Genetic introduction of thyroid hormone transporter into the Bood-Brain-Barrier

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It is very important to claify the transport system via blood-brain barrier system. Recently, we have identified that organic anion transporter oatp2 and oatp3 are expressed in the choroid plexus and it transport thyroid hormones.The aim of this study is to extablished an adenovirus-mediated gene transfer system into cells to deliver drugs in to brain. The coding region of LST-1 and LST-2, both of which have isolated by us have introduced into adenovirus vector, packaged, and transfected into HEK293 cell with adenovirus genomic DNA.The cells that produces homologous-recombinant shrieked and made plaque formation. After producing large scale infected medium, the recombinant adenovirus was purified by CsCl ultracentrifuge.The infection titer of the purified LST-1 and LST-2 adenoviruses were measured and the IC 50 of each virus was 1.26 and 1.48 X 10^<10> pfu, respectively. When recombinant virus was applied to cultured cell line, the LST-1 and LST-2 protein was strongly expressed within 24 hr after application. The size of the band was also confirmed by Western blot analysis. The LST-1 and LST-2 expressing mammalian cell transports various organic anions as substrates and the apparent Km value of taurocholate uptake is 24 micro M and 32 micro M, respectively and both of which is a comparative value of the native cells reported. Our study revaled that our system is very effective and useful tool to deliver drugs through cell membrane.