A High-Content Assay Enables the Automated Screening and Identification of Small Molecules with Specific ALDH1A1-Inhibitory Activity

• DOI: 10.1371/journal.pone.0170937
• Publication Date: 2017-01-27T13:33:20Z
• Authors: Adam Yasgar; Steven A. Titus; Yuhong Wang; Carina Danchik; Shyh-Ming Yang; Vasilis Vasiliou; Ajit Jadhav; David J. Maloney; Anton Simeonov; Natalia J. Martinez
• Keywords: High-Content Screening; High-Throughput Screening

Aldehyde dehydrogenase enzymes (ALDHs) have a broad spectrum of biological activities through the oxidation both endogenous and exogenous aldehydes. Increased expression ALDH1A1 has been identified in wide-range human cancer stem cells is associated with relapse poor prognosis, raising potential as therapeutic target. To facilitate quantitative high-throughput screening (qHTS) campaigns for discovery, characterization structure-activity-relationship (SAR) studies small molecule inhibitors cellular activity, we show herein miniaturization to 1536-well format automation high-content cell-based ALDEFLUOR assay. We demonstrate utility this assay by generating dose-response curves on comprehensive set prior art well hundreds synthesized house. Finally, established paradigm using pair cell lines low high expression, respectively, uncover novel cell-active ALDH1A1-specific from collection over 1,000 molecules.