Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment
ABSTRACT Urinary tract infection (UTI) is among the most common infections treated worldwide each year and is caused primarily by uropathogenic Escherichia coli (UPEC). Rising rates of antibiotic resistance among uropathogens have spurred a consideration of alternative treatment strategies, such as...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2022-08-01
|
| Series: | mSphere |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/msphere.00345-22 |
| _version_ | 1811214462554734592 |
|---|---|
| author | Jacob J. Zulk Justin R. Clark Samantha Ottinger Mallory B. Ballard Marlyd E. Mejia Vicki Mercado-Evans Emmaline R. Heckmann Belkys C. Sanchez Barbara W. Trautner Anthony W. Maresso Kathryn A. Patras |
| author_facet | Jacob J. Zulk Justin R. Clark Samantha Ottinger Mallory B. Ballard Marlyd E. Mejia Vicki Mercado-Evans Emmaline R. Heckmann Belkys C. Sanchez Barbara W. Trautner Anthony W. Maresso Kathryn A. Patras |
| author_sort | Jacob J. Zulk |
| collection | DOAJ |
| description | ABSTRACT Urinary tract infection (UTI) is among the most common infections treated worldwide each year and is caused primarily by uropathogenic Escherichia coli (UPEC). Rising rates of antibiotic resistance among uropathogens have spurred a consideration of alternative treatment strategies, such as bacteriophage (phage) therapy; however, phage-bacterial interactions within the urinary environment are poorly defined. Here, we assess the activity of two phages, namely, HP3 and ES17, against clinical UPEC isolates using in vitro and in vivo models of UTI. In both bacteriologic medium and pooled human urine, we identified phage resistance arising within the first 6 to 8 h of coincubation. Whole-genome sequencing revealed that UPEC strains resistant to HP3 and ES17 harbored mutations in genes involved in lipopolysaccharide (LPS) biosynthesis. Phage-resistant strains displayed several in vitro phenotypes, including alterations to adherence to and invasion of human bladder epithelial HTB-9 cells and increased biofilm formation in some isolates. Interestingly, these phage-resistant UPEC isolates demonstrated reduced growth in pooled human urine, which could be partially rescued by nutrient supplementation and were more sensitive to several outer membrane-targeting antibiotics than parental strains. Additionally, phage-resistant UPEC isolates were attenuated in bladder colonization in a murine UTI model. In total, our findings suggest that while resistance to phages, such as HP3 and ES17, may arise readily in the urinary environment, phage resistance is accompanied by fitness costs which may render UPEC more susceptible to host immunity or antibiotics. IMPORTANCE UTI is one of the most common causes of outpatient antibiotic use, and rising antibiotic resistance threatens the ability to control UTI unless alternative treatments are developed. Bacteriophage (phage) therapy is gaining renewed interest; however, much like with antibiotics, bacteria can readily become resistant to phages. For successful UTI treatment, we must predict how bacteria will evade killing by phage and identify the downstream consequences of phage resistance during bacterial infection. In our current study, we found that while phage-resistant bacteria quickly emerged in vitro, these bacteria were less capable of growing in human urine and colonizing the murine bladder. These results suggest that phage therapy poses a viable UTI treatment if phage resistance confers fitness costs for the uropathogen. These results have implications for developing cocktails of phage with multiple different bacterial targets, of which each is evaded only at the cost of bacterial fitness. |
| first_indexed | 2024-04-12T06:05:01Z |
| format | Article |
| id | doaj.art-6a92d0cf7da14d0d80830753163d2b33 |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-04-12T06:05:01Z |
| publishDate | 2022-08-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-6a92d0cf7da14d0d80830753163d2b332022-12-22T03:44:54ZengAmerican Society for MicrobiologymSphere2379-50422022-08-017410.1128/msphere.00345-22Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary EnvironmentJacob J. Zulk0Justin R. Clark1Samantha Ottinger2Mallory B. Ballard3Marlyd E. Mejia4Vicki Mercado-Evans5Emmaline R. Heckmann6Belkys C. Sanchez7Barbara W. Trautner8Anthony W. Maresso9Kathryn A. Patras10Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USACenter for Innovations in Quality, Effectiveness, and Safety (IQuESt), Michael E. DeBakey Veterans Affairs Medical Center, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USADepartment of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USAABSTRACT Urinary tract infection (UTI) is among the most common infections treated worldwide each year and is caused primarily by uropathogenic Escherichia coli (UPEC). Rising rates of antibiotic resistance among uropathogens have spurred a consideration of alternative treatment strategies, such as bacteriophage (phage) therapy; however, phage-bacterial interactions within the urinary environment are poorly defined. Here, we assess the activity of two phages, namely, HP3 and ES17, against clinical UPEC isolates using in vitro and in vivo models of UTI. In both bacteriologic medium and pooled human urine, we identified phage resistance arising within the first 6 to 8 h of coincubation. Whole-genome sequencing revealed that UPEC strains resistant to HP3 and ES17 harbored mutations in genes involved in lipopolysaccharide (LPS) biosynthesis. Phage-resistant strains displayed several in vitro phenotypes, including alterations to adherence to and invasion of human bladder epithelial HTB-9 cells and increased biofilm formation in some isolates. Interestingly, these phage-resistant UPEC isolates demonstrated reduced growth in pooled human urine, which could be partially rescued by nutrient supplementation and were more sensitive to several outer membrane-targeting antibiotics than parental strains. Additionally, phage-resistant UPEC isolates were attenuated in bladder colonization in a murine UTI model. In total, our findings suggest that while resistance to phages, such as HP3 and ES17, may arise readily in the urinary environment, phage resistance is accompanied by fitness costs which may render UPEC more susceptible to host immunity or antibiotics. IMPORTANCE UTI is one of the most common causes of outpatient antibiotic use, and rising antibiotic resistance threatens the ability to control UTI unless alternative treatments are developed. Bacteriophage (phage) therapy is gaining renewed interest; however, much like with antibiotics, bacteria can readily become resistant to phages. For successful UTI treatment, we must predict how bacteria will evade killing by phage and identify the downstream consequences of phage resistance during bacterial infection. In our current study, we found that while phage-resistant bacteria quickly emerged in vitro, these bacteria were less capable of growing in human urine and colonizing the murine bladder. These results suggest that phage therapy poses a viable UTI treatment if phage resistance confers fitness costs for the uropathogen. These results have implications for developing cocktails of phage with multiple different bacterial targets, of which each is evaded only at the cost of bacterial fitness.https://journals.asm.org/doi/10.1128/msphere.00345-22antimicrobial resistancebacteriophage therapyurinary tract infectionuropathogenic E. coli |
| spellingShingle | Jacob J. Zulk Justin R. Clark Samantha Ottinger Mallory B. Ballard Marlyd E. Mejia Vicki Mercado-Evans Emmaline R. Heckmann Belkys C. Sanchez Barbara W. Trautner Anthony W. Maresso Kathryn A. Patras Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment mSphere antimicrobial resistance bacteriophage therapy urinary tract infection uropathogenic E. coli |
| title | Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment |
| title_full | Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment |
| title_fullStr | Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment |
| title_full_unstemmed | Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment |
| title_short | Phage Resistance Accompanies Reduced Fitness of Uropathogenic Escherichia coli in the Urinary Environment |
| title_sort | phage resistance accompanies reduced fitness of uropathogenic escherichia coli in the urinary environment |
| topic | antimicrobial resistance bacteriophage therapy urinary tract infection uropathogenic E. coli |
| url | https://journals.asm.org/doi/10.1128/msphere.00345-22 |
| work_keys_str_mv | AT jacobjzulk phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT justinrclark phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT samanthaottinger phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT mallorybballard phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT marlydemejia phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT vickimercadoevans phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT emmalinerheckmann phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT belkyscsanchez phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT barbarawtrautner phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT anthonywmaresso phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment AT kathrynapatras phageresistanceaccompaniesreducedfitnessofuropathogenicescherichiacoliintheurinaryenvironment |