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

Abstract

SUMMARY Given the complex nature of Alzheimer’s disease (AD), a cell-based model that recapitulates physiological properties of the target neuronal population carries significant value in discovering improved drug candidates and chemical probes for uncovering disease mechanisms. We established phenotypic neuronal assays for biogenesis and synaptic action of amyloid β-peptide (Aβ) based on embryonic stem (ES) cell-derived neurons (ESNs). ESNs enriched with pyramidal neurons were robust, scalable and amenable to a small molecule screening assay, overcoming 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.

Drug Repositioning Approaches for the Discovery of New Therapeutics for Alzheimer's Disease

“…The Lilly Initiative uses an astrocytoma cell line, CCF-STTG1 (vs primary astrocytes) [31,32]. It is conceivable that the screen identified false positive hits or missed a number of hits (false negatives) that may be preferentially active in primary cells and more physiological cell models, as certain compounds have been found to exert biological activities only in physiologically relevant cells but not in transformed cell lines [33].…”

“…As physiological context is crucial for establishing predictive in vitro models for AD phenotypic screening, there is an increasing need for more physiological models, such as primary brain cells or stem cell-derived neurons or glial cells, which are scalable and robust for high throughput screening (HTS) assays [15,20,33,38]. Small molecule screenings for Aβ biogenesis and Aβ toxicity have been described in mouse embryonic stem cellderived pyramidal neurons and commercial human induced pluripotent stem (iPS) cell-derived neurons [33,39].…”

“…Small molecule screenings for Aβ biogenesis and Aβ toxicity have been described in mouse embryonic stem cellderived pyramidal neurons and commercial human induced pluripotent stem (iPS) cell-derived neurons [33,39]. While human patient-derived stem cells (e.g., iPS cells) carry great promise [40,41], their productive and reliable use for drug screening applications such as HTS remains challenging owing to laborious maintenance, which makes automation impractical [42][43][44].…”

Hit Triage and Validation in Phenotypic Screening: Considerations and Strategies

“…We acknowledge and thank colleagues from internal phenotypic programs as well as external scientists in the field for contributing to them through engaged scientific discussions. ll Review Gutierrez et al, 2014;Hadida et al, 2014;Huang et al, 2009;Jentzsch et al, 2012;Kato et al, 2016;Khare et al, 2016;Laraia et al, 2019;Lemm et al, 2010;Madoux et al, 2010;Mcintire et al, 2013;Peppard et al, 2014;Peppard et al, 2015;Sharabi et al, 2017;Su et al, 2010;Theriault et al, 2012;Titus et al, 2012;Wang et al, 2015;Warashina et al, 2006;Williams et al, 2003;Xiao et al, 2011;Xu et al, 2016.…”

The 14th International Conference on Alzheimer's Drug Discovery, 9 – 10 September 2013, in Jersey City, New Jersey

“…Tae-Wan Kim from Columbia University Medical Center talked on "Development of screening assays for tauopathy in stem cell derived neurons" [14]. Dr. Kim's approach is to use phenotypic drug discovery using mouse embryonic stem-cell-derived neurons to screen for small molecules to reduce the expression of tau and reverse the mis-localization of tau to dendrites.…”