Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance

Kirsten Leeten; Nicolas Jacques; Lidia Alejo Esquembre; Dana C. Schneider; Jan Straetener; Camilla Henriksen; Lucia Musumeci; Florence Putters; Sofia Melo; Elena Sánchez-López; Martin Giera; Noémie Penoy; Géraldine Piel; Olivier Verlaine; Ana Amoroso; Bernard Joris; Christoph J. Slavetinsky; Eric Goffin; Bernard Pirotte; Dorte Frees; Heike Brötz-Oesterhelt; Patrizio Lancellotti; Cécile Oury

doi:10.1128/mbio.01322-24

Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance

Kirsten Leeten, Nicolas Jacques, Lidia Alejo Esquembre, Dana C. Schneider, Jan Straetener, Camilla Henriksen, Lucia Musumeci, Florence Putters, Sofia Melo, Elena Sánchez-López, Martin Giera, Noémie Penoy, Géraldine Piel, Olivier Verlaine, Ana Amoroso, Bernard Joris, Christoph J. Slavetinsky, Eric Goffin, Bernard Pirotte, Dorte FreesHeike Brötz-Oesterhelt^*, Patrizio Lancellotti, Cécile Oury

^*Corresponding author af dette arbejde

Sektion, Food Safety and Zoonoses

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Abstract

Infections with multidrug-resistant bacteria pose a major healthcare problem which urges the need for novel treatment options. Besides its potent antiplatelet properties, ticagrelor has antibacterial activity against Gram-positive bacteria, including methicillin- and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA). Several retrospective studies in cardiovascular patients support an antibacterial effect of this drug which is not related to its antiplatelet activity. We investigated the mechanism of action of ticagrelor in Staphylococcus aureus and model Bacillus subtilis, and assessed cross-resistance with two conventional anti-MRSA antibiotics, vancomycin and daptomycin. Bacillus subtilis bioreporter strains revealed ticagrelor-induced cell envelope-related stress responses. Sub-inhibitory drug concentrations caused membrane depolarization, impaired positioning of both the peripheral membrane protein MinD and the peptidoglycan precursor lipid II, and it affected cell shape. At the MIC, ticagrelor destroyed membrane integrity, indicated by the influx of membrane impermeable dyes, and lipid aggregate formation. Whole-genome sequencing of in vitro-generated ticagrelor-resistant MRSA clones revealed mutations in genes encoding ClpP, ClpX, and YjbH. Lipidomic analysis of resistant clones displayed changes in levels of the most abundant lipids of the Staphylococcus aureus membrane, for example, cardiolipins, phosphatidylglycerols, and diacylglycerols. Exogeneous cardiolipin, phosphatidylglycerol, or diacylglycerol antagonized the antibacterial properties of ticagrelor. Ticagrelor enhanced MRSA growth inhibition and killing by vancomycin and daptomycin in both exponential and stationary phases. Finally, no cross-resistance was observed between ticagrelor and daptomycin, or vancomycin. Our study demonstrates that ticagrelor targets multiple lipids in the cytoplasmic membrane of Gram-positive bacteria, thereby retaining activity against multidrug-resistant staphylococci including daptomycin- and vancomycin-resistant strains.

Originalsprog	Engelsk
Artikelnummer	e01322-24
Tidsskrift	mBio
Vol/bind	15
Udgave nummer	10
Antal sider	20
ISSN	2161-2129
DOI	https://doi.org/10.1128/mbio.01322-24
Status	Udgivet - 2024

Bibliografisk note

Funding Information:
The authors thank the GIGA technological platforms, particularly GIGA-Genomics. C.O. is a Research Director at the Belgium National Funds for Scientific Research (F.R.S.-FNRS). Research from the University of Li\u00E8ge (ULi\u00E8ge) is funded by F.R.S.-FNRS (grant number PDR T.0190.20) to C.O., ULi\u00E8ge internal Fund grant numbers FSR-SSS-19/18 and FSR-S-SS-21/41 to C.O., and European Research Council Consolidator grant (grant number 647197) to P.L. S.M. has a fellowship from the FRIA (Belgium National Funds for Scientific Research). This work was supported by funding from the German Center for Infection Research, DZIF (D.C.S. and H.B.-O.) via project TTU 09.826 (PAACT), from the Deutsche Forschungsgemeinschaft (German Research Foundation, DFG) (L.A.E. and H.B.-O.) via TRR 261 (project ID 398967434). H.B.-O. and D.C.S. acknowledge infrastructural support from the Cluster of Excellence EXC 2124: Controlling Microbes to Fight Infection (CMFI, project ID 390838134) and L.A.E. from the Max-Planck Research School \u201CFrom Molecules to Organisms,\u201D Tuebingen. F\u00E9d\u00E9ration Wallonie-Bruxelles (French FSR-S-SS-19/18, C\u00E9cile Oury Community of Belgium) FSR-S-SS-21/41 EC | European Research Council (ERC) 647197 Patrizio Lancellotti Deutsches Zentrum f\u00FCr Infektionsforschung TTU 09.826 Heike Br\u00F6tz-Oester(DZIF) helt Deutsche Forschungsgemeinschaft (DFG) TRR 261 project Heike Br\u00F6tz-Oester398967434 helt

Funding Information:
C.O. is a Research Director at the Belgium National Funds for Research (F.R.S.-FNRS). Research from the University of Li\u00E8ge (ULi\u00E8ge) is funded by F.R.S.-FNRS (grant number PDR T.0190.20) to C.O., ULi\u00E8ge internal Fund grant numbers FSR-SSS-19/18 and FSR-S-SS-21/41 to C.O., and European Research Council Consolidator grant (grant number 647197) to P.L. S.M. has a fellowship from the FRIA (Belgium National Funds for Research). This work was supported by funding from the German Center for Infection Research, DZIF (D.C.S. and H.B.-O.) via project TTU 09.826 (PAACT), from the Deutsche Forschungsgemeinschaft (German Research Foundation, DFG) (L.A.E. and H.B.-O.) via TRR 261 (project ID 398967434). H.B.-O. and D.C.S. acknowledge infrastructural support from the Cluster of Excellence EXC 2124: Controlling Microbes to Fight Infection (CMFI, project ID 390838134) and L.A.E. from the Max-Planck Research School \u201CFrom Molecules to Organisms,\u201D Tuebingen.

Publisher Copyright:
Copyright © 2024 Leeten et al.

Adgang til dokumentet

10.1128/mbio.01322-24Licens: CC BY

FulltextForlagets udgivne version, 7,16 MBLicens: CC BY

Citationsformater

Leeten, K., Jacques, N., Esquembre, L. A., Schneider, D. C., Straetener, J., Henriksen, C., Musumeci, L., Putters, F., Melo, S., Sánchez-López, E., Giera, M., Penoy, N., Piel, G., Verlaine, O., Amoroso, A., Joris, B., Slavetinsky, C. J., Goffin, E., Pirotte, B., ... Oury, C. (2024). Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance. mBio, 15(10), [e01322-24]. https://doi.org/10.1128/mbio.01322-24

Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance. / Leeten, Kirsten; Jacques, Nicolas; Esquembre, Lidia Alejo; Schneider, Dana C.; Straetener, Jan; Henriksen, Camilla; Musumeci, Lucia; Putters, Florence; Melo, Sofia; Sánchez-López, Elena; Giera, Martin; Penoy, Noémie; Piel, Géraldine; Verlaine, Olivier; Amoroso, Ana; Joris, Bernard; Slavetinsky, Christoph J.; Goffin, Eric; Pirotte, Bernard; Frees, Dorte; Brötz-Oesterhelt, Heike; Lancellotti, Patrizio; Oury, Cécile.

I: mBio, Bind 15, Nr. 10, e01322-24, 2024.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Leeten, K, Jacques, N, Esquembre, LA, Schneider, DC, Straetener, J, Henriksen, C, Musumeci, L, Putters, F, Melo, S, Sánchez-López, E, Giera, M, Penoy, N, Piel, G, Verlaine, O, Amoroso, A, Joris, B, Slavetinsky, CJ, Goffin, E, Pirotte, B, Frees, D, Brötz-Oesterhelt, H, Lancellotti, P & Oury, C 2024, 'Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance', mBio, bind 15, nr. 10, e01322-24. https://doi.org/10.1128/mbio.01322-24

Leeten, Kirsten ; Jacques, Nicolas ; Esquembre, Lidia Alejo ; Schneider, Dana C. ; Straetener, Jan ; Henriksen, Camilla ; Musumeci, Lucia ; Putters, Florence ; Melo, Sofia ; Sánchez-López, Elena ; Giera, Martin ; Penoy, Noémie ; Piel, Géraldine ; Verlaine, Olivier ; Amoroso, Ana ; Joris, Bernard ; Slavetinsky, Christoph J. ; Goffin, Eric ; Pirotte, Bernard ; Frees, Dorte ; Brötz-Oesterhelt, Heike ; Lancellotti, Patrizio ; Oury, Cécile. / Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance. I: mBio. 2024 ; Bind 15, Nr. 10.

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abstract = "Infections with multidrug-resistant bacteria pose a major healthcare problem which urges the need for novel treatment options. Besides its potent antiplatelet properties, ticagrelor has antibacterial activity against Gram-positive bacteria, including methicillin- and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA). Several retrospective studies in cardiovascular patients support an antibacterial effect of this drug which is not related to its antiplatelet activity. We investigated the mechanism of action of ticagrelor in Staphylococcus aureus and model Bacillus subtilis, and assessed cross-resistance with two conventional anti-MRSA antibiotics, vancomycin and daptomycin. Bacillus subtilis bioreporter strains revealed ticagrelor-induced cell envelope-related stress responses. Sub-inhibitory drug concentrations caused membrane depolarization, impaired positioning of both the peripheral membrane protein MinD and the peptidoglycan precursor lipid II, and it affected cell shape. At the MIC, ticagrelor destroyed membrane integrity, indicated by the influx of membrane impermeable dyes, and lipid aggregate formation. Whole-genome sequencing of in vitro-generated ticagrelor-resistant MRSA clones revealed mutations in genes encoding ClpP, ClpX, and YjbH. Lipidomic analysis of resistant clones displayed changes in levels of the most abundant lipids of the Staphylococcus aureus membrane, for example, cardiolipins, phosphatidylglycerols, and diacylglycerols. Exogeneous cardiolipin, phosphatidylglycerol, or diacylglycerol antagonized the antibacterial properties of ticagrelor. Ticagrelor enhanced MRSA growth inhibition and killing by vancomycin and daptomycin in both exponential and stationary phases. Finally, no cross-resistance was observed between ticagrelor and daptomycin, or vancomycin. Our study demonstrates that ticagrelor targets multiple lipids in the cytoplasmic membrane of Gram-positive bacteria, thereby retaining activity against multidrug-resistant staphylococci including daptomycin- and vancomycin-resistant strains.",

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author = "Kirsten Leeten and Nicolas Jacques and Esquembre, {Lidia Alejo} and Schneider, {Dana C.} and Jan Straetener and Camilla Henriksen and Lucia Musumeci and Florence Putters and Sofia Melo and Elena S{\'a}nchez-L{\'o}pez and Martin Giera and No{\'e}mie Penoy and G{\'e}raldine Piel and Olivier Verlaine and Ana Amoroso and Bernard Joris and Slavetinsky, {Christoph J.} and Eric Goffin and Bernard Pirotte and Dorte Frees and Heike Br{\"o}tz-Oesterhelt and Patrizio Lancellotti and C{\'e}cile Oury",

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doi = "10.1128/mbio.01322-24",

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T1 - Ticagrelor alters the membrane of Staphylococcus aureus and enhances the activity of vancomycin and daptomycin without eliciting cross-resistance

AU - Leeten, Kirsten

AU - Jacques, Nicolas

AU - Esquembre, Lidia Alejo

AU - Schneider, Dana C.

AU - Straetener, Jan

AU - Henriksen, Camilla

AU - Musumeci, Lucia

AU - Putters, Florence

AU - Melo, Sofia

AU - Sánchez-López, Elena

AU - Giera, Martin

AU - Penoy, Noémie

AU - Piel, Géraldine

AU - Verlaine, Olivier

AU - Amoroso, Ana

AU - Joris, Bernard

AU - Slavetinsky, Christoph J.

AU - Goffin, Eric

AU - Pirotte, Bernard

AU - Frees, Dorte

AU - Brötz-Oesterhelt, Heike

AU - Lancellotti, Patrizio

AU - Oury, Cécile

PY - 2024

Y1 - 2024

N2 - Infections with multidrug-resistant bacteria pose a major healthcare problem which urges the need for novel treatment options. Besides its potent antiplatelet properties, ticagrelor has antibacterial activity against Gram-positive bacteria, including methicillin- and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA). Several retrospective studies in cardiovascular patients support an antibacterial effect of this drug which is not related to its antiplatelet activity. We investigated the mechanism of action of ticagrelor in Staphylococcus aureus and model Bacillus subtilis, and assessed cross-resistance with two conventional anti-MRSA antibiotics, vancomycin and daptomycin. Bacillus subtilis bioreporter strains revealed ticagrelor-induced cell envelope-related stress responses. Sub-inhibitory drug concentrations caused membrane depolarization, impaired positioning of both the peripheral membrane protein MinD and the peptidoglycan precursor lipid II, and it affected cell shape. At the MIC, ticagrelor destroyed membrane integrity, indicated by the influx of membrane impermeable dyes, and lipid aggregate formation. Whole-genome sequencing of in vitro-generated ticagrelor-resistant MRSA clones revealed mutations in genes encoding ClpP, ClpX, and YjbH. Lipidomic analysis of resistant clones displayed changes in levels of the most abundant lipids of the Staphylococcus aureus membrane, for example, cardiolipins, phosphatidylglycerols, and diacylglycerols. Exogeneous cardiolipin, phosphatidylglycerol, or diacylglycerol antagonized the antibacterial properties of ticagrelor. Ticagrelor enhanced MRSA growth inhibition and killing by vancomycin and daptomycin in both exponential and stationary phases. Finally, no cross-resistance was observed between ticagrelor and daptomycin, or vancomycin. Our study demonstrates that ticagrelor targets multiple lipids in the cytoplasmic membrane of Gram-positive bacteria, thereby retaining activity against multidrug-resistant staphylococci including daptomycin- and vancomycin-resistant strains.

AB - Infections with multidrug-resistant bacteria pose a major healthcare problem which urges the need for novel treatment options. Besides its potent antiplatelet properties, ticagrelor has antibacterial activity against Gram-positive bacteria, including methicillin- and vancomycin-resistant Staphylococcus aureus (MRSA and VRSA). Several retrospective studies in cardiovascular patients support an antibacterial effect of this drug which is not related to its antiplatelet activity. We investigated the mechanism of action of ticagrelor in Staphylococcus aureus and model Bacillus subtilis, and assessed cross-resistance with two conventional anti-MRSA antibiotics, vancomycin and daptomycin. Bacillus subtilis bioreporter strains revealed ticagrelor-induced cell envelope-related stress responses. Sub-inhibitory drug concentrations caused membrane depolarization, impaired positioning of both the peripheral membrane protein MinD and the peptidoglycan precursor lipid II, and it affected cell shape. At the MIC, ticagrelor destroyed membrane integrity, indicated by the influx of membrane impermeable dyes, and lipid aggregate formation. Whole-genome sequencing of in vitro-generated ticagrelor-resistant MRSA clones revealed mutations in genes encoding ClpP, ClpX, and YjbH. Lipidomic analysis of resistant clones displayed changes in levels of the most abundant lipids of the Staphylococcus aureus membrane, for example, cardiolipins, phosphatidylglycerols, and diacylglycerols. Exogeneous cardiolipin, phosphatidylglycerol, or diacylglycerol antagonized the antibacterial properties of ticagrelor. Ticagrelor enhanced MRSA growth inhibition and killing by vancomycin and daptomycin in both exponential and stationary phases. Finally, no cross-resistance was observed between ticagrelor and daptomycin, or vancomycin. Our study demonstrates that ticagrelor targets multiple lipids in the cytoplasmic membrane of Gram-positive bacteria, thereby retaining activity against multidrug-resistant staphylococci including daptomycin- and vancomycin-resistant strains.

KW - drug discovery

KW - drug interactions

KW - Gram-positive bacteria

KW - platelets

U2 - 10.1128/mbio.01322-24

DO - 10.1128/mbio.01322-24

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AN - SCOPUS:85206959108

VL - 15

JO - mBio

JF - mBio

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