EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition
Abstract Virulence factors of pathogens, such as toxin production and biofilm formation, often exhibit a public character, providing benefits to nearby non-producers. Consequently, anti-virulence drugs targeting these public traits may not select for resistance, as resistant mutants that resume prod...
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Nature Portfolio
2025-05-01
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| Series: | npj Biofilms and Microbiomes |
| Online Access: | https://doi.org/10.1038/s41522-025-00693-y |
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| author | Mathieu Joos Sybren Van Ginneken Xabier Villanueva Marie Dijkmans Guglielmo A. Coppola Camilo Andres Pérez-Romero Thijs Vackier Erik Van der Eycken Kathleen Marchal Bram Lories Hans P. Steenackers |
| author_facet | Mathieu Joos Sybren Van Ginneken Xabier Villanueva Marie Dijkmans Guglielmo A. Coppola Camilo Andres Pérez-Romero Thijs Vackier Erik Van der Eycken Kathleen Marchal Bram Lories Hans P. Steenackers |
| author_sort | Mathieu Joos |
| collection | DOAJ |
| description | Abstract Virulence factors of pathogens, such as toxin production and biofilm formation, often exhibit a public character, providing benefits to nearby non-producers. Consequently, anti-virulence drugs targeting these public traits may not select for resistance, as resistant mutants that resume production of the virulence factor share the benefits of their resistance with surrounding sensitive cells. In agreement with this, we show that even after long-term treatment with a 2-amino-imidazole (2-AI) biofilm inhibitor, Salmonella populations remained as susceptible to biofilm inhibition as the ancestral populations. Nonetheless, further genotypic and phenotypic analysis revealed that the Salmonella populations did adapt to the treatment and accumulated mutations in efflux pump regulators and alternative sigma factors. These mutations resulted in a reduced biofilm-forming capacity and increased efflux activity. Their selection was due to a growth delaying side effect of the biofilm inhibitor. Enhanced efflux activity helped overcome this growth delay, providing a fitness advantage over the ancestor. Finally, we demonstrate that chemical modification of the inhibitor enhances its specificity by partially alleviating the unintended growth delay while retaining the anti-biofilm activity, which in turn eliminated the selection pressure for increased efflux. Overall, our findings highlight that while unintended side effects can complicate anti-virulence strategies, adaptation to these effects does not necessarily restore the inhibited virulence trait. Moreover, chemical modification can mitigate these unintended side effects and enhance drug specificity. |
| format | Article |
| id | doaj-art-dd3cd89477f646ffb3e7a904341a2749 |
| institution | OA Journals |
| issn | 2055-5008 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Biofilms and Microbiomes |
| spelling | doaj-art-dd3cd89477f646ffb3e7a904341a27492025-08-20T02:15:00ZengNature Portfolionpj Biofilms and Microbiomes2055-50082025-05-0111111510.1038/s41522-025-00693-yEPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibitionMathieu Joos0Sybren Van Ginneken1Xabier Villanueva2Marie Dijkmans3Guglielmo A. Coppola4Camilo Andres Pérez-Romero5Thijs Vackier6Erik Van der Eycken7Kathleen Marchal8Bram Lories9Hans P. Steenackers10KU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsKU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsKU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsKU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsKU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsDepartment of Plant Biotechnology and Bioinformatics, UGent – Internet Technology and Data Science Lab (IDLab)KU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsDepartment of Chemistry, KU Leuven - Laboratory for Organic & Microwave-Assisted Chemistry (LOMAC)Department of Plant Biotechnology and Bioinformatics, UGent – Internet Technology and Data Science Lab (IDLab)KU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsKU Leuven - MiCA Lab, Centre of Microbial and Plant GeneticsAbstract Virulence factors of pathogens, such as toxin production and biofilm formation, often exhibit a public character, providing benefits to nearby non-producers. Consequently, anti-virulence drugs targeting these public traits may not select for resistance, as resistant mutants that resume production of the virulence factor share the benefits of their resistance with surrounding sensitive cells. In agreement with this, we show that even after long-term treatment with a 2-amino-imidazole (2-AI) biofilm inhibitor, Salmonella populations remained as susceptible to biofilm inhibition as the ancestral populations. Nonetheless, further genotypic and phenotypic analysis revealed that the Salmonella populations did adapt to the treatment and accumulated mutations in efflux pump regulators and alternative sigma factors. These mutations resulted in a reduced biofilm-forming capacity and increased efflux activity. Their selection was due to a growth delaying side effect of the biofilm inhibitor. Enhanced efflux activity helped overcome this growth delay, providing a fitness advantage over the ancestor. Finally, we demonstrate that chemical modification of the inhibitor enhances its specificity by partially alleviating the unintended growth delay while retaining the anti-biofilm activity, which in turn eliminated the selection pressure for increased efflux. Overall, our findings highlight that while unintended side effects can complicate anti-virulence strategies, adaptation to these effects does not necessarily restore the inhibited virulence trait. Moreover, chemical modification can mitigate these unintended side effects and enhance drug specificity.https://doi.org/10.1038/s41522-025-00693-y |
| spellingShingle | Mathieu Joos Sybren Van Ginneken Xabier Villanueva Marie Dijkmans Guglielmo A. Coppola Camilo Andres Pérez-Romero Thijs Vackier Erik Van der Eycken Kathleen Marchal Bram Lories Hans P. Steenackers EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition npj Biofilms and Microbiomes |
| title | EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition |
| title_full | EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition |
| title_fullStr | EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition |
| title_full_unstemmed | EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition |
| title_short | EPS inhibitor treatment of Salmonella impacts evolution without selecting for resistance to biofilm inhibition |
| title_sort | eps inhibitor treatment of salmonella impacts evolution without selecting for resistance to biofilm inhibition |
| url | https://doi.org/10.1038/s41522-025-00693-y |
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