Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides

ABSTRACT The mycobacterial cell wall affords natural resistance to antibiotics. Antimicrobial peptides (AMPs) modify the surface properties of mycobacteria and can act synergistically with antibiotics from differing classes. Here, we investigate the response of Mycobacterium smegmatis to the presenc...

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Autores principales:	DeDe Kwun-Wai Man, Tokuwa Kanno, Giorgia Manzo, Brian D. Robertson, Jenny K. W. Lam, A. James Mason
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	mycobacteria NMR metabolomics capreomycin rifampin synergy tyloxapol Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/30e2623b09474f2a90ca386387fab3dd
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spelling	oai:doaj.org-article:30e2623b09474f2a90ca386387fab3dd2021-11-15T15:25:50ZRifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides10.1128/mSphere.00218-182379-5042https://doaj.org/article/30e2623b09474f2a90ca386387fab3dd2018-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00218-18https://doaj.org/toc/2379-5042ABSTRACT The mycobacterial cell wall affords natural resistance to antibiotics. Antimicrobial peptides (AMPs) modify the surface properties of mycobacteria and can act synergistically with antibiotics from differing classes. Here, we investigate the response of Mycobacterium smegmatis to the presence of rifampin or capreomycin, either alone or in combination with two synthetic, cationic, α-helical AMPs that are distinguished by the presence (D-LAK120-HP13) or absence (D-LAK120-A) of a kink-inducing proline. Using a combination of high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) metabolomics, diphenylhexatriene (DPH) fluorescence anisotropy measurements, and laurdan emission spectroscopy, we show that M. smegmatis responds to challenge with rifampin or capreomycin by substantially altering its metabolism and, in particular, by remodeling the cell envelope. Overall, the changes are consistent with a reduction of trehalose dimycolate and an increase of trehalose monomycolate and are associated with increased rigidity of the mycolic acid layer observed following challenge by capreomycin but not rifampin. Challenge with D-LAK120-A or D-LAK120-HP13 induced no or modest changes, respectively, in mycomembrane metabolites and did not induce a significant increase in the rigidity of the mycolic acid layer. Furthermore, the response to rifampin or capreomycin was significantly reduced when these were combined with D-LAK120-HP13 and D-LAK120-A, respectively, suggesting a possible mechanism for the synergy of these combinations. The remodeling of the mycomembrane in M. smegmatis is therefore identified as an important countermeasure deployed against rifampin or capreomycin, but this can be mitigated and the efficacy of rifampin or capreomycin potentiated by combining the drug with AMPs. IMPORTANCE We have used a combined NMR metabolomics/biophysical approach to better understand differences in the mechanisms of two closely related antimicrobial peptides, as well as the response of the model organism Mycobacterium smegmatis to challenge with first- or second-line antibiotics used against mycobacterial pathogens. We show that, in addition to membrane damage, the triggering of oxidative stress may be an important part of the mechanism of action of one AMP. The metabolic shift that accompanied rifampin and, particularly, capreomycin challenge was associated with modest and more dramatic changes, respectively, in the mycomembrane, providing a rationale for how the response to one antibiotic may affect bacterial penetration and, hence, the action of another. This study presents the first insights into how antimicrobial peptides may operate synergistically with existing antibiotics whose efficacy is waning or sensitize MDR mycobacteria and/or latent mycobacterial infections to them, prolonging the useful life of these drugs.DeDe Kwun-Wai ManTokuwa KannoGiorgia ManzoBrian D. RobertsonJenny K. W. LamA. James MasonAmerican Society for MicrobiologyarticlemycobacteriaNMR metabolomicscapreomycinrifampinsynergytyloxapolMicrobiologyQR1-502ENmSphere, Vol 3, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	mycobacteria NMR metabolomics capreomycin rifampin synergy tyloxapol Microbiology QR1-502
spellingShingle	mycobacteria NMR metabolomics capreomycin rifampin synergy tyloxapol Microbiology QR1-502 DeDe Kwun-Wai Man Tokuwa Kanno Giorgia Manzo Brian D. Robertson Jenny K. W. Lam A. James Mason Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides
description	ABSTRACT The mycobacterial cell wall affords natural resistance to antibiotics. Antimicrobial peptides (AMPs) modify the surface properties of mycobacteria and can act synergistically with antibiotics from differing classes. Here, we investigate the response of Mycobacterium smegmatis to the presence of rifampin or capreomycin, either alone or in combination with two synthetic, cationic, α-helical AMPs that are distinguished by the presence (D-LAK120-HP13) or absence (D-LAK120-A) of a kink-inducing proline. Using a combination of high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) metabolomics, diphenylhexatriene (DPH) fluorescence anisotropy measurements, and laurdan emission spectroscopy, we show that M. smegmatis responds to challenge with rifampin or capreomycin by substantially altering its metabolism and, in particular, by remodeling the cell envelope. Overall, the changes are consistent with a reduction of trehalose dimycolate and an increase of trehalose monomycolate and are associated with increased rigidity of the mycolic acid layer observed following challenge by capreomycin but not rifampin. Challenge with D-LAK120-A or D-LAK120-HP13 induced no or modest changes, respectively, in mycomembrane metabolites and did not induce a significant increase in the rigidity of the mycolic acid layer. Furthermore, the response to rifampin or capreomycin was significantly reduced when these were combined with D-LAK120-HP13 and D-LAK120-A, respectively, suggesting a possible mechanism for the synergy of these combinations. The remodeling of the mycomembrane in M. smegmatis is therefore identified as an important countermeasure deployed against rifampin or capreomycin, but this can be mitigated and the efficacy of rifampin or capreomycin potentiated by combining the drug with AMPs. IMPORTANCE We have used a combined NMR metabolomics/biophysical approach to better understand differences in the mechanisms of two closely related antimicrobial peptides, as well as the response of the model organism Mycobacterium smegmatis to challenge with first- or second-line antibiotics used against mycobacterial pathogens. We show that, in addition to membrane damage, the triggering of oxidative stress may be an important part of the mechanism of action of one AMP. The metabolic shift that accompanied rifampin and, particularly, capreomycin challenge was associated with modest and more dramatic changes, respectively, in the mycomembrane, providing a rationale for how the response to one antibiotic may affect bacterial penetration and, hence, the action of another. This study presents the first insights into how antimicrobial peptides may operate synergistically with existing antibiotics whose efficacy is waning or sensitize MDR mycobacteria and/or latent mycobacterial infections to them, prolonging the useful life of these drugs.
format	article
author	DeDe Kwun-Wai Man Tokuwa Kanno Giorgia Manzo Brian D. Robertson Jenny K. W. Lam A. James Mason
author_facet	DeDe Kwun-Wai Man Tokuwa Kanno Giorgia Manzo Brian D. Robertson Jenny K. W. Lam A. James Mason
author_sort	DeDe Kwun-Wai Man
title	Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides
title_short	Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides
title_full	Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides
title_fullStr	Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides
title_full_unstemmed	Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides
title_sort	rifampin- or capreomycin-induced remodeling of the <italic toggle="yes">mycobacterium smegmatis</italic> mycolic acid layer is mitigated in synergistic combinations with cationic antimicrobial peptides
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/30e2623b09474f2a90ca386387fab3dd
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Rifampin- or Capreomycin-Induced Remodeling of the <italic toggle="yes">Mycobacterium smegmatis</italic> Mycolic Acid Layer Is Mitigated in Synergistic Combinations with Cationic Antimicrobial Peptides

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