NGN2 induces diverse neuron types from human pluripotency.

Area of Science:

• Neuroscience
• Stem Cell Biology
• Genetics

Background:

• Induced pluripotent stem cells (iPSCs) are crucial for modeling neurological diseases.
• Neurogenin 2 (NGN2) overexpression drives iPSC differentiation into neurons, but their heterogeneity is not fully understood.

Purpose of the Study:

• To investigate the cell states and molecular heterogeneity during NGN2-induced neuronal differentiation.
• To explore the impact of NGN2 expression levels and duration on neuron fate acquisition.

Main Methods:

• Single-cell transcriptomics was employed across a time course of NGN2 overexpression.
• Analysis spanned from pluripotency to functional neuronal maturation.

Main Results:

• Significant molecular heterogeneity was observed in NGN2-engineered neurons.
• At least two distinct neuron populations with peripheral nervous system gene expression profiles were identified.
• Neuron heterogeneity was consistent across multiple iPSC clones and lines.
• Neuron fate acquisition was sensitive to NGN2 expression level and duration.

Conclusions:

• NGN2 dosage is a critical regulator of neuron fate during iPSC differentiation.
• The observed heterogeneity in NGN2-induced neurons (NGN2-iNs) can confound studies sensitive to specific neuron types.