Scalable Production of iPSC-Derived Human Neurons to Identify Tau-Lowering Compounds by High-Content Screening

• Published in: Stem cell reports Vol. 9; no. 4; pp. 1221 - 1233
• Main Authors: Wang, Chao, Ward, Michael E., Chen, Robert, Liu, Kai, Tracy, Tara E., Chen, Xu, Xie, Min, Sohn, Peter Dongmin, Ludwig, Connor, Meyer-Franke, Anke, Karch, Celeste M., Ding, Sheng, Gan, Li
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 10-10-2017; Elsevier
• Subjects: adrenergic receptor; Alzheimer’s disease; Cell Differentiation; Cell Line; Cell Survival; Cells, Cultured; Drug Discovery - methods; Drug Evaluation, Preclinical; frontotemporal dementia; Gene Expression; Gene Expression Regulation - drug effects; Gene Order; Genetic Vectors - genetics; Glutamine - metabolism; high-content screening; High-Throughput Screening Assays; human induced pluripotent stem cells; human neurons; Humans; Induced Pluripotent Stem Cells - cytology; Membrane Potentials; neurodegeneration; neurogenin 2; Neurons - cytology; Neurons - drug effects; Neurons - metabolism; tau; tau Proteins - genetics; tau Proteins - metabolism; Tau-lowering; human induced pluripotent stem cells; human neurons; Tau-lowering; Alzheimer’s disease; adrenergic receptor; neurodegeneration; tau; frontotemporal dementia; neurogenin 2; high-content screening
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2017.08.019

Lowering total tau levels is an attractive therapeutic strategy for Alzheimer's disease and other tauopathies. High-throughput screening in neurons derived from human induced pluripotent stem cells (iPSCs) is a powerful tool to identify tau-targeted therapeutics. However, such screens have been hampered by heterogeneous neuronal production, high cost and low yield, and multi-step differentiation procedures. We engineered an isogenic iPSC line that harbors an inducible neurogenin 2 transgene, a transcription factor that rapidly converts iPSCs to neurons, integrated at the AAVS1 locus. Using a simplified two-step protocol, we differentiated these iPSCs into cortical glutamatergic neurons with minimal well-to-well variability. We developed a robust high-content screening assay to identify tau-lowering compounds in LOPAC and identified adrenergic receptors agonists as a class of compounds that reduce endogenous human tau. These techniques enable the use of human neurons for high-throughput screening of drugs to treat neurodegenerative disease.
•Inducible, isogenic, and integrated Ngn2 iPSCs were differentiated to pure neurons•A simple and scalable two-step protocol ensures minimal differentiation variability•An HCS assay was established to screen for tau-lowering compounds in LOPAC•AR agonists were identified as a class of compounds that reduce human tau
Gan and colleagues developed a simple and scalable technology to generate a large quantity of homogeneous glutamatergic cortical neurons by engineering a neurogenin 2-expressing cassette to the AAVS1 locus of iPSCs. They developed a high-content screening assay and identified adrenergic receptor agonists as a class of compounds that lower endogenous human tau, a key pathogenic factor in Alzheimer's disease.