Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>

ABSTRACT Staphylococcus aureus is a major concern in human health care, mostly due to the increasing prevalence of antibiotic resistance. Intracellular localization of S. aureus plays a key role in recurrent infections by protecting the pathogens from antibiotics and immune responses. Peptidoglycan...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Christian Röhrig, Markus Huemer, Dominique Lorgé, Samuel Luterbacher, Preeda Phothaworn, Christopher Schefer, Anna M. Sobieraj, Léa V. Zinsli, Srikanth Mairpady Shambat, Nadja Leimer, Anja P. Keller, Fritz Eichenseher, Yang Shen, Sunee Korbsrisate, Annelies S. Zinkernagel, Martin J. Loessner, Mathias Schmelcher
Formato: article
Lenguaje:EN
Publicado: American Society for Microbiology 2020
Materias:
Acceso en línea:https://doaj.org/article/d5ce57eae2424797bb7ba736b3e716b5
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:d5ce57eae2424797bb7ba736b3e716b5
record_format dspace
spelling oai:doaj.org-article:d5ce57eae2424797bb7ba736b3e716b52021-11-15T15:57:03ZTargeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>10.1128/mBio.00209-202150-7511https://doaj.org/article/d5ce57eae2424797bb7ba736b3e716b52020-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00209-20https://doaj.org/toc/2150-7511ABSTRACT Staphylococcus aureus is a major concern in human health care, mostly due to the increasing prevalence of antibiotic resistance. Intracellular localization of S. aureus plays a key role in recurrent infections by protecting the pathogens from antibiotics and immune responses. Peptidoglycan hydrolases (PGHs) are highly specific bactericidal enzymes active against both drug-sensitive and -resistant bacteria. However, PGHs able to effectively target intracellular S. aureus are not yet available. To overcome this limitation, we first screened 322 recombineered PGHs for staphylolytic activity under conditions found inside eukaryotic intracellular compartments. The most active constructs were modified by fusion to different cell-penetrating peptides (CPPs), resulting in increased uptake and enhanced intracellular killing (reduction by up to 4.5 log units) of various S. aureus strains (including methicillin-resistant S. aureus [MRSA]) in different tissue culture infection models. The combined application of synergistic PGH-CPP constructs further enhanced their intracellular efficacy. Finally, synergistically active PGH-CPP cocktails reduced the total S. aureus by more than 2.2 log units in a murine abscess model after peripheral injection. Significantly more intracellular bacteria were killed by the PGH-CPPs than by the PGHs alone. Collectively, our findings show that CPP-fused PGHs are effective novel protein therapeutics against both intracellular and drug-resistant S. aureus. IMPORTANCE The increasing prevalence of antibiotic-resistant bacteria is one of the most urgent problems of our time. Staphylococcus aureus is an important human pathogen that has acquired several mechanisms to evade antibiotic treatment. In addition, S. aureus is able to invade and persist within human cells, hiding from the immune response and antibiotic therapies. For these reasons, novel antibacterial strategies against these pathogens are needed. Here, we developed lytic enzymes which are able to effectively target drug-resistant and intracellular S. aureus. Fusion of these so-called enzybiotics to cell-penetrating peptides enhanced their uptake and intracellular bactericidal activity in cell culture and in an abscess mouse model. Our results suggest that cell-penetrating enzybiotics are a promising new class of therapeutics against staphylococcal infections.Christian RöhrigMarkus HuemerDominique LorgéSamuel LuterbacherPreeda PhothawornChristopher ScheferAnna M. SobierajLéa V. ZinsliSrikanth Mairpady ShambatNadja LeimerAnja P. KellerFritz EichenseherYang ShenSunee KorbsrisateAnnelies S. ZinkernagelMartin J. LoessnerMathias SchmelcherAmerican Society for MicrobiologyarticleendolysinMRSAStaphylococcus aureusantibiotic resistancebacteriophagescell-penetrating peptideMicrobiologyQR1-502ENmBio, Vol 11, Iss 2 (2020)
institution DOAJ
collection DOAJ
language EN
topic endolysin
MRSA
Staphylococcus aureus
antibiotic resistance
bacteriophages
cell-penetrating peptide
Microbiology
QR1-502
spellingShingle endolysin
MRSA
Staphylococcus aureus
antibiotic resistance
bacteriophages
cell-penetrating peptide
Microbiology
QR1-502
Christian Röhrig
Markus Huemer
Dominique Lorgé
Samuel Luterbacher
Preeda Phothaworn
Christopher Schefer
Anna M. Sobieraj
Léa V. Zinsli
Srikanth Mairpady Shambat
Nadja Leimer
Anja P. Keller
Fritz Eichenseher
Yang Shen
Sunee Korbsrisate
Annelies S. Zinkernagel
Martin J. Loessner
Mathias Schmelcher
Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>
description ABSTRACT Staphylococcus aureus is a major concern in human health care, mostly due to the increasing prevalence of antibiotic resistance. Intracellular localization of S. aureus plays a key role in recurrent infections by protecting the pathogens from antibiotics and immune responses. Peptidoglycan hydrolases (PGHs) are highly specific bactericidal enzymes active against both drug-sensitive and -resistant bacteria. However, PGHs able to effectively target intracellular S. aureus are not yet available. To overcome this limitation, we first screened 322 recombineered PGHs for staphylolytic activity under conditions found inside eukaryotic intracellular compartments. The most active constructs were modified by fusion to different cell-penetrating peptides (CPPs), resulting in increased uptake and enhanced intracellular killing (reduction by up to 4.5 log units) of various S. aureus strains (including methicillin-resistant S. aureus [MRSA]) in different tissue culture infection models. The combined application of synergistic PGH-CPP constructs further enhanced their intracellular efficacy. Finally, synergistically active PGH-CPP cocktails reduced the total S. aureus by more than 2.2 log units in a murine abscess model after peripheral injection. Significantly more intracellular bacteria were killed by the PGH-CPPs than by the PGHs alone. Collectively, our findings show that CPP-fused PGHs are effective novel protein therapeutics against both intracellular and drug-resistant S. aureus. IMPORTANCE The increasing prevalence of antibiotic-resistant bacteria is one of the most urgent problems of our time. Staphylococcus aureus is an important human pathogen that has acquired several mechanisms to evade antibiotic treatment. In addition, S. aureus is able to invade and persist within human cells, hiding from the immune response and antibiotic therapies. For these reasons, novel antibacterial strategies against these pathogens are needed. Here, we developed lytic enzymes which are able to effectively target drug-resistant and intracellular S. aureus. Fusion of these so-called enzybiotics to cell-penetrating peptides enhanced their uptake and intracellular bactericidal activity in cell culture and in an abscess mouse model. Our results suggest that cell-penetrating enzybiotics are a promising new class of therapeutics against staphylococcal infections.
format article
author Christian Röhrig
Markus Huemer
Dominique Lorgé
Samuel Luterbacher
Preeda Phothaworn
Christopher Schefer
Anna M. Sobieraj
Léa V. Zinsli
Srikanth Mairpady Shambat
Nadja Leimer
Anja P. Keller
Fritz Eichenseher
Yang Shen
Sunee Korbsrisate
Annelies S. Zinkernagel
Martin J. Loessner
Mathias Schmelcher
author_facet Christian Röhrig
Markus Huemer
Dominique Lorgé
Samuel Luterbacher
Preeda Phothaworn
Christopher Schefer
Anna M. Sobieraj
Léa V. Zinsli
Srikanth Mairpady Shambat
Nadja Leimer
Anja P. Keller
Fritz Eichenseher
Yang Shen
Sunee Korbsrisate
Annelies S. Zinkernagel
Martin J. Loessner
Mathias Schmelcher
author_sort Christian Röhrig
title Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>
title_short Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>
title_full Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>
title_fullStr Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>
title_full_unstemmed Targeting Hidden Pathogens: Cell-Penetrating Enzybiotics Eradicate Intracellular Drug-Resistant <named-content content-type="genus-species">Staphylococcus aureus</named-content>
title_sort targeting hidden pathogens: cell-penetrating enzybiotics eradicate intracellular drug-resistant <named-content content-type="genus-species">staphylococcus aureus</named-content>
publisher American Society for Microbiology
publishDate 2020
url https://doaj.org/article/d5ce57eae2424797bb7ba736b3e716b5
work_keys_str_mv AT christianrohrig targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT markushuemer targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT dominiquelorge targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT samuelluterbacher targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT preedaphothaworn targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT christopherschefer targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT annamsobieraj targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT leavzinsli targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT srikanthmairpadyshambat targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT nadjaleimer targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT anjapkeller targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT fritzeichenseher targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT yangshen targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT suneekorbsrisate targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT anneliesszinkernagel targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT martinjloessner targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
AT mathiasschmelcher targetinghiddenpathogenscellpenetratingenzybioticseradicateintracellulardrugresistantnamedcontentcontenttypegenusspeciesstaphylococcusaureusnamedcontent
_version_ 1718427056873668608