Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons

Nat Commun. 2013;4:2183. doi: 10.1038/ncomms3183.

Abstract

Cell fate can be reprogrammed by modifying intrinsic and extrinsic cues. Here we show that two small molecules (forskolin and dorsomorphin) enable the transcription factor Neurogenin 2 (NGN2) to convert human fetal lung fibroblasts into cholinergic neurons with high purity (>90%) and efficiency (up to 99% of NGN2-expressing cells). The conversion is direct without passing through a proliferative progenitor state. These human induced cholinergic neurons (hiCN) show mature electrophysiological properties and exhibit motor neuron-like features, including morphology, gene expression and the formation of functional neuromuscular junctions. Inclusion of an additional transcription factor, SOX11, also efficiently converts postnatal and adult skin fibroblasts from healthy and diseased human patients to cholinergic neurons. Taken together, this study identifies a simple and highly efficient strategy for reprogramming human fibroblasts to subtype-specific neurons. These findings offer a unique venue for investigating the molecular mechanisms underlying cellular plasticity and human neurodegenerative diseases.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / drug effects
  • Action Potentials / physiology
  • Adult
  • Basic Helix-Loop-Helix Transcription Factors / genetics*
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Differentiation
  • Cholinergic Neurons / cytology
  • Cholinergic Neurons / drug effects*
  • Cholinergic Neurons / metabolism
  • Colforsin / pharmacology*
  • Fetus
  • Fibroblasts / cytology
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Gene Expression
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Lung / cytology
  • Lung / metabolism
  • Nerve Tissue Proteins / genetics*
  • Nerve Tissue Proteins / metabolism
  • Neuromuscular Junction / cytology
  • Neuromuscular Junction / metabolism
  • Primary Cell Culture
  • Pyrazoles / pharmacology*
  • Pyrimidines / pharmacology*
  • SOXC Transcription Factors / genetics*
  • SOXC Transcription Factors / metabolism
  • Time Factors

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • NEUROG2 protein, human
  • Nerve Tissue Proteins
  • Pyrazoles
  • Pyrimidines
  • SOX11 protein, human
  • SOXC Transcription Factors
  • dorsomorphin
  • Green Fluorescent Proteins
  • Colforsin

Associated data

  • GEO/GSE45954