title: The Basic Helix-Loop-Helix Transcription Factor NEUROG3 Is Required for Development of the Human Endocrine Pancreas.
abstract: Neurogenin3 (NEUROG3) is a basic helix-loop-helix transcription factor required for development of the endocrine pancreas in mice. In contrast, humans with NEUROG3 mutations are born with endocrine pancreas function, calling into question whether NEUROG3 is required for human endocrine pancreas development. To test this directly, we generated human embryonic stem cell (hESC) lines where both alleles of NEUROG3 were disrupted using CRISPR/Cas9-mediated gene targeting. NEUROG3(-/-) hESC lines efficiently formed pancreatic progenitors but lacked detectible NEUROG3 protein and did not form endocrine cells in vitro. Moreover, NEUROG3(-/-) hESC lines were unable to form mature pancreatic endocrine cells after engraftment of PDX1(+)/NKX6.1(+) pancreatic progenitors into mice. In contrast, a 75-90% knockdown of NEUROG3 caused a reduction, but not a loss, of pancreatic endocrine cell development. We conclude that NEUROG3 is essential for endocrine pancreas development in humans and that as little as 10% NEUROG3 is sufficient for formation of pancreatic endocrine cells.
authors: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH.
authors: Patrick S
authors: PS
authors: McGrath
authors: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH Department of General Surgery, University of Cincinnati, Cincinnati, OH.
authors: Carey L
authors: CL
authors: Watson
authors: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH.
authors: Cameron
authors: C
authors: Ingram
authors: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, OH Department of General Surgery, University of Cincinnati, Cincinnati, OH.
authors: Michael A
authors: MA
authors: Helmrath
authors: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH Division of Endocrinology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH james.wells@cchmc.org.
authors: James M
authors: JM
authors: Wells
mesh_terms: D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000196:embryology*; D008969:Molecular Sequence Data; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; C498695:NEUROG3 protein, human
references: Nat Biotechnol. 2005 Dec;23(12):1534-41
references: 16258519
references: Dev Biol. 2007 Sep 15;309(2):298-305
references: 17706959
references: N Engl J Med. 2007 Apr 26;356(17):1781-2; author reply 1782
references: 17460236
references: EMBO J. 2002 Dec 2;21(23):6338-47
references: 12456641
references: Biochem J. 2007 Oct 15;407(2):277-84
references: 17623011
references: Cell. 2008 Jan 25;132(2):197-207
references: 18243096
references: Nat Biotechnol. 2008 Apr;26(4):443-52
references: 18288110
references: Dev Biol. 2008 May 15;317(2):531-40
references: 18394599
references: Nat Med. 2009 Jun;15(6):701-6
references: 19398967
references: Hum Pathol. 2009 Jul;40(7):1006-14
references: 19297006
references: Development. 2009 Nov;136(21):3567-74
references: 19793886
references: Development. 2010 Jan;137(2):203-12
references: 20040487
references: Dev Biol. 2010 Mar 1;339(1):26-37
references: 20025861
references: Nature. 2011 Feb 3;470(7332):105-9
references: 21151107
references: Diabetes. 2011 Apr;60(4):1349-53
references: 21378176
references: J Clin Endocrinol Metab. 2011 Jul;96(7):1960-5
references: 21490072
references: ScientificWorldJournal. 2012;2012:365104
references: 22778697
references: Diabetes. 2012 Aug;61(8):2016-29
references: 22740171
references: Cell Stem Cell. 2013 Apr 4;12(4):393-4
references: 23561441
references: J Biol Chem. 2013 Apr 26;288(17):11705-17
references: 23471965
references: Development. 2013 Jun;140(12):2472-83
references: 23715541
references: Diabetes. 2013 Oct;62(10):3514-22
references: 23630303
references: Pediatr Int. 2013 Oct;55(5):661-3
references: 24134759
references: Nat Protoc. 2013 Nov;8(11):2281-308
references: 24157548
references: Diabetologia. 2014 Nov;57(11):2421-4
references: 25120094
references: Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1607-11
references: 10677506
references: Genes Dev. 2003 Oct 15;17(20):2591-603
references: 14561778
references: Nature. 1997 Mar 27;386(6623):399-402
references: 9121556
references: Nature. 1999 Aug 26;400(6747):877-81
references: 10476967
references: Development. 2005 Jul;132(13):3139-49
references: 15944193
references: Mol Cell Biol. 2000 May;20(9):3292-307
references: 10757813
references: Development. 2000 Jun;127(12):2617-27
references: 10821760
references: Development. 2000 Aug;127(16):3533-42
references: 10903178
references: Biotechniques. 2000 Jul;29(1):52, 54
references: 10907076
references: Development. 2000 Dec;127(24):5533-40
references: 11076772
references: Diabetes. 2001 May;50(5):928-36
references: 11334435
references: Development. 2002 May;129(10):2447-57
references: 11973276
references: Genes Dev. 2002 Jun 15;16(12):1488-97
references: 12080087
references: EMBO J. 2006 Mar 22;25(6):1344-52
references: 16511571