Human iPSC Derived 2D & 3D Neuronal Models for Antisense Oligo Nucleotide Associated Acute Neurotoxicity Screening

Authors

• - Jyoti Roy
• - William Borschel
• - Elia R Lopez
• - Atena Farkhondeh Kalat
• - Edward Wu
• - Caroline Strong
• - Emily Lee
• - Marc Ferrer
• - Elizabeth A Ottinger
• - Bryan J Traynor

Abstract

Over the past decade, antisense oligonucleotides (ASOs) have been approved by FDA to treat rare neurological disorders. However, acute neurotoxicity has been observed in vivo for many ASOs during preclinical development. While animal models are an established method to assess the neurotoxic effects of ASOs at the pre-clinical stage, there is a need for in vitro assays that can reliably predict in vivo neurotoxicity with higher throughput and lower cost. We have established a platform of functional assays using human induced pluripotent stem cell (hiPSC) derived neurons for acute neurotoxicity screening of ASOs. This assay platform includes 2D & 3D neuronal models. The 2D neural model uses spinal motor neuron (SMN) and astrocyte co-cultures with a multi-electrode array (MEA) assay readout. We also use 3D neural spheroid models generated using astrocytes and GABAergic neurons mixed with either glutamatergic or dopaminergic neurons to mimic the ventral tegmental area (VTA)-like or pre-frontal cortex (PFC)-like resembling different brain regions. The 3D neural spheroids show spontaneous and synchronized calcium oscillations that can be measured in high throughput format using calcium fluorescence dyes. ASOs with known in vivo toxicity were tested on the cultured 2D or 3D neuronal models across multiple concentrations and we observed the changes in assay end points of our model that coincides with the in vivo toxicity score. Therefore, both these 2D and 3D human neuronal models have good in vitro-in vivo correlation and can be used to predict acute toxicity of ASOs in vitro.

Scientific Focus Area: Clinical Research

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