title: Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons.
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.
authors.affiliation: Department of Molecular Biology, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard, Dallas, TX 75390, USA.
authors.forename: Meng-Lu
authors.initials: ML
authors.lastname: Liu
authors.forename: Tong
authors.initials: T
authors.lastname: Zang
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Zou
authors.forename: Joshua C
authors.initials: JC
authors.lastname: Chang
authors.forename: Jay R
authors.initials: JR
authors.lastname: Gibson
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Huber
authors.forename: Chun-Li
authors.initials: CL
authors.lastname: Zhang
mesh_terms: D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000328:Adult; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation; D059329:Cholinergic Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology*; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000166:cytology / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D009469:Neuromuscular Junction / Q000166:cytology / Q000378:metabolism; D061251:Primary Cell Culture; D011720:Pyrazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D055751:SOXC Transcription Factors / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005576:Colforsin; D049452:Green Fluorescent Proteins; D009419:Nerve Tissue Proteins; D011720:Pyrazoles; D011743:Pyrimidines; D055751:SOXC Transcription Factors; C491183:NEUROG2 protein, human; C097473:SOX11 protein, human; C516138:dorsomorphin