A mesenchymal-to-epithelial transition initiates and is required for the nuclear reprogramming of mouse fibroblasts

Ronghui Li; Jialiang Liang; Su Ni; Ting Zhou; Xiaobing Qing; Huapeng Li; Wenzhi He; Jiekai Chen; Feng Li; Qiang Zhuang; Baoming Qin; Jianyong Xu; Wen Li; Jiayin Yang; Yi Gan; Dajiang Qin; Shipeng Feng; Hong Song; Dongshan Yang; Biliang Zhang; Lingwen Zeng; Liangxue Lai; Miguel Angel Esteban; Duanqing Pei

doi:10.1016/j.stem.2010.04.014

A mesenchymal-to-epithelial transition initiates and is required for the nuclear reprogramming of mouse fibroblasts

Cell Stem Cell. 2010 Jul 2;7(1):51-63. doi: 10.1016/j.stem.2010.04.014. Epub 2010 Jun 17.

Affiliation

¹ Stem Cell and Cancer Biology Group, Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.

PMID: 20621050
DOI: 10.1016/j.stem.2010.04.014

Abstract

Epithelial-to-mesenchymal transition (EMT) is a developmental process important for cell fate determination. Fibroblasts, a product of EMT, can be reset into induced pluripotent stem cells (iPSCs) via exogenous transcription factors but the underlying mechanism is unclear. Here we show that the generation of iPSCs from mouse fibroblasts requires a mesenchymal-to-epithelial transition (MET) orchestrated by suppressing pro-EMT signals from the culture medium and activating an epithelial program inside the cells. At the transcriptional level, Sox2/Oct4 suppress the EMT mediator Snail, c-Myc downregulates TGF-beta1 and TGF-beta receptor 2, and Klf4 induces epithelial genes including E-cadherin. Blocking MET impairs the reprogramming of fibroblasts whereas preventing EMT in epithelial cells cultured with serum can produce iPSCs without Klf4 and c-Myc. Our work not only establishes MET as a key cellular mechanism toward induced pluripotency, but also demonstrates iPSC generation as a cooperative process between the defined factors and the extracellular milieu. PAPERCLIP:

MeSH terms

Animals
Cadherins / genetics
Cadherins / metabolism
Cells, Cultured
Cellular Reprogramming / genetics
Cellular Reprogramming / physiology*
Chromatin Immunoprecipitation
Enzyme-Linked Immunosorbent Assay
Epithelial Cells / metabolism
Epithelial Cells / pathology*
Fibroblasts / cytology
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / metabolism
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors / genetics
Kruppel-Like Transcription Factors / metabolism
Mesoderm / metabolism
Mesoderm / pathology*
Mice
Models, Biological
Octamer Transcription Factor-3 / genetics
Octamer Transcription Factor-3 / metabolism
Proto-Oncogene Proteins c-myc / genetics
Proto-Oncogene Proteins c-myc / metabolism
Reverse Transcriptase Polymerase Chain Reaction
SOXB1 Transcription Factors / genetics
SOXB1 Transcription Factors / metabolism

Substances

Cadherins
Klf4 protein, mouse
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors
Myc protein, mouse
Octamer Transcription Factor-3
Pou5f1 protein, mouse
Proto-Oncogene Proteins c-myc
SOXB1 Transcription Factors
Sox2 protein, mouse

Associated data

GEO/GSE15267
GEO/GSE21062
GEO/GSE21064
GEO/GSE21067