MDD489 - MULTIPLEXED AUTOMATED ASSAYS FOR NEUROTOXICITY EVALUATION USING INDUCED PLURIPOTENT STEM CELL-DERIVED NEURAL 3D CELL MODELS
Thursday, June 25, 2020
Theme: Modeling Development and Disease
Sirenko, Oksana - R&D, Molecular Devices, San Jose, CA, USA Zanella, Fabian - R&D, Stemonix, San Diego, CA, USA Crittenden, Carole - Drug Discovery, Molecular Devices, San Jose, CA, USA Chapa, Jared - Communication, Molecular Devices, San Jose, CA, USA Carromeu, Cassiano - R&D, Stemonix, San Diego, CA, USA
– Sr. Scientist, Molecular Devices, San Jose, California
Abstract: Cell-based phenotypic assays have become an increasingly attractive alternative to traditional in vitro and in vivo testing in pharmaceutical drug development and toxicological safety assessment. The effectiveness of automated imaging assays combined with the organotypic nature of human induced pluripotent stem cell (iPSC)-derived cells opens new opportunities to employ physiologically relevant in vitro model systems to improve screening for new drugs or potential chemical toxicities. In our studies, we used human iPSC-derived neural cultures to test functional and morphological end points for toxicity evaluation in a multi-parametric assay format.
For neurotoxicity assessment we employed iPSC-based 3D neural platform composed of mature cortical neurons and astrocytes. Neural spheroids generated spontaneous synchronized calcium oscillations. We used fast fluorescence kinetic imaging to measure the patterns and frequencies of the Ca2+ oscillations. Characterization of oscillation profiles was recorded by performed through multi-parametric analysis presenting more than 15 read-outs including frequency, amplitude, characterized primary and secondary peaks, peak width, and waveform irregularities. In addition, cellular and mitochondrial toxicities were assessed by high-content imaging. The assay was optimized for high throughput screening in 384-well plates and displayed a highly consistent performance. We evaluated neuroactive and neurotoxic profiles of more than 120 compounds, including set of neuromodulators with known mechanisms of action, set of various pharmaceutical drugs, sample library of cannabinoids, and a library of neurotoxic chemicals including flame retardants, pesticides, and poly-aromatic hydrocarbons.
Our results show that the iPSC-derived 3D neurospheroid assay platform is a promising biologically-relevant tool to assess the neuroactive and neurotoxic potential of pharmaceutical drugs and environmental toxicants.