Poly (acetyl, arginyl) glucosamine disrupts Pseudomonas aeruginosa biofilms and enhances bacterial clearance in a rat lung infection model

Bryan A. Garcia; Melissa S. McDaniel; Allister J. Loughran; J. Dixon Johns; Vidya Narayanaswamy; Courtney Fernandez Petty; Susan E. Birket; Shenda M. Baker; Roxanna Barnaby; Bruce A. Stanton; Jeremy B. Foote; Steven M. Rowe; W. Edward Swords

doi:10.1099/mic.0.001121

Volume 168, Issue 1

Research Article

Open Access

Poly (acetyl, arginyl) glucosamine disrupts Pseudomonas aeruginosa biofilms and enhances bacterial clearance in a rat lung infection model

Bryan A. Garcia^1,2,3, Melissa S. McDaniel^3,4, Allister J. Loughran^5,6, J. Dixon Johns^2,3, Vidya Narayanaswamy⁵, Courtney Fernandez Petty², Susan E. Birket^2,3,4,7, Shenda M. Baker^5,8, Roxanna Barnaby⁹, Bruce A. Stanton⁹, Jeremy B. Foote¹⁰, Steven M. Rowe^2,3,4,7 and W. Edward Swords^2,3,4
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Affiliations: ¹ Department of Medicine, Division of Pulmonary, Critical Care, and Sleep Medicine, Medical University of South Carolina, Charleston, SC, USA ² Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA ³ Division of Pulmonary, Allergy, and Critical Care Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA ⁴ Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, Alabama, USA ⁵ Synedgen, Inc., Claremont, CA, USA ⁶ St Jude Children’s Research Hospital, Memphis, TN, USA ⁷ Department of Cell Developmental and Integrative Physiology, University of Alabama at Birmingham, Birmingham, Alabama, USA ⁸ Synspira Therapeutics, Inc., Framingham, MA, USA ⁹ Dartmouth Cystic Fibrosis Research Center, New Hanover, NH, USA ¹⁰ Comparative Pathology Laboratory, at University of Alabama at Birmingham, Birmingham, Alabama, USA
*Correspondence: W. Edward Swords, [email protected]
Published: 25 January 2022 https://doi.org/10.1099/mic.0.001121

Abstract

Pseudomonas aeruginosa is a common opportunistic pathogen that can cause chronic infections in multiple disease states, including respiratory infections in patients with cystic fibrosis (CF) and non-CF bronchiectasis. Like many opportunists, P. aeruginosa forms multicellular biofilm communities that are widely thought to be an important determinant of bacterial persistence and resistance to antimicrobials and host immune effectors during chronic/recurrent infections. Poly (acetyl, arginyl) glucosamine (PAAG) is a glycopolymer that has antimicrobial activity against a broad range of bacterial species, and also has mucolytic activity, which can normalize the rheological properties of cystic fibrosis mucus. In this study, we sought to evaluate the effect of PAAG on P. aeruginosa bacteria within biofilms in vitro, and in the context of experimental pulmonary infection in a rodent infection model. PAAG treatment caused significant bactericidal activity against P. aeruginosa biofilms, and a reduction in the total biomass of preformed P. aeruginosa biofilms on abiotic surfaces, as well as on the surface of immortalized cystic fibrosis human bronchial epithelial cells. Studies of membrane integrity indicated that PAAG causes changes to P. aeruginosa cell morphology and dysregulates membrane polarity. PAAG treatment reduced infection and consequent tissue inflammation in experimental P. aeruginosa rat infections. Based on these findings we conclude that PAAG represents a novel means to combat P. aeruginosa infection, and may warrant further evaluation as a therapeutic.

Received: 28/05/2021
Accepted: 29/10/2021
Published Online: 25/01/2022

Keyword(s): biofilm , cystic fibrosis , mucus and Pseudomonas aeruginosa

Funding

This study was supported by the:

National Heart, Lung, and Blood Institute (Award R35 HL135816)
- Principle Award Recipient: StevenM. Rowe
National Institute of Diabetes and Digestive and Kidney Diseases (Award P30DK072482)
- Principle Award Recipient: StevenM. Rowe
Cystic Fibrosis Foundation (Award CFF SWORDS20G0)
- Principle Award Recipient: Edward Edward SwordsW.
Cystic Fibrosis Foundation (Award CFF SWORDS1810)
- Principle Award Recipient: Edward Edward SwordsW.
Cystic Fibrosis Foundation (Award RDP R15RO)
- Principle Award Recipient: StevenM. Rowe
Synedgen
- Principle Award Recipient: StevenM. Rowe

This is an open-access article distributed under the terms of the Creative Commons Attribution License.

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References

Carmeli Y, Troillet N, Eliopoulos GM, Samore MH. Emergence of antibiotic-resistant Pseudomonas aeruginosa: comparison of risks associated with different antipseudomonal agents. Antimicrob Agents Chemother 1999; 43:1379–1382 [View Article] [PubMed]
[Google Scholar]
Planquette B, Timsit J-F, Misset BY, Schwebel C, Azoulay E et al. Pseudomonas aeruginosa ventilator-associated pneumonia. predictive factors of treatment failure. Am J Respir Crit Care Med 2013; 188:69–76 [View Article] [PubMed]
[Google Scholar]
Sadikot RT, Blackwell TS, Christman JW, Prince AS. Pathogen-host interactions in Pseudomonas aeruginosa pneumonia. Am J Respir Crit Care Med 2005; 171:1209–1223 [View Article] [PubMed]
[Google Scholar]
Trecarichi EM, Tumbarello M. Antimicrobial-resistant Gram-negative bacteria in febrile neutropenic patients with cancer: current epidemiology and clinical impact. Curr Opin Infect Dis 2014; 27:200–210 [View Article] [PubMed]
[Google Scholar]
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: [View Article] [PubMed]
[Google Scholar]
Wilson R, Aksamit T, Aliberti S, De Soyza A, Elborn JS et al. Challenges in managing Pseudomonas aeruginosa in non-cystic fibrosis bronchiectasis. Respir Med 2016; 117:179–189 [View Article] [PubMed]
[Google Scholar]
Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M et al. Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis 2018; 18:318–327 [View Article] [PubMed]
[Google Scholar]
Livermore DM. Multiple mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst nightmare?. Clin Infect Dis 2002; 34:634–640 [View Article] [PubMed]
[Google Scholar]
Bhagirath AY, Li Y, Somayajula D, Dadashi M, Badr S et al. Cystic fibrosis lung environment and Pseudomonas aeruginosa infection. BMC Pulm Med 2016; 16: [View Article] [PubMed]
[Google Scholar]
Maurice NM, Bedi B, Sadikot RT. Pseudomonas aeruginosa Biofilms: Host response and clinical implications in lung infections. Am J Respir Cell Mol Biol 2018; 58:428–439 [View Article] [PubMed]
[Google Scholar]
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 [View Article] [PubMed]
[Google Scholar]
Ciofu O, Tolker-Nielsen T. Tolerance and resistance of Pseudomonas aeruginosa biofilms to antimicrobial agents-how P. aeruginosa can escape antibiotics. Front Microbiol 2019; 10: [View Article] [PubMed]
[Google Scholar]
Flemming HC, Wingender J. The biofilm matrix. Nat Rev Microbiol 2010; 8:623–633 [View Article] [PubMed]
[Google Scholar]
Mann EE, Wozniak DJ. Pseudomonas biofilm matrix composition and niche biology. FEMS Microbiol Rev 2012; 36:893–916 [View Article] [PubMed]
[Google Scholar]
Mack D, Fischer W, Krokotsch A, Leopold K, Hartmann R et al. The intercellular adhesin involved in biofilm accumulation of Staphylococcus epidermidis is a linear beta-1,6-linked glucosaminoglycan: purification and structural analysis. J Bacteriol 1996; 178:175–183 [View Article] [PubMed]
[Google Scholar]
Wei Q, Ma LZ. Biofilm matrix and its regulation in Pseudomonas aeruginosa. Int J Mol Sci 2013; 14:20983–21005 [View Article] [PubMed]
[Google Scholar]
Fernandez-Petty CM, Hughes GW, Bowers HL, Watson JD, Rosen BH et al. A glycopolymer improves vascoelasticity and mucociliary transport of abnormal cystic fibrosis mucus. JCI Insight 20194 4.: [View Article]
[Google Scholar]
Quinton PM. Cystic fibrosis: impaired bicarbonate secretion and mucoviscidosis. Lancet 2008; 372:415–417 [View Article] [PubMed]
[Google Scholar]
Gustafsson JK, Ermund A, Ambort D, Johansson MEV, Nilsson HE et al. Bicarbonate and functional CFTR channel are required for proper mucin secretion and link cystic fibrosis with its mucus phenotype. J Exp Med 2012; 209:1263–1272 [View Article] [PubMed]
[Google Scholar]
Kesimer M, Makhov AM, Griffith JD, Verdugo P, Sheehan JK. Unpacking a gel-forming mucin: a view of MUC5B organization after granular release. Am J Physiol Lung Cell Mol Physiol 2010; 298:L15–22 [View Article] [PubMed]
[Google Scholar]
Clifton LA, Skoda MWA, Le Brun AP, Ciesielski F, Kuzmenko I et al. Effect of divalent cation removal on the structure of Gram-negative bacterial outer membrane models. Langmuir 2015; 31:404–412 [View Article] [PubMed]
[Google Scholar]
Nikaido H, Vaara M. Molecular basis of bacterial outer membrane permeability. Microbiol Rev 1985; 49:1–32 [View Article]
[Google Scholar]
Wang T, Flint S, Palmer J. Magnesium and calcium ions: roles in bacterial cell attachment and biofilm structure maturation. Biofouling 2019; 35:959–974 [View Article] [PubMed]
[Google Scholar]
Das T, Sehar S, Koop L, Wong YK, Ahmed S et al. Influence of calcium in extracellular DNA mediated bacterial aggregation and biofilm formation. PLoS One 2014; 9:e91935 [View Article] [PubMed]
[Google Scholar]
Banin E, Brady KM, Greenberg EP. Chelator-induced dispersal and killing of Pseudomonas aeruginosa cells in a biofilm. Appl Environ Microbiol 2006; 72:2064–2069 [View Article] [PubMed]
[Google Scholar]
Narayanaswamy VP, Giatpaiboon S, Baker SM, Wiesmann WP, LiPuma JJ et al. Novel glycopolymer sensitizes Burkholderia cepacia complex isolates from cystic fibrosis patients to tobramycin and meropenem. PLoS One 2017; 12:e0179776 [View Article] [PubMed]
[Google Scholar]
Narayanaswamy VP, Duncan AP, LiPuma JJ, Wiesmann WP, Baker SM et al. In vitro activity of a novel Glycopolymer against biofilms of Burkholderia cepacia complex cystic fibrosis clinical isolates. Antimicrob Agents Chemother 201963 63.: [View Article]
[Google Scholar]
Bloemberg GV, O’Toole GA, Lugtenberg BJ, Kolter R. Green fluorescent protein as a marker for Pseudomonas spp. Appl Environ Microbiol 1997; 63:4543–4551 [View Article] [PubMed]
[Google Scholar]
Suzuki H, Wang ZY, Yamakoshi M, Kobayashi M, Nozawa T. Probing the transmembrane potential of bacterial cells by voltage-sensitive dyes. Anal Sci 2003; 19:1239–1242 [View Article] [PubMed]
[Google Scholar]
Nikaido H. Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 2003; 67:593–656 [View Article] [PubMed]
[Google Scholar]
Grant MM, Niederman MS, Poehlman MA, Fein AM. Characterization of Pseudomonas aeruginosa adherence to cultured hamster tracheal epithelial cells. Am J Respir Cell Mol Biol 1991; 5:563–570 [View Article] [PubMed]
[Google Scholar]
Stover CK, Pham XQ, Erwin AL, Mizoguchi SD, Warrener P et al. Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 2000; 406:959–964 [View Article] [PubMed]
[Google Scholar]
Rowe SM, Miller S, Sorscher EJ. Cystic fibrosis. N Engl J Med 2005; 352:1992–2001 [View Article] [PubMed]
[Google Scholar]

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Poly (acetyl, arginyl) glucosamine disrupts Pseudomonas aeruginosa biofilms and enhances bacterial clearance in a rat lung infection model

Microbiology 168, 001121 (2022); https://doi.org/10.1099/mic.0.001121

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Volume 168, Issue 1

Research Article

Open Access

Poly (acetyl, arginyl) glucosamine disrupts Pseudomonas aeruginosa biofilms and enhances bacterial clearance in a rat lung infection model

Abstract

Funding

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