Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons

Houbo Jiang; Zhimin Xu; Ping Zhong; Yong Ren; Gaoyang Liang; Haley A Schilling; Zihua Hu; Yi Zhang; Xiaomin Wang; Shengdi Chen; Zhen Yan; Jian Feng

doi:10.1038/ncomms10100

Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons

Nat Commun. 2015 Dec 7:6:10100. doi: 10.1038/ncomms10100.

Authors

Houbo Jiang^{1

2}, Zhimin Xu^{2

3}, Ping Zhong^{1

2}, Yong Ren², Gaoyang Liang⁴, Haley A Schilling², Zihua Hu⁵, Yi Zhang⁴, Xiaomin Wang⁶, Shengdi Chen³, Zhen Yan^{1

2}, Jian Feng^{1

2

6}

Affiliations

¹ Veterans Affairs Western New York Healthcare System, Buffalo, New York 14215, USA.
² Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214, USA.
³ Department of Neurology and Institute of Neurology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
⁴ Howard Hughes Medical Institute, Departments of Genetics &Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA.
⁵ 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.
⁶ Department of Neurobiology, Key Laboratory for Neurodegenerative Disorders of the Ministry of Education, Beijing Institute for Brain Disorders, Capital Medical University, Beijing 100069, China.

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.

Publication types

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

MeSH terms

Basic Helix-Loop-Helix Transcription Factors / genetics
Basic Helix-Loop-Helix Transcription Factors / metabolism
Cell Cycle
Cell Cycle Checkpoints
Cell Line
Cell Transdifferentiation / genetics*
Cellular Reprogramming
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism
Dopaminergic Neurons / cytology*
Fibroblasts / cytology*
G1 Phase Cell Cycle Checkpoints / genetics*
Gene Knockdown Techniques
Humans
LIM-Homeodomain Proteins / genetics
LIM-Homeodomain Proteins / metabolism
Mesencephalon
MicroRNAs / genetics
Mixed Function Oxygenases
Nuclear Receptor Subfamily 4, Group A, Member 2 / genetics
Nuclear Receptor Subfamily 4, Group A, Member 2 / metabolism
Proto-Oncogene Proteins / genetics
Proto-Oncogene Proteins / metabolism
Transcription Factors / genetics
Transcription Factors / metabolism
Tumor Suppressor Protein p53 / genetics*

Substances

ASCL1 protein, human
Basic Helix-Loop-Helix Transcription Factors
DNA-Binding Proteins
LIM-Homeodomain Proteins
LMX1A protein, human
MIRN124 microRNA, human
MicroRNAs
NR4A2 protein, human
Nuclear Receptor Subfamily 4, Group A, Member 2
Proto-Oncogene Proteins
TP53 protein, human
Transcription Factors
Tumor Suppressor Protein p53
Mixed Function Oxygenases
TET1 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding