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

Chao Wang; Michael E Ward; Robert Chen; Kai Liu; Tara E Tracy; Xu Chen; Min Xie; Peter Dongmin Sohn; Connor Ludwig; Anke Meyer-Franke; Celeste M Karch; Sheng Ding; Li Gan

doi:10.1016/j.stemcr.2017.08.019

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

Stem Cell Reports. 2017 Oct 10;9(4):1221-1233. doi: 10.1016/j.stemcr.2017.08.019. Epub 2017 Sep 28.

Authors

Chao Wang¹, Michael E Ward², Robert Chen³, Kai Liu⁴, Tara E Tracy¹, Xu Chen¹, Min Xie⁴, Peter Dongmin Sohn¹, Connor Ludwig³, Anke Meyer-Franke⁵, Celeste M Karch⁶, Sheng Ding⁴, Li Gan⁷

Affiliations

¹ Gladstone Institute of Neurological Disease, 1650 Owens Street, San Francisco, CA 94158, USA; Department of Neurology, University of California, San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA.
² Gladstone Institute of Neurological Disease, 1650 Owens Street, San Francisco, CA 94158, USA; Department of Neurology, University of California, San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA; National Institute of Neurological Disorders and Stroke, National Institutes of Health, 35 Convent Drive, Bethesda, MD 20892, USA.
³ Department of Neurology, University of California, San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA.
⁴ Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA; Department of Pharmaceutical Chemistry, University of California, San Francisco, 600 16th Street, San Francisco, CA 94158, USA.
⁵ Gladstone Institute of Neurological Disease, 1650 Owens Street, San Francisco, CA 94158, USA.
⁶ Department of Psychiatry, Washington University School of Medicine, 425 South Euclid Avenue, St. Louis, MO 63110, USA.
⁷ Gladstone Institute of Neurological Disease, 1650 Owens Street, San Francisco, CA 94158, USA; Department of Neurology, University of California, San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: lgan@gladstone.ucsf.edu.

Abstract

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.

Keywords: Alzheimer’s disease; Tau-lowering; adrenergic receptor; frontotemporal dementia; high-content screening; human induced pluripotent stem cells; human neurons; neurodegeneration; neurogenin 2; tau.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Cell Differentiation*
Cell Line
Cell Survival
Cells, Cultured
Drug Discovery* / methods
Drug Evaluation, Preclinical
Gene Expression
Gene Expression Regulation / drug effects*
Gene Order
Genetic Vectors / genetics
Glutamine / metabolism
High-Throughput Screening Assays*
Humans
Induced Pluripotent Stem Cells / cytology*
Membrane Potentials
Neurons / cytology
Neurons / drug effects*
Neurons / metabolism*
tau Proteins / genetics*
tau Proteins / metabolism

Substances

tau Proteins
Glutamine

Abstract

Publication types

MeSH terms

Substances

Grants and funding