title: Direct gene transfer into human epileptogenic hippocampal tissue with an adeno-associated virus vector: implications for a gene therapy approach to epilepsy.
abstract: OBJECTIVE
  Virus vectors capable of transferring genetic information into human cells provide hope for improved therapy in several neurological diseases, including epilepsy. We evaluated the ability of an adeno-associated virus (AAV) vector to transfer and cause expression of a lacZ marker gene in brain slices obtained from patients undergoing temporal lobectomy for control of medically intractable seizures.
  
  
  METHODS
  Human brain slices were injected with an AAV vector (AAVlacZ) encoding Escherichia coli beta-galactosidase and incubated for as long as 24 h. The presence of lacZ mRNA. beta-galactosidase protein and enzymatic activity were assayed by reverse transcriptase polymerase chain reaction (rtPCR), immunocytochemistry, and the X-Gal technique, respectively.
  
  
  RESULTS
  AAVlacZ directed the expression in human epileptogenic brain of E. coli beta-galactosidase that had functional activity. Expression was observed in < or =5 h and was sustained for as long as the slices were viable. Morphological analysis indicated that neurons were preferentially transfected, and there was no evidence of cytotoxicity.
  
  
  CONCLUSIONS
  Our results confirm the feasibility of using AAV vectors to transfer genes into the human CNS and in particular, into neurons. Replacement of the lacZ gene with a functional gene modulating hippocampal neuronal physiology, might allow a localized genetic intervention for focal seizures based on the stereotaxic or endovascular delivery of such a vector system into the appropriate brain region.
authors.affiliation: Division of Neurosurgery, University of Pennsylvania School of Medicine, Philadelphia 19104-6380, USA.
authors.forename: A
authors.initials: A
authors.lastname: Freese
authors.forename: M G
authors.initials: MG
authors.lastname: Kaplitt
authors.forename: W M
authors.initials: WM
authors.lastname: O'Connor
authors.forename: M
authors.initials: M
authors.lastname: Abbey
authors.forename: D
authors.initials: D
authors.lastname: Langer
authors.forename: P
authors.initials: P
authors.lastname: Leone
authors.forename: M J
authors.initials: MJ
authors.lastname: O'Connor
authors.forename: M J
authors.initials: MJ
authors.lastname: During
mesh_terms: D000229:Dependovirus / Q000235:genetics*; D004827:Epilepsy / Q000628:therapy*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D007763:Lac Operon / Q000235:genetics; D014162:Transfection; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001616:beta-Galactosidase