title: AMPK governs lineage specification through Tfeb-dependent regulation of lysosomes.
abstract: Faithful execution of developmental programs relies on the acquisition of unique cell identities from pluripotent progenitors, a process governed by combinatorial inputs from numerous signaling cascades that ultimately dictate lineage-specific transcriptional outputs. Despite growing evidence that metabolism is integrated with many molecular networks, how pathways that control energy homeostasis may affect cell fate decisions is largely unknown. Here, we show that AMP-activated protein kinase (AMPK), a central metabolic regulator, plays critical roles in lineage specification. Although AMPK-deficient embryonic stem cells (ESCs) were normal in the pluripotent state, these cells displayed profound defects upon differentiation, failing to generate chimeric embryos and preferentially adopting an ectodermal fate at the expense of the endoderm during embryoid body (EB) formation. AMPK(-/-) EBs exhibited reduced levels of Tfeb, a master transcriptional regulator of lysosomes, leading to diminished endolysosomal function. Remarkably, genetic loss of Tfeb also yielded endodermal defects, while AMPK-null ESCs overexpressing this transcription factor normalized their differential potential, revealing an intimate connection between Tfeb/lysosomes and germ layer specification. The compromised endolysosomal system resulting from AMPK or Tfeb inactivation blunted Wnt signaling, while up-regulating this pathway restored expression of endodermal markers. Collectively, these results uncover the AMPK pathway as a novel regulator of cell fate determination during differentiation.
authors.affiliation: Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA;
authors.forename: Nathan P
authors.initials: NP
authors.lastname: Young
authors.affiliation: Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA;
authors.forename: Anwesh
authors.initials: A
authors.lastname: Kamireddy
authors.affiliation: Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA;
authors.forename: Jeanine L
authors.initials: JL
authors.lastname: Van Nostrand
authors.affiliation: Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA;
authors.forename: Lillian J
authors.initials: LJ
authors.lastname: Eichner
authors.affiliation: Integrative Genomics and Bioinformatics Core, The Salk Institute for Biological Studies, La Jolla, California 92037, USA;
authors.forename: Maxim Nikolaievich
authors.initials: MN
authors.lastname: Shokhirev
authors.affiliation: Transgenic Core Facility, The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
authors.forename: Yelena
authors.initials: Y
authors.lastname: Dayn
authors.affiliation: Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA;
authors.forename: Reuben J
authors.initials: RJ
authors.lastname: Shaw
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation; D019070:Cell Lineage / Q000235:genetics*; D053595:Embryonic Stem Cells; D004707:Endoderm / Q000473:pathology; D018507:Gene Expression Regulation, Developmental; D008247:Lysosomes / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D015398:Signal Transduction / Q000235:genetics; D060449:Wnt Signaling Pathway / Q000235:genetics
chemical_list: D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; C484068:Tcfeb protein, mouse; D055372:AMP-Activated Protein Kinases
keywords: AMPK; Tfeb; Wnt; embryonic stem cells; endoderm; germ layer specification; lysosomes