Abstract

Cystic fibrosis (CF) is a genetic disease caused by a defective cystic fibrosis transmembrane conductance regulator (CFTR) gene. Because the most serious clinical manifestations of the disease are due to defects in airway epithelial cells, it is believed that it may be possible to use viral mediated gene therapy directed at airway epithelial cells to correct the problem. Adeno-associated virus (AAV) is a non-pathogenic human DNA virus which can integrate into human cells with high efficiency. Since AAV is normally found associated with adenovirus, airway epithelial cells may be a normal target for AAV integration. Therefore, in this application we propose to test the feasibility of using AAV as a viral vector for the delivery and integration of a functional CFTR gene into airway epithelial cells. Specifically, we propose to: 1) isolate and test AAV recombinant viruses that contain the CFTR CDNA, 2) make an AAV recombinant virus carrying a marker gene to measure the frequency of transduction in primary airway epithelial cells, and 30 develop new methods for growing pure AAV recombinant viruses.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL050257-02
Application #
3369205
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Project Start
1992-09-30
Project End
1994-08-31
Budget Start
1993-09-30
Budget End
1994-08-31
Support Year
2
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
State University New York Stony Brook
Department
Type
Schools of Medicine
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794

  Publications

Klein, R L; McNamara, R K; King, M A et al. (1999) Generation of aberrant sprouting in the adult rat brain by GAP-43 somatic gene transfer. Brain Res 832:136-44
Klein, R L; Muir, D; King, M A et al. (1999) Long-term actions of vector-derived nerve growth factor or brain-derived neurotrophic factor on choline acetyltransferase and Trk receptor levels in the adult rat basal forebrain. Neuroscience 90:815-21
Klein, R L; Lewis, M H; Muzyczka, N et al. (1999) Prevention of 6-hydroxydopamine-induced rotational behavior by BDNF somatic gene transfer. Brain Res 847:314-20
Pereira, D J; Muzyczka, N (1997) The cellular transcription factor SP1 and an unknown cellular protein are required to mediate Rep protein activation of the adeno-associated virus p19 promoter. J Virol 71:1747-56
Peel, A L; Zolotukhin, S; Schrimsher, G W et al. (1997) Efficient transduction of green fluorescent protein in spinal cord neurons using adeno-associated virus vectors containing cell type-specific promoters. Gene Ther 4:16-24
Gnatenko, D; Arnold, T E; Zolotukhin, S et al. (1997) Characterization of recombinant adeno-associated virus-2 as a vehicle for gene delivery and expression into vascular cells. J Investig Med 45:87-98
Pereira, D J; Muzyczka, N (1997) The adeno-associated virus type 2 p40 promoter requires a proximal Sp1 interaction and a p19 CArG-like element to facilitate Rep transactivation. J Virol 71:4300-9
Ryan, J H; Zolotukhin, S; Muzyczka, N (1996) Sequence requirements for binding of Rep68 to the adeno-associated virus terminal repeats. J Virol 70:1542-53
Fasbender, A J; Zabner, J; Welsh, M J (1995) Optimization of cationic lipid-mediated gene transfer to airway epithelia. Am J Physiol 269:L45-51
Einerhand, M P; Antoniou, M; Zolotukhin, S et al. (1995) Regulated high-level human beta-globin gene expression in erythroid cells following recombinant adeno-associated virus-mediated gene transfer. Gene Ther 2:336-43