The University of Southampton
University of Southampton Institutional Repository

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
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.
0066-4804
e02086-16
Collins, Samuel A.
3c35238c-dbbd-4021-b7fa-c5b89e471981
Kelso, Michael J.
6afd2b75-c893-44cf-8b86-93e89e21f894
Rineh, Ardeshir
9b3b5d87-023c-49a3-9290-c793e4fa07c2
Yepuri, Nageshwar R.
74fb7761-9b7e-43ec-b5df-274c2458eb78
Coles, Janice
fb9d20aa-93b9-42b3-9b9e-bab2f565ea60
Jackson, Claire L.
64cdd6fa-74c3-4ac6-94ef-070620a6efd9
Halladay, Georgia D.
cf225cb0-b2d2-4d40-9ed2-cc6a9b63b6e0
Walker, Woolf T.
58aae223-5b0e-4f34-9ee7-58bb68278c3a
Webb, Jeremy S.
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Connett, Gary J.
55d5676c-90d8-46bf-a508-62eded276516
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Faust, Saul N.
f97df780-9f9b-418e-b349-7adf63e150c1
Lucas, Jane S.A.
5cb3546c-87b2-4e59-af48-402076e25313
Allan, Raymond N.
390a7d0a-38e1-410a-8dfe-c8ef8408f5e1
Collins, Samuel A.
3c35238c-dbbd-4021-b7fa-c5b89e471981
Kelso, Michael J.
6afd2b75-c893-44cf-8b86-93e89e21f894
Rineh, Ardeshir
9b3b5d87-023c-49a3-9290-c793e4fa07c2
Yepuri, Nageshwar R.
74fb7761-9b7e-43ec-b5df-274c2458eb78
Coles, Janice
fb9d20aa-93b9-42b3-9b9e-bab2f565ea60
Jackson, Claire L.
64cdd6fa-74c3-4ac6-94ef-070620a6efd9
Halladay, Georgia D.
cf225cb0-b2d2-4d40-9ed2-cc6a9b63b6e0
Walker, Woolf T.
58aae223-5b0e-4f34-9ee7-58bb68278c3a
Webb, Jeremy S.
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Hall-Stoodley, Luanne
94ebdc00-b549-4488-b15f-5310fb965f5b
Connett, Gary J.
55d5676c-90d8-46bf-a508-62eded276516
Feelisch, Martin
8c1b9965-8614-4e85-b2c6-458a2e17eafd
Faust, Saul N.
f97df780-9f9b-418e-b349-7adf63e150c1
Lucas, Jane S.A.
5cb3546c-87b2-4e59-af48-402076e25313
Allan, Raymond N.
390a7d0a-38e1-410a-8dfe-c8ef8408f5e1

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), e02086-16. (doi:10.1128/AAC.02086-16). (PMID:27919896)

Record type: Article

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.

Text
Final revised manuscript.pdf - Accepted Manuscript
Download (3MB)

More information

Accepted/In Press date: 26 November 2016
e-pub ahead of print date: 5 December 2016
Published date: February 2017
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 403594
URI: https://eprints.soton.ac.uk/id/eprint/403594
ISSN: 0066-4804
PURE UUID: 6eb2bda0-0626-4fde-94db-faf8e4d5f051
ORCID for Jeremy S. Webb: ORCID iD orcid.org/0000-0003-2068-8589
ORCID for Gary J. Connett: ORCID iD orcid.org/0000-0003-1310-3239
ORCID for Martin Feelisch: ORCID iD orcid.org/0000-0003-2320-1158
ORCID for Saul N. Faust: ORCID iD orcid.org/0000-0003-3410-7642

Catalogue record

Date deposited: 06 Dec 2016 16:57
Last modified: 03 Dec 2019 01:46

Export record

Altmetrics

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: Jeremy S. Webb ORCID iD
Author: Luanne Hall-Stoodley
Author: Gary J. Connett ORCID iD
Author: Martin Feelisch ORCID iD
Author: Saul N. Faust ORCID iD
Author: Jane S.A. Lucas
Author: Raymond N. Allan

University divisions

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of https://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×