A Whole-Cell-Based High-Throughput Screening Method to Identify Molecules Targeting Pseudomonas aeruginosa Persister Cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1333; p. 113
• Main Authors: Liebens, Veerle, Defraine, Valerie, Fauvart, Maarten
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Anti-Bacterial Agents - pharmacology; High-Throughput Screening Assays - methods; Kinetics; Microbial Viability - drug effects; Pseudomonas aeruginosa - drug effects; Pseudomonas aeruginosa - growth & development; Quorum Sensing - drug effects; Small Molecule Libraries - chemistry; Small Molecule Libraries - pharmacology; High-throughput screening; Pseudomonas aeruginosa; Small molecules; Combination therapy; Growth kinetics
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-2854-5_10

Despite its clinical relevance and the fact that the phenomenon of persistence was discovered in the 1940s, little is known about the mechanisms behind persister cell formation. Research in this field has mainly focused on the model organism Escherichia coli and few genetic determinants of persistence have been described in other bacterial species, impairing the development of target-based strategies to combat these antibiotic-tolerant cells. In this chapter we describe a top-down large-scale screening method capable of specifically identifying small molecule compounds that, in combination with conventional antibiotics, significantly reduce the persister fraction in Pseudomonas aeruginosa. The method is readily adaptable for other species. Further characterization and analysis of the mode of action of the identified compounds can provide additional insight into the mechanisms behind persister formation and can guide the development of future anti-persister therapies.