title: Glutamine and alanyl-glutamine promote crypt expansion and mTOR signaling in murine enteroids.
abstract: L-glutamine (Gln) is a key metabolic fuel for intestinal epithelial cell proliferation and survival and may be conditionally essential for gut homeostasis during catabolic states. We show that L-alanyl-L-glutamine (Ala-Gln), a stable Gln dipeptide, protects mice against jejunal crypt depletion in the setting of dietary protein and fat deficiency. Separately, we show that murine crypt cultures (enteroids) derived from the jejunum require Gln or Ala-Gln for maximal expansion. Once expanded, enteroids deprived of Gln display a gradual atrophy of cryptlike domains, with decreased epithelial proliferation, but stable proportions of Paneth and goblet cell differentiation, at 24 h. Replenishment of enteroid medium with Gln selectively activates mammalian target of rapamycin (mTOR) signaling pathways, rescues proliferation, and promotes crypt regeneration. Gln deprivation beyond 48 h leads to destabilization of enteroids but persistence of EGFP-Lgr5-positive intestinal stem cells with the capacity to regenerate enteroids upon Gln rescue. Collectively, these findings indicate that Gln deprivation induces a reversible quiescence of intestinal stem cells and provides new insights into nutritional regulation of intestinal epithelial homeostasis.
authors: Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio; sean.moore@cchmc.org.
authors: Sean R
authors: SR
authors: Moore
authors: Department of Physiology and Pharmacology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceará, Fortaleza, Ceará, Brazil;
authors: Marjorie M
authors: MM
authors: Guedes
authors: Department of Morphology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceará, Fortaleza, Ceará, Brazil;
authors: Tie B
authors: TB
authors: Costa
authors: Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio;
authors: Jefferson
authors: J
authors: Vallance
authors: Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio;
authors: Elizabeth A
authors: EA
authors: Maier
authors: Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio;
authors: Kristina J
authors: KJ
authors: Betz
authors: Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, Ohio; and.
authors: Eitaro
authors: E
authors: Aihara
authors: Division of Pediatric Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio.
authors: Maxime M
authors: MM
authors: Mahe
authors: Department of Physiology and Pharmacology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceará, Fortaleza, Ceará, Brazil;
authors: Aldo A M
authors: AA
authors: Lima
authors: Department of Physiology and Pharmacology, Clinical Research Unit and Institute of Biomedicine/Center for Global Health, Faculty of Medicine, Federal University of Ceará, Fortaleza, Ceará, Brazil;
authors: Reinaldo B
authors: RB
authors: Oriá
authors: Division of Gastroenterology, Hepatology, and Nutrition, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio;
authors: Noah F
authors: NF
authors: Shroyer
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004151:Dipeptides / Q000378:metabolism*; D004847:Epithelial Cells; D005260:Female; D005973:Glutamine / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015398:Signal Transduction / Q000502:physiology; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*
chemical_list: D004151:Dipeptides; D005973:Glutamine; C546843:mTOR protein, mouse; D058570:TOR Serine-Threonine Kinases; C054122:alanylglutamine
keywords: ERK; intestinal organoids; l-alanyl-l-glutamine; l-glutamine; mammalian target of rapamycin