Lipid A in outer membrane vesicles shields bacteria from polymyxins

Journal of Extracellular Vesicles, vol. 13, pp. e12447

Abstract

The continuous emergence of multidrug-resistant bacterial pathogens poses a major global healthcare challenge, with Klebsiella pneumoniae being a prominent threat. We conducted a comprehensive study on K. pneumoniae's antibiotic resistance mechanisms, focusing on outer membrane vesicles (OMVs) and polymyxin, a last-resort antibiotic. Our research demonstrates that OMVs protect bacteria from polymyxins. OMVs derived from Polymyxin B (PB)-stressed K. pneumoniae exhibited heightened protective efficacy due to increased vesiculation, compared to OMVs from unstressed Klebsiella. OMVs also shield bacteria from different bacterial families. This was validated ex vivo and in vivo using precision cut lung slices (PCLS) and Galleria mellonella. In all models, OMVs protected K. pneumoniae from PB and reduced the associated stress response on protein level. We observed significant changes in the lipid composition of OMVs upon PB treatment, affecting their binding capacity to PB. The altered binding capacity of single OMVs from PB stressed K. pneumoniae could be linked to a reduction in the lipid A amount of their released vesicles. Although the amount of lipid A per vesicle is reduced, the overall increase in the number of vesicles results in an increased protection because the sum of lipid A and therefore PB binding sites have increased. This unravels the mechanism of the altered PB protective efficacy of OMVs from PB stressed K. pneumoniae compared to control OMVs. The lipid A-dependent protective effect against PB was confirmed in vitro using artificial vesicles. Moreover, artificial vesicles successfully protected Klebsiella from PB ex vivo and in vivo. The findings indicate that OMVs act as protective shields for bacteria by binding to polymyxins, effectively serving as decoys and preventing antibiotic interaction with the cell surface. Our findings provide valuable insights into the mechanisms underlying antibiotic cross-protection and offer potential avenues for the development of novel therapeutic interventions to address the escalating threat of multidrug-resistant bacterial infections.

Authors 20

1. Marie Burt

   Philipps University of Marburg · German Center for Lung Research · Universities of Giessen and Marburg Lung Center

   Affiliation as printed

   Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL) Philipps‐University Marburg Marburg Germany

   Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL), Philipps-University Marburg, Marburg, Germany

2. Georgia Angelidou

   Max Planck Institute for Terrestrial Microbiology

   Affiliation as printed

   Core Facility for Mass Spectrometry and Proteomics Max Planck Institute for terrestrial Microbiology Marburg Germany

   Core Facility for Metabolomics and Small Molecules Mass Spectrometry Max Planck Institute for Terrestrial Microbiology Marburg Germany

   Core Facility for Mass Spectrometry and Proteomics, Max Planck Institute for terrestrial Microbiology, Marburg, Germany

   Core Facility for Metabolomics and Small Molecules Mass Spectrometry, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany

3. Christopher-Nils Mais

   Philipps University of Marburg · Loewe Center for Synthetic Microbiology

   Affiliation as printed

   Center for Synthetic Microbiology (SYNMIKRO) Philipps‐University Marburg Marburg Germany

   Center for Synthetic Microbiology (SYNMIKRO), Philipps-University Marburg, Marburg, Germany

4. Christian Preußer

   Philipps University of Marburg

   Affiliation as printed

   Core Facility ‐ Extracellular Vesicles Philipps‐University Marburg Marburg Germany

   Institute for Tumor Immunology Philipps‐University Marburg Marburg Germany

   Core Facility - Extracellular Vesicles, Philipps-University Marburg, Marburg, Germany

   Institute for Tumor Immunology, Philipps-University Marburg, Marburg, Germany

5. Timo Glatter

   Max Planck Institute for Terrestrial Microbiology

   Affiliation as printed

   Core Facility for Mass Spectrometry and Proteomics Max Planck Institute for terrestrial Microbiology Marburg Germany

   Core Facility for Mass Spectrometry and Proteomics, Max Planck Institute for terrestrial Microbiology, Marburg, Germany

6. Thomas Heimerl

   Philipps University of Marburg · Loewe Center for Synthetic Microbiology

   Affiliation as printed

   Center for Synthetic Microbiology (SYNMIKRO) Philipps‐University Marburg Marburg Germany

   Center for Synthetic Microbiology (SYNMIKRO), Philipps-University Marburg, Marburg, Germany

7. Rüdiger M. Groß

   Universität Ulm · University Hospital Ulm

   Affiliation as printed

   Institute of Molecular Virology Ulm University Medical Center Ulm Germany

   Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany

8. Javier Serranía

   Philipps University of Marburg · Loewe Center for Synthetic Microbiology

   Affiliation as printed

   Center for Synthetic Microbiology (SYNMIKRO) Philipps‐University Marburg Marburg Germany

   Center for Synthetic Microbiology (SYNMIKRO), Philipps-University Marburg, Marburg, Germany

9. Gowtham Boosarpu

   Philipps University of Marburg · German Center for Lung Research · Universities of Giessen and Marburg Lung Center

   Affiliation as printed

   Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL) Philipps‐University Marburg Marburg Germany

   Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL), Philipps-University Marburg, Marburg, Germany

10. Elke Pogge von Strandmann

    Philipps University of Marburg

    Affiliation as printed

    Core Facility ‐ Extracellular Vesicles Philipps‐University Marburg Marburg Germany

    Institute for Tumor Immunology Philipps‐University Marburg Marburg Germany

    Institute for Tumor Immunology, Philipps-University Marburg, Marburg, Germany

11. Janis A. Müller

    Philipps University of Marburg

    Affiliation as printed

    Institute of Virology Philipps‐University Marburg Marburg Germany

    Institute of Virology, Philipps-University Marburg, Marburg, Germany

12. Gert Bange

    Philipps University of Marburg · Loewe Center for Synthetic Microbiology

    Affiliation as printed

    Center for Synthetic Microbiology (SYNMIKRO) Philipps‐University Marburg Marburg Germany

    Center for Synthetic Microbiology (SYNMIKRO), Philipps-University Marburg, Marburg, Germany

13. Anke Becker

    Philipps University of Marburg · Loewe Center for Synthetic Microbiology

    Affiliation as printed

    Center for Synthetic Microbiology (SYNMIKRO) Philipps‐University Marburg Marburg Germany

    Center for Synthetic Microbiology (SYNMIKRO), Philipps-University Marburg, Marburg, Germany

14. Mareike Lehmann

    Philipps University of Marburg · Helmholtz Munich · German Center for Lung Research · Universities of Giessen and Marburg Lung Center

    Affiliation as printed

    Comprehensive Pneumology Center (CPC), Institute of Lung Health and Immunity Helmholtz Zentrum München German Center for Lung Research (DZL) Munich Germany

    Institute for Lung Health (ILH) Giessen Germany

    Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL) Philipps‐University Marburg Marburg Germany

    Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL), Philipps-University Marburg, Marburg, Germany

15. Danny David Jonigk Aachen Institute of Pathology, University Medical Center

    Universitätsklinikum Aachen · German Center for Lung Research · RWTH Aachen University

    Affiliation as printed

    Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) German Center of Lung Research (DZL) Hannover Germany

    Institute of Pathology University Medical Center RWTH University of Aachen Aachen Germany

    Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center of Lung Research (DZL), Hannover, Germany

    Institute of Pathology, University Medical Center RWTH University of Aachen, Aachen, Germany

16. Lavinia Neubert

    Medizinische Hochschule Hannover · German Center for Lung Research

    Affiliation as printed

    Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) German Center of Lung Research (DZL) Hannover Germany

    Institute of Pathology Hannover Medical School Hannover Germany

    Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center of Lung Research (DZL), Hannover, Germany

    Institute of Pathology, Hannover Medical School, Hannover, Germany

17. Hinrich Freitag

    Medizinische Hochschule Hannover · German Center for Lung Research

    Affiliation as printed

    Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH) German Center of Lung Research (DZL) Hannover Germany

    Institute of Pathology Hannover Medical School Hannover Germany

    Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), German Center of Lung Research (DZL), Hannover, Germany

    Institute of Pathology, Hannover Medical School, Hannover, Germany

18. Nicole Paczia

    Max Planck Institute for Terrestrial Microbiology

    Affiliation as printed

    Core Facility for Metabolomics and Small Molecules Mass Spectrometry Max Planck Institute for Terrestrial Microbiology Marburg Germany

    Core Facility for Metabolomics and Small Molecules Mass Spectrometry, Max Planck Institute for Terrestrial Microbiology, Marburg, Germany

19. Bernd Schmeck

    Philipps University of Marburg · Universitätsklinikum Gießen und Marburg · German Center for Infection Research · German Center for Lung Research · Universities of Giessen and Marburg Lung Center · Loewe Center for Synthetic Microbiology

    Affiliation as printed

    Center for Synthetic Microbiology (SYNMIKRO) Philipps‐University Marburg Marburg Germany

    Department of Medicine, Pulmonary and Critical Care Medicine University Medical Center Marburg Universities of Giessen and Marburg Lung Center Philipps‐University Marburg Marburg Germany

    Institute for Lung Health (ILH) Giessen Germany

    Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL) Philipps‐University Marburg Marburg Germany

    Member of the German Center for Infectious Disease Research (DZIF) Marburg Germany

    Center for Synthetic Microbiology (SYNMIKRO), Philipps-University Marburg, Marburg, Germany

    Department of Medicine, Pulmonary and Critical Care Medicine, University Medical Center Marburg, Universities of Giessen and Marburg Lung Center, Philipps-University Marburg, Marburg, Germany

    Institute for Lung Health (ILH), Giessen, Germany

    Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL), Philipps-University Marburg, Marburg, Germany

    Member of the German Center for Infectious Disease Research (DZIF), Marburg, Germany

20. Anna Lena Jung corresponding

    Philipps University of Marburg · German Center for Lung Research · Universities of Giessen and Marburg Lung Center

    Affiliation as printed

    Core Facility Flow Cytometry – Bacterial Vesicles Philipps‐University Marburg Marburg Germany

    Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL) Philipps‐University Marburg Marburg Germany

    Institute for Lung Research, Universities of Giessen and Marburg Lung Center, German Center for Lung Research (DZL), Philipps-University Marburg, Marburg, Germany