TY - JOUR
T1 - Lysosomal Signaling Licenses Embryonic Stem Cell Differentiation via Inactivation of Tfe3
AU - Villegas, Florian
AU - Lehalle, Daphné
AU - Mayer, Daniela
AU - Rittirsch, Melanie
AU - Stadler, Michael B.
AU - Zinner, Marietta
AU - Olivieri, Daniel
AU - Vabres, Pierre
AU - Duplomb-Jego, Laurence
AU - De Bont, Eveline S.J.M.
AU - Duffourd, Yannis
AU - Duijkers, Floor
AU - Avila, Magali
AU - Geneviève, David
AU - Houcinat, Nada
AU - Jouan, Thibaud
AU - Kuentz, Paul
AU - Lichtenbelt, Klaske D.
AU - Thauvin-Robinet, Christel
AU - St-Onge, Judith
AU - Thevenon, Julien
AU - van Gassen, Koen L.I.
AU - van Haelst, Mieke
AU - van Koningsbruggen, Silvana
AU - Hess, Daniel
AU - Smallwood, Sebastien A.
AU - Rivière, Jean Baptiste
AU - Faivre, Laurence
AU - Betschinger, Joerg
N1 - Funding Information:
We would like to thank the patients and families who participated in this study. We also thank the University of Burgundy Centre de Calcul (CCuB) for technical support and management of the informatics platform; S. Dessus-Babus, K. Jacobeit, and T. Roloff (FMI) for processing sequencing samples; L. Gelman (FMI) for imaging assistance; H. Gut (FMI) for structural modelling; M. Flemr (FMI) for advice regarding genome editing; J. Chao (FMI), A. Smith (University of Cambridge), F. Stewart (University of Dresden), K. Yusa (Wellcome Trust Sanger Institute), and M. Leeb (MFPL) for providing reagents; and S. Gasser, P. Liberali, A. Peters, D. Schuebeler, N. Thomae (FMI), S. Stricker (Helmholtz Zentrum), and D. Teis (University of Innsbruck) for comments on the manuscript. This work was funded by EMBO and Marie Curie Actions ( ALTF 1632-2014 to D.O.), the Programme Hospitalier de Recherche Clinique (PHRC) National (to P.V.), the Regional Council of Burgundy and Centre Hospitalo-Universitaire de Dijon PARI 2015 (to L.F.), and the Novartis Research Foundation (to J.B.).
Funding Information:
We would like to thank the patients and families who participated in this study. We also thank the University of Burgundy Centre de Calcul (CCuB) for technical support and management of the informatics platform; S. Dessus-Babus, K. Jacobeit, and T. Roloff (FMI) for processing sequencing samples; L. Gelman (FMI) for imaging assistance; H. Gut (FMI) for structural modelling; M. Flemr (FMI) for advice regarding genome editing; J. Chao (FMI), A. Smith (University of Cambridge), F. Stewart (University of Dresden), K. Yusa (Wellcome Trust Sanger Institute), and M. Leeb (MFPL) for providing reagents; and S. Gasser, P. Liberali, A. Peters, D. Schuebeler, N. Thomae (FMI), S. Stricker (Helmholtz Zentrum), and D. Teis (University of Innsbruck) for comments on the manuscript. This work was funded by EMBO and Marie Curie Actions (ALTF 1632-2014 to D.O.), the Programme Hospitalier de Recherche Clinique (PHRC) National (to P.V.), the Regional Council of Burgundy and Centre Hospitalo-Universitaire de Dijon PARI 2015 (to L.F.), and the Novartis Research Foundation (to J.B.).
Publisher Copyright:
© 2018 Elsevier Inc.
PY - 2019/2/7
Y1 - 2019/2/7
N2 - Self-renewal and differentiation of pluripotent murine embryonic stem cells (ESCs) is regulated by extrinsic signaling pathways. It is less clear whether cellular metabolism instructs developmental progression. In an unbiased genome-wide CRISPR/Cas9 screen, we identified components of a conserved amino-acid-sensing pathway as critical drivers of ESC differentiation. Functional analysis revealed that lysosome activity, the Ragulator protein complex, and the tumor-suppressor protein Folliculin enable the Rag GTPases C and D to bind and seclude the bHLH transcription factor Tfe3 in the cytoplasm. In contrast, ectopic nuclear Tfe3 represses specific developmental and metabolic transcriptional programs that are associated with peri-implantation development. We show differentiation-specific and non-canonical regulation of Rag GTPase in ESCs and, importantly, identify point mutations in a Tfe3 domain required for cytoplasmic inactivation as potentially causal for a human developmental disorder. Our work reveals an instructive and biomedically relevant role of metabolic signaling in licensing embryonic cell fate transitions.
AB - Self-renewal and differentiation of pluripotent murine embryonic stem cells (ESCs) is regulated by extrinsic signaling pathways. It is less clear whether cellular metabolism instructs developmental progression. In an unbiased genome-wide CRISPR/Cas9 screen, we identified components of a conserved amino-acid-sensing pathway as critical drivers of ESC differentiation. Functional analysis revealed that lysosome activity, the Ragulator protein complex, and the tumor-suppressor protein Folliculin enable the Rag GTPases C and D to bind and seclude the bHLH transcription factor Tfe3 in the cytoplasm. In contrast, ectopic nuclear Tfe3 represses specific developmental and metabolic transcriptional programs that are associated with peri-implantation development. We show differentiation-specific and non-canonical regulation of Rag GTPase in ESCs and, importantly, identify point mutations in a Tfe3 domain required for cytoplasmic inactivation as potentially causal for a human developmental disorder. Our work reveals an instructive and biomedically relevant role of metabolic signaling in licensing embryonic cell fate transitions.
KW - developmental disorder
KW - differentiation
KW - embryonic stem cell
KW - Flcn
KW - mTOR
KW - pluripotency
KW - Rag GTPases
KW - Ragulator
KW - Tfe3
UR - https://www.scopus.com/pages/publications/85060114283
U2 - 10.1016/j.stem.2018.11.021
DO - 10.1016/j.stem.2018.11.021
M3 - Article
C2 - 30595499
AN - SCOPUS:85060114283
SN - 1934-5909
VL - 24
SP - 257-270.e8
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 2
ER -