LKB1 specifies neural crest cell fates through pyruvate-alanine cycling

Glial specification of neural crest cells requires the tumor suppressor LKB1-mediated action on alanine biosynthesis. Metabolic processes underlying the development of the neural crest, an embryonic population of multipotent migratory cells, are poorly understood. Here, we report that conditional ablation of the Lkb1 tumor suppressor kinase in mouse neural crest stem cells led to intestinal pseudo-obstruction and hind limb paralysis. This phenotype originated from a postnatal degeneration of the enteric nervous ganglia and from a defective differentiation of Schwann cells. Metabolomic profiling revealed that pyruvate-alanine conversion is enhanced in the absence of Lkb1. Mechanistically, inhibition of alanine transaminases restored glial differentiation in an mTOR-dependent manner, while increased alanine level directly inhibited the glial commitment of neural crest cells. Treatment with the metabolic modulator AICAR suppressed mTOR signaling and prevented Schwann cell and enteric defects of Lkb1 mutant mice. These data uncover a link between pyruvate-alanine cycling and the specification of glial cell fate with potential implications in the understanding of the molecular pathogenesis of neural crest diseases.

[1]  P. Trainor,et al.  Faculty Opinions recommendation of LKB1 specifies neural crest cell fates through pyruvate-alanine cycling. , 2020 .

[2]  A. Furlan,et al.  Schwann cell precursor: a neural crest cell in disguise? , 2018, Developmental biology.

[3]  J. Mao,et al.  mTOR acts as a pivotal signaling hub for neural crest cells during craniofacial development , 2018, PLoS genetics.

[4]  U. Suter,et al.  mTORC1 Is Transiently Reactivated in Injured Nerves to Promote c-Jun Elevation and Schwann Cell Dedifferentiation , 2018, The Journal of Neuroscience.

[5]  Sheng-Cai Lin,et al.  AMPK: Sensing Glucose as well as Cellular Energy Status. , 2017, Cell metabolism.

[6]  U. Suter,et al.  Dual function of the PI3K-Akt-mTORC1 axis in myelination of the peripheral nervous system. , 2017 .

[7]  J. Milbrandt,et al.  mTORC1 promotes proliferation of immature Schwann cells and myelin growth of differentiated Schwann cells , 2017, Proceedings of the National Academy of Sciences.

[8]  M. Bronner,et al.  Ancient evolutionary origin of vertebrate enteric neurons from trunk-derived neural crest , 2017, Nature.

[9]  D. Hayes,et al.  LKB1 loss links serine metabolism to DNA methylation and tumorigenesis , 2016, Nature.

[10]  D. Bouvard,et al.  LKB1 signaling in cephalic neural crest cells is essential for vertebrate head development. , 2016, Developmental biology.

[11]  H. Land,et al.  Addiction to Coupling of the Warburg Effect with Glutamine Catabolism in Cancer Cells , 2016, Cell reports.

[12]  M. Cobb,et al.  Amino Acids Regulate mTORC1 by an Obligate Two-step Mechanism* , 2016, The Journal of Biological Chemistry.

[13]  S. Gygi,et al.  Differential Glutamate Metabolism in Proliferating and Quiescent Mammary Epithelial Cells. , 2016, Cell metabolism.

[14]  J. Suttles,et al.  AMPK-dependent and independent effects of AICAR and compound C on T-cell responses , 2016, Oncotarget.

[15]  A. Clarke,et al.  Energy sensing and cancer: LKB1 function and lessons learnt from Peutz-Jeghers syndrome. , 2016, Seminars in cell & developmental biology.

[16]  F. Polleux,et al.  AMP-activated protein kinase mediates mitochondrial fission in response to energy stress , 2016, Science.

[17]  Xuesong Yang,et al.  High glucose environment inhibits cranial neural crest survival by activating excessive autophagy in the chick embryo , 2015, Scientific Reports.

[18]  F. Cagampang,et al.  AMPK Activation via Modulation of De Novo Purine Biosynthesis with an Inhibitor of ATIC Homodimerization. , 2015, Chemistry & biology.

[19]  Hideki Enomoto,et al.  Neuronal Differentiation in Schwann Cell Lineage Underlies Postnatal Neurogenesis in the Enteric Nervous System , 2015, The Journal of Neuroscience.

[20]  C. Wessig,et al.  mTORC1 controls PNS myelination along the mTORC1-RXRγ-SREBP-lipid biosynthesis axis in Schwann cells. , 2014, Cell reports.

[21]  Prof Vikas Kumar,et al.  The tumour suppressor LKB1 regulates myelination through mitochondrial metabolism , 2014, Nature Communications.

[22]  Takla Griss,et al.  Differential effects of AMPK agonists on cell growth and metabolism , 2014, Oncogene.

[23]  C. Glorieux,et al.  AICAR induces Nrf2 activation by an AMPK-independent mechanism in hepatocarcinoma cells. , 2014, Biochemical pharmacology.

[24]  T. Weichhart,et al.  mTORC1 Is Essential for Early Steps during Schwann Cell Differentiation of Amniotic Fluid Stem Cells and Regulates Lipogenic Gene Expression , 2014, PloS one.

[25]  J. Milbrandt,et al.  Metabolic regulator LKB1 is crucial for Schwann cell–mediated axon maintenance , 2014, Nature Neuroscience.

[26]  E. Ullian,et al.  Phosphorylation of LKB1/Par-4 Establishes Schwann Cell Polarity to Initiate and Control Myelin Extent , 2014, Nature Communications.

[27]  R. Morrison,et al.  p53 and mitochondrial function in neurons. , 2014, Biochimica et biophysica acta.

[28]  D. Sabatini,et al.  Regulation of mTORC1 by amino acids. , 2014, Trends in cell biology.

[29]  P. Codogno,et al.  Regulation of autophagy by amino acids and MTOR-dependent signal transduction , 2014, Amino Acids.

[30]  Shailendra Giri,et al.  Discrete mechanisms of mTOR and cell cycle regulation by AMPK agonists independent of AMPK , 2014, Proceedings of the National Academy of Sciences.

[31]  J. Ochocki,et al.  Nutrient-sensing pathways and metabolic regulation in stem cells , 2013, The Journal of cell biology.

[32]  D. Hayes,et al.  The LKB1 Tumor Suppressor as a Biomarker in Mouse and Human Tissues , 2013, PloS one.

[33]  D. Shackelford.  Unravelling the connection between metabolism and tumorigenesis through studies of the liver kinase B1 tumour suppressor , 2013, Journal of carcinogenesis.

[34]  R. Mayor,et al.  The neural crest , 2013, Development.

[35]  D. Hardie,et al.  LKB1 and AMPK and the cancer-metabolism link - ten years after , 2013, BMC Biology.

[36]  M. Mark,et al.  A Subpopulation of Smooth Muscle Cells, Derived from Melanocyte-Competent Precursors, Prevents Patent Ductus Arteriosus , 2013, PloS one.

[37]  C. Perou,et al.  LKB1/STK11 inactivation leads to expansion of a prometastatic tumor subpopulation in melanoma. , 2012, Cancer cell.

[38]  F. Fauvelle,et al.  Prediction of neuroprotective treatment efficiency using a HRMAS NMR-based statistical model of refractory status epilepticus on mouse: a metabolomic approach supported by histology. , 2012, Journal of proteome research.

[39]  L. Sommer,et al.  Neural crest progenitors and stem cells: from early development to adulthood. , 2012, Developmental biology.

[40]  B. Daignan-Fornier,et al.  5-Aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-Monophosphate (AICAR), a Highly Conserved Purine Intermediate with Multiple Effects , 2012, Metabolites.

[41]  K. Nave,et al.  Arrest of Myelination and Reduced Axon Growth When Schwann Cells Lack mTOR , 2012, The Journal of Neuroscience.

[42]  E. Huang,et al.  Homeodomain Interacting Protein Kinase 2 Regulates Postnatal Development of Enteric Dopaminergic Neurons and Glia via BMP Signaling , 2011, The Journal of Neuroscience.

[43]  P. Park,et al.  The Lkb1 metabolic sensor maintains haematopoietic stem cell survival , 2011, Nature.

[44]  M. Wegner,et al.  Sox10 is required for Schwann‐cell homeostasis and myelin maintenance in the adult peripheral nerve , 2011, Glia.

[45]  S. Schuster,et al.  Inhibition of Alanine Aminotransferase in Silico and in Vivo Promotes Mitochondrial Metabolism to Impair Malignant Growth* , 2011, The Journal of Biological Chemistry.

[46]  A. Clarke,et al.  LKB1 loss of function studied in vivo , 2011, FEBS letters.

[47]  P. Park,et al.  The Lkb1 metabolic sensor maintains haematopoietic stem cell survival , 2010, Nature.

[48]  V. Pachnis,et al.  Prospective Identification and Isolation of Enteric Nervous System Progenitors Using Sox2 , 2010, Stem cells.

[49]  N. deSouza,et al.  Detection of cancer in cervical tissue biopsies using mobile lipid resonances measured with diffusion‐weighted 1H magnetic resonance spectroscopy , 2010, NMR in biomedicine.

[50]  C. Lobe,et al.  Genomic localization of the Z/EG transgene in the mouse genome , 2009, Genesis.

[51]  S. Lyonnet,et al.  Deletion of Pten in the mouse enteric nervous system induces ganglioneuromatosis and mimics intestinal pseudoobstruction. , 2009, The Journal of clinical investigation.

[52]  R. Shaw,et al.  The LKB1–AMPK pathway: metabolism and growth control in tumour suppression , 2009, Nature Reviews Cancer.

[53]  L. Larue,et al.  The tyrosinase promoter is active in a subset of vagal neural crest cells during early development in mice , 2009, Pigment cell & melanoma research.

[54]  A. Hezel,et al.  LKB1; linking cell structure and tumor suppression , 2008, Oncogene.

[55]  H. Enomoto,et al.  Diminished Ret expression compromises neuronal survival in the colon and causes intestinal aganglionosis in mice. , 2008, The Journal of clinical investigation.

[56]  B. Turk,et al.  AMPK phosphorylation of raptor mediates a metabolic checkpoint. , 2008, Molecular cell.

[57]  M. Simon,et al.  The role of oxygen availability in embryonic development and stem cell function , 2008, Nature Reviews Molecular Cell Biology.

[58]  F. Nan,et al.  AICAR Induces Astroglial Differentiation of Neural Stem Cells via Activating the JAK/STAT3 Pathway Independently of AMP-activated Protein Kinase* , 2008, Journal of Biological Chemistry.

[59]  S. Fuchs,et al.  Establishment and controlled differentiation of neural crest stem cell lines using conditional transgenesis. , 2007, Differentiation; research in biological diversity.

[60]  J. Milbrandt,et al.  Conditional ablation of GFRα1 in postmigratory enteric neurons triggers unconventional neuronal death in the colon and causes a Hirschsprung's disease phenotype , 2007, Development.

[61]  B. Viollet,et al.  5′-AMP-Activated Protein Kinase (AMPK) Is Induced by Low-Oxygen and Glucose Deprivation Conditions Found in Solid-Tumor Microenvironments , 2006, Molecular and Cellular Biology.

[62]  Kei Sakamoto,et al.  LKB1-dependent signaling pathways. , 2006, Annual review of biochemistry.

[63]  V. Pachnis,et al.  Maintenance of mammalian enteric nervous system progenitors by SOX10 and endothelin 3 signalling , 2006, Development.

[64]  Shailendra Giri,et al.  5-Aminoimidazole-4-carboxamide-1-β-D-ribofuranoside Inhibits Cancer Cell Proliferation in Vitro and in Vivo via AMP-activated Protein Kinase* , 2005, Journal of Biological Chemistry.

[65]  R. Mirsky,et al.  The origin and development of glial cells in peripheral nerves , 2005, Nature Reviews Neuroscience.

[66]  U. Rutishauser,et al.  Adherens Junctions in Myelinating Schwann Cells Stabilize Schmidt-Lanterman Incisures via Recruitment of p120 Catenin to E-Cadherin , 2005, The Journal of Neuroscience.

[67]  R. DePinho,et al.  The LKB1 tumor suppressor negatively regulates mTOR signaling. , 2004, Cancer cell.

[68]  Hans C Clevers,et al.  Complete Polarization of Single Intestinal Epithelial Cells upon Activation of LKB1 by STRAD , 2004, Cell.

[69]  M. Rossel,et al.  Stability of the Peutz–Jeghers syndrome kinase LKB1 requires its binding to the molecular chaperones Hsp90/Cdc37 , 2003, Oncogene.

[70]  K. Inoki,et al.  TSC2 Mediates Cellular Energy Response to Control Cell Growth and Survival , 2003, Cell.

[71]  H C Clevers,et al.  Activation of the tumour suppressor kinase LKB1 by the STE20‐like pseudokinase STRAD , 2003, The EMBO journal.

[72]  L. Larue,et al.  SP-14 Cre-mediated recombination in the skin melanocyte lineage , 2003 .

[73]  Ronald A. DePinho,et al.  Loss of the Lkb1 tumour suppressor provokes intestinal polyposis but resistance to transformation , 2002, Nature.

[74]  Caiying Guo,et al.  Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon cre‐mediated excision , 2000, Genesis.

[75]  V. Govindaraju,et al.  Proton NMR chemical shifts and coupling constants for brain metabolites , 2000, NMR in biomedicine.

[76]  Chaya Kalcheim,et al.  The Neural Crest: Author Index , 1999 .

[77]  J. Milbrandt,et al.  GFRα1-Deficient Mice Have Deficits in the Enteric Nervous System and Kidneys , 1998, Neuron.

[78]  J. Avruch,et al.  Amino Acid Sufficiency and mTOR Regulate p70 S6 Kinase and eIF-4E BP1 through a Common Effector Mechanism* , 1998, The Journal of Biological Chemistry.

[79]  D. Hardie,et al.  AICA riboside increases AMP-activated protein kinase, fatty acid oxidation, and glucose uptake in rat muscle. , 1997, American journal of physiology. Endocrinology and metabolism.

[80]  David J. Anderson,et al.  Isolation of a stem cell for neurons and glia from the mammalian neural crest , 1992, Cell.

[81]  U. Suter,et al.  Dual function of the PI 3 K-Akt-mTORC 1 axis in myelination of the peripheral nervous system , 2018 .

[82]  D. Hayes,et al.  The LKB 1 Tumor Suppressor as a Biomarker in Mouse and Human Tissues , 2013 .

[83]  Philippe Soriano.  Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[84]  R. Polakiewicz,et al.  Lipopolysaccharide induces proenkephalin gene expression in rat lymph nodes and adrenal glands. , 1994, Endocrinology.

[85]  A. Sim,et al.  Location and function of three sites phosphorylated on rat acetyl-CoA carboxylase by the AMP-activated protein kinase. , 1990, European journal of biochemistry.