Cephalosporin-3’ -diazeniumdiolate NO-donor prodrug PYRRO-C3D enhances azithromycin susceptibility of Non-typeable Haemophilus influenzae biofilms
Cephalosporin-3’ -diazeniumdiolate NO-donor prodrug PYRRO-C3D enhances azithromycin susceptibility of Non-typeable Haemophilus influenzae biofilms
Objectives: PYRRO-C3D is a cephalosporin-3-diazeniumdiolate nitric oxide (NO)-donor prodrug designed to selectively deliver NO to bacterial infection sites. The objective of this study was to assess the activity of PYRRO-C3D against non-typeable Haemophilus influenzae (NTHi) biofilms and examine the role of NO in reducing biofilm-associated antibiotic tolerance.
Methods: The activity of PYRRO-C3D on in vitro NTHi biofilms was assessed through CFU enumeration and confocal microscopy. NO release measurements were performed using an ISO-NO probe. NTHi biofilms grown on primary ciliated respiratory epithelia at an air-liquid interface were used to investigate the effects of PYRRO-C3D in the presence of host tissue. Label-free LC/MS proteomic analyses were performed to identify differentially expressed proteins following NO treatment.
Results: PYRRO-C3D specifically released NO in the presence of NTHi, while no evidence of spontaneous NO release was observed when the compound was exposed to primary epithelial cells. NTHi lacking ?-lactamase activity failed to trigger NO release. Treatment significantly increased the susceptibility of in vitro NTHi biofilms to azithromycin, causing a log-fold reduction in viability (p<0.05) relative to azithromycin alone. The response was more pronounced for biofilms grown on primary respiratory epithelia, where a 2-log reduction was observed (p<0.01). Label-free proteomics showed that NO increased expression of sixteen proteins involved in metabolic and transcriptional/translational functions.
Conclusions: NO release from PYRRO-C3D enhances the efficacy of azithromycin against NTHi biofilms, putatively via modulation of NTHi metabolic activity. Adjunctive therapy with NO mediated through PYRRO-C3D represents a promising approach for reducing biofilm-associated antibiotic tolerance.
e02086-16
Collins, Samuel A.
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Kelso, Michael J.
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Rineh, Ardeshir
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Yepuri, Nageshwar R.
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Coles, Janice
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Jackson, Claire L.
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Halladay, Georgia D.
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Walker, Woolf T.
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Webb, Jeremy S.
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Hall-Stoodley, Luanne
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Connett, Gary J.
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Feelisch, Martin
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Faust, Saul N.
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Lucas, Jane S.A.
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Allan, Raymond N.
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February 2017
Collins, Samuel A.
3c35238c-dbbd-4021-b7fa-c5b89e471981
Kelso, Michael J.
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Rineh, Ardeshir
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Yepuri, Nageshwar R.
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Coles, Janice
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Jackson, Claire L.
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Halladay, Georgia D.
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Walker, Woolf T.
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Webb, Jeremy S.
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Hall-Stoodley, Luanne
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Connett, Gary J.
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Feelisch, Martin
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Faust, Saul N.
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Lucas, Jane S.A.
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Allan, Raymond N.
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Collins, Samuel A., Kelso, Michael J., Rineh, Ardeshir, Yepuri, Nageshwar R., Coles, Janice, Jackson, Claire L., Halladay, Georgia D., Walker, Woolf T., Webb, Jeremy S., Hall-Stoodley, Luanne, Connett, Gary J., Feelisch, Martin, Faust, Saul N., Lucas, Jane S.A. and Allan, Raymond N.
(2017)
Cephalosporin-3’ -diazeniumdiolate NO-donor prodrug PYRRO-C3D enhances azithromycin susceptibility of Non-typeable Haemophilus influenzae biofilms.
Antimicrobial Agents and Chemotherapy, 61 (2), .
(doi:10.1128/AAC.02086-16).
(PMID:27919896)
Abstract
Objectives: PYRRO-C3D is a cephalosporin-3-diazeniumdiolate nitric oxide (NO)-donor prodrug designed to selectively deliver NO to bacterial infection sites. The objective of this study was to assess the activity of PYRRO-C3D against non-typeable Haemophilus influenzae (NTHi) biofilms and examine the role of NO in reducing biofilm-associated antibiotic tolerance.
Methods: The activity of PYRRO-C3D on in vitro NTHi biofilms was assessed through CFU enumeration and confocal microscopy. NO release measurements were performed using an ISO-NO probe. NTHi biofilms grown on primary ciliated respiratory epithelia at an air-liquid interface were used to investigate the effects of PYRRO-C3D in the presence of host tissue. Label-free LC/MS proteomic analyses were performed to identify differentially expressed proteins following NO treatment.
Results: PYRRO-C3D specifically released NO in the presence of NTHi, while no evidence of spontaneous NO release was observed when the compound was exposed to primary epithelial cells. NTHi lacking ?-lactamase activity failed to trigger NO release. Treatment significantly increased the susceptibility of in vitro NTHi biofilms to azithromycin, causing a log-fold reduction in viability (p<0.05) relative to azithromycin alone. The response was more pronounced for biofilms grown on primary respiratory epithelia, where a 2-log reduction was observed (p<0.01). Label-free proteomics showed that NO increased expression of sixteen proteins involved in metabolic and transcriptional/translational functions.
Conclusions: NO release from PYRRO-C3D enhances the efficacy of azithromycin against NTHi biofilms, putatively via modulation of NTHi metabolic activity. Adjunctive therapy with NO mediated through PYRRO-C3D represents a promising approach for reducing biofilm-associated antibiotic tolerance.
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Accepted/In Press date: 26 November 2016
e-pub ahead of print date: 5 December 2016
Published date: February 2017
Organisations:
Clinical & Experimental Sciences
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Local EPrints ID: 403594
URI: http://eprints.soton.ac.uk/id/eprint/403594
ISSN: 0066-4804
PURE UUID: 6eb2bda0-0626-4fde-94db-faf8e4d5f051
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Date deposited: 06 Dec 2016 16:57
Last modified: 16 Mar 2024 04:09
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Contributors
Author:
Samuel A. Collins
Author:
Michael J. Kelso
Author:
Ardeshir Rineh
Author:
Nageshwar R. Yepuri
Author:
Janice Coles
Author:
Claire L. Jackson
Author:
Georgia D. Halladay
Author:
Woolf T. Walker
Author:
Luanne Hall-Stoodley
Author:
Gary J. Connett
Author:
Raymond N. Allan
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