A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants
ABSTRACT Drug repurposing offers an expedited and economical route to develop new clinical therapeutics in comparison to traditional drug development. Growth-based high-throughput screening is concomitant with drug repurposing and enables rapid identification of new therapeutic uses for investigated...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2018-10-01
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| Series: | mSphere |
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| Online Access: | https://journals.asm.org/doi/10.1128/mSphere.00422-18 |
| _version_ | 1830408813726924800 |
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| author | Ryan P. Trombetta Paul M. Dunman Edward M. Schwarz Stephen L. Kates Hani A. Awad |
| author_facet | Ryan P. Trombetta Paul M. Dunman Edward M. Schwarz Stephen L. Kates Hani A. Awad |
| author_sort | Ryan P. Trombetta |
| collection | DOAJ |
| description | ABSTRACT Drug repurposing offers an expedited and economical route to develop new clinical therapeutics in comparison to traditional drug development. Growth-based high-throughput screening is concomitant with drug repurposing and enables rapid identification of new therapeutic uses for investigated drugs; however, this traditional method is not compatible with microorganisms with abnormal growth patterns such as Staphylococcus aureus small-colony variants (SCV). SCV subpopulations are auxotrophic for key compounds in biosynthetic pathways, which result in low growth rate. SCV formation is also associated with reduced antibiotic susceptibility, and the SCV’s ability to revert to the normal cell growth state is thought to contribute to recurrence of S. aureus infections. Thus, there is a critical need to identify antimicrobial agents that are potent against SCV in order to effectively treat chronic infections. Accordingly, here we describe adapting an adenylate kinase (AK)-based cell death reporter assay to identify members of a Food and Drug Administration (FDA)-approved drug library that display bactericidal activity against S. aureus SCV. Four library members, daunorubicin, ketoconazole, rifapentine, and sitafloxacin, exhibited potent SCV bactericidal activity against a stable S. aureus SCV. Further investigation showed that sitafloxacin was potent against methicillin-susceptible and -resistant S. aureus, as well as S. aureus within an established biofilm. Taken together, these results demonstrate the ability to use the AK assay to screen small-molecule libraries for SCV bactericidal agents and highlight the therapeutic potential of sitafloxacin to be repurposed to treat chronic S. aureus infections associated with SCV and/or biofilm growth states. IMPORTANCE Conventional antibiotics fail to successfully treat chronic osteomyelitis, endocarditis, and device-related and airway infections. These recurring infections are associated with the emergence of SCV, which are recalcitrant to conventional antibiotics. Studies have investigated antibiotic therapies to treat SCV-related infections but have had little success, emphasizing the need to identify novel antimicrobial drugs. However, drug discovery is a costly and time-consuming process. An alternative strategy is drug repurposing, which could identify FDA-approved and well-characterized drugs that could have off-label utility in treating SCV. In this study, we adapted a high-throughput AK-based assay to identify 4 FDA-approved drugs, daunorubicin, ketoconazole, rifapentine, and sitafloxacin, which display antimicrobial activity against S. aureus SCV, suggesting an avenue for drug repurposing in order to effectively treat SCV-related infections. Additionally, this screening paradigm can easily be adapted for other drug/chemical libraries to identify compounds bactericidal against SCV. |
| first_indexed | 2024-12-20T18:58:43Z |
| format | Article |
| id | doaj.art-1e0a50069cf744c4848bf014e566bb50 |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-12-20T18:58:43Z |
| publishDate | 2018-10-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-1e0a50069cf744c4848bf014e566bb502022-12-21T19:29:26ZengAmerican Society for MicrobiologymSphere2379-50422018-10-013510.1128/mSphere.00422-18A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony VariantsRyan P. Trombetta0Paul M. Dunman1Edward M. Schwarz2Stephen L. Kates3Hani A. Awad4Department of Biomedical Engineering, University of Rochester, Rochester, New York, USADepartment of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USADepartment of Biomedical Engineering, University of Rochester, Rochester, New York, USACenter for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York, USADepartment of Biomedical Engineering, University of Rochester, Rochester, New York, USAABSTRACT Drug repurposing offers an expedited and economical route to develop new clinical therapeutics in comparison to traditional drug development. Growth-based high-throughput screening is concomitant with drug repurposing and enables rapid identification of new therapeutic uses for investigated drugs; however, this traditional method is not compatible with microorganisms with abnormal growth patterns such as Staphylococcus aureus small-colony variants (SCV). SCV subpopulations are auxotrophic for key compounds in biosynthetic pathways, which result in low growth rate. SCV formation is also associated with reduced antibiotic susceptibility, and the SCV’s ability to revert to the normal cell growth state is thought to contribute to recurrence of S. aureus infections. Thus, there is a critical need to identify antimicrobial agents that are potent against SCV in order to effectively treat chronic infections. Accordingly, here we describe adapting an adenylate kinase (AK)-based cell death reporter assay to identify members of a Food and Drug Administration (FDA)-approved drug library that display bactericidal activity against S. aureus SCV. Four library members, daunorubicin, ketoconazole, rifapentine, and sitafloxacin, exhibited potent SCV bactericidal activity against a stable S. aureus SCV. Further investigation showed that sitafloxacin was potent against methicillin-susceptible and -resistant S. aureus, as well as S. aureus within an established biofilm. Taken together, these results demonstrate the ability to use the AK assay to screen small-molecule libraries for SCV bactericidal agents and highlight the therapeutic potential of sitafloxacin to be repurposed to treat chronic S. aureus infections associated with SCV and/or biofilm growth states. IMPORTANCE Conventional antibiotics fail to successfully treat chronic osteomyelitis, endocarditis, and device-related and airway infections. These recurring infections are associated with the emergence of SCV, which are recalcitrant to conventional antibiotics. Studies have investigated antibiotic therapies to treat SCV-related infections but have had little success, emphasizing the need to identify novel antimicrobial drugs. However, drug discovery is a costly and time-consuming process. An alternative strategy is drug repurposing, which could identify FDA-approved and well-characterized drugs that could have off-label utility in treating SCV. In this study, we adapted a high-throughput AK-based assay to identify 4 FDA-approved drugs, daunorubicin, ketoconazole, rifapentine, and sitafloxacin, which display antimicrobial activity against S. aureus SCV, suggesting an avenue for drug repurposing in order to effectively treat SCV-related infections. Additionally, this screening paradigm can easily be adapted for other drug/chemical libraries to identify compounds bactericidal against SCV.https://journals.asm.org/doi/10.1128/mSphere.00422-18Staphylococcus aureussmall-colony variantshigh-throughput screendrug repurposingchronic infection |
| spellingShingle | Ryan P. Trombetta Paul M. Dunman Edward M. Schwarz Stephen L. Kates Hani A. Awad A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants mSphere Staphylococcus aureus small-colony variants high-throughput screen drug repurposing chronic infection |
| title | A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants |
| title_full | A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants |
| title_fullStr | A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants |
| title_full_unstemmed | A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants |
| title_short | A High-Throughput Screening Approach To Repurpose FDA-Approved Drugs for Bactericidal Applications against <named-content content-type="genus-species">Staphylococcus aureus</named-content> Small-Colony Variants |
| title_sort | high throughput screening approach to repurpose fda approved drugs for bactericidal applications against named content content type genus species staphylococcus aureus named content small colony variants |
| topic | Staphylococcus aureus small-colony variants high-throughput screen drug repurposing chronic infection |
| url | https://journals.asm.org/doi/10.1128/mSphere.00422-18 |
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