Although long term gene transfer and expression have been demonstrated within the central nervous system, it is important to validate specific gene transfer and expression strategies in animal models before attempting application to the treatment of chronic neurological disorders, such as epilepsy. Using an adeno-associated virus (AAV) vector with a cytomegalovirus promoter (CMV) we have demonstrated stable, long term (3 months) expression of a foreign reporter gene in the rat inferior colliculus, a site of seizure genesis (McCown et al., Brain Res. 713:99, 1996). Therefore, it is hypothesized that AAV-CMV vectors will transfer and express the genes for specific receptor subunit proteins in the inferior colliculus, causing the assembly of functional neurotransmitter receptors, or in the case of anti sense constructs, disrupt assembly of specific receptors. To test this hypothesis, AAV-CMV vectors will be constructed with one of three GABA receptor subunit cassettes, or an NMDA receptor subunit cassette. These vectors will be infused into the inferior colliculus of rats. Seven days and 3 months later, we will determine if the vector-mediated gene delivery 1) increases the amount of specific subunit mRNA by quantitative RT-PGR, 2) increases the amount of receptor subunit protein using quantitative immunohistochemistry and 3) increases functional GABA receptor assembly using a chloride up take measure or functional NMDA receptor assembly using NMDA specific [3H]glutamate binding. Because the inferior colliculus is central to a number of seizure models, the same vectors will be infused into the inferior colliculus of normal or genetic epilepsy-prone rats, and in vivo seizure sensitivity will be evaluated. These studies represent a unique opportunity to evaluate gene delivery strategies in vivo. Moreover, since we are targeting receptors that modulate seizure sensitivity, the findings may have direct application to the treatment of focal seizure disorders.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS035633-05
Application #
6393826
Study Section
Special Emphasis Panel (ZRG1-NLS-3 (01))
Program Officer
Fureman, Brandy E
Project Start
1997-04-01
Project End
2002-09-29
Budget Start
2001-04-01
Budget End
2002-09-29
Support Year
5
Fiscal Year
2001
Total Cost
$218,678
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Psychiatry
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Weinberg, Marc S; Blake, Bonita L; McCown, Thomas J (2013) Opposing actions of hippocampus TNF? receptors on limbic seizure susceptibility. Exp Neurol 247:429-37
Gray, S J; Nagabhushan Kalburgi, S; McCown, T J et al. (2013) Global CNS gene delivery and evasion of anti-AAV-neutralizing antibodies by intrathecal AAV administration in non-human primates. Gene Ther 20:450-9
Hayes, Dayna M; Fee, Jon R; McCown, Thomas J et al. (2012) Neuropeptide Y signaling modulates the expression of ethanol-induced behavioral sensitization in mice. Addict Biol 17:338-50
Gray, Steven J; Foti, Stacey B; Schwartz, Joel W et al. (2011) Optimizing promoters for recombinant adeno-associated virus-mediated gene expression in the peripheral and central nervous system using self-complementary vectors. Hum Gene Ther 22:1143-53
Weinberg, M S; Blake, B L; Samulski, R J et al. (2011) The influence of epileptic neuropathology and prior peripheral immunity on CNS transduction by rAAV2 and rAAV5. Gene Ther 18:961-8
McCown, Thomas J (2011) Adeno-Associated Virus (AAV) Vectors in the CNS. Curr Gene Ther 11:181-8
Johnson, Jarrod S; Li, Chengwen; DiPrimio, Nina et al. (2010) Mutagenesis of adeno-associated virus type 2 capsid protein VP1 uncovers new roles for basic amino acids in trafficking and cell-specific transduction. J Virol 84:8888-902
Koh, Ming Teng; Haberman, Rebecca P; Foti, Stacey et al. (2010) Treatment strategies targeting excess hippocampal activity benefit aged rats with cognitive impairment. Neuropsychopharmacology 35:1016-25
McCown, Thomas J (2010) The future of epilepsy treatment: focus on adeno-associated virus vector gene therapy. Drug News Perspect 23:281-6
McCown, Thomas J (2009) Adeno-associated virus vector-mediated expression and constitutive secretion of galanin suppresses limbic seizure activity. Neurotherapeutics 6:307-11

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