1887

Abstract

Non-typeable (NTHi) is an opportunistic pathogen that plays a major role in a number of respiratory tract infections, including otitis media, cystic fibrosis and chronic obstructive pulmonary disease. Biofilm formation has been implicated in both NTHi colonization and disease, and is responsible for the increased tolerance of this pathogen towards antibiotic treatment. Targeting metabolic pathways that are important in NTHi biofilm formation represents a potential strategy to combat this antibiotic recalcitrance. A previous investigation demonstrated increased expression of a putative -methionine uptake protein following exposure of NTHi biofilms to the ubiquitous signalling molecule, nitric oxide. We therefore hypothesized that treatment with exogenous -methionine would impact on NTHi biofilm formation and increase antibiotic sensitivity. Treatment of NTHi during the process of biofilm formation resulted in a reduction in biofilm viability, increased biomass, changes in the overall biofilm architecture and the adoption of an amorphous cellular morphology. Quantitative proteomic analyses identified 124 proteins that were differentially expressed following -methionine treatment, of which 51 (41 %) were involved in metabolic and transport processes. Nine proteins involved in peptidoglycan synthesis and cell division showed significantly increased expression. Furthermore, -methionine treatment augmented the efficacy of azithromycin treatment and highlighted the potential of -methionine as an adjunctive therapeutic approach for NTHi biofilm-associated infections.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000491
2017-07-01
2020-01-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/163/7/1093.html?itemId=/content/journal/micro/10.1099/mic.0.000491&mimeType=html&fmt=ahah

References

  1. Halbert RJ, Natoli JL, Gano A, Badamgarav E, Buist AS et al. Global burden of COPD: systematic review and meta-analysis. Eur Respir J 2006;28:523–532 [CrossRef][PubMed]
    [Google Scholar]
  2. Ramsey KA, Ranganathan S, Park J, Skoric B, Adams AM et al. Early respiratory infection is associated with reduced spirometry in children with cystic fibrosis. Am J Respir Crit Care Med 2014;190:1111–1116 [CrossRef][PubMed]
    [Google Scholar]
  3. Murphy TF, Faden H, Bakaletz LO, Kyd JM, Forsgren A et al. Nontypeable Haemophilus influenzae as a pathogen in children. Pediatr Infect Dis J 2009;28:43–48 [CrossRef][PubMed]
    [Google Scholar]
  4. Clementi CF, Murphy TF. Non-typeable Haemophilus influenzae invasion and persistence in the human respiratory tract. Front Cell Infect Microbiol 2011;1:1 [CrossRef][PubMed]
    [Google Scholar]
  5. Hall-Stoodley L, Hu FZ, Gieseke A, Nistico L, Nguyen D et al. Direct detection of bacterial biofilms on the middle-ear mucosa of children with chronic otitis media. JAMA 2006;296:202–211 [CrossRef][PubMed]
    [Google Scholar]
  6. Van Acker H, Van Dijck P, Coenye T. Molecular mechanisms of antimicrobial tolerance and resistance in bacterial and fungal biofilms. Trends Microbiol 2014;22:326–333 [CrossRef][PubMed]
    [Google Scholar]
  7. Wood TK. Combatting bacterial persister cells. Biotechnol Bioeng 2016;113:476–483 [CrossRef][PubMed]
    [Google Scholar]
  8. Jakubovics NS, Robinson JC, Samarian DS, Kolderman E, Yassin SA et al. Critical roles of arginine in growth and biofilm development by Streptococcus gordonii. Mol Microbiol 2015;97:281–300 [CrossRef][PubMed]
    [Google Scholar]
  9. He J, Hwang G, Liu Y, Gao L, Kilpatrick-Liverman L et al. l-Arginine modifies the Exopolysaccharide matrix and Thwarts Streptococcus mutans outgrowth within mixed-species oral biofilms. J Bacteriol 2016;198:2651–2661 [CrossRef][PubMed]
    [Google Scholar]
  10. Kolderman E, Bettampadi D, Samarian D, Dowd SE, Foxman B et al. L-arginine destabilizes oral multi-species biofilm communities developed in human saliva. PLoS One 2015;10:e0121835 [CrossRef][PubMed]
    [Google Scholar]
  11. Gnanadhas DP, Elango M, Datey A, Chakravortty D. Chronic lung infection by Pseudomonas aeruginosa biofilm is cured by L-Methionine in combination with antibiotic therapy. Sci Rep 2015;5:16043 [CrossRef][PubMed]
    [Google Scholar]
  12. Garlick PJ. Toxicity of methionine in humans. J Nutr 2006;136:1722S–1725S[PubMed]
    [Google Scholar]
  13. Collins SA, Kelso MJ, Rineh A, Yepuri NR, Coles J et al. Cephalosporin-3′-diazeniumdiolate NO donor prodrug PYRRO-C3D enhances azithromycin susceptibility of nontypeable Haemophilus influenzae biofilms. Antimicrob Agents Chemother 2017;61:e02086-16 [CrossRef][PubMed]
    [Google Scholar]
  14. Allan RN, Morgan S, Brito-Mutunayagam S, Skipp P, Feelisch M et al. Low concentrations of nitric oxide modulate Streptococcus pneumoniae biofilm metabolism and antibiotic tolerance. Antimicrob Agents Chemother 2016;60:2456–2466 [CrossRef][PubMed]
    [Google Scholar]
  15. Heydorn A, Nielsen AT, Hentzer M, Sternberg C, Givskov M et al. Quantification of biofilm structures by the novel computer program COMSTAT. Microbiology 2000;146:2395–2407 [CrossRef][PubMed]
    [Google Scholar]
  16. Allan RN, Skipp P, Jefferies J, Clarke SC, Faust SN et al. Pronounced metabolic changes in adaptation to biofilm growth by Streptococcus pneumoniae. PLoS One 2014;9:e107015 [CrossRef][PubMed]
    [Google Scholar]
  17. Wiśniewski JR, Zougman A, Nagaraj N, Mann M. Universal sample preparation method for proteome analysis. Nat Methods 2009;6:359–362 [CrossRef][PubMed]
    [Google Scholar]
  18. Snyder LA, Saunders NJ, Shafer WM. A putatively phase variable gene (dca) required for natural competence in Neisseria gonorrhoeae but not Neisseria meningitidis is located within the division cell wall (dcw) gene cluster. J Bacteriol 2001;183:1233–1241 [CrossRef][PubMed]
    [Google Scholar]
  19. Tamames J, González-Moreno M, Mingorance J, Valencia A, Vicente M. Bringing gene order into bacterial shape. Trends Genet 2001;17:124–126 [CrossRef][PubMed]
    [Google Scholar]
  20. Mingorance J, Tamames J, Vicente M. Genomic channeling in bacterial cell division. J Mol Recognit 2004;17:481–487 [CrossRef][PubMed]
    [Google Scholar]
  21. Liberek K, Marszalek J, Ang D, Georgopoulos C, Zylicz M. Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK. Proc Natl Acad Sci USA 1991;88:2874–2878 [CrossRef][PubMed]
    [Google Scholar]
  22. Gentry DR, Cashel M. Mutational analysis of the Escherichia coli spoT gene identifies distinct but overlapping regions involved in ppGpp synthesis and degradation. Mol Microbiol 1996;19:1373–1384 [CrossRef][PubMed]
    [Google Scholar]
  23. Sugisaki K, Hanawa T, Yonezawa H, Osaki T, Fukutomi T et al. Role of (p)ppGpp in biofilm formation and expression of filamentous structures in Bordetella pertussis. Microbiology 2013;159:1379–1389 [CrossRef][PubMed]
    [Google Scholar]
  24. Harrington JC, Wong SM, Rosadini CV, Garifulin O, Boyartchuk V et al. Resistance of Haemophilus influenzae to reactive nitrogen donors and gamma interferon-stimulated macrophages requires the formate-dependent nitrite reductase regulator-activated ytfE gene. Infect Immun 2009;77:1945–1958 [CrossRef][PubMed]
    [Google Scholar]
  25. Marti S, Puig C, Merlos A, Vinas M, de Jonge MI et al. Bacterial lysis through interference with peptidoglycan synthesis increases Biofilm formation by nontypeable Haemophilus influenzae. mSphere 2017;2:e00329-16 [CrossRef][PubMed]
    [Google Scholar]
  26. Caparrós M, Pisabarro AG, de Pedro MA. Effect of D-amino acids on structure and synthesis of peptidoglycan in Escherichia coli. J Bacteriol 1992;174:5549–5559 [CrossRef][PubMed]
    [Google Scholar]
  27. Cava F, de Pedro MA, Lam H, Davis BM, Waldor MK. Distinct pathways for modification of the bacterial cell wall by non-canonical D-amino acids. EMBO J 2011;30:3442–3453 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000491
Loading
/content/journal/micro/10.1099/mic.0.000491
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error