Anticonvulsant effects and behavioural outcomes of rAAV serotype 1 vector-mediated neuropeptide Y overexpression in rat hippocampus.
Neuropeptide Y (NPY) is an endogenous peptide with powerful anticonvulsant properties. Its overexpression in the rat hippocampus, mediated by the local application of recombinant adeno-associated viral (rAAV) vectors carrying the human NPY gene, results in significant reduction of seizures in acute and chronic seizure models. In this study, we characterized a more efficient rAAV-NPY vector to improve cell transfection in the injected area. The changes included pseudotyping with the AAV vector serotype 1 (rAAV1), and using the strong constitutive hybrid CBA promoter, which contains a cytomegalovirus enhancer and chicken beta-actin promoter sequences. We compared NPY expression and the associated anticonvulsant effects of this new vector, with those mediated by the former rAAV vector with chimeric serotype 1/2 (rAAV1/2). In addition, we investigated whether rAAV serotype 1 vector-mediated chronic NPY overexpression causes behavioural deficits that may detract from the clinical utility of this therapeutic approach. We report that rAAV-NPY serotype 1 vector has significantly improved anticonvulsant activity when compared with serotype 1/2 vector, as assessed by measuring EEG seizure activity in kainic acid treated rats. rAAV1-mediated NPY overexpression in naive rats did not result in alterations of physiological functions such as learning and memory, anxiety and locomotor activity. In addition, we did not observe glia activation, or humoral immune responses against serotype 1 vector, which could inactivate gene expression. Our findings show that rAAV1-NPY vector with the CBA promoter mediates powerful anticonvulsant effects and seems to be safe in rodents, thus it may be considered a vector of choice for possible clinical applications.
Noe
F
F
Laboratory of Experimental Neurology, Mario Negri Institute for Pharmacological Research, Milano, Italy.
Vaghi
V
V
Balducci
C
C
Fitzsimons
H
H
Bland
R
R
Zardoni
D
D
Sperk
G
G
Carli
M
M
During
M J
MJ
Vezzani
A
A
D000199:Actins / Q000235:genetics; D000818:Animals; D000229:Dependovirus; D004833:Epilepsy, Temporal Lobe / Q000503:physiopathology / Q000628:therapy*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006624:Hippocampus / Q000378:metabolism*; D056724:Immunity, Humoral; D007608:Kainic Acid / Q000009:adverse effects; D007858:Learning; D008297:Male; D008568:Memory; D009043:Motor Activity; D009478:Neuropeptide Y / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D012640:Seizures / Q000503:physiopathology / Q000628:therapy*; D014161:Transduction, Genetic / Q000379:methods*
D000199:Actins; D009478:Neuropeptide Y; D007608:Kainic Acid