title: A High-Content Assay Enables the Automated Screening and Identification of Small Molecules with Specific ALDH1A1-Inhibitory Activity.
abstract: Aldehyde dehydrogenase enzymes (ALDHs) have a broad spectrum of biological activities through the oxidation of both endogenous and exogenous aldehydes. Increased expression of ALDH1A1 has been identified in a wide-range of human cancer stem cells and is associated with cancer relapse and poor prognosis, raising the potential of ALDH1A1 as a therapeutic target. To facilitate quantitative high-throughput screening (qHTS) campaigns for the discovery, characterization and structure-activity-relationship (SAR) studies of small molecule ALDH1A1 inhibitors with cellular activity, we show herein the miniaturization to 1536-well format and automation of a high-content cell-based ALDEFLUOR assay. We demonstrate the utility of this assay by generating dose-response curves on a comprehensive set of prior art inhibitors as well as hundreds of ALDH1A1 inhibitors synthesized in house. Finally, we established a screening paradigm using a pair of cell lines with low and high ALDH1A1 expression, respectively, to uncover novel cell-active ALDH1A1-specific inhibitors from a collection of over 1,000 small molecules.
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Adam
authors.initials: A
authors.lastname: Yasgar
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Titus
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Carina
authors.initials: C
authors.lastname: Danchik
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Shyh-Ming
authors.initials: SM
authors.lastname: Yang
authors.affiliation: Department of Environmental Health Sciences, Yale School of Public Health, New Haven, CT, United States of America.
authors.forename: Vasilis
authors.initials: V
authors.lastname: Vasiliou
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Ajit
authors.initials: A
authors.lastname: Jadhav
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: David J
authors.initials: DJ
authors.lastname: Maloney
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Anton
authors.initials: A
authors.lastname: Simeonov
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, United States of America.
authors.forename: Natalia J
authors.identifier: http://orcid.org/0000-0002-0663-9011
authors.initials: NJ
authors.lastname: Martinez
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D000080924:Aldehyde Dehydrogenase 1 Family; D000447:Aldehydes / Q000378:metabolism; D001681:Biological Assay; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D014411:Neoplastic Stem Cells / Q000187:drug effects; D010084:Oxidation-Reduction; D050697:Retinal Dehydrogenase; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D013329:Structure-Activity Relationship
chemical_list: D000447:Aldehydes; D004791:Enzyme Inhibitors; D054852:Small Molecule Libraries; D000080924:Aldehyde Dehydrogenase 1 Family; D000444:Aldehyde Dehydrogenase; C510223:ALDH1A1 protein, human; D050697:Retinal Dehydrogenase