Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency

  • Nat Commun. 2019 Feb 7;10(1):632. doi: 10.1038/s41467-018-08020-0.
J Mathieu  1  2  3 ,  D Detraux  1  2  4 ,  D Kuppers  5 ,  Y Wang  2  6 ,  C Cavanaugh  2  3 ,  S Sidhu  1  2 ,  S Levy  1  2 ,  A M Robitaille  2  7 ,  A Ferreccio  1  2 ,  T Bottorff  1  2 ,  A McAlister  1  2 ,  L Somasundaram  1  2 ,  F Artoni  1  2 ,  S Battle  2  8 ,  R D Hawkins  2  8 ,  R T Moon  2  7 ,  C B Ware  2  3 ,  P J Paddison  9  10 ,  H Ruohola-Baker  11  12
Affiliations
  • 1. Department of Biochemistry, University of Washington, Seattle, WA, 98195, USA.
  • 2. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, 98109, USA.
  • 3. Department of Comparative Medicine, University of Washington, Seattle, WA, 98109, USA.
  • 4. Laboratory of Cellular Biochemistry and Biology (URBC), University of Namur, Namur, 5000, Belgium.
  • 5. Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA, 98109, USA.
  • 6. Paul G. Allen School of Computer Science & Engineering, University of Washington, Seattle, WA, 98109, USA.
  • 7. Department of Pharmacology, University of Washington, Seattle, WA, 98195, USA.
  • 8. Department of Medical Genetics & Genome Sciences, University of Washington, Seattle, WA, 98195, USA.
  • 9. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, 98109, USA. [email protected].
  • 10. Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA, 98109, USA. [email protected].
  • 11. Department of Biochemistry, University of Washington, Seattle, WA, 98195, USA. [email protected].
  • 12. Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, 98109, USA. [email protected].
PMID: 30733432 DOI: 10.1038/s41467-018-08020-0
Abstract

To reveal how cells exit human pluripotency, we designed a CRISPR-Cas9 screen exploiting the metabolic and epigenetic differences between naïve and primed pluripotent cells. We identify the tumor suppressor, Folliculin(FLCN) as a critical gene required for the exit from human pluripotency. Here we show that FLCN Knock-out (KO) hESCs maintain the naïve pluripotent state but cannot exit the state since the critical transcription factor TFE3 remains active in the nucleus. TFE3 targets up-regulated in FLCN KO exit assay are members of Wnt pathway and ESRRB. Treatment of FLCN KO hESC with a Wnt Inhibitor, but not ESRRB/FLCN double mutant, rescues the cells, allowing the exit from the naïve state. Using co-immunoprecipitation and mass spectrometry analysis we identify unique FLCN binding partners. The interactions of FLCN with components of the mTOR pathway (mTORC1 and mTORC2) reveal a mechanism of FLCN function during exit from naïve pluripotency.