Fast bacterial growth reduces antibiotic accumulation and efficacy
Phenotypic variations between individual microbial cells play a key role in the resistance of microbial pathogens to pharmacotherapies. Nevertheless, little is known about cell individuality in antibiotic accumulation. Here, we hypothesise that phenotypic diversification can be driven by fundamental...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2022-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/74062 |
| _version_ | 1811203373057179648 |
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| author | Urszula Łapińska Margaritis Voliotis Ka Kiu Lee Adrian Campey M Rhia L Stone Brandon Tuck Wanida Phetsang Bing Zhang Krasimira Tsaneva-Atanasova Mark AT Blaskovich Stefano Pagliara |
| author_facet | Urszula Łapińska Margaritis Voliotis Ka Kiu Lee Adrian Campey M Rhia L Stone Brandon Tuck Wanida Phetsang Bing Zhang Krasimira Tsaneva-Atanasova Mark AT Blaskovich Stefano Pagliara |
| author_sort | Urszula Łapińska |
| collection | DOAJ |
| description | Phenotypic variations between individual microbial cells play a key role in the resistance of microbial pathogens to pharmacotherapies. Nevertheless, little is known about cell individuality in antibiotic accumulation. Here, we hypothesise that phenotypic diversification can be driven by fundamental cell-to-cell differences in drug transport rates. To test this hypothesis, we employed microfluidics-based single-cell microscopy, libraries of fluorescent antibiotic probes and mathematical modelling. This approach allowed us to rapidly identify phenotypic variants that avoid antibiotic accumulation within populations of Escherichia coli, Pseudomonas aeruginosa, Burkholderia cenocepacia, and Staphylococcus aureus. Crucially, we found that fast growing phenotypic variants avoid macrolide accumulation and survive treatment without genetic mutations. These findings are in contrast with the current consensus that cellular dormancy and slow metabolism underlie bacterial survival to antibiotics. Our results also show that fast growing variants display significantly higher expression of ribosomal promoters before drug treatment compared to slow growing variants. Drug-free active ribosomes facilitate essential cellular processes in these fast-growing variants, including efflux that can reduce macrolide accumulation. We used this new knowledge to eradicate variants that displayed low antibiotic accumulation through the chemical manipulation of their outer membrane inspiring new avenues to overcome current antibiotic treatment failures. |
| first_indexed | 2024-04-12T02:54:40Z |
| format | Article |
| id | doaj.art-f9443964bba24775899537b4edeb0b4a |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:54:40Z |
| publishDate | 2022-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-f9443964bba24775899537b4edeb0b4a2022-12-22T03:50:53ZengeLife Sciences Publications LtdeLife2050-084X2022-06-011110.7554/eLife.74062Fast bacterial growth reduces antibiotic accumulation and efficacyUrszula Łapińska0https://orcid.org/0000-0003-3593-9248Margaritis Voliotis1https://orcid.org/0000-0001-6488-7198Ka Kiu Lee2Adrian Campey3M Rhia L Stone4Brandon Tuck5Wanida Phetsang6Bing Zhang7Krasimira Tsaneva-Atanasova8https://orcid.org/0000-0002-6294-7051Mark AT Blaskovich9Stefano Pagliara10https://orcid.org/0000-0001-9796-1956Living Systems Institute, University of Exeter, Exeter, United Kingdom; Biosciences, University of Exeter, Exeter, United KingdomLiving Systems Institute, University of Exeter, Exeter, United Kingdom; Department of Mathematics, University of Exeter, Exeter, United KingdomLiving Systems Institute, University of Exeter, Exeter, United Kingdom; Biosciences, University of Exeter, Exeter, United KingdomLiving Systems Institute, University of Exeter, Exeter, United Kingdom; Biosciences, University of Exeter, Exeter, United KingdomCentre for Superbug Solutions, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia; Department of Chemistry and Chemical Biology, Rutgers, the State University of New Jersey, Piscataway, United StatesLiving Systems Institute, University of Exeter, Exeter, United Kingdom; Biosciences, University of Exeter, Exeter, United KingdomCentre for Superbug Solutions, Institute for Molecular Bioscience, The University of Queensland, Brisbane, AustraliaCentre for Superbug Solutions, Institute for Molecular Bioscience, The University of Queensland, Brisbane, AustraliaLiving Systems Institute, University of Exeter, Exeter, United Kingdom; Department of Mathematics, University of Exeter, Exeter, United Kingdom; EPSRC Hub for Quantitative Modelling in Healthcare, University of Exeter, Exeter, United Kingdom; Department of Bioinformatics and Mathematical Modelling, Institute of Biophysics and Biomedical Engineering, Bulgarian Academy of Sciences, Sofia, BulgariaCentre for Superbug Solutions, Institute for Molecular Bioscience, The University of Queensland, Brisbane, AustraliaLiving Systems Institute, University of Exeter, Exeter, United Kingdom; Biosciences, University of Exeter, Exeter, United KingdomPhenotypic variations between individual microbial cells play a key role in the resistance of microbial pathogens to pharmacotherapies. Nevertheless, little is known about cell individuality in antibiotic accumulation. Here, we hypothesise that phenotypic diversification can be driven by fundamental cell-to-cell differences in drug transport rates. To test this hypothesis, we employed microfluidics-based single-cell microscopy, libraries of fluorescent antibiotic probes and mathematical modelling. This approach allowed us to rapidly identify phenotypic variants that avoid antibiotic accumulation within populations of Escherichia coli, Pseudomonas aeruginosa, Burkholderia cenocepacia, and Staphylococcus aureus. Crucially, we found that fast growing phenotypic variants avoid macrolide accumulation and survive treatment without genetic mutations. These findings are in contrast with the current consensus that cellular dormancy and slow metabolism underlie bacterial survival to antibiotics. Our results also show that fast growing variants display significantly higher expression of ribosomal promoters before drug treatment compared to slow growing variants. Drug-free active ribosomes facilitate essential cellular processes in these fast-growing variants, including efflux that can reduce macrolide accumulation. We used this new knowledge to eradicate variants that displayed low antibiotic accumulation through the chemical manipulation of their outer membrane inspiring new avenues to overcome current antibiotic treatment failures.https://elifesciences.org/articles/74062membrane transportantibioticssingle-cell analysisantibiotic resistancephenotypic heterogeneityantibiotic uptake |
| spellingShingle | Urszula Łapińska Margaritis Voliotis Ka Kiu Lee Adrian Campey M Rhia L Stone Brandon Tuck Wanida Phetsang Bing Zhang Krasimira Tsaneva-Atanasova Mark AT Blaskovich Stefano Pagliara Fast bacterial growth reduces antibiotic accumulation and efficacy eLife membrane transport antibiotics single-cell analysis antibiotic resistance phenotypic heterogeneity antibiotic uptake |
| title | Fast bacterial growth reduces antibiotic accumulation and efficacy |
| title_full | Fast bacterial growth reduces antibiotic accumulation and efficacy |
| title_fullStr | Fast bacterial growth reduces antibiotic accumulation and efficacy |
| title_full_unstemmed | Fast bacterial growth reduces antibiotic accumulation and efficacy |
| title_short | Fast bacterial growth reduces antibiotic accumulation and efficacy |
| title_sort | fast bacterial growth reduces antibiotic accumulation and efficacy |
| topic | membrane transport antibiotics single-cell analysis antibiotic resistance phenotypic heterogeneity antibiotic uptake |
| url | https://elifesciences.org/articles/74062 |
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