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

Abstract

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.

Long-Term Adult Feline Liver Organoid Cultures for Disease Modeling of Hepatic Steatosis

“…Feline liver organoids were mainly negative for ZO1, but in some organoids a weak membranous staining was observed in small clusters of cells within single organoids. Metaphase spread analysis showed a normal chromosome count in low and high passages, indicating long-term genetic stability of the cells similarly to liver organoids from other species (Figure 2D; Huch et al., 2013, Huch et al., 2015, Nantasanti et al., 2015). …”

Modeling Development and Disease with Organoids

“…Again, the cells could be converted into functional hepatocytes in vitro and upon transplantation into mice . The same protocols allowed long-term expansion of canine liver progenitor cells that could be differentiated toward functional hepatocytes (Nantasanti et al, 2015).…”

“…Liver organoids from dogs deficient in the copper-transporter COMMD1 mimicked the disease by accumulating toxic levels of copper, which could be salvaged by re-expression of wild-type COMMD1 protein (Nantasanti et al, 2015).…”

Intestinal Organoids—Current and Future Applications

“…Organoids have bene described in numerous tissues form men and mice [35], regarding companion animals only one paper described canine liver organoids [36]. Since growth factor dependency for the various organ/species derived organoids various greatly, the time discover the optimal culture conditions can be cumbersome.…”