Production and analyses of diverse models for human Wison's disease

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The LEC rat, an animal model of Wilson's disease, has a deletion in Atp7b gene homologous to the ATP7B and shows some of the clinical features similar to Wilson's disease. To further ascertain the function of ATP7B gene in copper metabolism and its relation to hepatic disease in vivo, human ATP7B cDNA with CXN promoter was constructed and introduced into the LEC rats by prenuclear microinjection. The transgenic rats (Tg) expressed human ATP7B transcripts and protein in their multiple tissues, including liver, kidney, brain and muscle etc. The elevated ceruloplasmin ferroxidase activity and copper concentration in Tg plasma was confirmed, corresponding to the appearance of holoceruloplasmin (Holo-CPN) detected by western blot analysis. Copper content in Tg bile increased by nearly 2 fold of that in non-Tg littermates from 16 weeks of age. GOT and GPT activities in Tg much decreased compared with those in non-TG littermate. Long-term survival in Tg rats was up approximately to 97%, in co … More ntrast with 26.3% survival in non-Tg littermates. Long-team survival in Tg rats was up approximately to 97%, in contrast with 26.3% survival in non-Tg littermates. These findings and histological examination indicate that Tg rats were rescued from fulminant hepatitis and exhibited late onset of hepatic fibrosis. In addition, the products derived from ATP7B involve in intercellular copper transport by incorporation with CPN and by biliary excretion pathway. These results indicated that human ATP7B could partially complement the function of rat atp7b by gene transfer in vivo. Significant deprivation of hepatic iron from the Tg liver was also found, indicating that ATP7B may also act function in iron metabolism, and iron probably plays an essential role in the development of hepatic abnormalities in Wilson's disease. This successful rescue of Tg rats has important implication for the gene therapy for Wilson's disease and the Tg system will be useful to further study the function of ATP7B, its relation to copper or iron metabolism and the development of hepatic disease. Less