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 diseases. 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 GAD gene may be beneficial. GAD gene expression may be useful in supplying the inhibitory neurotransmitter GABA to particular critical brain territories. We show herein that a nonreplicative recombinant adenovirus carrying the GAD67 gene under the control of Rous sarcoma virus long terminal repeat promoter is able to express the transgene in primary cultures of neurons and glial cells. Expression of the GAD67 gene was assessed by immunoblotting and immunohistochemical analysis. We demonstrated the functionality of the transgene, the expression of which resulted in production of large amounts of GABA in neuronal and glial cell cultures. Substantial production of the enzyme was also detected for several weeks in infected organotypic slices cultured from new-born rat hippocampal tissues. The virally encoded GAD67 was also expressed in vivo in various brain areas involved in various neurological disorders and thus may be of value for the development of gene therapies.
Robert
J J
JJ
Laboratoire mixte Rhône-Poulenc-Rorer/CNRS, Génétique Moléculaire de la Neurotransmission et des Processus Neurodégénératifs, Hôpital de la Pitié Salpétrière, Paris, France.
Bouilleret
V
V
Ridoux
V
V
Valin
A
A
Geoffroy
M C
MC
Mallet
J
J
Le Gal La Salle
G
G
D000256:Adenoviridae; D000818:Animals; D001358:Avian Sarcoma Viruses; D002478:Cells, Cultured; D004827:Epilepsy / Q000628:therapy*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006624:Hippocampus / Q000201:enzymology; D015151:Immunoblotting; D007150:Immunohistochemistry; D009457:Neuroglia / Q000201:enzymology; D009474:Neurons / Q000201:enzymology; D009924:Organ Culture Techniques; D051381:Rats; D017208:Rats, Wistar; D019076:Transgenes; D005680:gamma-Aminobutyric Acid / Q000032:analysis / Q000235:genetics*
D005680:gamma-Aminobutyric Acid