title: Proneural transcription factor Atoh1 drives highly efficient differentiation of human pluripotent stem cells into dopaminergic neurons.
abstract: Human pluripotent stem cells (PSCs) are a promising cell resource for various applications in regenerative medicine. Highly efficient approaches that differentiate human PSCs into functional lineage-specific neurons are critical for modeling neurological disorders and testing potential therapies. Proneural transcription factors are crucial drivers of neuron development and hold promise for driving highly efficient neuronal conversion in PSCs. Here, we study the functions of proneural transcription factor Atoh1 in the neuronal differentiation of PSCs. We show that Atoh1 is induced during the neuronal conversion of PSCs and that ectopic Atoh1 expression is sufficient to drive PSCs into neurons with high efficiency. Atoh1 induction, in combination with cell extrinsic factors, differentiates PSCs into functional dopaminergic (DA) neurons with >80% purity. Atoh1-induced DA neurons recapitulate key biochemical and electrophysiological features of midbrain DA neurons, the degeneration of which is responsible for clinical symptoms in Parkinson's disease (PD). Atoh1-induced DA neurons provide a reliable disease model for studying PD pathogenesis, such as neurotoxin-induced neurodegeneration in PD. Overall, our results determine the role of Atoh1 in regulating neuronal differentiation and neuron subtype specification of human PSCs. Our Atoh1-mediated differentiation approach will enable large-scale applications of PD patient-derived midbrain DA neurons in mechanistic studies and drug screening for both familial and sporadic PD.
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Sagal
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Xiping
authors.initials: X
authors.lastname: Zhan
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Jinchong
authors.initials: J
authors.lastname: Xu
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Tilghman
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Senthilkumar S
authors.initials: SS
authors.lastname: Karuppagounder
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Chen
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Valina L
authors.initials: VL
authors.lastname: Dawson
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: Ted M
authors.initials: TM
authors.lastname: Dawson
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
authors.forename: John
authors.initials: J
authors.lastname: Laterra
authors.affiliation: Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA ying@kennedykrieger.org.
authors.forename: Mingyao
authors.initials: M
authors.lastname: Ying
mesh_terms: D000200:Action Potentials; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D059290:Dopaminergic Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008636:Mesencephalon / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D058953:Neural Stem Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D055495:Neurogenesis / Q000187:drug effects; D016627:Oxidopamine / Q000633:toxicity; D013997:Time Factors; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D016627:Oxidopamine; C500942:ATOH1 protein, human
keywords: Basic helix-loop-helix transcription factors; Embryonic stem cells; Induced pluripotent stem cells; Parkinson’s disease; Tet-On