mTOR-mediated activation of p70 S6K induces differentiation of pluripotent human embryonic stem cells

Cell Reprogram. 2010 Jun;12(3):263-73. doi: 10.1089/cell.2010.0011.

Abstract

Deciding to exit pluripotency and undergo differentiation is of singular importance for pluripotent cells, including embryonic stem cells (ESCs). The molecular mechanisms for these decisions to differentiate, as well as reversing those decisions during induced pluripotency (iPS), have focused largely on transcriptomic controls. Here, we explore the role of translational control for the maintenance of pluripotency and the decisions to differentiate. Global protein translation is significantly reduced in hESCs compared to their differentiated progeny. Furthermore, p70 S6K activation is restricted in hESCs compared to differentiated fibroblast-like cells. Disruption of p70 S6K-mediated translation by rapamycin or siRNA knockdown in undifferentiated hESCs does not alter cell viability or expression of the pluripotency markers Oct4 and Nanog. However, expression of constitutively active p70 S6K, but not wild-type p70 S6K, induces differentiation. Additionally, hESCs exhibit high levels of the mTORC1/p70 S6K inhibitory complex TSC1/TSC2 and preferentially express more rapamycin insensitive mTORC2 compared to differentiated cells. siRNA-mediated knockdown of both TSC2 and Rictor elevates p70 S6K activation and induces differentiation of hESCs. These results suggest that hESCs tightly regulate mTORC1/p70 S6K-mediated protein translation to maintain a pluripotent state as well as implicate a novel role for protein synthesis as a driving force behind hESC differentiation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Differentiation / physiology*
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / drug effects
  • Enzyme Activation
  • Humans
  • Microscopy, Electron, Transmission
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects
  • RNA Interference
  • Ribosomal Protein S6 Kinases, 70-kDa / metabolism*
  • Sirolimus / pharmacology
  • TOR Serine-Threonine Kinases / physiology*

Substances

  • MTOR protein, human
  • Ribosomal Protein S6 Kinases, 70-kDa
  • TOR Serine-Threonine Kinases
  • Sirolimus