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

J. Mathieu; D. Detraux; D. Kuppers; Y. Wang; C. Cavanaugh; S. Sidhu; S. Levy; A. M. Robitaille; A. Ferreccio; T. Bottorff; A. McAlister; L. Somasundaram; F. Artoni; S. Battle; R. D. Hawkins; R. T. Moon; C. B. Ware; P. J. Paddison; H. Ruohola-Baker

doi:10.1038/s41467-018-08020-0

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

J. Mathieu, D. Detraux, D. Kuppers, Y. Wang, C. Cavanaugh, S. Sidhu, S. Levy, A. M. Robitaille, A. Ferreccio, T. Bottorff, A. McAlister, L. Somasundaram, F. Artoni, S. Battle, R. D. Hawkins, R. T. Moon, C. B. Ware, P. J. Paddison, H. Ruohola-Baker

Laboratory of Cellular Biochemistry and Biology

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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.

Original language	English
Article number	632
Pages (from-to)	632
Journal	Nature Communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-018-08020-0
Publication status	Published - 7 Feb 2019

Keywords

Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics
CRISPR-Cas Systems/genetics
Cell Line
Estrone/genetics
Humans
Immunoprecipitation
Mechanistic Target of Rapamycin Complex 1/genetics
Mechanistic Target of Rapamycin Complex 2/genetics
Proteomics
Receptors, Estrogen/genetics
Wnt Signaling Pathway/genetics

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Mathieu, J., Detraux, D., Kuppers, D., Wang, Y., Cavanaugh, C., Sidhu, S., Levy, S., Robitaille, A. M., Ferreccio, A., Bottorff, T., McAlister, A., Somasundaram, L., Artoni, F., Battle, S., Hawkins, R. D., Moon, R. T., Ware, C. B., Paddison, P. J., & Ruohola-Baker, H. (2019). Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency. Nature Communications, 10(1), 632. Article 632. https://doi.org/10.1038/s41467-018-08020-0

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title = "Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency",

abstract = "To reveal how cells exit human pluripotency, we designed a CRISPR-Cas9 screen exploiting the metabolic and epigenetic differences between na{\"i}ve 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 na{\"i}ve 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 na{\"i}ve 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 na{\"i}ve pluripotency.",

keywords = "Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics, CRISPR-Cas Systems/genetics, Cell Line, Estrone/genetics, Humans, Immunoprecipitation, Mechanistic Target of Rapamycin Complex 1/genetics, Mechanistic Target of Rapamycin Complex 2/genetics, Proteomics, Receptors, Estrogen/genetics, Wnt Signaling Pathway/genetics",

author = "J. Mathieu and D. Detraux and D. Kuppers and Y. Wang and C. Cavanaugh and S. Sidhu and S. Levy and Robitaille, {A. M.} and A. Ferreccio and T. Bottorff and A. McAlister and L. Somasundaram and F. Artoni and S. Battle and Hawkins, {R. D.} and Moon, {R. T.} and Ware, {C. B.} and Paddison, {P. J.} and H. Ruohola-Baker",

year = "2019",

month = feb,

day = "7",

doi = "10.1038/s41467-018-08020-0",

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journal = "Nature Communications",

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Mathieu, J, Detraux, D, Kuppers, D, Wang, Y, Cavanaugh, C, Sidhu, S, Levy, S, Robitaille, AM, Ferreccio, A, Bottorff, T, McAlister, A, Somasundaram, L, Artoni, F, Battle, S, Hawkins, RD, Moon, RT, Ware, CB, Paddison, PJ & Ruohola-Baker, H 2019, 'Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency', Nature Communications, vol. 10, no. 1, 632, pp. 632. https://doi.org/10.1038/s41467-018-08020-0

TY - JOUR

T1 - Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency

AU - Mathieu, J.

AU - Detraux, D.

AU - Kuppers, D.

AU - Wang, Y.

AU - Cavanaugh, C.

AU - Sidhu, S.

AU - Levy, S.

AU - Robitaille, A. M.

AU - Ferreccio, A.

AU - Bottorff, T.

AU - McAlister, A.

AU - Somasundaram, L.

AU - Artoni, F.

AU - Battle, S.

AU - Hawkins, R. D.

AU - Moon, R. T.

AU - Ware, C. B.

AU - Paddison, P. J.

AU - Ruohola-Baker, H.

PY - 2019/2/7

Y1 - 2019/2/7

N2 - 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.

AB - 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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KW - CRISPR-Cas Systems/genetics

KW - Cell Line

KW - Estrone/genetics

KW - Humans

KW - Immunoprecipitation

KW - Mechanistic Target of Rapamycin Complex 1/genetics

KW - Mechanistic Target of Rapamycin Complex 2/genetics

KW - Proteomics

KW - Receptors, Estrogen/genetics

KW - Wnt Signaling Pathway/genetics

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U2 - 10.1038/s41467-018-08020-0

DO - 10.1038/s41467-018-08020-0

M3 - Article

C2 - 30733432

AN - SCOPUS:85061262323

SN - 2041-1723

VL - 10

SP - 632

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 632

ER -

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

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Keywords

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