Longitudinal Evolution of the Pseudomonas-Derived Cephalosporinase (PDC) Structure and Activity in a Cystic Fibrosis Patient Treated with β-Lactams.
| Title: | Longitudinal Evolution of the Pseudomonas-Derived Cephalosporinase (PDC) Structure and Activity in a Cystic Fibrosis Patient Treated with β-Lactams. |
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| Authors: | Colque CA; Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Córdoba, Argentina.; CONICET, Universidad Nacional de Córdoba, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), Córdoba, Argentina.; Albarracín Orio AG; Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Córdoba, Argentina.; CONICET, Universidad Nacional de Córdoba, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), Córdoba, Argentina.; IRNASUS, Universidad Católica de Córdoba, CONICET, Facultad de Ciencias Agropecuarias, Córdoba, Argentina.; Tomatis PE; Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Universidad Nacional de Rosario, Rosario, Argentina.; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.; Dotta G; Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Universidad Nacional de Rosario, Rosario, Argentina.; Moreno DM; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.; IQUIR, Instituto de Química de Rosario, CONICET, Universidad Nacional de Rosario, Rosario, Argentina.; Hedemann LG; Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Córdoba, Argentina.; CONICET, Universidad Nacional de Córdoba, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), Córdoba, Argentina.; Hickman RA; Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.; Novo Nordisk Foundation Centre for Biosustainability, Technical University of Denmarkgrid.5170.3, Lyngby, Denmark.; Sommer LM; Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.; Novo Nordisk Foundation Centre for Biosustainability, Technical University of Denmarkgrid.5170.3, Lyngby, Denmark.; Feliziani S; Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Córdoba, Argentina.; CONICET, Universidad Nacional de Córdoba, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), Córdoba, Argentina.; Moyano AJ; Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Córdoba, Argentina.; CONICET, Universidad Nacional de Córdoba, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), Córdoba, Argentina.; Bonomo RA; Departments of Molecular Biology and Microbiology, Medicine, Biochemistry, Pharmacology, and Proteomics and Bioinformatics, Case Western Reserve University, Cleveland, Ohio, USA.; Senior Clinical Scientist Investigator, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA.; K Johansen H; Department of Clinical Microbiology, Rigshospitalet, Copenhagen, Denmark.; Novo Nordisk Foundation Centre for Biosustainability, Technical University of Denmarkgrid.5170.3, Lyngby, Denmark.; Department of Clinical Medicine, University of Copenhagen, Copenhagen, Denmark.; Molin S; Novo Nordisk Foundation Centre for Biosustainability, Technical University of Denmarkgrid.5170.3, Lyngby, Denmark.; Vila AJ; Instituto de Biología Molecular y Celular de Rosario (IBR), CONICET, Universidad Nacional de Rosario, Rosario, Argentina.; Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.; Smania AM; Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Química Biológica Ranwel Caputto, Córdoba, Argentina.; CONICET, Universidad Nacional de Córdoba, Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), Córdoba, Argentina. |
| Source: | MBio [mBio] 2022 Oct 26; Vol. 13 (5), pp. e0166322. Date of Electronic Publication: 2022 Sep 08. |
| Publication Type: | Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 101519231 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2150-7511 (Electronic) NLM ISO Abbreviation: mBio Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: Washington, D.C. : American Society for Microbiology |
| MeSH Terms: | Cystic Fibrosis*/microbiology ; Pseudomonas Infections*/microbiology; Cephalosporinase/genetics ; Ceftazidime/pharmacology ; Pseudomonas/metabolism ; beta-Lactamases/metabolism ; Cephalosporins/pharmacology ; Cephalosporins/therapeutic use ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/therapeutic use ; Humans ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa |
| Abstract: | Traditional studies on the evolution of antibiotic resistance development use approaches that can range from laboratory-based experimental studies, to epidemiological surveillance, to sequencing of clinical isolates. However, evolutionary trajectories also depend on the environment in which selection takes place, compelling the need to more deeply investigate the impact of environmental complexities and their dynamics over time. Herein, we explored the within-patient adaptive long-term evolution of a Pseudomonas aeruginosa hypermutator lineage in the airways of a cystic fibrosis (CF) patient by performing a chronological tracking of mutations that occurred in different subpopulations; our results demonstrated parallel evolution events in the chromosomally encoded class C β-lactamase (blaPDC). These multiple mutations within blaPDC shaped diverse coexisting alleles, whose frequency dynamics responded to the changing antibiotic selective pressures for more than 26 years of chronic infection. Importantly, the combination of the cumulative mutations in blaPDC provided structural and functional protein changes that resulted in a continuous enhancement of its catalytic efficiency and high level of cephalosporin resistance. This evolution was linked to the persistent treatment with ceftazidime, which we demonstrated selected for variants with robust catalytic activity against this expanded-spectrum cephalosporin. A "gain of function" of collateral resistance toward ceftolozane, a more recently introduced cephalosporin that was not prescribed to this patient, was also observed, and the biochemical basis of this cross-resistance phenomenon was elucidated. This work unveils the evolutionary trajectories paved by bacteria toward a multidrug-resistant phenotype, driven by decades of antibiotic treatment in the natural CF environmental setting. IMPORTANCE Antibiotics are becoming increasingly ineffective to treat bacterial infections. It has been consequently predicted that infectious diseases will become the biggest challenge to human health in the near future. Pseudomonas aeruginosa is considered a paradigm in antimicrobial resistance as it exploits intrinsic and acquired resistance mechanisms to resist virtually all antibiotics known. AmpC β-lactamase is the main mechanism driving resistance in this notorious pathogen to β-lactams, one of the most widely used classes of antibiotics for cystic fibrosis infections. Here, we focus on the β-lactamase gene as a model resistance determinant and unveil the trajectory P. aeruginosa undertakes on the path toward a multidrug-resistant phenotype during the course of two and a half decades of chronic infection in the airways of a cystic fibrosis patient. Integrating genetic and biochemical studies in the natural environment where evolution occurs, we provide a unique perspective on this challenging landscape, addressing fundamental molecular mechanisms of resistance. |
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| Grant Information: | I01 BX001974 United States BX BLRD VA; R01 AI063517 United States AI NIAID NIH HHS; R01 AI100560 United States AI NIAID NIH HHS |
| Contributed Indexing: | Keywords: Pseudomonas aeruginosa; ceftolozane resistance; cystic fibrosis; hypermutability; β-lactamase evolution |
| Substance Nomenclature: | EC 3.5.2.- (Cephalosporinase); 9M416Z9QNR (Ceftazidime); EC 3.5.2.6 (beta-Lactamases); 0 (Cephalosporins); 0 (Anti-Bacterial Agents) |
| Entry Date(s): | Date Created: 20220908 Date Completed: 20221028 Latest Revision: 20240524 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC9600753 |
| DOI: | 10.1128/mbio.01663-22 |
| PMID: | 36073814 |
| Database: | MEDLINE |
Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, Non-U.S. Gov't