UC San Diego

Approximately 30% of epilepsy patients experience intractable epilepsy, where traditional pharmacological therapies fail to provide seizure relief. Although challenging to sustain, patients often rely on the high-fat, low-carbohydrate ketogenic diet (KD) as a viable alternative for managing seizures. Despite its clinical success, the exact anti-epileptic mechanisms of the KD, remain unclear. Mitochondria have emerged as central players in this mechanism, with studies highlighting them as both targets through which the KD protects against seizures and key regulators of neuronal health and excitability. This research centers on leveraging human cerebral organoids (COs) and CRISPR-Cas9 genome editing to develop an in vitro epilepsy model. Utilizing advanced lattice light-sheet microscopy (LLSM), this study captures live 4D (x, y, z, time) morphological and dynamic data of mitochondria in COs, allowing for a comprehensive analysis of mitochondrial network behavior and its response to KD treatment. The study aims to elucidate the mechanisms through which the KD modulates mitochondrial structure and function, potentially guiding the development of novel therapeutics for drug-resistant epilepsy. By integrating cutting-edge in vitro human neural tissue models, CRISPR-Cas9 genome editing, and innovative live tissue 4D microscopy techniques, this thesis seeks to advance our understanding of the KD's anti-seizure mechanisms and contribute to the broader field of epilepsy research.