Demonstrating Potential of Cell Therapy for Wilson's Disease with the Long-Evans Cinnamon Rat Model.

Wilson's disease (WD) is characterized by the inability to excrete copper (Cu) from the body with progressive tissue injury, especially in liver and brain. The molecular defect in WD concerns mutations in ATP7B gene leading to loss of Cu transport from the hepatocyte to the bile canaliculus. While drugs, e.g., Cu chelators, have been available for several decades, these must be taken lifelong, which can be difficult due to issues of compliance or side effects. Many individuals may require liver transplantation, which can also be difficult due to donor organ shortages. Therefore, achieving permanent cures via cell or gene therapy are of great interest for WD. Cell therapy is feasible because transplanted hepatocytes can integrate in liver parenchyma and restore deficient functions, including transport of Cu into bile. The availability of authentic animal models that recapitulate hepatic WD, especially the Long-Evans Cinnamon (LEC) rat, has advanced cell transplantation research in WD. We describe requirements for cell therapy in animal models with several standardized methods for studies to test or refine cell therapy strategies in WD.

The Long Evans Cinnamon (LEC) rat develops hepatocellular damage in the absence of antimicrosomal antibodies.

Long Evans Cinnamon (LEC) rats are an inbred strain with a mutation affecting a copper transporter. As a result, hepatic copper levels rise and the rats spontaneously develop hepatitis that is fatal in about 40% of the rats. The rats that die have been reported to develop anti-microsomal antibodies, most frequently against protein disulfide isomerase (PDI), prior to the onset of liver damage. The association between the presence of antibodies and death of the LEC rats, along with the detection of antibodies prior to the detection of liver damage suggested that the antibodies may have a role in the pathogenesis of liver damage. The objective of this study was to more clearly delineate the temporal relationship between antibody production and the onset of liver damage and copper accumulation. Serum was screened for the presence of anti-microsomal antibodies by immunoblotting. Liver damage was assessed by serum biochemistry and histological examination on rats between 6 and 12 weeks of age (four per group). Copper accumulation in the liver was determined by biochemistry and histological examination. Evidence of liver damage was detectable by serum biochemistry and histopathology by 11.5 weeks. Copper was rarely detected in hepatocytes, although it was detected in macrophages. Sera from only one of seven rats with evidence of liver damage had detectable anti-microsomal protein antibodies. The protein recognized was not PDI. The development of anti-microsomal autoantibodies did not precede the development of significant liver damage, suggesting that they play only a secondary role, if any, in the pathogenesis of hepatitis in this rat strain.