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Cephalosporin-3’ -diazeniumdiolate NO-donor prodrug PYRRO-C3D enhances azithromycin susceptibility of Non-typeable Haemophilus influenzae biofilms

Journal Article


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Abstract


  • 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 nontypeable Haemophilus influenzae (NTHi) biofilms and examine the role of NO in reducing biofilm-associated antibiotic tolerance. 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 liquid chromatography-mass spectrometry (LC/MS) proteomic analyses were performed to identify differentially expressed proteins following NO treatment. 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 β-lactamase activity failed to trigger NO release. Treatment significantly increased the susceptibility of in vitro NTHi biofilms to azithromycin, causing a log fold reduction (10-fold reduction or 1-log-unit 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-unit reduction was observed (P < 0.01). Label-free proteomics showed that NO increased expression of 16 proteins involved in metabolic and transcriptional/translational functions. 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.

Authors


  •   Collins, Samuel (external author)
  •   Kelso, Michael J.
  •   Rineh, Ardeshir (external author)
  •   Yepuri, Rao (external author)
  •   Coles, Janice (external author)
  •   Jackson, Claire (external author)
  •   Halladay, Georgia (external author)
  •   Walker, Woolf (external author)
  •   Webb, Jeremy (external author)
  •   Hall-Stoodley, Luanne (external author)
  •   Connett, Gary (external author)
  •   Feelisch, Martin (external author)
  •   Faust, Saul (external author)
  •   Lucas, Jane (external author)
  •   Allan, Raymond (external author)

Publication Date


  • 2017

Citation


  • Collins, S. A., Kelso, M. J., Rineh, A., Yepuri, N. R., Coles, J., Jackson, C. L., Halladay, G. D., Walker, W. T., Webb, J. S., Hall-Stoodley, L., Connett, G. J., Feelisch, M., Faust, S. N., Lucas, J. S. A. & Allan, R. 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), e02086-16-1-e02086-16-12.

Scopus Eid


  • 2-s2.0-85011096744

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=2075&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1049

Has Global Citation Frequency


Start Page


  • e02086-16-1

End Page


  • e02086-16-12

Volume


  • 61

Issue


  • 2

Place Of Publication


  • United States

Abstract


  • 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 nontypeable Haemophilus influenzae (NTHi) biofilms and examine the role of NO in reducing biofilm-associated antibiotic tolerance. 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 liquid chromatography-mass spectrometry (LC/MS) proteomic analyses were performed to identify differentially expressed proteins following NO treatment. 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 β-lactamase activity failed to trigger NO release. Treatment significantly increased the susceptibility of in vitro NTHi biofilms to azithromycin, causing a log fold reduction (10-fold reduction or 1-log-unit 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-unit reduction was observed (P < 0.01). Label-free proteomics showed that NO increased expression of 16 proteins involved in metabolic and transcriptional/translational functions. 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.

Authors


  •   Collins, Samuel (external author)
  •   Kelso, Michael J.
  •   Rineh, Ardeshir (external author)
  •   Yepuri, Rao (external author)
  •   Coles, Janice (external author)
  •   Jackson, Claire (external author)
  •   Halladay, Georgia (external author)
  •   Walker, Woolf (external author)
  •   Webb, Jeremy (external author)
  •   Hall-Stoodley, Luanne (external author)
  •   Connett, Gary (external author)
  •   Feelisch, Martin (external author)
  •   Faust, Saul (external author)
  •   Lucas, Jane (external author)
  •   Allan, Raymond (external author)

Publication Date


  • 2017

Citation


  • Collins, S. A., Kelso, M. J., Rineh, A., Yepuri, N. R., Coles, J., Jackson, C. L., Halladay, G. D., Walker, W. T., Webb, J. S., Hall-Stoodley, L., Connett, G. J., Feelisch, M., Faust, S. N., Lucas, J. S. A. & Allan, R. 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), e02086-16-1-e02086-16-12.

Scopus Eid


  • 2-s2.0-85011096744

Ro Full-text Url


  • http://ro.uow.edu.au/cgi/viewcontent.cgi?article=2075&context=ihmri

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1049

Has Global Citation Frequency


Start Page


  • e02086-16-1

End Page


  • e02086-16-12

Volume


  • 61

Issue


  • 2

Place Of Publication


  • United States