(PTFS07-03-24) Investigating Specific Molecular and Functional Impacts of Citicoline on Brain Health

Objectives: Cytidine 5’-diphosphocholine (citicoline) is an endogenously generated nucleotide that serves as an essential intermediate in the synthesis of phosphatidylcholine (PC), the major phospholipid component of human cell membranes. Cognizin®, a citicoline supplement is widely used as a dietary supplement, and supplementation of citicoline has shown beneficial effects on cognitive function and behavior clinically. Despite these encouraging results, the biological mechanisms by which citicoline exerts a neuroprotective effect on brain function and ageing are not completely understood. The objective of this study was to perform an integrative multiomic and predictive modelling analysis to characterize the impacts of citicoline administration on brain health.

Methods: The experiments were performed with a human iPSC-derived tri-culture system consisting of cortical neurons, cortical astrocytes, and microglia cells from BrainXell Inc. Cells were cultivated on MEA plates to have a consistent substrate. The tri-cultures were cultured on 48 well MEAs (Axion Biosystems) for 3-4 weeks until maturation. Maturated cells were treated with citicoline then performed activity recording and RNA Sequencing experiments. The effect of the compounds on the cell activity were assessed using microelectrode arrays.

Results: Multi-parametric characterization of cell activity after treatment with citicoline or control were recorded. There was a statistically significant dose-dependent response in parameters for cell activity with the treatment of citicoline compared to control. The generation and interrogation of RNA-seq citicoline treatment signatures, accompanied by the investigation of comprehensive network model of neuronal cells revealed the predicted effects of citicoline.

Conclusions: This is the first study to investigate the comprehensive molecular mechanism of citicoline in an in-vitro neuronal tri-culture system with iPSC derived cells. The findings identified the potential impacts and relevance of citicoline supplementation in brain health. However, further analysis is required to decipher the molecular underpinnings of its mechanism of action.

Funding Sources: KIRIN Holdings Company, Limited