ADENOVIRUS-MEDIATED TRANSFER OF A FUNCTIONAL GAD GENE INTO NERVE-CELLS - POTENTIAL FOR THE TREATMENT OF NEUROLOGICAL DISEASES

The recent development of efficient virus-mediated gene transfer into nerve cells allows the prospect of new strategies for the treatment of drug-resistant neurological disorders. Some forms of epilepsy may be amenable to gene therapy. Although there is no obvious candidate gene, the consensual GABA hypothesis of epilepsy suggests that the gene may be beneficial. GAD gene expression may be useful in supplying the inh ibitory neurotransmitter GABA to particular critical brain territories . We show herein that a nonreplicative recombinant adenovirus carrying the 67 gene under the control of Rous sarcoma virus long terminal rep eat promoter is able to express the transgene in primary cultures of n eurons and glial cells. Expression of the GAD67 gene was assessed by i mmunoblotting and immunohistochemical analysis. We demonstrated the fu nctionality of the transgene, the expression of which resulted in prod uction of large amounts of GABA in neuronal and glial cell cultures. S ubstantial production of the enzyme was also detected for several week s in infected organotypic slices cultured from new-born rat hippocampa l tissues. The virally encoded GAD67 was also expressed in vivo in var ious brain areas involved in various neurological disorders and thus m ay be of value for the development of gene therapies.