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Abstract

Bacterial persisters form a subpopulation of cells that survive lethal concentrations of antibiotics without being genetically different from the susceptible population. They are generally considered to be phenotypic variants that spontaneously have entered a dormant state with low ATP levels or reduced membrane potential. In , a serious opportunistic human pathogen, persisters are believed to contribute to chronic infections that are a major global healthcare problem. While persisters have mostly been studied in laboratory strains, we have here investigated the ability of clinical strains to form persisters. For 44 clinical strains belonging to the major clonal complexes CC5, CC8, CC30 or CC45, we examined persister cell formation in stationary phase when exposed to 100 times the MIC of ciprofloxacin, an antibiotic that targets DNA replication. We find that while all strains are able to form persisters, those belonging to CC30 displayed on average 100-fold higher persister cell frequencies when compared to strains of other CCs. Importantly, there was no correlation between persister formation and the cellular ATP content of the individual strains, but the group of CC30 strains displayed slightly lower membrane potential compared to the non-CC30 group. CC30 strains have previously been associated with chronic and reoccuring infections and we hypothesize that there could be a correlation between lineage-specific characteristics displayed via persister assays and the observed clinical spectrum of disease.

Funding
This study was supported by the:
  • Hanne Ingmer , Danmarks Grundforskningsfond , (Award 120)
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/content/journal/micro/10.1099/mic.0.000926
2020-05-19
2020-06-04
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References

  1. Bigger JW. Treatment of staphylococcal infections with penicillin. Lancet 1944; 2:497–500
    [Google Scholar]
  2. Lewis K, cells P. Dormancy and infectious disease. Nat Rev Microbiol 2007; 5:48–56
    [Google Scholar]
  3. Balaban NQ, Merrin J, Chait R, Kowalik L, Leibler S. Bacterial persistence as a phenotypic switch. Science 2004; 305:1622–1625 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  4. Amato SM, Orman MA, Brynildsen MP. Metabolic control of persister formation in Escherichia coli. Mol Cell 2013; 50:475–487 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  5. Wood TK, Knabel SJ, Kwan BW. Bacterial persister cell formation and dormancy. Appl Environ Microbiol 2013; 79:7116–7121 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  6. Korch SB, Henderson TA, Hill TM. Characterization of the hipA7 allele of Escherichia coli and evidence that high persistence is governed by (p)ppGpp synthesis. Mol Microbiol 2003; 50:1199–1213 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  7. Conlon BP, Rowe SE, Gandt AB, Nuxoll AS, Donegan NP et al. Persister formation in Staphylococcus aureus is associated with ATP depletion. Nature Microbiology 2016; 1:16051 [CrossRef]
    [Google Scholar]
  8. Shan Y, Brown Gandt A, Rowe SE, Deisinger JP, Conlon BP et al. ATP-dependent persister formation in Escherichia coli. mBio 2017; 8:e02267-16 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  9. Wang Y, Bojer MS, George SE, Wang Z, Jensen PR et al. Inactivation of TCA cycle enhances Staphylococcus aureus persister cell formation in stationary phase. Sci Rep 2018; 8:10849 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  10. Pontes MH, Groisman EA. Slow growth determines nonheritable antibiotic resistance in Salmonella enterica. Sci Signal 2019; 12::eaax3938 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  11. Lopatkin AJ, Stokes JM, Zheng EJ, Yang JH, Takahashi MK et al. Bacterial metabolic state more accurately predicts antibiotic lethality than growth rate. Nat Microbiol 2019; 4:2109–2117 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  12. Fauvart M, De Groote VN, Michiels J. Role of persister cells in chronic infections: clinical relevance and perspectives on anti-persister therapies. J Med Microbiol 2011; 60:699–709 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  13. Lafleur MD, Qi Q, Lewis K. Patients with long-term oral carriage harbor high-persister mutants of Candida albicans. Antimicrob Agents Chemother 2010; 54:39–44 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  14. Mulcahy LR, Burns JL, Lory S, Lewis K. Emergence of Pseudomonas aeruginosa strains producing high levels of persister cells in patients with cystic fibrosis. J Bacteriol 2010; 192:6191–6199 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  15. Sharma-Kuinkel BK, Mongodin EF, Myers JR, Vore KL, Canfield GS et al. Potential influence of Staphylococcus aureus clonal complex 30 genotype and transcriptome on hematogenous infections. Open Forum Infect Dis 2015; 2:ofv093 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  16. Fowler VG, Nelson CL, McIntyre LM, Kreiswirth BN, Monk A et al. Potential associations between hematogenous complications and bacterial genotype in Staphylococcus aureus infection. J Infect Dis 2007; 196:738–747 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  17. Pérez-Montarelo D, Viedma E, Murcia M, Muñoz-Gallego I, Larrosa N et al. Pathogenic Characteristics of Staphylococcus aureus Endovascular Infection Isolates from Different Clonal Complexes. Front Microbiol 2017; 8:917 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  18. Stegger M, Driebe EM, Roe C, Lemmer D, Bowers JR et al. Genome sequence of Staphylococcus aureus strain CA-347, a USA600 methicillin-resistant isolate. Genome Announc 2013; 1:e00517-13 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  19. Keren I, Kaldalu N, Spoering A, Wang Y, Lewis K. Persister cells and tolerance to antimicrobials. FEMS Microbiol Lett 2004; 230:13–18 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  20. Grassi L, Di Luca M, Maisetta G, Rinaldi AC, Esin S et al. Generation of Persister Cells of Pseudomonas aeruginosa and Staphylococcus aureus by Chemical Treatment and Evaluation of Their Susceptibility to Membrane-Targeting Agents. Front Microbiol 2017; 8:8 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  21. Bojer MS, Lindemose S, Vestergaard M, Ingmer H. Quorum Sensing-Regulated Phenol-Soluble Modulins Limit Persister Cell Populations in Staphylococcus aureus. Front Microbiol 2018; 9:255 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  22. Chatterjee I, Herrmann M, Proctor RA, Peters G, Kahl BC. Enhanced post-stationary-phase survival of a clinical thymidine-dependent small-colony variant of Staphylococcus aureus results from lack of a functional tricarboxylic acid cycle. J Bacteriol 2007; 189:2936–2940 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  23. Romaniuk JAH, Cegelski L. Bacterial cell wall composition and the influence of antibiotics by cell-wall and whole-cell NMR. Philos Trans R Soc Lond B Biol Sci 2015; 370:20150024 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  24. Cho H, Uehara T, Bernhardt TG. Beta-Lactam antibiotics induce a lethal malfunctioning of the bacterial cell wall synthesis machinery. Cell 2014; 159:1300–1311 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  25. Kamble E, Pardesi K. Antibiotic Tolerance in Biofilm and Stationary-Phase Planktonic Cells of Staphylococcus aureus. Microbial Drug Resistance 2020 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  26. Harms A, Maisonneuve E, Gerdes K. Mechanisms of bacterial persistence during stress and antibiotic exposure. Science 2016; 354:aaf4268 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  27. Dörr T, Vulić M, Lewis K. Ciprofloxacin causes persister formation by inducing the TisB toxin in Escherichia coli. PLoS Biol 2010; 8:e1000317 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  28. Dimroth P, Kaim G, Matthey U. Crucial role of the membrane potential for ATP synthesis by F(1)F(o) ATP synthases. J Exp Biol 2000; 203:51–59[PubMed][PubMed]
    [Google Scholar]
  29. Kwan BW, Valenta JA, Benedik MJ, Wood TK. Arrested protein synthesis increases persister-like cell formation. Antimicrob Agents Chemother 2013; 57:1468–1473 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  30. Monecke S, Müller E, Büchler J, Stieber B, Ehricht R. Staphylococcus aureus in vitro secretion of alpha toxin (hla) correlates with the affiliation to clonal complexes. PLoS One 2014; 9:e100427 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  31. Cheung GYC, Kretschmer D, Duong AC, Yeh AJ, Ho TV et al. Production of an attenuated phenol-soluble modulin variant unique to the MRSA clonal complex 30 increases severity of bloodstream infection. PLoS Pathog 2014; 10:e1004298 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  32. Mok WWK, Brynildsen MP. Timing of DNA damage responses impacts persistence to fluoroquinolones. Proc Natl Acad Sci U S A 2018; 115:E6301–E6309 [CrossRef][PubMed][PubMed]
    [Google Scholar]
  33. Hong Y, Li Q, Gao Q, Xie J, Huang H et al. Reactive oxygen species play a dominant role in all pathways of rapid quinolone-mediated killing. J Antimicrob Chemother 2019
    [Google Scholar]
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