Functional expression of Wilson's disease gene in the LEC rats by adenovirus-mediated gene delivery.

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WND gene (officially designated ATP7B), which encodes a putative copper transporting P-type ATPase, is defective in the patients with Wilson's disease, resulting in the excessive accumulation of copper in the liver. The Long-Evans Cinnamon (LEC) rat, known as a rodent model for Wilson's disease, shows some of clinical features similar to Wilson's disease. Atp7b, the rat gene homologue to WND,is also defective in the rat. To investigate the in vivo function of WND protein as well as its intracellular localization, WND cDNA was introduced to the LEC rats by recombinant adenovirus mediated gene delivery. The recombinant adenoviruses containing WND cDNA,1X10^<10> plaque forming units, were administered to 4-6 week old LEC rats by tail vein injection. Immunofluorescent study and subcellular fractionation study revealed the transgene expression in liver and its localization to the Golgi apparatus. Moreover, since the synthesis of holoceruloplasmin is disturbed in the LEC rat, the plasma level of holoceruloplasmin, oxidase active and copper bound form, was examined to evaluate the function of WND protein with respect to the copper transport. Consequently, the appearance of holoceruloplasmin in plasma was confirmed by Western blot analysis and plasma measurements for the oxidase activity and the copper content. Conclusions : 1) Introduced WND protein may function in the copper transport coupled with the synthesis of ceruloplasmin. 2) The Golgi apparatus is the likely site for WND protein to manifest its function. 3) Adenovirus mediated gene delivery could be a possible treatment for Wilson's disease.