Lysosomal Signaling Licenses Embryonic Stem Cell Differentiation via Inactivation of Tfe3

Florian Villegas (1, 2) , Daphne Lehalle (3, 4, 5, 6) , Daniela Mayer (1, 2) , Melanie Rittirsch (1) , Michael Stadler (7, 1) , Marietta Zinner (1, 2) , Daniel Olivieri (1) , Pierre Vabres (3, 5, 6, 8) , Laurence Duplomb-Jego (5, 6) , Eveline S.J.M de Bont (9, 10) , Yannis Duffourd (5, 6) , Floor Duijkers (11, 12) , Magali Avila (5, 6) , David Geneviève (13) , Nada Houcinat (3, 5, 6, 4) , Thibaud Jouan (5, 6) , Paul Kuentz (3, 5, 6, 4) , Klaske D. Lichtenbelt (14) , Christel Thauvin-Robinet (3, 5, 6, 4) , Judith St-Onge (5, 6, 15, 16) , Julien Thevenon (3, 5, 6, 4) , Koen L van Gassen (14) , Mieke M. van Haelst (11, 12) , Silvana van Koningsbruggen (11, 12) , Daniel Hess (1) , Sébastien A. Smallwood (1) , Jean-Baptiste Rivière (5, 6) , Laurence Faivre (3, 5, 6, 4) , Joerg Betschinger (1)

1 FMI - Friedrich Miescher Institute for Biomedical Research
2 Unibas - University of Basel
3 FHU TRANSLAD (CHU de Dijon)
4 CHU Dijon - Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand
5 Equipe GAD (LNC - U1231)
6 UBFC - Université Bourgogne Franche-Comté [COMUE]
7 SIB - Swiss Institute of Bioinformatics [Lausanne]
8 Service de Dermatologie (CHU de Dijon)
9 Beatrix Children's Hospital
10 UMCG - University Medical Center Groningen [Groningen]
11 Department of Clinical Genetics (Academic Medical Center, University of Amsterdam)
12 VU University Medical Center [Amsterdam]
13 Département de génétique médicale, maladies rares et médecine personnalisée [CHRU Montpellier]
14 University Medical Center [Utrecht]
15 Research Institute of the McGill University Hospital Centre
16 MUHC - McGill University Health Center [Montreal]

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.

Mots clés: embryonic stem cell; differentiation; pluripotency; mTOR; Flcn; Rag GTPases; Ragulator; Tfe3; developmental disorder

Domaines: Sciences du Vivant [q-bio]; Génétique; Génétique humaine

Florian Villegas, Daphne Lehalle, Daniela Mayer, Melanie Rittirsch, Michael Stadler, et al.. Lysosomal Signaling Licenses Embryonic Stem Cell Differentiation via Inactivation of Tfe3. Cell Stem Cell, 2019, 24 (2), pp.257-270.e8. ⟨10.1016/j.stem.2018.11.021⟩. ⟨hal-02005576⟩