High-throughput screen for compounds that modulate neurite growth of human induced pluripotent stem cell-derived neurons

Abstract

Development of technology platforms to perform compound screens of human induced pluripotent stem cell (hiPSC)-derived neurons with relatively high throughput is essential to realize their potential for drug discovery. Here, we demonstrate the feasibility of high-throughput screening of hiPSC-derived neurons using a high-content, image-based approach focused on neurite growth, a process that is fundamental to formation of neural networks and nerve regeneration. From a collection of 4421 bioactive small molecules, we identified 108 hit compounds, including 37 approved drugs, that target molecules or pathways known to regulate neurite growth, as well as those not previously associated with this process. These data provide evidence that many pathways and targets known to play roles in neurite growth have similar activities in hiPSC-derived neurons that can be identified in an unbiased phenotypic screen. The data also suggest that hiPSC-derived neurons provide a useful system to study the mechanisms of action and off-target activities of the approved drugs identified as hits, leading to a better understanding of their clinical efficacy and toxicity, especially in the context of specific human genetic backgrounds. Finally, the hit set we report constitutes a sublibrary of approved drugs and tool compounds that modulate neurites. This sublibrary will be invaluable for phenotypic analyses and interrogation of hiPSC-based disease models as probes for defining phenotypic differences and cellular vulnerabilities in patient versus control cells, as well as for investigations of the molecular mechanisms underlying human neurite growth in development and maintenance of neuronal networks, and nerve regeneration.

Human in vitro models for understanding mechanisms of autism spectrum disorder

“…Stem cell-based models can also be used to screen drugs for treatment of neurodevelopmental disorders including ASD [91,104]. For example, one study screened 4421 unique compounds and identified 108 compounds that regulate neurite growth [105], a process which has been variably linked to some forms of ASD [106]. Another study screened a set of 50,000 compounds in neural stem cells to find activators of FMR1, a gene silenced in fragile X syndrome, which increases risk for ASD [107].…”

Human Induced Pluripotent Stem Cell Derived Sensory Neurons are Sensitive to the Neurotoxic Effects of Paclitaxel

“…In recent years, human induced pluripotent stem cell (iPSC)-derived neurons have been used for the study of CIPN. Commercially available iPSC-derived neurons (e.g., iCell Neurons ® and Peri.4U Neurons ® ) have been evaluated as a model of neurotoxicity 9 , used to screen for neurotoxic compounds [10][11][12][13] , and utilized for functional validation of genes identified in human genome wide association studies of CIPN 9, [14][15][16] . The use of human iPSC-derived neurons affords an advantage over rodent DRG neurons in their human origin and the potential to differentiate into specific peripheral sensory neuron populations.…”

Unprecedented Potential for Neural Drug Discovery Based on Self-Organizing hiPSC Platforms

“…The prior example explains the feasibility of testing new compounds predicted by AI in hiPSCsderived neurons, while other groups demonstrated the utility of hiPSCs derived neurons as platforms for testing neurite growth related drugs, which is highly relevant for neural repair [34]. In this context, high throughput screening of drugs on human iPSC differentiated neurons proved useful for assessing toxicity of compounds, determining off-target effects of compounds already in use and to identify new mechanisms and specific molecular pathways to target.…”