohanizadeh R, Young PM, Traini D, Cavaliere R, Whitchurch CB, Lee WH (2016) Combination of silver nanoparticles and curcumin nanoparticles for enhanced anti-biofilm activities. J Agric Food Chem 64(12):2513–2522. https://doi.org/10.1021/acs.jafc.5b04559
Macedo AJ, Abraham WR (2009) Can infectious biofilm be controlled by blocking bacterial communication? Med Chem 5(6):517–528. https://doi.org/10.2174/157340609790170515
Nadell CD, Xavier JB, Levin SA, Foster KR (2008) The evolution of quorum sensing in bacterial biofilms. PLoS Biol 6:e14. https://doi.org/10.1371/journal.pbio.0060014
Okusanya OO, Bhavnani SM, Hammel J, Minic P, Dupont LJ, Forrest A, Mulder GJ, Mackinson C, Ambrose PG, Gupta R (2009) Pharmacokinetic and pharmacodynamic evaluation of liposomal amikacin for inhalation in cystic fibrosis patients with chronic pseudomonal infection. Antimicrob Agents Chemother 53(9):3847–3854. https://doi.org/10.1128/AAC.00872-08
Poole K (2005) Aminoglycoside resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother 49(2):479–487. https://doi.org/10.1128/AAC.49.2.479-487.2005
Reffuveille F, de la Fuente-Nunez C, Mansour S, Hancock RE (2014) A broad-spectrum antibiofilm peptide enhances antibiotic action against bacterial biofilms. Antimicrob Agents Chemother 58(9):5363–5371. https://doi.org/10.1128/AAC.03163-14
Rutherford ST, Bassler BL (2012) Bacterial quorum sensing: its role in virulence and possibilities for its control. Cold Spring Harb Perspect Med 2(11). https://doi.org/10.1101/cshperspect.a012427
Sarabhai S, Harjai K, Sharma P, Capalash N (2015) Ellagic acid derivatives from Terminalia chebula Retz. Increase the susceptibility of Pseudomonas aeruginosa to stress by inhibiting polyphosphate kinase. J Appl Microbiol 118(4):817–825. https://doi.org/10.1111/jam.12733
Sawicki GS, Signorovitch JE, Zhang J, Latremouille-Viau D, von Wartburg M, Wu EQ, Shi L (2012) Reduced mortality in cystic fibrosis patients treated with tobramycin inhalation solution. Pediatr Pulmonol 47(1):44–52. https://doi.org/10.1002/ppul.21521
Siarkou VI, Vitti D, Protonotariou E, Ikonomidis A, Sofianou D (2009) Molecular epidemiology of outbreak-related Pseudomonas aeruginosa strains carrying the novel variant blaVIM-17 metallo-beta-lactamase gene. Antimicrob Agents Chemother 53(4):1325–1330. https://doi.org/10.1128/AAC.01230-08
Skindersoe ME, Alhede M, Phipps R, Yang L, Jensen PO, Rasmussen TB, Bjarnsholt T, Tolker-Nielsen T, Hoiby N, Givskov M (2008) Effects of antibiotics on quorum sensing in Pseudomonas aeruginosa. Antimicrob Agents Chemother 52(10):3648–3663. https://doi.org/10.1128/AAC.01230-07
Sopirala MM, Mangino JE, Gebreyes WA, Biller B, Bannerman T, Balada-Llasat JM, Pancholi P (2010) Synergy testing by Etest, microdilution checkerboard, and time-kill methods for pan-drug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 54(11):4678–4683. https://doi.org/10.1128/AAC.00497-10
Suci PA, Mittelman M, Yu FP, Geesey GG (1994) Investigation of ciprofloxacin penetration into Pseudomonas aeruginosa biofilms. Antimicrob Agents Chemother 38(9):2125–2133
Szaff M, Hoiby N, Flensborg EW (1983) Frequent antibiotic therapy improves survival of cystic fibrosis patients with chronic Pseudomonas aeruginosa infection. Acta Paediatr Scand 72(5):651–657
Todd DA, Parlet CP, Crosby HA, Malone CL, Heilmann KP, Horswill AR, Cech NB (2017) Signal biosynthesis inhibition with ambuic acid as a strategy to target antibiotic-resistant infections. Antimicrob Agents Chemother 61(8). https://doi.org/10.1128/AAC.00263-17
Wang X, Yao X, Zhu Z, Tang T, Dai K, Sadovskaya I, Flahaut S, Jabbouri S (2009) Effect of berberine on Staphylococcus epidermidis biofilm formation. Int J Antimicrob Agents 34(1):60–66. https://doi.org/10.1016/j.ijantimicag.2008.10.033
Yang L, Rybtke MT, Jakobsen TH, Hentzer M, Bjarnsholt T, Givskov M, Tolker-Nielsen T (2009) Computer-aided identification of recognized drugs as Pseudomonas aeruginosaquorum-sensing inhibitors. Antimicrob Agents Chemother 53(6):2432–2443. https://doi.org/10.1128/Aac.01283-08
Zhou JW, Luo HZ, Jiang H, Jian TK, Chen ZQ, Jia AQ (2018) Hordenine: a novel quorum sensing inhibitor and antibiofilm agent against Pseudomonas aeruginosa. J Agric Food Chem 66(7):1620–1628. https://doi.org/10.1021/acs.jafc.7b05035
Funding
This work was supported by grants from the National Key Research and Development Program of China (2017YFD0201401); National Natural Science Foundation of China (41766006); Natural Science Foundation of Jiangsu Province, China (BK20170859); Science and Technology Development Program of Modern Agriculture, Nanjing (201608052); Six Talent Peaks Project in Jiangsu Province; and Fundamental Research Funds for the Central Universities (30916011307).
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Zhou, JW., Hou, B., Liu, GY. et al. Attenuation of Pseudomonas aeruginosa biofilm by hordenine: a combinatorial study with aminoglycoside antibiotics. Appl Microbiol Biotechnol 102, 9745–9758 (2018). https://doi.org/10.1007/s00253-018-9315-8
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DOI: https://doi.org/10.1007/s00253-018-9315-8