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Robust antibiotic sensitization of pathogenic Pseudomonas aeruginosa via negative hysteresis in the cell envelope
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  • Published: 26 March 2026

Robust antibiotic sensitization of pathogenic Pseudomonas aeruginosa via negative hysteresis in the cell envelope

  • Florian Buchholz  ORCID: orcid.org/0000-0003-3882-91321 na1,
  • Lina M. Upterworth  ORCID: orcid.org/0009-0000-2632-39571 na1,
  • Leif Tueffers1,2,
  • Espen E. Groth  ORCID: orcid.org/0000-0003-0883-63663,4,5,
  • Kira Haas1,
  • Daniel Schütz1,
  • Abigail Savietto Scholz6,
  • Aditi Batra1,
  • Surajit Pal1,
  • Samarpita Banerjee1,
  • Badri N. Dubey  ORCID: orcid.org/0000-0002-8626-89187,8,
  • Sören Franzenburg  ORCID: orcid.org/0000-0001-6374-49109,
  • Barbara Kalsdorf10,11,12,
  • Klaus F. Rabe3,4,5,
  • Dennis Nurjadi2,
  • Jan Rupp  ORCID: orcid.org/0000-0001-8722-12332,11,13,
  • Dan I. Andersson  ORCID: orcid.org/0000-0001-6640-217414,
  • Holger Sondermann  ORCID: orcid.org/0000-0003-2211-62347,
  • Marc Bramkamp  ORCID: orcid.org/0000-0002-7704-32666,
  • Roderich Roemhild  ORCID: orcid.org/0000-0001-9480-52611,14,15,16 na2 &
  • …
  • Hinrich Schulenburg  ORCID: orcid.org/0000-0002-1413-913X1,16 na2 

Nature Communications , Article number:  (2026) Cite this article

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We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Antibiotics
  • Antimicrobial resistance
  • Bacterial genetics

Abstract

Antibiotic combination in time and space is a key strategy to combat antimicrobial resistance. The success of such treatment designs requires their robust efficacy across treatment conditions and a pathogen’s genomic diversity. This study found that an initial treatment with a β-lactam antibiotic causes robust cellular sensitization towards an aminoglycoside antibiotic across the high-risk human pathogen Pseudomonas aeruginosa, including resistant strains. This phenomenon of cellular sensitization, termed negative hysteresis, is modulated by the Cpx envelope stress response system and linked to membrane stress during growth. The increase in efficacy is achieved through a β-lactam induced elevated cellular uptake of the subsequently administered aminoglycoside. Negative hysteresis and the Cpx system are linked in several cases to the expression of synergistic drug interactions, thus enhancing efficacy of antibiotic combinations. Overall, our study identifies the phenomenon of negative hysteresis as a robustly inducible phenotype and thus a unique focus for optimizing antimicrobial therapy.

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Data availability

The source data of this study is provided in a supplementary Source Data file. The statistical data generated, as well as Key Resources used, are provided in the Supplementary Data files and as Supplementary Tables. The transcriptomic data generated in this study have been deposited in the NCBI’s Gene Expression Omnibus under GEO Series accession number GSE290299. Source data are provided with this paper.

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Acknowledgements

We are very grateful to S. Hernando-Amado (Madrid, Spain), C. Pál (Szeged, Hungary), and T. Bollenbach (Cologne, Germany) for critical comments and advice on the manuscript. We further thank D. Rogers, J. Summers (Ploen, Germany) for guidance in allelic exchange, P. Rainey (Ploen, Germany) for providing the plasmids and strains, then J. Lorenzen, K. Flinder, N. Steinbach, S. Butze (all Schulenburg lab), and L. Kirchhoff (Rupp lab) for supporting the experimental work, and also the Rupp and Schulenburg groups for general feedback. We are grateful for financial support from the German Research Foundation within the Research and Training Group 2501 (RTG 2501) on Translational Evolutionary Research (project 4.2 to H.S.), within the Excellence cluster Precision Medicine in chronic Inflammation (PMI; funding under Germany’s Excellence Strategy EXC 2167-390884018, to B.K., K.R., J.R., H.S.), within the Clinician Scientist Program in Evolutionary Medicine (CSEM) – project number 413490537 (to EEG), and as part of the individual grants SCHU 1415/12-2 (to H.S.) and BR-2915/7-1 (to M.B.). We are grateful for financial support from the Swedish Research Council, project number 2021-02091 (to D.I.A.). We are also grateful for financial support from the Max-Planck Society (Fellowship to H.S.), the Leibniz Association within the Leibniz Science-Campus Evolutionary Medicine of the Lung (EvoLUNG, to H.S.), and the project SKILLED funded by the DAMP foundation (to J.R., H.S.). This work was also supported by the ZMB Young Scientist award and the FWF grant 10.55776/ESP219 (to R.R.) and the TransEvo Innovation prize (to F.B.). The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Funding

Open Access funding enabled and organized by Projekt DEAL.

Author information

Author notes

  1. These authors contributed equally: Florian Buchholz, Lina M. Upterworth.

  2. These authors jointly supervised this work: Roderich Roemhild, Hinrich Schulenburg.

Authors and Affiliations

  1. Department of Evolutionary Ecology and Genetics, University of Kiel, Kiel, Germany

    Florian Buchholz, Lina M. Upterworth, Leif Tueffers, Kira Haas, Daniel Schütz, Aditi Batra, Surajit Pal, Samarpita Banerjee, Roderich Roemhild & Hinrich Schulenburg

  2. Institute of Medical Microbiology, University Hospital Schleswig-Holstein, Lübeck, Germany

    Leif Tueffers, Dennis Nurjadi & Jan Rupp

  3. LungenClinic Grosshansdorf, Großhansdorf, Germany

    Espen E. Groth & Klaus F. Rabe

  4. Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL), Großhansdorf, Germany

    Espen E. Groth & Klaus F. Rabe

  5. Department of Medicine, University of Kiel, Kiel, Germany

    Espen E. Groth & Klaus F. Rabe

  6. Institute for General Microbiology, University of Kiel, Kiel, Germany

    Abigail Savietto Scholz & Marc Bramkamp

  7. CSSB Centre for Structural Systems Biology, Deutsches Elektronen-Synchrotron DESY, Hamburg, Germany

    Badri N. Dubey & Holger Sondermann

  8. Department of Biosciences and Bioengineering, Indian Institute of Technology Dharwad, Dharwad, India

    Badri N. Dubey

  9. Competence Centre for Genomic Analysis Kiel, University of Kiel, Kiel, Germany

    Sören Franzenburg

  10. Clinical Infectious Disease, Research Center Borstel, Leibniz Lung Center, Borstel, Germany

    Barbara Kalsdorf

  11. German Center for Infection Research (DZIF), Hamburg-Lübeck-Borstel-Riems, Germany

    Barbara Kalsdorf & Jan Rupp

  12. Respiratory Medicine & International Health, University of Lübeck, Lübeck, Germany

    Barbara Kalsdorf

  13. Infectious Diseases Clinic, University Hospital Schleswig-Holstein, Lübeck, Germany

    Jan Rupp

  14. Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden

    Dan I. Andersson & Roderich Roemhild

  15. Institute of Science and Technology Austria, Klosterneuburg, Austria

    Roderich Roemhild

  16. Max Planck Institute for Evolutionary Biology, Ploen, Germany

    Roderich Roemhild & Hinrich Schulenburg

Authors
  1. Florian Buchholz
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  2. Lina M. Upterworth
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  3. Leif Tueffers
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  4. Espen E. Groth
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  5. Kira Haas
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  6. Daniel Schütz
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  7. Abigail Savietto Scholz
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  8. Aditi Batra
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  9. Surajit Pal
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  10. Samarpita Banerjee
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  11. Badri N. Dubey
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  12. Sören Franzenburg
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  13. Barbara Kalsdorf
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  14. Klaus F. Rabe
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  15. Dennis Nurjadi
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  16. Jan Rupp
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  17. Dan I. Andersson
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  18. Holger Sondermann
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  19. Marc Bramkamp
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  20. Roderich Roemhild
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  21. Hinrich Schulenburg
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Contributions

Conceptualization: F.B., L.U., L.T., E.E.G., J.R., D.I.A., H.So, M.B., R.R., H.Schu. Formal analysis: F.B., L.U., L.T., E.E.G., D.S., A.S.S., R.R.; Funding acquisition: F.B., K.F.R., J.R., R.R., H.Schu. Investigation and methodology: F.B., L.U., L.T., E.E.G., K.H., D.S., A.S.S., A.B., S.P., S.B., D.N., B.N.D., S.F., B.K., R.R.; Resources: E.E.G., B.K., K.F.R., J.R., H.Schu. Supervision: F.B., L.U., L.T., E.E.G., D.N., J.R., D.I.A., H.So, M.B., R.R., H.Schu. Writing of original draft: F.B., L.U., R.R., H.Schu. Review and editing: all authors.

Corresponding authors

Correspondence to Roderich Roemhild or Hinrich Schulenburg.

Ethics declarations

Competing interests

The authors declare the following competing interest: Access to the CombiANT technology was provided by Rx Dynamics AB as a product BETA test. R.R. and D.I.A. are co-founders of Rx Dynamics AB. The company had no influence on study design, investigation, data analysis, manuscript writing and decision to publish. Apart from this, the authors declare no conflict of interest.

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Supplementary information

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Supplementary Dataset 1 (download XLSX )

Supplementary Dataset 2 (download XLSX )

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Supplementary Dataset 13 (download XLSX )

Reporting summary (download PDF )

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Source data

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Buchholz, F., Upterworth, L.M., Tueffers, L. et al. Robust antibiotic sensitization of pathogenic Pseudomonas aeruginosa via negative hysteresis in the cell envelope. Nat Commun (2026). https://doi.org/10.1038/s41467-026-71178-5

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  • Received: 26 March 2025

  • Accepted: 13 March 2026

  • Published: 26 March 2026

  • DOI: https://doi.org/10.1038/s41467-026-71178-5

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