title: Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system.
abstract: The enteric nervous system (ENS) of the gastrointestinal tract controls many diverse functions, including motility and epithelial permeability. Perturbations in ENS development or function are common, yet there is no human model for studying ENS-intestinal biology and disease. We used a tissue-engineering approach with embryonic and induced pluripotent stem cells (PSCs) to generate human intestinal tissue containing a functional ENS. We recapitulated normal intestinal ENS development by combining human-PSC-derived neural crest cells (NCCs) and developing human intestinal organoids (HIOs). NCCs recombined with HIOs in vitro migrated into the mesenchyme, differentiated into neurons and glial cells and showed neuronal activity, as measured by rhythmic waves of calcium transients. ENS-containing HIOs grown in vivo formed neuroglial structures similar to a myenteric and submucosal plexus, had functional interstitial cells of Cajal and had an electromechanical coupling that regulated waves of propagating contraction. Finally, we used this system to investigate the cellular and molecular basis for Hirschsprung's disease caused by a mutation in the gene PHOX2B. This is, to the best of our knowledge, the first demonstration of human-PSC-derived intestinal tissue with a functional ENS and how this system can be used to study motility disorders of the human gastrointestinal tract.
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Workman
authors.affiliation: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Maxime M
authors.identifier: 0000-0003-2560-193X
authors.initials: MM
authors.lastname: Mahe
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: Trisno
authors.affiliation: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Holly M
authors.initials: HM
authors.lastname: Poling
authors.affiliation: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Carey L
authors.initials: CL
authors.lastname: Watson
authors.affiliation: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Nambirajan
authors.initials: N
authors.lastname: Sundaram
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.|Division of Plastic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Ching-Fang
authors.initials: CF
authors.lastname: Chang
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Schiesser
authors.affiliation: INSERM UMR 913, University of Nantes, CHU de Nantes-Institut des Maladies de l'Appareil Digestif, Nantes, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Aubert
authors.affiliation: Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, Victoria, Australia.|Department of Pediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria, Australia.|Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, Victoria, Australia.
authors.forename: Edouard G
authors.initials: EG
authors.lastname: Stanley
authors.affiliation: Murdoch Children's Research Institute, The Royal Children's Hospital, Parkville, Victoria, Australia.|Department of Pediatrics, Faculty of Medicine, Dentistry and Health Sciences, University of Melbourne, Parkville, Victoria, Australia.|Faculty of Medicine, Nursing and Health Sciences, Monash University, Clayton, Victoria, Australia.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Elefanty
authors.affiliation: Gladstone Institutes, San Francisco, California, USA.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Miyaoka
authors.affiliation: Gladstone Institutes, San Francisco, California, USA.
authors.forename: Mohammad A
authors.initials: MA
authors.lastname: Mandegar
authors.affiliation: Gladstone Institutes, San Francisco, California, USA.|Departments of Medicine and Molecular and Cellular Pharmacology, University of California at San Francisco, San Francisco, California, USA.
authors.forename: Bruce R
authors.initials: BR
authors.lastname: Conklin
authors.affiliation: INSERM UMR 913, University of Nantes, CHU de Nantes-Institut des Maladies de l'Appareil Digestif, Nantes, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Neunlist
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.|Division of Plastic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Samantha A
authors.initials: SA
authors.lastname: Brugmann
authors.affiliation: Division of Pediatric General and Thoracic Surgery, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Helmrath
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.|Division of Endocrinology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Wells
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002642:Chick Embryo; D017615:Enteric Nervous System / Q000502:physiology* / Q000503:physiopathology; D005769:Gastrointestinal Motility; D006627:Hirschsprung Disease / Q000235:genetics / Q000503:physiopathology; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D057026:Induced Pluripotent Stem Cells; D056885:Interstitial Cells of Cajal / Q000502:physiology; D007422:Intestines / Q000502:physiology* / Q000503:physiopathology; D051379:Mice; D016513:Mice, SCID; D018613:Microscopy, Confocal; D008954:Models, Biological; D009154:Mutation; D009197:Myenteric Plexus / Q000502:physiology / Q000503:physiopathology; D009432:Neural Crest; D055495:Neurogenesis / Q000502:physiology; D009457:Neuroglia / Q000502:physiology; D009474:Neurons / Q000502:physiology; D009940:Organoids; D010539:Permeability; D060888:Real-Time Polymerase Chain Reaction; D013368:Submucous Plexus / Q000502:physiology / Q000503:physiopathology; D023822:Tissue Engineering / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics
chemical_list: D018398:Homeodomain Proteins; C109220:NBPhox protein; D014157:Transcription Factors; D002118:Calcium