Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection

ABSTRACT Staphylococcus aureus is a human pathogen causing life-threatening diseases. The increasing prevalence of multidrug-resistant S. aureus infections is a global health concern, requiring development of novel therapeutic options. Peptidoglycan-degrading enzymes (peptidoglycan hydrolases, PGHs)...

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Autores principales:	Anna M. Sobieraj, Markus Huemer, Léa V. Zinsli, Susanne Meile, Anja P. Keller, Christian Röhrig, Fritz Eichenseher, Yang Shen, Annelies S. Zinkernagel, Martin J. Loessner, Mathias Schmelcher
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	endolysin protein therapeutic antibiotic resistance MRSA circulation half-life Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/db6d42fc26334af090638c286d488157
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spelling	oai:doaj.org-article:db6d42fc26334af090638c286d4881572021-11-15T16:19:08ZEngineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection10.1128/mBio.01781-202150-7511https://doaj.org/article/db6d42fc26334af090638c286d4881572020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01781-20https://doaj.org/toc/2150-7511ABSTRACT Staphylococcus aureus is a human pathogen causing life-threatening diseases. The increasing prevalence of multidrug-resistant S. aureus infections is a global health concern, requiring development of novel therapeutic options. Peptidoglycan-degrading enzymes (peptidoglycan hydrolases, PGHs) have emerged as a highly effective class of antimicrobial proteins against S. aureus and other pathogens. When applied to Gram-positive bacteria, PGHs hydrolyze bonds within the peptidoglycan layer, leading to rapid bacterial death by lysis. This activity is highly specific and independent of the metabolic activity of the cell or its antibiotic resistance patterns. However, systemic application of PGHs is limited by their often low activity in vivo and by an insufficient serum circulation half-life. To address this problem, we aimed to extend the half-life of PGHs selected for high activity against S. aureus in human serum. Half-life extension and increased serum circulation were achieved through fusion of PGHs to an albumin-binding domain (ABD), resulting in high-affinity recruitment of human serum albumin and formation of large protein complexes. Importantly, the ABD-fused PGHs maintained high killing activity against multiple drug-resistant S. aureus strains, as determined by ex vivo testing in human blood. The top candidate, termed ABD_M23, was tested in vivo to treat S. aureus-induced murine bacteremia. Our findings demonstrate a significantly higher efficacy of ABD_M23 than of the parental M23 enzyme. We conclude that fusion with ABD represents a powerful approach for half-life extension of PGHs, expanding the therapeutic potential of these enzybiotics for treatment of multidrug-resistant bacterial infections. IMPORTANCE Life-threatening infections with Staphylococcus aureus are often difficult to treat due to the increasing prevalence of antibiotic-resistant bacteria and their ability to persist in protected niches in the body. Bacteriolytic enzymes are promising new antimicrobials because they rapidly kill bacteria, including drug-resistant and persisting cells, by destroying their cell wall. However, when injected into the bloodstream, these enzymes are not retained long enough to clear an infection. Here, we describe a modification to increase blood circulation time of the enzymes and enhance treatment efficacy against S. aureus-induced bloodstream infections. This was achieved by preselecting enzyme candidates for high activity in human blood and coupling them to serum albumin, thereby preventing their elimination by kidney filtration and blood vessel cells.Anna M. SobierajMarkus HuemerLéa V. ZinsliSusanne MeileAnja P. KellerChristian RöhrigFritz EichenseherYang ShenAnnelies S. ZinkernagelMartin J. LoessnerMathias SchmelcherAmerican Society for Microbiologyarticleendolysinprotein therapeuticantibiotic resistanceMRSAcirculation half-lifeMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	endolysin protein therapeutic antibiotic resistance MRSA circulation half-life Microbiology QR1-502
spellingShingle	endolysin protein therapeutic antibiotic resistance MRSA circulation half-life Microbiology QR1-502 Anna M. Sobieraj Markus Huemer Léa V. Zinsli Susanne Meile Anja P. Keller Christian Röhrig Fritz Eichenseher Yang Shen Annelies S. Zinkernagel Martin J. Loessner Mathias Schmelcher Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection
description	ABSTRACT Staphylococcus aureus is a human pathogen causing life-threatening diseases. The increasing prevalence of multidrug-resistant S. aureus infections is a global health concern, requiring development of novel therapeutic options. Peptidoglycan-degrading enzymes (peptidoglycan hydrolases, PGHs) have emerged as a highly effective class of antimicrobial proteins against S. aureus and other pathogens. When applied to Gram-positive bacteria, PGHs hydrolyze bonds within the peptidoglycan layer, leading to rapid bacterial death by lysis. This activity is highly specific and independent of the metabolic activity of the cell or its antibiotic resistance patterns. However, systemic application of PGHs is limited by their often low activity in vivo and by an insufficient serum circulation half-life. To address this problem, we aimed to extend the half-life of PGHs selected for high activity against S. aureus in human serum. Half-life extension and increased serum circulation were achieved through fusion of PGHs to an albumin-binding domain (ABD), resulting in high-affinity recruitment of human serum albumin and formation of large protein complexes. Importantly, the ABD-fused PGHs maintained high killing activity against multiple drug-resistant S. aureus strains, as determined by ex vivo testing in human blood. The top candidate, termed ABD_M23, was tested in vivo to treat S. aureus-induced murine bacteremia. Our findings demonstrate a significantly higher efficacy of ABD_M23 than of the parental M23 enzyme. We conclude that fusion with ABD represents a powerful approach for half-life extension of PGHs, expanding the therapeutic potential of these enzybiotics for treatment of multidrug-resistant bacterial infections. IMPORTANCE Life-threatening infections with Staphylococcus aureus are often difficult to treat due to the increasing prevalence of antibiotic-resistant bacteria and their ability to persist in protected niches in the body. Bacteriolytic enzymes are promising new antimicrobials because they rapidly kill bacteria, including drug-resistant and persisting cells, by destroying their cell wall. However, when injected into the bloodstream, these enzymes are not retained long enough to clear an infection. Here, we describe a modification to increase blood circulation time of the enzymes and enhance treatment efficacy against S. aureus-induced bloodstream infections. This was achieved by preselecting enzyme candidates for high activity in human blood and coupling them to serum albumin, thereby preventing their elimination by kidney filtration and blood vessel cells.
format	article
author	Anna M. Sobieraj Markus Huemer Léa V. Zinsli Susanne Meile Anja P. Keller Christian Röhrig Fritz Eichenseher Yang Shen Annelies S. Zinkernagel Martin J. Loessner Mathias Schmelcher
author_facet	Anna M. Sobieraj Markus Huemer Léa V. Zinsli Susanne Meile Anja P. Keller Christian Röhrig Fritz Eichenseher Yang Shen Annelies S. Zinkernagel Martin J. Loessner Mathias Schmelcher
author_sort	Anna M. Sobieraj
title	Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection
title_short	Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection
title_full	Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection
title_fullStr	Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection
title_full_unstemmed	Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection
title_sort	engineering of long-circulating peptidoglycan hydrolases enables efficient treatment of systemic <named-content content-type="genus-species">staphylococcus aureus</named-content> infection
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/db6d42fc26334af090638c286d488157
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Engineering of Long-Circulating Peptidoglycan Hydrolases Enables Efficient Treatment of Systemic <named-content content-type="genus-species">Staphylococcus aureus</named-content> Infection

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