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Assay development and high-throughput screening for small molecule inhibitors of a Vibrio cholerae stress response
Laura Stanbery1, Jyl S Matson1
1Department of Medical Microbiology and Immunology, College of Medicine and Life Sciences, The University of Toledo, Toledo, OH, USA.
Abstract:
Antibiotics are important adjuncts to oral rehydration therapy in cholera disease management. However, due to the rapid emergence of resistance to the antibiotics used to treat cholera, therapeutic options are becoming limited. Therefore, there is a critical need to develop additional therapeutics to aid in the treatment of cholera. Previous studies showed that the extracytoplasmic stress response (σE) pathway of Vibrio cholerae is required for full virulence of the organism. The pathway is also required for bacterial growth in the presence of ethanol. Therefore, we exploited this ethanol sensitivity phenotype in order to develop a screen for inhibitors of the pathway, with the aim of also inhibiting virulence of the pathogen. Here we describe the optimization and implementation of our high-throughput screening strategy. From a primary screen of over 100,000 compounds, we have identified seven compounds that validated the growth phenotypes from the primary and counterscreens. These compounds have the potential to be developed into therapeutic agents for cholera and will also be valuable probes for uncovering basic molecular mechanisms of an important cause of diarrheal disease.
Insights
New cholera therapeutics are needed due to antibiotic resistance. Researchers screened compounds targeting the Vibrio cholerae extracytoplasmic stress response (σE) pathway, identifying seven potential drug candidates.
Area of Science:
- Microbiology
- Pathogen Biology
- Drug Discovery
Background:
- Antibiotic resistance in Vibrio cholerae limits treatment options for cholera.
- The extracytoplasmic stress response (σE) pathway is crucial for Vibrio cholerae virulence.
- Vibrio cholerae's reliance on the σE pathway for growth in ethanol presents a therapeutic vulnerability.
Purpose of the Study:
- To develop a high-throughput screening strategy to identify inhibitors of the Vibrio cholerae σE pathway.
- To discover novel therapeutic agents for cholera treatment by targeting bacterial virulence mechanisms.
- To find chemical probes for studying the σE pathway in Vibrio cholerae.
Main Methods:
- Optimization and implementation of a high-throughput screening assay based on ethanol sensitivity.
- Screening of over 100,000 chemical compounds.
- Validation of hit compounds through primary and counter-screening assays to confirm growth inhibition phenotypes.
Main Results:
- Identification of seven validated compounds from a screen of over 100,000 molecules.
- These compounds demonstrated efficacy in inhibiting the targeted growth phenotype.
- The identified compounds show potential as starting points for cholera drug development.
Conclusions:
- The developed high-throughput screen effectively identified inhibitors of the Vibrio cholerae σE pathway.
- The validated compounds represent promising candidates for new cholera therapeutics.
- These findings offer valuable tools for further research into Vibrio cholerae pathogenesis and treatment.