title: Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons.
abstract: The direct conversion of fibroblasts to induced dopaminergic (iDA) neurons and other cell types demonstrates the plasticity of cell fate. The low efficiency of these relatively fast conversions suggests that kinetic barriers exist to safeguard cell-type identity. Here we show that suppression of p53, in conjunction with cell cycle arrest at G1 and appropriate extracellular environment, markedly increase the efficiency in the transdifferentiation of human fibroblasts to iDA neurons by Ascl1, Nurr1, Lmx1a and miR124. The conversion is dependent on Tet1, as G1 arrest, p53 knockdown or expression of the reprogramming factors induces Tet1 synergistically. Tet1 knockdown abolishes the transdifferentiation while its overexpression enhances the conversion. The iDA neurons express markers for midbrain DA neurons and have active dopaminergic transmission. Our results suggest that overcoming these kinetic barriers may enable highly efficient epigenetic reprogramming in general and will generate patient-specific midbrain DA neurons for Parkinson's disease research and therapy.
authors.affiliation: Veterans Affairs Western New York Healthcare System, Buffalo, New York 14215, USA.|Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
authors.forename: Houbo
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.|Department of Neurology and Institute of Neurology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Xu
authors.affiliation: Veterans Affairs Western New York Healthcare System, Buffalo, New York 14215, USA.|Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhong
authors.affiliation: Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ren
authors.affiliation: Howard Hughes Medical Institute, Departments of Genetics &Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Gaoyang
authors.initials: G
authors.lastname: Liang
authors.affiliation: Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
authors.forename: Haley A
authors.initials: HA
authors.lastname: Schilling
authors.affiliation: Center for Computational Research, New York State Center of Excellence in Bioinformatics &Life Sciences, State University of New York at Buffalo, Buffalo, New York 14260, USA.
authors.forename: Zihua
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Howard Hughes Medical Institute, Departments of Genetics &Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Neurology and Institute of Neurology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Shengdi
authors.initials: S
authors.lastname: Chen
authors.affiliation: Veterans Affairs Western New York Healthcare System, Buffalo, New York 14215, USA.|Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Yan
authors.affiliation: Veterans Affairs Western New York Healthcare System, Buffalo, New York 14215, USA.|Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.|Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Feng
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D059447:Cell Cycle Checkpoints; D002460:Cell Line; D054338:Cell Transdifferentiation / Q000235:genetics*; D065150:Cellular Reprogramming; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D059585:G1 Phase Cell Cycle Checkpoints / Q000235:genetics*; D055785:Gene Knockdown Techniques; D006801:Humans; D060850:LIM-Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D008636:Mesencephalon; D035683:MicroRNAs / Q000235:genetics; D006899:Mixed Function Oxygenases; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2 / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D060850:LIM-Homeodomain Proteins; D035683:MicroRNAs; D006899:Mixed Function Oxygenases; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D011518:Proto-Oncogene Proteins; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; C065500:ASCL1 protein, human; C078021:LMX1A protein, human; C509207:MIRN124 microRNA, human; C539882:NR4A2 protein, human; C495901:TP53 protein, human; C462539:TET1 protein, human