Altered Protein Phosphorylation in a Novel Midbrain Organoid Model for Bipolar Disorder

Bipolar disorder (BD) is a severe psychiatric condition marked by episodes of mania and depression, with neurotransmitter imbalance in the midbrain believed to play a critical role in its pathophysiology. Despite this, there is currently no validated midbrain model for examining BD-associated molecular changes available. Leveraging recent advances in stem cell technology, we developed a midbrain organoid model using human induced pluripotent stem cells (hiPSCs) from BD patients and healthy controls (CTR). To address issues of variability and enhance the throughput in organoid production, we implemented liquid handling and high-content imaging techniques. Quality control metrics were established to identify organoids unsuitable for further study. Electrophysiological analysis via high-density microelectrode arrays (MEAs) revealed significantly elevated neuronal properties in individual BD organoids, including increased mean amplitude, conduction velocity, and extended axonal and dendritic growth. Transcriptome and proteome analyses indicated significant dysregulation of BD-relevant signaling pathways—such as those involving phosphatidylinositol, glycogen synthase kinase-3 beta, and AKT. Notably, we identified dysregulated casein kinase 2 (CSNK2A1) and calmodulin 3 (CALM3) in BD organoids, which were reversed by lithium treatment, highlighting potential novel targets for therapeutic intervention. This study validates the midbrain organoid model as a valuable tool for exploring the molecular underpinnings of BD and identifying new treatment avenues.