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Nat Commun. 2019 Feb 7;10(1):632. doi: 10.1038/s41467-018-08020-0.

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

Author information

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. paddison@fredhutch.org.
10
Human Biology Division, Fred Hutchinson Cancer Research Center, Seattle, WA, 98109, USA. paddison@fredhutch.org.
11
Department of Biochemistry, University of Washington, Seattle, WA, 98195, USA. hannele@uw.edu.
12
Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, 98109, USA. hannele@uw.edu.

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.

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