1887

Abstract

Purpose. In the cystic fibrosis (CF) airways, Pseudomonas aeruginosa undergoes diverse physiological changes in response to inflammation, antibiotic pressure, oxidative stress and a dynamic bioavailable nutrient pool. These include loss-of-function mutations that result in reduced virulence, altered metabolism and other phenotypes that are thought to confer a selective advantage for long-term persistence. Recently, clinical isolates of P. aeruginosa that hyperproduce agmatine (decarboxylated arginine) were cultured from individuals with CF. Sputum concentrations of this metabolite were also shown to correlate with disease severity. This raised the question of whether agmatine accumulation might also confer a selective advantage for P. aeruginosa during chronic colonization of the lung.

Methodology and Results. We screened a library of P. aeruginosa CF clinical isolates and found that ~5 % of subjects harboured isolates with an agmatine hyperproducing phenotype. Agmatine accumulation was a direct result of mutations in aguA, encoding the arginine deiminase that catalyses the conversion of agmatine into various polyamines. We also found that agmatine hyperproducing isolates (aguA-) had increased tolerance to the cationic antibiotics gentamicin, tobramycin and colistin relative to their chromosomally complemented strains (aguA+). Finally, we revealed that agmatine diminishes IL-8 production by airway epithelial cells in response to bacterial infection, with a consequent decrease in neutrophil recruitment to the murine airways in an acute pneumonia model.

Conclusion. These data highlight a potential new role for bacterial-derived agmatine that may have important consequences for the long-term persistence of P. aeruginosa in the CF airways.

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2019-01-22
2019-12-15
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References

  1. Kerem B, Rommens JM, Buchanan JA, Markiewicz D, Cox TK et al. Identification of the cystic fibrosis gene: genetic analysis. Science 1989;245:1073–1080 [CrossRef]
    [Google Scholar]
  2. Hauser AR, Jain M, Bar-Meir M, McColley SA. Clinical significance of microbial infection and adaptation in cystic fibrosis. Clin Microbiol Rev 2011;24:29–70 [CrossRef]
    [Google Scholar]
  3. Sibley CD, Surette MG. The polymicrobial nature of airway infections in cystic fibrosis: Cangene gold medal Lecture. Can J Microbiol 2011;57:69–77
    [Google Scholar]
  4. Price KE, Hampton TH, Gifford AH, Dolben EL, Hogan DA et al. Unique microbial communities persist in individual cystic fibrosis patients throughout a clinical exacerbation. Microbiome 2013;1:27 [CrossRef]
    [Google Scholar]
  5. Zhao J, Schloss PD, Kalikin LM, Carmody LA, Foster BK et al. Decade-long bacterial community dynamics in cystic fibrosis airways. Proc Natl Acad Sci USA 2012;109:5809–5814 [CrossRef]
    [Google Scholar]
  6. Zemanick ET, Wagner BD, Robertson CE, Stevens MJ, Szefler SJ et al. Assessment of airway microbiota and inflammation in cystic fibrosis using multiple sampling methods. Annals Am Thorac Soc 2015;12:221–229 [CrossRef]
    [Google Scholar]
  7. Lyczak JB, Cannon CL, Pier GB. Establishment of Pseudomonas aeruginosa infection: lessons from a versatile opportunist. Microb Infect 2000;2:1051–1060
    [Google Scholar]
  8. Cystic Fibrosis Foundation 2016;Cystic fibrosis foundation patient registry 2016 annual data report Bethesda, MD: Cystic Fibrosis Foundation;
  9. Ciofu O, Riis B, Pressler T, Poulsen HE, Hoiby N. Occurrence of hypermutable Pseudomonas aeruginosa in cystic fibrosis patients. Antimicrob Agents Chemother 2005;49:2276–2282
    [Google Scholar]
  10. Gibson RL, Burns JL, Ramsey BW. Pathophysiology and management of pulmonary infections in cystic fibrosis. Am J Respir Crit Care Med 2003;168:918–951 [CrossRef]
    [Google Scholar]
  11. Kosorok MR, Zeng L, West SE, Rock MJ, Splaingard ML et al. Acceleration of lung disease in children with cystic fibrosis after Pseudomonas aeruginosa acquisition. Pediatr Pulmonol 2001;32:277–287 [CrossRef]
    [Google Scholar]
  12. Hoffman LR, Kulasekara HD, Emerson J, Houston LS, Burns JL et al. Pseudomonas aeruginosa lasR mutants are associated with cystic fibrosis lung disease progression. J Cyst Fib 2009;8:66–70 [CrossRef]
    [Google Scholar]
  13. D’Argenio DA, Wu M, Hoffman LR, Kulasekara HD, Déziel E et al. Growth phenotypes of Pseudomonas aeruginosalasR mutants adapted to the airways of cystic fibrosis patients. Mol Microbiol 2007;64:512–533 [CrossRef]
    [Google Scholar]
  14. De Vos D, De Chial M, Cochez C, Jansen S, Tümmler B et al. Study of pyoverdine type and production by Pseudomonas aeruginosa isolated from cystic fibrosis patients: prevalence of type II pyoverdine isolates and accumulation of pyoverdine-negative mutations. Arch Microbiol 2001;175:384–388 [CrossRef]
    [Google Scholar]
  15. Smith EE, Buckley DG, Wu Z, Saenphimmachak C, Hoffman LR et al. Genetic adaptation by Pseudomonas aeruginosa to the airways of cystic fibrosis patients. Proc Nat Acad Sci USA 2006;103:8487–8492 [CrossRef]
    [Google Scholar]
  16. Cabeen MT. Stationary phase-specific virulence factor overproduction by a lasR mutant of Pseudomonas aeruginosa. PLoS One 2014;9:e88743 [CrossRef]
    [Google Scholar]
  17. Römling U, Fiedler B, Bosshammer J, Grothues D, Greipel J et al. Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. J Infect Dis 1994;170:1616–1621 [CrossRef]
    [Google Scholar]
  18. Govan JR, Deretic V. Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepecia. Microbiological Reviews 1996;50:539–574
    [Google Scholar]
  19. Mahenthiralingam E, Campbell ME, Speert DP. Nonmotility and phagocytic resistance of Pseudomonas aeruginosa from chronically colonized patients with cystic fibrosis. Infect Immun 1994;62:596–605
    [Google Scholar]
  20. Taylor RF, Hodson ME, Pitt TL. Auxotrophy of Pseudomonas aeruginosa in cystic fibrosis. FEMS Microbiol Lett 1992;92:243–246 [CrossRef]
    [Google Scholar]
  21. Hancock RE, Mutharia LM, Chan L, Darveau RP, Speert DP et al. Pseudomonas aeruginosa isolates from patients with cystic fibrosis: a class of serum-sensitive, nontypable strains deficient in lipopolysaccharide O side chains. Infect Immun 1983;42:170–177
    [Google Scholar]
  22. Spencer DH, Kas A, Smith EE, Raymond CK, Sims EH et al. Whole-genome sequence variation among multiple isolates of Pseudomonas aeruginosa. J Bacteriol 2003;185:1316–1325 [CrossRef]
    [Google Scholar]
  23. Ernst RK, Moskowitz SM, Emerson JC, Kraig GM, Adams KN et al. Unique lipid A modifications in Pseudomonas aeruginosa isolated from the airways of patients with cystic fibrosis. J Infect Dis 2007;196:1088–1092 [CrossRef]
    [Google Scholar]
  24. Oliver A, Cantón R, Campo P, Baquero F, Blázquez J. High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science 2000;288:1251–1253 [CrossRef]
    [Google Scholar]
  25. Hogardt M, Heesemann J.Microevolution of Pseudomonas aeruginosa to a chronic pathogen of the cystic fibrosis lung In Dobrindt U, Hacker JH, Svanborg C. (editors) Between Pathogenicity and Commensalism Berlin, Heidelberg: Springer; 2012;91–118
    [Google Scholar]
  26. Paulson NB, Gilbertsen AJ, Dalluge JJ, Welchlin CW, Hughes J et al. The arginine decarboxylase pathways of host and pathogen interact to impact inflammatory pathways in the lung. PLoS One 2014;9:e111441 [CrossRef]
    [Google Scholar]
  27. Esther CR, Coakley RD, Henderson AG, Zhou YH, Wright FA et al. Metabolomic evaluation of neutrophilic airway inflammation in cystic fibrosis. Chest 2015;148:507–515 [CrossRef]
    [Google Scholar]
  28. Polineni D, Dang H, Gallins PJ, Jones LC, Pace RG et al. Airway mucosal host defense is key to genomic regulation of cystic fibrosis lung disease severity. Am J Respir Crit Care Med 2018;197:79–93 [CrossRef]
    [Google Scholar]
  29. Johnson L, Mulcahy H, Kanevets U, Shi Y, Lewenza S. Surface-localized spermidine protects the Pseudomonas aeruginosa outer membrane from antibiotic treatment and oxidative stress. J Bacteriol 2012;194:813–826 [CrossRef]
    [Google Scholar]
  30. Rahme LG, Stevens EJ, Wolfort SF, Shao J, Tompkins RG et al. Common virulence factors for bacterial pathogenicity in plants and animals. Science 1995;268:1899–1902 [CrossRef]
    [Google Scholar]
  31. Gilbertsen A, Williams B. Development of a Pseudomonas aeruginosa agmatine biosensor. Biosensors 2014;4:387–402 [CrossRef]
    [Google Scholar]
  32. Williams BJ, Du RH, Calcutt MW, Abdolrasulnia R, Christman BW et al. Discovery of an operon that participates in agmatine metabolism and regulates biofilm formation in Pseudomonas aeruginosa. Mol Microbiol 2010;76:104–119 [CrossRef]
    [Google Scholar]
  33. de Lorenzo V, Timmis KN. Analysis and construction of stable phenotypes in gram-negative bacteria with Tn5- and Tn10-derived minitransposons. Meth Enzymol 1994;235:386–405
    [Google Scholar]
  34. Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP. A broad-host-range Flp-FRT recombination system for site-specific excision of chromosomally-located DNA sequences: application for isolation of unmarked Pseudomonas aeruginosa mutants. Gene 1998;212:77–86 [CrossRef]
    [Google Scholar]
  35. Kwon DH, Lu CD. Polyamines induce resistance to cationic peptide, aminoglycoside, and quinolone antibiotics in Pseudomonas aeruginosa PAO1. Antimicrob Agents Chemother 2006;50:1615–1622 [CrossRef]
    [Google Scholar]
  36. Soothill JS, Ward R, Girling AJ. The IC50: an exactly defined measure of antibiotic sensitivity. J Antimicrob Chemother 1992;29:137–139 [CrossRef]
    [Google Scholar]
  37. Rodriguez-Rojas A, Mena A, Martin S, Borrell N, Oliver A et al. Inactivation of the hmgA gene of Pseudomonas aeruginosa leads to pyomelanin hyperproduction, stress resistance and increased persistence in chronic lung infection. Microbiol 2009;155:1050–1057 [CrossRef]
    [Google Scholar]
  38. Gordon CA, Hodges NA, Marriott C. Antibiotic interaction and diffusion through alginate and exopolysaccharide of cystic fibrosis-derived Pseudomonas aeruginosa. J Antimicrob Chemother 1988;22:667–674 [CrossRef]
    [Google Scholar]
  39. Hodges NA, Gordon CA. Protection of Pseudomonas aeruginosa against ciprofloxacin and beta-lactams by homologous alginate. Antimicrob Agents Chemother 1991;35:2450–2452 [CrossRef]
    [Google Scholar]
  40. Li X, Liu Z, Jin H, Fan X, Yang X et al. Agmatine protects against zymosan-induced acute lung injury in mice by inhibiting NF-κB-mediated inflammatory response. Biomed Res Int 2014;583736 [CrossRef]
    [Google Scholar]
  41. Bragonzi A. Murine models of acute and chronic lung infection with cystic fibrosis pathogens. Int J Med Microbiol 2010;300:584–593 [CrossRef]
    [Google Scholar]
  42. Harada A, Sekido N, Akahoshi T, Wada T, Mukaida N et al. Essential involvement of interleukin-8 (IL-8) in acute inflammation. J Leukoc Biol 1994;56:559–564 [CrossRef]
    [Google Scholar]
  43. Kunkel SL, Standiford T, Kasahara K, Strieter RM. Interleukin-8 (IL-8): the major neutrophil chemotactic factor in the lung. Exp Lung Res 1991;17:17–23 [CrossRef]
    [Google Scholar]
  44. Van Zee KJ, DeForge LE, Fischer E, Marano MA, Kenney J et al. IL-8 in septic shock, endotoxemia, and after IL-1 administation. J Immunol 1991;146:3478–3482
    [Google Scholar]
  45. Miller-Fleming L, Olin-Sandoval V, Campbell K, Ralser M. Remaining mysteries of molecular biology: the role of polyamines in the cell. J Mol Biol 2015;427:3389–3406 [CrossRef]
    [Google Scholar]
  46. Pegg AE. Functions of polyamines in mammals. J Biol Chem 2016;291:14904–14912 [CrossRef]
    [Google Scholar]
  47. Piletz JE, Aricioglu F, Cheng JT, Fairbanks CA, Gilad VH et al. Agmatine: clinical applications after 100 years in translation. Drug Disc Today 2013;18:880–893 [CrossRef]
    [Google Scholar]
  48. Wade CL, Eskridge LL, Nguyen HOX, Kitto KF, Stone LS et al. Immunoneutralization of agmatine sensitized mice to μ-Opioid receptor tolerance. J Pharm Exp Therapeutics 2009;331:539–546 [CrossRef]
    [Google Scholar]
  49. Laube G, Bernstein HG. Agmatine: multifunctional arginine metabolite and magic bullet in clinical neuroscience?. Biochem J 2017;474:2619–2640 [CrossRef]
    [Google Scholar]
  50. Balasubramanian D, Kumari H, Mathee K. Pseudomonas aeruginosa AmpR: an acute-chronic switch regulator. Pathog Dis 2015;73:1–14
    [Google Scholar]
  51. Oliver A, Mena A. Bacterial hypermutation in cystic fibrosis, not only for antibiotic resistance. Clin Microbiol Infect 2010;16:798–808 [CrossRef]
    [Google Scholar]
  52. Winstanley C, O’Brien S, Brockhurst MA. Pseudomonas aeruginosa evolutionary adaptation and diversification in cystic fibrosis chronic lung infections. Trends in Microbiology 2016;24:327–337 [CrossRef]
    [Google Scholar]
  53. Ciofu O, Mandsberg LF, Bjarnsholt T, Wassermann T, Hoiby N. Genetic adaptation of Pseudomonas aeruginosa during chronic lung infection of patients with cystic fibrosis: strong and weak mutators with heterogeneous genetic backgrounds emerge in mucA and/or lasR mutants. Microbiol 2010;156:1108–1119 [CrossRef]
    [Google Scholar]
  54. Lutz L, Leão RS, Ferreira AG, Pereira DC, Raupp C et al. Hypermutable Pseudomonas aeruginosa in cystic fibrosis patients from two Brazilian cities. J Clin Microbiol 2013;51:927–930 [CrossRef]
    [Google Scholar]
  55. Crull MR, Ramos KJ, Caldwell E, Mayer-Hamblett N, Aitken ML et al. Change in Pseudomonas aeruginosa prevalence in cystic fibrosis adults over time. BMC Pulm Med 2016;16:176 [CrossRef]
    [Google Scholar]
  56. Storm DR, Rosenthal KS, Swanson PE. Polymyxin and related peptide antibiotics. Annu Rev Biochem 1977;46:723–763 [CrossRef]
    [Google Scholar]
  57. Hancock RE, Wong PG. Compounds which increase the permeability of the Pseudomonas aeruginosa outer membrane. Antimicrob Agents Chemother 1984;26:48–52 [CrossRef]
    [Google Scholar]
  58. Peterson AA, Hancock REW, McGroarty EJ. Binding of polycationic antibiotics and polyamines to lipopolysaccharides of Pseudomonas aeruginosa. J Bacteriol 1985;164:1256–1261
    [Google Scholar]
  59. Mager J. The stabilizing effect of spermine and related polyamines and bacterial protoplasts. Biochim Biophys Acta 1959;36:529–531 [CrossRef]
    [Google Scholar]
  60. Stevens L. Studies on the interaction of homologues of spermine with deoxyribonucleic acid and with bacterial protoplasts. Biochem J 1967;103:811–815 [CrossRef]
    [Google Scholar]
  61. Tannenbaum CS, Hastie AT, Higgins ML, Kueppers F, Weinbaum G. Inability of purified Pseudomonas aeruginosa exopolysaccharide to bind selected antibiotics. Antimicrob Agents Chemother 1984;25:673–675 [CrossRef]
    [Google Scholar]
  62. Jennings LK, Storek KM, Ledvina HE, Coulon C, Marmont LS et al. Pel is a cationic exopolysaccharide that cross-links extracellular DNA in the Pseudomonas aeruginosa biofilm matrix. Proc Natl Acad Sci USA 2015;112:11353–11358 [CrossRef]
    [Google Scholar]
  63. Kadurugamuwa JL, Lam JS, Beveridge TJ. Interaction of gentamicin with the A band and B band lipopolysaccharides of Pseudomonas aeruginosa and its possible lethal effect. Antimicrob Agents Chemother 1993;37:715–721 [CrossRef]
    [Google Scholar]
  64. Hunter RC, Newman DK. A putative ABC transporter, hatABCDE, is among molecular determinants of pyomelanin production in Pseudomonas aeruginosa. J Bacteriol 2010;192:5962–5971 [CrossRef]
    [Google Scholar]
  65. Cohen T. S, Parker D, Prince A.Pseudomonas aeruginosa host immune evasion In: Ramos JL. (editor) Pseudomonas Dordrecht: Springer; 2015;3–23
    [Google Scholar]
  66. LaFayette SL, Houle D, Beaudoin T, Wojewodka G, Radzioch D et al. Cystic fibrosis–adapted Pseudomonas aeruginosa quorum sensing lasR mutants cause hyperinflammatory responses. Sci Adv 2015;1:e1500199 [CrossRef]
    [Google Scholar]
  67. Scott MG, Vreugdenhil ACE, Buurman WA, Hancock REW, Gold MR. Cutting edge: cationic antimicrobial peptides block the binding of lipopolysaccharide (LPS) to LPS binding protein. J Immunol 2000;164:549–553 [CrossRef]
    [Google Scholar]
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