Abstract

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The discovery of therapeutics for neurological diseases is hindered by the lack of predictive in vitro and in vivo models. Traditional in vitro assays rely on 2D-engineered cell lines, which are suitable for large-scale screening but lack physiological relevance. Human induced pluripotent stem cell (iPSC)-derived 3D neural spheroids offer a more physiologically relevant alternative, incorporating complex neuronal and glial cell populations that exhibit synchronous activity. This study evaluates the potential of iPSC-derived cortical neural spheroids as a high-throughput screening (HTS) platform for drug discovery by assessing a library of 687 neuroactive compounds. Calcium activity, measured as a functional biomarker, was analyzed using multiparametric data analysis, demonstrating the robustness and applicability of this approach for pharmacological screening.

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Key Learnings

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• iPSC-derived cortical spheroids provide a physiologically relevant 3D model for high-throughput drug screening

• Calcium fluorescence imaging allows for real-time assessment of compound-induced neural activity changes

• Multiparametric analysis enables classification of compound responses based on calcium signal characteristics

• The screening approach successfully identified pharmacological activity profiles that could aid in neurological disease modeling and therapeutic development

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Methods

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• iPSC-derived cortical neural spheroids were obtained and cultured for 10.5 weeks before screening

• A library of 687 neuroactive compounds was tested using a calcium fluorescence assay

• Calcium activity was measured using a Fluorescent Imaging Plate Reader (FLIPR), and data was analyzed using principal component analysis (PCA) and Sammon mapping

• Compounds were classified based on their impact on seven peak characteristics of calcium activity

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Conclusions

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This study demonstrates that iPSC-derived cortical neural spheroids serve as a robust, physiologically relevant platform for high-throughput pharmacological screening. The integration of multiparametric calcium fluorescence analysis allows for detailed classification of neuroactive compound responses, offering a powerful tool for advancing neurological disease research and drug discovery efforts.

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Reference

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Boutin, M. E., Strong, C. E., Van Hese, B., Hu, X., Itkin, Z., Chen, Y.-C., LaCroix, A., Gordon, R., Guicherit, O., Carromeu, C., Kundu, S., Lee, E., & Ferrer, M. (2022). A multiparametric calcium signal screening platform using iPSC-derived cortical neural spheroids. SLAS Discovery, 27(3), 209–218.

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