Nicotinamide restricts neural precursor proliferation to enhance catecholaminergic neuronal subtype differentiation from mouse embryonic stem cells

Abstract

Emerging evidence indicates that a strong relationship exists between brain regenerative therapies and nutrition. Early life nutrition plays an important role during embryonic brain development, and there are clear consequences to an imbalance in nutritional factors on both the production and survival of mature neuronal populations and the infant’s risk of diseases in later life. Our research and that of others suggest that vitamins play a fundamental role in the formation of neurons and their survival. There is a growing body of evidence that nicotinamide, the water-soluble amide form of vitamin B 3 , is implicated in the conversion of pluripotent stem cells to clinically relevant cells for regenerative therapies. This study investigated the ability of nicotinamide to promote the development of mature catecholaminergic neuronal populations (associated with Parkinson’s disease) from mouse embryonic stem cells, as well as investigating the underlying mechanisms of nicotinamide’s action. Nicotinamide selectively enhanced the production of tyrosine hydroxylase-expressing neurons and serotonergic neurons from mouse embryonic stem cell cultures ( Sox1 GFP knock-in 46C cell line). A 5-Ethynyl-2´-deoxyuridine (EdU) assay ascertained that nicotinamide, when added in the initial phase, reduced cell proliferation. Nicotinamide drove tyrosine hydroxylase-expressing neuron differentiation as effectively as an established cocktail of signalling factors, reducing the proliferation of neural progenitors and accelerating neuronal maturation, neurite outgrowth and neurotransmitter expression. These novel findings show that nicotinamide enhanced and enriched catecholaminergic differentiation and inhibited cell proliferation by directing cell cycle arrest in mouse embryonic stem cell cultures, thus driving a critical neural proliferation-to-differentiation switch from neural progenitors to neurons. Further research into the role of vitamin metabolites in embryogenesis will significantly advance cell-based regenerative medicine, and help realize their role as crucial developmental signalling molecules in brain development.

Differential responses of primary neuron-secreted MCP-1 and IL-9 to type 2 diabetes and Alzheimer’s disease-associated metabolites

“…L3224) in the nano and micromolar range. We established the range we tested for our live/dead assay based on the ranges tested in these metabolites with different cell lines in prior studies 103 – 111 (Supplementary Table S3 ). Like the primary neuron culturing methods, one pregnant CD1 mouse at gestational day 15 (n = 12 embryos) was sacrificed and plated, with the adjustment of 175,000 cells/well in a 96-well plate.…”

Neonatal hair metabolome of birthweight discordant twins is associated with neurobehavioral impairments at 2-3 years of age

“…47 Nicotinamide has been identified to be associated with neuron differentiation by reducing the proliferation of neural progenitors and accelerating neuronal maturation, neurite outgrowth, and neurotransmitter expression. 48,49 We speculate that nicotinamide has a protective effect on fetal neurodevelopment during gestation, especially aiding the differentiation and maturation of nerve cells involved in motor and neurocognitive development.…”