A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili
ABSTRACT The increasing threat of antibiotic resistance underscores the urgent need for innovative strategies to combat infectious diseases, including the development of antivirulants. Microbial pathogens rely on their virulence factors to initiate and sustain infections. Antivirulants are small mol...
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| Format: | Article |
| Language: | English |
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Wiley
2025-01-01
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| Series: | Microbial Biotechnology |
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| Online Access: | https://doi.org/10.1111/1751-7915.70081 |
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| author | Tori M. Shimozono Nancy J. Vogelaar Megan T. O'Hara Zhaomin Yang |
| author_facet | Tori M. Shimozono Nancy J. Vogelaar Megan T. O'Hara Zhaomin Yang |
| author_sort | Tori M. Shimozono |
| collection | DOAJ |
| description | ABSTRACT The increasing threat of antibiotic resistance underscores the urgent need for innovative strategies to combat infectious diseases, including the development of antivirulants. Microbial pathogens rely on their virulence factors to initiate and sustain infections. Antivirulants are small molecules designed to target virulence factors, thereby attenuating the virulence of infectious microbes. The bacterial type IV pilus (T4P), an extracellular protein filament that depends on the T4P machinery (T4PM) for its biogenesis, dynamics and function, is a key virulence factor in many significant bacterial pathogens. While the T4PM presents a promising antivirulence target, the systematic identification of inhibitors for its multiple protein constituents remains a considerable challenge. Here we report a novel high‐throughput screening (HTS) approach for discovering T4P inhibitors. It uses Pseudomonas aeruginosa, a high‐priority pathogen, in combination with its T4P‐targeting phage, φKMV. Screening of a library of 2168 compounds using an optimised protocol led to the identification of tuspetinib, based on its deterrence of the lysis of P. aeruginosa by φKMV. Our findings show that tuspetinib also inhibits two additional T4P‐targeting phages, while having no effect on a phage that recognises lipopolysaccharides as its receptor. Additionally, tuspetinib impedes T4P‐mediated motility in P. aeruginosa and Acinetobacter species without impacting growth or flagellar motility. This bacterium‐phage pairing approach is applicable to a broad range of virulence factors that are required for phage infection, paving ways for the development of advanced chemotherapeutics against antibiotic‐resistant infections. |
| format | Article |
| id | doaj-art-6ef5503fec39413db856944413066697 |
| institution | Kabale University |
| issn | 1751-7915 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| spelling | doaj-art-6ef5503fec39413db8569444130666972025-01-31T06:26:35ZengWileyMicrobial Biotechnology1751-79152025-01-01181n/an/a10.1111/1751-7915.70081A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV PiliTori M. Shimozono0Nancy J. Vogelaar1Megan T. O'Hara2Zhaomin Yang3Department of Biological Sciences Virginia Tech Blacksburg Virginia USAVirginia Tech Center for Drug Discovery Virginia Tech Blacksburg Virginia USADepartment of Biological Sciences Virginia Tech Blacksburg Virginia USADepartment of Biological Sciences Virginia Tech Blacksburg Virginia USAABSTRACT The increasing threat of antibiotic resistance underscores the urgent need for innovative strategies to combat infectious diseases, including the development of antivirulants. Microbial pathogens rely on their virulence factors to initiate and sustain infections. Antivirulants are small molecules designed to target virulence factors, thereby attenuating the virulence of infectious microbes. The bacterial type IV pilus (T4P), an extracellular protein filament that depends on the T4P machinery (T4PM) for its biogenesis, dynamics and function, is a key virulence factor in many significant bacterial pathogens. While the T4PM presents a promising antivirulence target, the systematic identification of inhibitors for its multiple protein constituents remains a considerable challenge. Here we report a novel high‐throughput screening (HTS) approach for discovering T4P inhibitors. It uses Pseudomonas aeruginosa, a high‐priority pathogen, in combination with its T4P‐targeting phage, φKMV. Screening of a library of 2168 compounds using an optimised protocol led to the identification of tuspetinib, based on its deterrence of the lysis of P. aeruginosa by φKMV. Our findings show that tuspetinib also inhibits two additional T4P‐targeting phages, while having no effect on a phage that recognises lipopolysaccharides as its receptor. Additionally, tuspetinib impedes T4P‐mediated motility in P. aeruginosa and Acinetobacter species without impacting growth or flagellar motility. This bacterium‐phage pairing approach is applicable to a broad range of virulence factors that are required for phage infection, paving ways for the development of advanced chemotherapeutics against antibiotic‐resistant infections.https://doi.org/10.1111/1751-7915.70081antivirulencehigh‐throughput screenphagePseudomonas aeruginosatype 4 pilus (T4P) |
| spellingShingle | Tori M. Shimozono Nancy J. Vogelaar Megan T. O'Hara Zhaomin Yang A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili Microbial Biotechnology antivirulence high‐throughput screen phage Pseudomonas aeruginosa type 4 pilus (T4P) |
| title | A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili |
| title_full | A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili |
| title_fullStr | A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili |
| title_full_unstemmed | A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili |
| title_short | A Phage‐Based Approach to Identify Antivirulence Inhibitors of Bacterial Type IV Pili |
| title_sort | phage based approach to identify antivirulence inhibitors of bacterial type iv pili |
| topic | antivirulence high‐throughput screen phage Pseudomonas aeruginosa type 4 pilus (T4P) |
| url | https://doi.org/10.1111/1751-7915.70081 |
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