Abstract
The rapid emergence of antibiotic resistant bacterial pathogens constitutes a critical problem in healthcare and requires the development of novel treatments. Potential strategies include the exploitation of microbial social interactions based on public goods, which are produced at a fitness cost by cooperative microorganisms, but can be exploited by cheaters that do not produce these goods. Cheater invasion has been proposed as a ‘Trojan horse’ approach to infiltrate pathogen populations with strains deploying built-in weaknesses (e.g., sensitiveness to antibiotics). However, previous attempts have been often unsuccessful because population invasion by cheaters was prevented by various mechanisms including the presence of spatial structure (e.g., growth in biofilms), which limits the diffusion and exploitation of public goods. Here we followed an alternative approach and examined whether the manipulation of public good uptake and not its production could result in potential ‘Trojan horses’ suitable for population invasion. We focused on the siderophore pyoverdine produced by the human pathogen Pseudomonas aeruginosa MPAO1 and manipulated its uptake by deleting and/or overexpressing the pyoverdine primary (FpvA) and secondary (FpvB) receptors. We found that receptor synthesis feeds back on pyoverdine production and uptake rates, which led to strains with altered pyoverdine-associated costs and benefits. Moreover, we found that the receptor FpvB was advantageous under iron-limited conditions but revealed hidden costs in the presence of an antibiotic stressor (gentamicin). As a consequence, FpvB mutants became the fittest strain under gentamicin exposure, displacing the wildtype in liquid cultures, and in biofilms and during infections of the wax moth larvae Galleria mellonella, which both represent structured environments. Our findings reveal that an evolutionary trade-off associated with the costs and benefits of a versatile pyoverdine uptake strategy can be harnessed for devising a Trojan-horse candidate for medical interventions.
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Acknowledgements
The authors would like to thank Dr Melanie Ghoul, Prof. Stuart West and Prof. Craig McLean (University of Oxford), and Dr Jorge Gutiérrez (University of Surrey) for the insightful discussions on population invasion and their constructive feedback on the manuscript. The authors are indebted to Dr Helen King, Dr Mandy Fivian-Hughes and Anita Sicilia for their technical assistance. JG was the recipient of a PhD studentship of the University of Surrey and an EMBO Short-Term Fellowship (ASFT number: 8166). CAR and JIJ acknowledge the support received from the Biotechnology and Biological Sciences Research Council (BBSRC) (Grants BB/L02683X/1 and BB/T011289/1 from the ERA-Cobiotech programme of the EU). MSpi, CC and MJB acknowledge the support from the Engineering and Physical Sciences Research Council through a strategic equipment grant (EP/P001440/1). RK has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 681295). ÖÖ has received funding from the Forschungskredit by the University of Zurich.
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JG, CAR, RK and JIJ designed and supervised the study. JG, MSal, ÖÖ, KR, MSpi and CC conducted experimental work. All authors analysed the data and wrote the manuscript.
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González, J., Salvador, M., Özkaya, Ö. et al. Loss of a pyoverdine secondary receptor in Pseudomonas aeruginosa results in a fitter strain suitable for population invasion. ISME J 15, 1330–1343 (2021). https://doi.org/10.1038/s41396-020-00853-2
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DOI: https://doi.org/10.1038/s41396-020-00853-2
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