Introduction

The assessment of neurotoxicity in the context of pharmaceutical development presents a persistent set of challenges, particularly with respect to the limitations of traditional animal models and two-dimensional cell cultures. In response to these constraints, Takeda has engaged in a collaborative effort with 28bio to implement CNS-3D organoids for the evaluation of neurotoxic potential in candidate compounds.

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Background and Rationale

Takeda’s interest in the CNS-3D organoid model is rooted in the need for more physiologically relevant systems that can recapitulate human neurodevelopmental processes. The 28bio CNS-3D organoids, derived from human pluripotent stem cells, are engineered to self-organize into structures that reflect the cellular diversity and cytoarchitecture of the human cortex. This model offers the potential to detect neurotoxic effects that may not be apparent in conventional preclinical assays.

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Methodological Approach

Takeda adopted 28bio’s standardized protocol for the generation and maintenance of CNS-3D organoids. The process entailed the directed differentiation of pluripotent stem cells, followed by the cultivation of organoids under defined conditions to ensure reproducibility and fidelity to cortical identity.

For neurotoxicity assessment, organoids were exposed to a series of test articles, including both reference neurotoxicants and negative controls. The exposure regimens were selected to approximate clinically relevant concentrations and durations. Endpoints included quantitative measures of cell viability, neurite morphology, synaptic marker expression, and electrophysiological function, utilizing immunocytochemistry, high-content imaging, and multi-electrode array technology.

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Results

The CNS-3D organoid model demonstrated consistent reproducibility across independent preparations, with stable expression of cortical markers and preservation of three-dimensional architecture. Upon exposure to known neurotoxicants, the organoids exhibited concentration-dependent reductions in viability and neurite complexity, as well as discernible changes in synaptic protein localization. Electrophysiological analyses revealed alterations in network activity, including reduced spike frequency and modified burst patterns.

Organoids treated with negative controls maintained baseline viability and functional parameters, supporting the specificity of the observed neurotoxic responses. Notably, the sensitivity of the CNS-3D platform enabled the detection of neurotoxic effects at concentrations relevant to anticipated human exposures, thereby enhancing the translational value of the findings.

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Discussion

The results of Takeda’s work with CNS-3D organoids underscore the advantages of this model in preclinical neurotoxicity screening. CNS-3D organoids provide a human-relevant context for the detection of compound-induced neurotoxicity, supports multiplexed endpoint analysis, and reduces reliance on animal testing. The concordance between organoid-based readouts and established neurotoxicant profiles further substantiates the model’s applicability to pharmaceutical research.

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Conclusion

Takeda’s application of the CNS-3D technology represents a substantive advancement in the field of neurotoxicity assessment. The collaborative work has demonstrated the feasibility and value of integrating organoid assays into preclinical safety pipelines.

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References

Webinar: “Preclinical Neurotoxicity Predictions using a Cortical Brain Organoid Model,” Takeda.

Publication: “Assessment of a 3D Neural Spheroid Model to Detect Pharmaceutical-Induced Neurotoxicity,” Takeda.

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