Assay Development for High-Throughput Drug Screening Against Mycobacteria

Area of Science:

• Microbiology
• Drug Discovery
• Infectious Diseases

Background:

• Mycobacterium abscessus infections present significant treatment challenges due to intrinsic drug resistance, similar to multidrug-resistant tuberculosis.
• Current treatment regimens are often ineffective, leading to poor patient compliance and a critical need for novel therapeutic strategies.
• Drug repurposing offers a promising avenue for identifying new treatments by leveraging existing drugs, thus expediting the drug development process.

Purpose of the Study:

• To develop and validate a high-throughput drug screening assay for identifying novel therapeutic agents against Mycobacterium abscessus.
• To establish a rapid and reliable method for assessing bacterial viability in drug screening, reducing time and variability.
• To facilitate the discovery of effective drugs to combat drug-resistant Mycobacterium abscessus infections.

Main Methods:

• Development of an in-house double-reporter strain of Mycobacterium abscessus for enhanced screening capabilities.
• Utilization of liquid-handling robotics, automated microscopy, and analysis for rapid bacterial viability measurement via luminescence and fluorescence readouts.
• Validation of the assay by screening a library of 1280 compounds to assess its efficacy in detecting active agents.

Main Results:

• The developed assay enables rapid measurement of bacterial viability using dual luminescence and fluorescence readouts without additional reagents or steps.
• Screening of a 1280-compound library successfully identified active compounds, with results corroborated by existing literature.
• The assay demonstrated efficiency in detecting compounds active against Mycobacterium abscessus, confirming its utility.

Conclusions:

• A novel, efficient high-throughput screening assay has been successfully developed and validated for Mycobacterium abscessus drug discovery.
• This tool significantly reduces assay time and variability, offering a valuable resource for the scientific community in the fight against drug-resistant infections.
• The assay provides a much-needed platform to accelerate the identification of new drugs to combat challenging Mycobacterium abscessus infections.