NGN2 induces diverse neuron types from human pluripotency

Summary

Human neurons engineered from induced pluripotent stem cells (iPSCs) through neurogenin 2 ( NGN2) overexpression are widely used to study neuronal differentiation mechanisms and to model neurological diseases. However, the differentiation paths and heterogeneity of emerged neurons have not been fully explored. Here, we used single-cell transcriptomics to dissect the cell states that emerge during NGN2 overexpression across a time course from pluripotency to neuron functional maturation. We find a substantial molecular heterogeneity in the neuron types generated, with at least two populations that express genes associated with neurons of the peripheral nervous system. Neuron heterogeneity is observed across multiple iPSC clones and lines from different individuals. We find that neuron fate acquisition is sensitive to NGN2 expression level and the duration of NGN2-forced expression. Our data reveal that NGN2 dosage can regulate neuron fate acquisition, and that NGN2-iN heterogeneity can confound results that are sensitive to neuron type.

Highlights

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  NGN2-iNs are molecularly heterogeneous

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  NGN2-iNs subtypes have signatures of central and peripheral nervous system

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  Neural fate acquisition is sensitive to the level and duration of NGN2 expression

Abstract

In this article, Lin, He, and Ebert et al. show that neurons induced through forced expression of NGN2 in iPSCs ( NGN2-iNs) present substantial molecular heterogeneity. Using scRNA-seq, they found NGN2-iN subtypes that express genes associated with the peripheral nervous system, and that neural fate acquisition is sensitive to the level and duration of NGN2 expression.