Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa
ABSTRACT Antibiotic resistance is emerging as a significant global health crisis, necessitating the urgent development of novel antibiotics or alternative therapies. Although it is recognized that bacteria require multiple mutations to develop resistance levels exceeding the mutant prevention concen...
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| Language: | English |
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American Society for Microbiology
2025-04-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02981-24 |
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| author | Xiao-Quan Yu Hao Yang Han-Zhong Feng Jun Hou Jun-Qiang Tian Shao-Min Niu Chong-Ge You Xuan-Yu Tao Si-Ping Zhang Zhi-Ping Wang Yong-Xing He |
| author_facet | Xiao-Quan Yu Hao Yang Han-Zhong Feng Jun Hou Jun-Qiang Tian Shao-Min Niu Chong-Ge You Xuan-Yu Tao Si-Ping Zhang Zhi-Ping Wang Yong-Xing He |
| author_sort | Xiao-Quan Yu |
| collection | DOAJ |
| description | ABSTRACT Antibiotic resistance is emerging as a significant global health crisis, necessitating the urgent development of novel antibiotics or alternative therapies. Although it is recognized that bacteria require multiple mutations to develop resistance levels exceeding the mutant prevention concentration, the specific mutation combinations conferring high resistance have been largely undefined. Here, we investigated the multi-step evolution of fluoroquinolone resistance in Pseudomonas aeruginosa through experimental evolution and whole-genome sequencing coupled with proteomic approaches. We discovered that in low-dose and high-dose experimental evolution scenarios, combinations of mutations in the negative regulators of efflux pumps (nfxB/mexR) and DNA gyrases (gyrA/gyrB) contributed to the high-level resistance and some of these combinations were also prevalent in clinical isolates of P. aeruginosa. Notably, the selected nfxB mutation, which resulted in the overexpression of the MexCD-OprJ efflux pump, also exhibited collateral sensitivity to aminoglycosides and enhanced antibiotic tolerance. It was further revealed that the efflux pump inhibitor phenylalanine-arginine β-naphthylamide (PAβN) could effectively prevent evolution to high-level resistance for both laboratory and clinical P. aeruginosa strains. Our work highlights the critical role of efflux pump repressor-related mutations in the evolution of high-level antibiotic resistance and demonstrates the potential of targeting these mutations to impede the evolution toward high-level resistance.IMPORTANCEIn this study, we examined the stepwise evolution of fluoroquinolone resistance in Pseudomonas aeruginosa using experimental evolution, whole-genome sequencing, and proteomic analyses. Our findings revealed that under both low-dose and high-dose conditions, mutations in efflux pump regulators (nfxB/mexR) and DNA gyrase genes (gyrA/gyrB) synergistically contributed to high-level resistance. These mutation combinations were not only observed in experimental settings but also detected in clinical isolates of P. aeruginosa. This work underscores the pivotal role of efflux pump repressor-related mutations in the progression to high-level antibiotic resistance. It also highlights the promise of targeting efflux pumps as a strategy to prevent the multi-step evolution of resistance in P. aeruginosa. |
| format | Article |
| id | doaj-art-a33822d920004baeb9cbfbcfbb75cc69 |
| institution | DOAJ |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-a33822d920004baeb9cbfbcfbb75cc692025-08-20T02:53:34ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-04-0113410.1128/spectrum.02981-24Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosaXiao-Quan Yu0Hao Yang1Han-Zhong Feng2Jun Hou3Jun-Qiang Tian4Shao-Min Niu5Chong-Ge You6Xuan-Yu Tao7Si-Ping Zhang8Zhi-Ping Wang9Yong-Xing He10Institute of Urology, Gansu Province Clinical Research Center for urinary system disease, The Second Hospital & Clinical Medical School, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaInstitute of Urology, Gansu Province Clinical Research Center for urinary system disease, The Second Hospital & Clinical Medical School, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaInstitute of Urology, Gansu Province Clinical Research Center for urinary system disease, The Second Hospital & Clinical Medical School, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaLaboratory Medicine Center, The Second Hospital of Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaInstitute for Environmental Genomics, University of Oklahoma, Norman, Oklahoma, USAMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaInstitute of Urology, Gansu Province Clinical Research Center for urinary system disease, The Second Hospital & Clinical Medical School, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaMinistry of Education Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou, Gansu, People's Republic of ChinaABSTRACT Antibiotic resistance is emerging as a significant global health crisis, necessitating the urgent development of novel antibiotics or alternative therapies. Although it is recognized that bacteria require multiple mutations to develop resistance levels exceeding the mutant prevention concentration, the specific mutation combinations conferring high resistance have been largely undefined. Here, we investigated the multi-step evolution of fluoroquinolone resistance in Pseudomonas aeruginosa through experimental evolution and whole-genome sequencing coupled with proteomic approaches. We discovered that in low-dose and high-dose experimental evolution scenarios, combinations of mutations in the negative regulators of efflux pumps (nfxB/mexR) and DNA gyrases (gyrA/gyrB) contributed to the high-level resistance and some of these combinations were also prevalent in clinical isolates of P. aeruginosa. Notably, the selected nfxB mutation, which resulted in the overexpression of the MexCD-OprJ efflux pump, also exhibited collateral sensitivity to aminoglycosides and enhanced antibiotic tolerance. It was further revealed that the efflux pump inhibitor phenylalanine-arginine β-naphthylamide (PAβN) could effectively prevent evolution to high-level resistance for both laboratory and clinical P. aeruginosa strains. Our work highlights the critical role of efflux pump repressor-related mutations in the evolution of high-level antibiotic resistance and demonstrates the potential of targeting these mutations to impede the evolution toward high-level resistance.IMPORTANCEIn this study, we examined the stepwise evolution of fluoroquinolone resistance in Pseudomonas aeruginosa using experimental evolution, whole-genome sequencing, and proteomic analyses. Our findings revealed that under both low-dose and high-dose conditions, mutations in efflux pump regulators (nfxB/mexR) and DNA gyrase genes (gyrA/gyrB) synergistically contributed to high-level resistance. These mutation combinations were not only observed in experimental settings but also detected in clinical isolates of P. aeruginosa. This work underscores the pivotal role of efflux pump repressor-related mutations in the progression to high-level antibiotic resistance. It also highlights the promise of targeting efflux pumps as a strategy to prevent the multi-step evolution of resistance in P. aeruginosa.https://journals.asm.org/doi/10.1128/spectrum.02981-24antimicrobial resistance evolutionefflux pumpPseudomonas aeruginosa |
| spellingShingle | Xiao-Quan Yu Hao Yang Han-Zhong Feng Jun Hou Jun-Qiang Tian Shao-Min Niu Chong-Ge You Xuan-Yu Tao Si-Ping Zhang Zhi-Ping Wang Yong-Xing He Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa Microbiology Spectrum antimicrobial resistance evolution efflux pump Pseudomonas aeruginosa |
| title | Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa |
| title_full | Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa |
| title_fullStr | Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa |
| title_full_unstemmed | Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa |
| title_short | Targeting efflux pumps prevents the multi-step evolution of high-level resistance to fluoroquinolone in Pseudomonas aeruginosa |
| title_sort | targeting efflux pumps prevents the multi step evolution of high level resistance to fluoroquinolone in pseudomonas aeruginosa |
| topic | antimicrobial resistance evolution efflux pump Pseudomonas aeruginosa |
| url | https://journals.asm.org/doi/10.1128/spectrum.02981-24 |
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