Disease Modeling and Gene Therapy of Copper Storage Disease in Canine Hepatic Organoids

•Canine hepatic organoids can be grown long term with chromosomal stability•Organoids can be derived from liver tissues as small as fine-needle biopsies•Differentiation and proliferation of canine organoids involve Wnt and Notch•Successful gene supplementation proves potential for stem cell-based gene therapy

SummaryThe 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.