Document Detail


MicroRNA-302 increases reprogramming efficiency via repression of NR2F2.
MedLine Citation:
PMID:  23136034     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
MicroRNAs (miRNAs) have emerged as critical regulators of gene expression through translational inhibition and RNA decay and have been implicated in the regulation of cellular differentiation, proliferation, angiogenesis, and apoptosis. In this study, we analyzed global miRNA and mRNA microarrays to predict novel miRNA-mRNA interactions in human embryonic stem cells and induced pluripotent stem cells (iPSCs). In particular, we demonstrate a regulatory feedback loop between the miR-302 cluster and two transcription factors, NR2F2 and OCT4. Our data show high expression of miR-302 and OCT4 in pluripotent cells, while NR2F2 is expressed exclusively in differentiated cells. Target analysis predicts that NR2F2 is a direct target of miR-302, which we experimentally confirm by reporter luciferase assays and real-time polymerase chain reaction. We also demonstrate that NR2F2 directly inhibits the activity of the OCT4 promoter and thus diminishes the positive feedback loop between OCT4 and miR-302. Importantly, higher reprogramming efficiencies were obtained when we reprogrammed human adipose-derived stem cells into iPSCs using four factors (KLF4, C-MYC, OCT4, and SOX2) plus miR-302 (this reprogramming cocktail is hereafter referred to as "KMOS3") when compared to using four factors ("KMOS"). Furthermore, shRNA knockdown of NR2F2 mimics the over-expression of miR-302 by also enhancing reprogramming efficiency. Interestingly, we were unable to generate iPSCs from miR-302a/b/c/d alone, which is in contrast to previous publications that have reported that miR-302 by itself can reprogram human skin cancer cells and human hair follicle cells. Taken together, these findings demonstrate that miR-302 inhibits NR2F2 and promotes pluripotency through indirect positive regulation of OCT4. This feedback loop represents an important new mechanism for understanding and inducing pluripotency in somatic cells.
Authors:
Shijun Hu; Kitchener D Wilson; Zhumur Ghosh; Leng Han; Yongming Wang; Feng Lan; Katherine J Ransohoff; Paul Burridge; Joseph C Wu
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Stem cells (Dayton, Ohio)     Volume:  31     ISSN:  1549-4918     ISO Abbreviation:  Stem Cells     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-02-13     Completed Date:  2013-07-18     Revised Date:  2014-03-19    
Medline Journal Info:
Nlm Unique ID:  9304532     Medline TA:  Stem Cells     Country:  United States    
Other Details:
Languages:  eng     Pagination:  259-68     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 AlphaMed Press.
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MeSH Terms
Descriptor/Qualifier:
Adipocytes / cytology,  drug effects*,  metabolism
COUP Transcription Factor II / antagonists & inhibitors,  genetics*,  metabolism
Cell Differentiation / drug effects
Feedback, Physiological
Female
Gene Expression Regulation / drug effects
Genes, Reporter
Humans
Induced Pluripotent Stem Cells / cytology,  drug effects*,  metabolism
Kruppel-Like Transcription Factors / genetics,  metabolism,  pharmacology
Luciferases
MicroRNAs / genetics*,  metabolism,  pharmacology
Microarray Analysis
Nuclear Reprogramming / drug effects,  genetics
Octamer Transcription Factor-3 / genetics*,  metabolism,  pharmacology
Primary Cell Culture
Promoter Regions, Genetic
Proto-Oncogene Proteins c-myc / genetics,  metabolism,  pharmacology
RNA, Small Interfering / genetics
SOXB1 Transcription Factors / genetics,  metabolism,  pharmacology
Signal Transduction / drug effects
Grant Support
ID/Acronym/Agency:
DP2 OD004437/OD/NIH HHS; DP2OD004437/OD/NIH HHS; R01 HL093172/HL/NHLBI NIH HHS; R01 HL113006/HL/NHLBI NIH HHS; R01 HL113006/HL/NHLBI NIH HHS; RC1 AG036142/AG/NIA NIH HHS; RC1 AG036142/AG/NIA NIH HHS; U01 HL099776/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/COUP Transcription Factor II; 0/GKLF protein; 0/Kruppel-Like Transcription Factors; 0/MIRN302 microRNA, human; 0/MYC protein, human; 0/MicroRNAs; 0/NR2F2 protein, human; 0/Octamer Transcription Factor-3; 0/POU5F1 protein, human; 0/Proto-Oncogene Proteins c-myc; 0/RNA, Small Interfering; 0/SOX2 protein, human; 0/SOXB1 Transcription Factors; EC 1.13.12.-/Luciferases
Comments/Corrections

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