M.D. Undergraduate Central South University Changsha, Hunan, China (People's Republic)
Abstract: Developing effective organoid models for cerebellar diseases is crucial for understanding disease mechanisms and exploring new treatments. Spinocerebellar ataxia type 3 (SCA3) is a genetic neurodegenerative disease affecting the cerebellum with no effective treatment available. In our study, we used control and SCA3 induced-pluripotent stem cells to create human cerebellar organoids, following an updated 2024 protocol that improves cellular purity. These organoids were assessed for their development stages, cellular composition, and SCA3 pathology markers to evaluate their suitability for disease modeling. Through immunofluorescence staining, the organoids displayed markers of cerebellar neuron progenitors (PTF1A, ATOH1, SKOR2) and mature neurons (Calbindin, BARHL1, MAP2, NEUN, SYN1). Electrophysiological activity detected by multi-electrode array (MEA) revealed spikes and burst networks, indicating an integrated functional neuronal network. Crucially, immunofluorescence staining showed the presence of nuclear inclusion bodies, a hallmark of SCA3 pathology. Single-cell RNA sequencing is underway to further characterize the cellular composition and gene expression profiles within these organoids, providing deeper insights into cellular heterogeneity and mechanisms of neuronal vulnerability in early SCA3. Overall, our findings suggest that these organoids successfully replicate key aspects of SCA3 pathology and possess cerebellar characteristics, making them valuable for studying disease mechanisms and testing potential treatments for cerebellar diseases.
