title: Disease Modeling and Gene Therapy of Copper Storage Disease in Canine Hepatic Organoids.
abstract: The recent development of 3D-liver stem cell cultures (hepatic organoids) opens up new avenues for gene and/or stem cell therapy to treat liver disease. To test safety and efficacy, a relevant large animal model is essential but not yet established. Because of its shared pathologies and disease pathways, the dog is considered the best model for human liver disease. Here we report the establishment of a long-term canine hepatic organoid culture allowing undifferentiated expansion of progenitor cells that can be differentiated toward functional hepatocytes. We show that cultures can be initiated from fresh and frozen liver tissues using Tru-Cut or fine-needle biopsies. The use of Wnt agonists proved important for canine organoid proliferation and inhibition of differentiation. Finally, we demonstrate that successful gene supplementation in hepatic organoids of COMMD1-deficient dogs restores function and can be an effective means to cure copper storage disease.
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Sathidpak
authors.initials: S
authors.lastname: Nantasanti
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Bart
authors.initials: B
authors.lastname: Spee
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Hedwig S
authors.initials: HS
authors.lastname: Kruitwagen
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands; Hubrecht Institute and University Medical Centre, Utrecht, 3584 CT, the Netherlands.
authors.forename: Chen
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands; Hubrecht Institute and University Medical Centre, Utrecht, 3584 CT, the Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Geijsen
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Loes A
authors.initials: LA
authors.lastname: Oosterhoff
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Monique E
authors.initials: ME
authors.lastname: van Wolferen
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pelaez
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Hille
authors.initials: H
authors.lastname: Fieten
authors.affiliation: Centre for Cellular Imaging (CCI), Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CL, the Netherlands.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Wubbolts
authors.affiliation: Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CL, the Netherlands.
authors.forename: Guy C
authors.initials: GC
authors.lastname: Grinwis
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720-1460, USA.
authors.forename: Jefferson
authors.initials: J
authors.lastname: Chan
authors.affiliation: Hubrecht Institute and University Medical Centre, Utrecht, 3584 CT, the Netherlands.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Huch
authors.affiliation: Hubrecht Institute and University Medical Centre, Utrecht, 3584 CT, the Netherlands.
authors.forename: Robert R G
authors.initials: RRG
authors.lastname: Vries
authors.affiliation: Hubrecht Institute and University Medical Centre, Utrecht, 3584 CT, the Netherlands.
authors.forename: Hans
authors.initials: H
authors.lastname: Clevers
authors.affiliation: Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CL, the Netherlands; Department of Pediatrics, Division of Molecular Genetics, University Medical Center Groningen, University of Groningen, Groningen, 9713 AV, the Netherlands.
authors.forename: Alain
authors.initials: A
authors.lastname: de Bruin
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: Rothuizen
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands.
authors.forename: Louis C
authors.initials: LC
authors.lastname: Penning
authors.affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, 3584 CM, the Netherlands. Electronic address: b.a.schotanus@uu.nl.
authors.forename: Baukje A
authors.initials: BA
authors.lastname: Schotanus
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004285:Dogs; D015316:Genetic Therapy / Q000379:methods*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006527:Hepatolenticular Degeneration / Q000235:genetics / Q000628:therapy*; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism; D051153:Wnt Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051880:Receptors, Notch; D051153:Wnt Proteins