Abstract

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Early-onset epileptic encephalopathies are severe disorders often linked to specific genetic mutations. One such condition, CDKL5 deficiency disorder (CDD), is characterized by early-onset seizures, intellectual delay, and motor dysfunction. Despite the importance of CDKL5 in brain development, its precise targets and relation to patient symptoms remain unclear. This study utilizes induced pluripotent stem cells (iPSCs) from CDD patients to generate neural cells, revealing disruptions in microtubule-based processes and cytoskeleton organization. CDD-derived neural progenitor cells (NPCs) exhibited proliferation defects, while neurons displayed morphological alterations and impaired glutamatergic synaptogenesis. Hyperactivity in CDD cortical neurons led to an overly synchronized network, which was rescued by lead compounds identified through high-throughput drug screening. These findings illuminate cellular and molecular mechanisms of genetic epilepsy, offering new therapeutic possibilities.

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Key Learnings

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• CDKL5 deficiency disrupts neural development, impacting cytoskeleton organization and neural connectivity.

• CDD neurons exhibit hyperactivity and abnormal synaptic function, leading to network synchronization defects.

• High-throughput drug screening identified compounds capable of rescuing the hyperactive neural network, highlighting potential treatments for CDD and related epileptic syndromes.

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Methods

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• Patient-derived iPSCs were reprogrammed into NPCs and neurons.

• Proteomic and phosphoproteomic analyses were performed to assess cellular disruptions.

• Neural morphology, synaptogenesis, and network activity were analyzed through various imaging and electrophysiological techniques.

• A high-throughput drug screening platform was developed to identify compounds that could rescue the altered network activity in CDD neurons.

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Conclusions

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This study provides insights into the mechanisms underlying CDKL5 deficiency disorder, demonstrating how CDKL5 mutations lead to neural network hyperactivity. By leveraging human stem cell models and high-throughput drug screening, researchers identified potential therapeutic candidates capable of restoring neural function. These findings not only enhance our understanding of genetic epilepsy but also pave the way for novel treatment strategies for CDD and similar conditions.

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Reference

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Negraes, P. D., Kelahmet, S., Xie, Y., Siegler, M., Devido, D., Alsaqati, M., ... & Muotri, A. R. (2021). Altered network and rescue of human neurons derived from individuals with early-onset genetic epilepsy. Molecular Psychiatry.