MTOR-mediated Activation of P70 S6K Induces Differentiation of Pluripotent Human Embryonic Stem Cells

Overview

Journal Cell Reprogram

Date 2010 Aug 12

PMID 20698768

Citations 74

Authors

Charles A Easley 4th

Ahmi Ben-Yehudah

Carrie J Redinger

Stacie L Oliver

Sandra T Varum

Vonya M Eisinger

Diane L Carlisle

Peter J Donovan

Gerald P Schatten

Affiliations

Soon will be listed here.

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.

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References

Pullen N, Thomas G . The modular phosphorylation and activation of p70s6k. FEBS Lett. 1997; 410(1):78-82. DOI: 10.1016/s0014-5793(97)00323-2. View

Varas F, Stadtfeld M, de Andres-Aguayo L, Maherali N, Di Tullio A, Pantano L . Fibroblast-derived induced pluripotent stem cells show no common retroviral vector insertions. Stem Cells. 2008; 27(2):300-6. PMC: 2729671. DOI: 10.1634/stemcells.2008-0696. View

Lowry W, Richter L, Yachechko R, Pyle A, Tchieu J, Sridharan R . Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc Natl Acad Sci U S A. 2008; 105(8):2883-8. PMC: 2268554. DOI: 10.1073/pnas.0711983105. View

Ludwig T, Levenstein M, Jones J, Berggren W, Mitchen E, Frane J . Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol. 2006; 24(2):185-7. DOI: 10.1038/nbt1177. View

Schalm S, Blenis J . Identification of a conserved motif required for mTOR signaling. Curr Biol. 2002; 12(8):632-9. DOI: 10.1016/s0960-9822(02)00762-5. View

Navara C, Redinger C, Mich-Basso J, Oliver S, Ben-Yehudah A, Castro C . Derivation and characterization of nonhuman primate embryonic stem cells. Curr Protoc Stem Cell Biol. 2008; Chapter 1:Unit 1A.1. DOI: 10.1002/9780470151808.sc01a01s1. View

Sampath P, Pritchard D, Pabon L, Reinecke H, Schwartz S, Morris D . A hierarchical network controls protein translation during murine embryonic stem cell self-renewal and differentiation. Cell Stem Cell. 2008; 2(5):448-60. DOI: 10.1016/j.stem.2008.03.013. View

Stadtfeld M, Nagaya M, Utikal J, Weir G, Hochedlinger K . Induced pluripotent stem cells generated without viral integration. Science. 2008; 322(5903):945-9. PMC: 3987909. DOI: 10.1126/science.1162494. View

Navara C, Mich-Basso J, Redinger C, Ben-Yehudah A, Jacoby E, Kovkarova-Naumovski E . Pedigreed primate embryonic stem cells express homogeneous familial gene profiles. Stem Cells. 2007; 25(11):2695-2704. PMC: 4357318. DOI: 10.1634/stemcells.2007-0286. View

10.

Strome S, Lehmann R . Germ versus soma decisions: lessons from flies and worms. Science. 2007; 316(5823):392-3. DOI: 10.1126/science.1140846. View

11.

Sabers C, Martin M, Brunn G, Williams J, Dumont F, Wiederrecht G . Isolation of a protein target of the FKBP12-rapamycin complex in mammalian cells. J Biol Chem. 1995; 270(2):815-22. DOI: 10.1074/jbc.270.2.815. View

12.

Blagosklonny M . Aging, stem cells, and mammalian target of rapamycin: a prospect of pharmacologic rejuvenation of aging stem cells. Rejuvenation Res. 2008; 11(4):801-8. DOI: 10.1089/rej.2008.0722. View

13.

Gingras A, Raught B, Sonenberg N . Regulation of translation initiation by FRAP/mTOR. Genes Dev. 2001; 15(7):807-26. DOI: 10.1101/gad.887201. View

14.

Wernig M, Meissner A, Cassady J, Jaenisch R . c-Myc is dispensable for direct reprogramming of mouse fibroblasts. Cell Stem Cell. 2008; 2(1):10-2. DOI: 10.1016/j.stem.2007.12.001. View

15.

Liu H, Zhu F, Yong J, Zhang P, Hou P, Li H . Generation of induced pluripotent stem cells from adult rhesus monkey fibroblasts. Cell Stem Cell. 2008; 3(6):587-90. DOI: 10.1016/j.stem.2008.10.014. View

16.

Li W, Wei W, Zhu S, Zhu J, Shi Y, Lin T . Generation of rat and human induced pluripotent stem cells by combining genetic reprogramming and chemical inhibitors. Cell Stem Cell. 2008; 4(1):16-9. DOI: 10.1016/j.stem.2008.11.014. View

17.

Easley 4th C, Brown C, Horwitz A, Tombes R . CaMK-II promotes focal adhesion turnover and cell motility by inducing tyrosine dephosphorylation of FAK and paxillin. Cell Motil Cytoskeleton. 2008; 65(8):662-74. PMC: 2830206. DOI: 10.1002/cm.20294. View

18.

Raught B, Gingras A, Sonenberg N . The target of rapamycin (TOR) proteins. Proc Natl Acad Sci U S A. 2001; 98(13):7037-44. PMC: 34619. DOI: 10.1073/pnas.121145898. View

19.

Weng Q, Kozlowski M, Belham C, Zhang A, Comb M, Avruch J . Regulation of the p70 S6 kinase by phosphorylation in vivo. Analysis using site-specific anti-phosphopeptide antibodies. J Biol Chem. 1998; 273(26):16621-9. DOI: 10.1074/jbc.273.26.16621. View

20.

Park I, Zhao R, West J, Yabuuchi A, Huo H, Ince T . Reprogramming of human somatic cells to pluripotency with defined factors. Nature. 2007; 451(7175):141-6. DOI: 10.1038/nature06534. View