Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa
ABSTRACT In the current scenario of growing antibiotic resistance, understanding the interplay between resistance mechanisms and biological costs is crucial for designing therapeutic strategies. In this regard, intrinsic AmpC β-lactamase hyperproduction is probably the most important resistance mech...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2022-10-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02700-22 |
| _version_ | 1811257704211021824 |
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| author | Isabel M. Barceló Elena Jordana-Lluch María Escobar-Salom Gabriel Torrens Pablo A. Fraile-Ribot Gabriel Cabot Xavier Mulet Laura Zamorano Carlos Juan Antonio Oliver |
| author_facet | Isabel M. Barceló Elena Jordana-Lluch María Escobar-Salom Gabriel Torrens Pablo A. Fraile-Ribot Gabriel Cabot Xavier Mulet Laura Zamorano Carlos Juan Antonio Oliver |
| author_sort | Isabel M. Barceló |
| collection | DOAJ |
| description | ABSTRACT In the current scenario of growing antibiotic resistance, understanding the interplay between resistance mechanisms and biological costs is crucial for designing therapeutic strategies. In this regard, intrinsic AmpC β-lactamase hyperproduction is probably the most important resistance mechanism of Pseudomonas aeruginosa, proven to entail important biological burdens that attenuate virulence mostly under peptidoglycan recycling alterations. P. aeruginosa can acquire resistance to new β-lactam–β-lactamase inhibitor combinations (ceftazidime-avibactam and ceftolozane-tazobactam) through mutations affecting ampC and its regulatory genes, but the impact of these mutations on the associated biological cost and the role that β-lactamase activity plays per se in contributing to the above-mentioned virulence attenuation are unknown. The same questions remain unsolved for plasmid-encoded AmpC-type β-lactamases such as FOX enzymes, some of which also provide resistance to new β-lactam–β-lactamase inhibitor combinations. Here, we assessed from different perspectives the effects of changes in the active center and, thus, in the hydrolytic spectrum resistance to inhibitors of AmpC-type β-lactamases on the fitness and virulence of P. aeruginosa, using site-directed mutagenesis; the previously described AmpC variants T96I, G183D, and ΔG229-E247; and, finally, blaFOX-4 versus blaFOX-8. Our results indicate the essential role of AmpC activity per se in causing the reported full virulence attenuation (in terms of growth, motility, cytotoxicity, and Galleria mellonella larvae killing), although the biological cost of the above-mentioned AmpC-type variants was similar to that of the wild-type enzymes. This suggests that there is not an important biological burden that may limit the selection/spread of these variants, which could progressively compromise the future effectiveness of the above-mentioned drug combinations. IMPORTANCE The growing antibiotic resistance of the top nosocomial pathogen Pseudomonas aeruginosa pushes research to explore new therapeutic strategies, for which the resistance-versus-virulence balance is a promising source of targets. While resistance often entails significant biological costs, little is known about the bases of the virulence attenuations associated with a resistance mechanism as extraordinarily relevant as β-lactamase production. We demonstrate that besides potential energy and cell wall alterations, the enzymatic activity of the P. aeruginosa cephalosporinase AmpC is essential for causing the full attenuation associated with its hyperproduction by affecting different features related to pathogenesis, a fact exploitable from the antivirulence perspective. Less encouraging, we also show that the production of different chromosomal/plasmid-encoded AmpC derivatives conferring resistance to some of the newest antibiotic combinations causes no significantly increased biological burdens, which suggests a free way for the selection/spread of these types of variants, potentially compromising the future effectiveness of these antipseudomonal therapies. |
| first_indexed | 2024-04-12T18:01:36Z |
| format | Article |
| id | doaj.art-8f922c055f53469cbd8b38b88b446cbd |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-12T18:01:36Z |
| publishDate | 2022-10-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-8f922c055f53469cbd8b38b88b446cbd2022-12-22T03:22:09ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972022-10-0110510.1128/spectrum.02700-22Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosaIsabel M. Barceló0Elena Jordana-Lluch1María Escobar-Salom2Gabriel Torrens3Pablo A. Fraile-Ribot4Gabriel Cabot5Xavier Mulet6Laura Zamorano7Carlos Juan8Antonio Oliver9Research Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainResearch Unit, University Hospital Son Espases-Health Research Institute of the Balearic Islands (IdISBa), Palma, SpainABSTRACT In the current scenario of growing antibiotic resistance, understanding the interplay between resistance mechanisms and biological costs is crucial for designing therapeutic strategies. In this regard, intrinsic AmpC β-lactamase hyperproduction is probably the most important resistance mechanism of Pseudomonas aeruginosa, proven to entail important biological burdens that attenuate virulence mostly under peptidoglycan recycling alterations. P. aeruginosa can acquire resistance to new β-lactam–β-lactamase inhibitor combinations (ceftazidime-avibactam and ceftolozane-tazobactam) through mutations affecting ampC and its regulatory genes, but the impact of these mutations on the associated biological cost and the role that β-lactamase activity plays per se in contributing to the above-mentioned virulence attenuation are unknown. The same questions remain unsolved for plasmid-encoded AmpC-type β-lactamases such as FOX enzymes, some of which also provide resistance to new β-lactam–β-lactamase inhibitor combinations. Here, we assessed from different perspectives the effects of changes in the active center and, thus, in the hydrolytic spectrum resistance to inhibitors of AmpC-type β-lactamases on the fitness and virulence of P. aeruginosa, using site-directed mutagenesis; the previously described AmpC variants T96I, G183D, and ΔG229-E247; and, finally, blaFOX-4 versus blaFOX-8. Our results indicate the essential role of AmpC activity per se in causing the reported full virulence attenuation (in terms of growth, motility, cytotoxicity, and Galleria mellonella larvae killing), although the biological cost of the above-mentioned AmpC-type variants was similar to that of the wild-type enzymes. This suggests that there is not an important biological burden that may limit the selection/spread of these variants, which could progressively compromise the future effectiveness of the above-mentioned drug combinations. IMPORTANCE The growing antibiotic resistance of the top nosocomial pathogen Pseudomonas aeruginosa pushes research to explore new therapeutic strategies, for which the resistance-versus-virulence balance is a promising source of targets. While resistance often entails significant biological costs, little is known about the bases of the virulence attenuations associated with a resistance mechanism as extraordinarily relevant as β-lactamase production. We demonstrate that besides potential energy and cell wall alterations, the enzymatic activity of the P. aeruginosa cephalosporinase AmpC is essential for causing the full attenuation associated with its hyperproduction by affecting different features related to pathogenesis, a fact exploitable from the antivirulence perspective. Less encouraging, we also show that the production of different chromosomal/plasmid-encoded AmpC derivatives conferring resistance to some of the newest antibiotic combinations causes no significantly increased biological burdens, which suggests a free way for the selection/spread of these types of variants, potentially compromising the future effectiveness of these antipseudomonal therapies.https://journals.asm.org/doi/10.1128/spectrum.02700-22Pseudomonas aeruginosaAmpCceftolozane-tazobactamceftazidime-avibactamGalleria mellonellablaFOX β-lactamases |
| spellingShingle | Isabel M. Barceló Elena Jordana-Lluch María Escobar-Salom Gabriel Torrens Pablo A. Fraile-Ribot Gabriel Cabot Xavier Mulet Laura Zamorano Carlos Juan Antonio Oliver Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa Microbiology Spectrum Pseudomonas aeruginosa AmpC ceftolozane-tazobactam ceftazidime-avibactam Galleria mellonella blaFOX β-lactamases |
| title | Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa |
| title_full | Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa |
| title_fullStr | Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa |
| title_full_unstemmed | Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa |
| title_short | Role of Enzymatic Activity in the Biological Cost Associated with the Production of AmpC β-Lactamases in Pseudomonas aeruginosa |
| title_sort | role of enzymatic activity in the biological cost associated with the production of ampc β lactamases in pseudomonas aeruginosa |
| topic | Pseudomonas aeruginosa AmpC ceftolozane-tazobactam ceftazidime-avibactam Galleria mellonella blaFOX β-lactamases |
| url | https://journals.asm.org/doi/10.1128/spectrum.02700-22 |
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