Attenuation of seizures and neuronal death by adeno-associated virus vector galanin expression and secretion.
Seizure disorders present an attractive gene therapy target, particularly because viral vectors such as adeno-associated virus (AAV) and lentivirus can stably transduce neurons. When we targeted the N-methyl-D-aspartic acid (NMDA) excitatory amino acid receptor with an AAV-delivered antisense oligonucleotide, however, the promoter determined whether focal seizure sensitivity was significantly attenuated or facilitated. One potential means to circumvent this liability would be to express an inhibitory neuroactive peptide and constitutively secrete the peptide from the transduced cell. The neuropeptide galanin can modulate seizure activity in vivo, and the laminar protein fibronectin is usually secreted through a constitutive pathway. Initially, inclusion of the fibronectin secretory signal sequence (FIB) in an AAV vector caused significant gene product secretion in vitro. More importantly, the combination of this secretory signal with the coding sequence for the active galanin peptide significantly attenuated in vivo focal seizure sensitivity, even with different promoters, and prevented kainic acid-induced hilar cell death. Thus, neuroactive peptide expression and local secretion provides a new gene therapy platform for the treatment of neurological disorders.
Haberman
Rebecca P
RP
Gene Therapy Center, 7119 Thurston-Bowles Building, CB 7352, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599, USA.
Samulski
R Jude
RJ
McCown
Thomas J
TJ
D000818:Animals; D016923:Cell Death / Q000502:physiology*; D000229:Dependovirus / Q000235:genetics*; D004569:Electroencephalography; D018690:Excitatory Amino Acid Agonists / Q000378:metabolism; D019004:Galanin / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006624:Hippocampus / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007608:Kainic Acid / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008845:Microinjections; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012640:Seizures / Q000378:metabolism / Q000473:pathology / Q000628:therapy*
D018690:Excitatory Amino Acid Agonists; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D019004:Galanin; D007608:Kainic Acid