1887

Abstract

P128, a phage-derived lysin, exerts antibacterial activity on staphylococci by cleaving the pentaglycine-bridge of peptidoglycan. We sought to determine whether the presence of P128 could re-sensitize drug-resistant bacteria to antibiotics by virtue of its cell wall degrading property.

P128 was tested in combination with standard-of-care (SoC) drugs by chequerboard assays on planktonic cells and biofilms of strains individually resistant to these drugs. The bactericidal effect of P128 and drug combinations on planktonic cells and biofilms was measured by c.f.u. reduction assays. A mouse model of MRSA bacteraemia was used to test the efficacy of P128 and oxacillin in combination.

A combination of sub-MIC P128 (0.025–0.20 µg ml) and 0.5 µg ml of oxacillin resulted in inhibition of bacterial growth in four MRSA strains. Similar results were seen with all the other drugs tested, wherein sub-MIC of P128 re-sensitized and CoNS strains to SoC drugs. The chequerboard assays on strains of and CoNS showed that combinations of P128 and antibiotics consistently inhibited bacterial growth on biofilms. Data from scanning electron microscopy and c.f.u. reduction assays on drug-resistant and CoNS demonstrated that sub-MICs of P128 and SoC antibiotics could kill biofilm-embedded bacteria. , a combination of sub-therapeutic doses of P128 and oxacillin could help protect animals from fatal bacteraemia.

The ability of P128 to re-sensitize bacteria to SoC drugs suggests that combinations of P128 and SoC antibiotics can potentially be developed to treat infections caused by drug-resistant strains of staphylococci.

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2018-03-01
2024-03-29
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References

  1. Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev 2015; 28:603–661 [View Article][PubMed]
    [Google Scholar]
  2. Becker K, Heilmann C, Peters G. Coagulase-negative staphylococci. Clin Microbiol Rev 2014; 27:870–926 [View Article][PubMed]
    [Google Scholar]
  3. Kelley PG, Gao W, Ward PB, Howden BP. Daptomycin non-susceptibility in vancomycin-intermediate Staphylococcus aureus (VISA) and heterogeneous-VISA (hVISA): implications for therapy after vancomycin treatment failure. J Antimicrob Chemother 2011; 66:1057–1060 [View Article][PubMed]
    [Google Scholar]
  4. Gu B, Kelesidis T, Tsiodras S, Hindler J, Humphries RM. The emerging problem of linezolid-resistant Staphylococcus. J Antimicrob Chemother 2013; 68:4–11 [View Article][PubMed]
    [Google Scholar]
  5. May L, Klein EY, Rothman RE, Laxminarayan R. Trends in antibiotic resistance in coagulase-negative staphylococci in the United States, 1999 to 2012. Antimicrob Agents Chemother 2014; 58:1404–1409 [View Article][PubMed]
    [Google Scholar]
  6. Yayan J, Ghebremedhin B, Rasche K. No outbreak of vancomycin and linezolid resistance in staphylococcal pneumonia over a 10-year period. PLoS One 2015; 10:e0138895 [View Article][PubMed]
    [Google Scholar]
  7. Archer NK, Mazaitis MJ, Costerton JW, Leid JG, Powers ME et al. Staphylococcus aureus biofilms: properties, regulation, and roles in human disease. Virulence 2011; 2:445–459 [View Article][PubMed]
    [Google Scholar]
  8. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: a common cause of persistent infections. Science 1999; 284:1318–1322 [View Article][PubMed]
    [Google Scholar]
  9. Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 2002; 15:167–193 [View Article][PubMed]
    [Google Scholar]
  10. Otto M. Staphylococcal biofilms. Curr Top Microbiol Immunol 2008; 322:207–228[PubMed]
    [Google Scholar]
  11. Amorena B, Gracia E, Monzón M, Leiva J, Oteiza C et al. Antibiotic susceptibility assay for Staphylococcus aureus in biofilms developed in vitro. J Antimicrob Chemother 1999; 44:43–55 [View Article][PubMed]
    [Google Scholar]
  12. Chung PY, Toh YS. Anti-biofilm agents: recent breakthrough against multi-drug resistant Staphylococcus aureus. Pathog Dis 2014; 70:231–239 [View Article][PubMed]
    [Google Scholar]
  13. Kristiansen JE, Hendricks O, Delvin T, Butterworth TS, Aagaard L et al. Reversal of resistance in microorganisms by help of non-antibiotics. J Antimicrob Chemother 2007; 59:1271–1279 [View Article][PubMed]
    [Google Scholar]
  14. Worthington RJ, Melander C. Combination approaches to combat multidrug-resistant bacteria. Trends Biotechnol 2013; 31:177–184 [View Article][PubMed]
    [Google Scholar]
  15. Lee SH, Jarantow LW, Wang H, Sillaots S, Cheng H et al. Antagonism of chemical genetic interaction networks resensitize MRSA to β-lactam antibiotics. Chem Biol 2011; 18:1379–1389 [View Article][PubMed]
    [Google Scholar]
  16. Farha MA, Leung A, Sewell EW, D'Elia MA, Allison SE et al. Inhibition of WTA synthesis blocks the cooperative action of PBPs and sensitizes MRSA to β-lactams. ACS Chem Biol 2013; 8:226–233 [View Article][PubMed]
    [Google Scholar]
  17. Wang H, Gill CJ, Lee SH, Mann P, Zuck P et al. Discovery of wall teichoic acid inhibitors as potential anti-MRSA β-lactam combination agents. Chem Biol 2013; 20:272–284 [View Article][PubMed]
    [Google Scholar]
  18. Klitgaard JK, Skov MN, Kallipolitis BH, Kolmos HJ. Reversal of methicillin resistance in Staphylococcus aureus by thioridazine. J Antimicrob Chemother 2008; 62:1215–1221 [View Article][PubMed]
    [Google Scholar]
  19. Kiri N, Archer G, Climo MW. Combinations of lysostaphin with β-lactams are synergistic against oxacillin-resistant Staphylococcus epidermidis. Antimicrob Agents Chemother 2002; 46:2017–2020 [View Article][PubMed]
    [Google Scholar]
  20. Climo MW, Ehlert K, Archer GL. Mechanism and suppression of lysostaphin resistance in oxacillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2001; 45:1431–1437 [View Article][PubMed]
    [Google Scholar]
  21. Rodríguez-Rubio L, Martínez B, Donovan DM, Rodríguez A, García P. Bacteriophage virion-associated peptidoglycan hydrolases: potential new enzybiotics. Crit Rev Microbiol 2013; 39:427–434 [View Article][PubMed]
    [Google Scholar]
  22. Roach DR, Donovan DM. Antimicrobial bacteriophage-derived proteins and therapeutic applications. Bacteriophage 2015; 5:e1062590 [View Article][PubMed]
    [Google Scholar]
  23. Sharma U, Paul VD. Bacteriophage lysins as antibacterials. Crit Care 2017; 21:99 [View Article][PubMed]
    [Google Scholar]
  24. Paul VD, Rajagopalan SS, Sundarrajan S, George SE, Asrani JY et al. A novel bacteriophage tail-associated muralytic enzyme (TAME) from phage K and its development into a potent antistaphylococcal protein. BMC Microbiol 2011; 11:226 [View Article][PubMed]
    [Google Scholar]
  25. Schuch R, Lee HM, Schneider BC, Sauve KL, Law C et al. Combination therapy with lysin CF-301 and antibiotic is superior to antibiotic alone for treating methicillin-resistant Staphylococcus aureus-induced murine bacteremia. J Infect Dis 2014; 209:1469–1478 [View Article][PubMed]
    [Google Scholar]
  26. Nair S, Desai S, Poonacha N, Vipra A, Sharma U. Antibiofilm activity and synergistic inhibition of Staphylococcus aureus biofilms by bactericidal protein P128 in combination with antibiotics. Antimicrob Agents Chemother 2016; 60:7280–7289 [View Article][PubMed]
    [Google Scholar]
  27. Poonacha N, Nair S, Desai S, Tuppad D, Hiremath D et al. Efficient killing of planktonic and biofilm-embedded coagulase-negative staphylococci by bactericidal protein P128. Antimicrob Agents Chemother 2017; 61:e00457-17 [View Article][PubMed]
    [Google Scholar]
  28. Drilling AJ, Cooksley C, Chan C, Wormald PJ, Vreugde S. Fighting sinus-derived Staphylococcus aureus biofilms in vitro with a bacteriophage-derived muralytic enzyme. Int Forum Allergy Rhinol 2016; 6:349–355 [View Article][PubMed]
    [Google Scholar]
  29. Clinical and Laboratory Standards Institute Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard, 9th ed. CLSI document M07-A9 Wayne, PA: Clinical and Laboratory Standards Institute; 2012
    [Google Scholar]
  30. National Committee for Clinical Laboratory Standards Methods for Determining Bactericidal Activity of Antimicrobial Agents Approved Guideline M26-A Wayne, PA: NCCLS;
    [Google Scholar]
  31. Odds FC. Synergy, antagonism, and what the chequerboard puts between them. J Antimicrob Chemother 2003; 52:1 [View Article][PubMed]
    [Google Scholar]
  32. Jo DS, Montgomery CP, Yin S, Boyle-Vavra S, Daum RS. Improved oxacillin treatment outcomes in experimental skin and lung infection by a methicillin-resistant Staphylococcus aureus isolate with a vraSR operon deletion. Antimicrob Agents Chemother 2011; 55:2818–2823 [View Article][PubMed]
    [Google Scholar]
  33. Sundarrajan S, Raghupatil J, Vipra A, Narasimhaswamy N, Saravanan S et al. Bacteriophage-derived CHAP domain protein, P128, kills Staphylococcus cells by cleaving interpeptide cross-bridge of peptidoglycan. Microbiology 2014; 160:2157–2169 [View Article][PubMed]
    [Google Scholar]
  34. Bonde M, Højland DH, Kolmos HJ, Kallipolitis BH, Klitgaard JK. Thioridazine affects transcription of genes involved in cell wall biosynthesis in methicillin-resistant Staphylococcus aureus. FEMS Microbiol Lett 2011; 318:168–176 [View Article][PubMed]
    [Google Scholar]
  35. Gonzales PR, Pesesky MW, Bouley R, Ballard A, Biddy BA et al. Synergistic, collaterally sensitive β-lactam combinations suppress resistance in MRSA. Nat Chem Biol 2015; 11:855–861 [View Article][PubMed]
    [Google Scholar]
  36. Daniel A, Euler C, Collin M, Chahales P, Gorelick KJ et al. Synergism between a novel chimeric lysin and oxacillin protects against infection by methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2010; 54:1603–1612 [View Article][PubMed]
    [Google Scholar]
  37. Tan CM, Therien AG, Lu J, Lee SH, Caron A. Restoring methicillin-resistant Staphylococcus aureus susceptibility to β-lactam antibiotics. Sci Transl Med 2012; 4:126ra35 [View Article][PubMed]
    [Google Scholar]
  38. Singh R, Ray P, das A, Sharma M. Role of persisters and small-colony variants in antibiotic resistance of planktonic and biofilm-associated Staphylococcus aureus: an in vitro study. J Med Microbiol 2009; 58:1067–1073 [View Article][PubMed]
    [Google Scholar]
  39. Clinical and Laboratory Standards Institute Performance Standards for Antimicrobial Susceptibility Testing Twenty-second Informational Supplement CLSI document M100-S22 vol. 32 Wayne, PA: Clinical and Laboratory Standards Institute; 2012
    [Google Scholar]
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