Phenotypic Assays for β-Amyloid in Mouse Embryonic Stem Cell-Derived Neurons

•Scaling and optimizing primary neurons derived from mouse ESCs•Modeling phenotypic assays using mouse ESNs for biogenesis and synaptic action of Aβ•Isolation and use of ESCs originating from a mouse model of AD•Screen of clinical compounds and identification of Aβ-lowering small molecules

SummaryGiven the complex nature of Alzheimer’s disease (AD), a cell-based model that recapitulates the physiological properties of the target neuronal population would be extremely valuable for discovering improved drug candidates and chemical probes to uncover disease mechanisms. We established phenotypic neuronal assays for the biogenesis and synaptic action of amyloid β peptide (Aβ) based on embryonic stem cell-derived neurons (ESNs). ESNs enriched with pyramidal neurons were robust, scalable, and amenable to a small-molecule screening assay, overcoming the apparent limitations of neuronal models derived from human pluripotent cells. Small-molecule screening of clinical compounds identified four compounds capable of reducing Aβ levels in ESNs derived from the Tg2576 mouse model of AD. Our approach is therefore highly suitable for phenotypic screening in AD drug discovery and has the potential to identify therapeutic candidates with improved efficacy and safety potential.

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