Sirtuin 1 facilitates generation of induced pluripotent stem cells from mouse embryonic fibroblasts through the miR-34a and p53 pathways

PLoS One. 2012;7(9):e45633. doi: 10.1371/journal.pone.0045633. Epub 2012 Sep 21.

Abstract

Forced-expression of transcription factors can reprogram somatic cells into induced pluripotent stem cells (iPSC). Recent studies show that the reprogramming efficiency can be improved by inclusion of small molecules that regulate chromatin modifying enzymes. We report here that sirtuin 1 (SIRT1), a member of the sirtuin family of NAD(+)-dependent protein deacetylases, is involved in iPSC formation. By using an efficient mouse secondary fibroblast reprogramming system with doxycycline (DOX) inducible Yamanaka's transcription factors delivered by piggyBac (PB) transposition (2°F/1B MEF), we show that SIRT1 knockdown decreased while resveratrol (RSV) increased the efficiency of iPSC formation. The treatments were associated with altered acetylated p53 and its downstream Nanog but not p21 expression. The stimulatory effect was also confirmed by SIRT1 over-expression, which stimulated the formation of colonies with induced Nanog and reduced p21 expression. Furthermore, the effects of RSV and SIRT1 knockdown on reprogramming were most pronounced during the initiation phase of reprogramming. MicroRNA-34a is a known regulator of SIRT1. Its inhibitor increased, while its mimics reduced iPSC formation. The stimulatory effect of SIRT1 during reprogramming was also confirmed in the primary MEF. RSV increased while tenovin-6, a small molecule that activates p53 through SIRT1 inhibition, suppressed reprogramming. In conclusion, SIRT1 enhances iPSC generation, in part, through deacetylation of p53, inhibition of p21 and enhancement of Nanog expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Differentiation / physiology*
  • Cells, Cultured
  • Down-Regulation
  • Embryo, Mammalian / cytology*
  • Fibroblasts / cytology
  • Humans
  • Mice
  • MicroRNAs / metabolism*
  • Pluripotent Stem Cells / cytology*
  • Polymerase Chain Reaction
  • Sirtuin 1 / physiology*
  • Tumor Suppressor Protein p53 / metabolism*
  • Up-Regulation

Substances

  • MIRN34a microRNA, mouse
  • MicroRNAs
  • Tumor Suppressor Protein p53
  • Sirt1 protein, mouse
  • Sirtuin 1

Grants and funding

This work was supported by the General Research Fund (HKU775711M) from the Research Grant Council, Hong Kong (http://www.ugc.edu.hk/cgi-bin/ugc/search_project.pl). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.