The ability to deliver genes using cell surface receptors to achieve organ specific targeting in vivo has been demonstrated. This approach holds the promise of many exciting and effective therapies. The objective of this project is to develop chemically defined delivery systems that provide highly efficient delivery of genes into the nucleus of the targeted cells, first in vitro and then in vivo.
Specific Aim 1 is to prepare, by organic synthesis, DNA binding polypeptides containing high affinity, chemically defined carbohydrate receptor ligands for the asialoglycoprotein receptor in the liver and for the mannose-6-phosphate receptor in skeletal muscle. Physical and chemical methods will be used to characterize the structural, thermodynamic, and kinetic properties of the DNA complexes.
Specific Aim 2 is to determine, after endocytosis of the DNA:ligand complex and its release into the cytoplasm, what kind of structural and compositional changes in the complex are necessary for the DNA to move through the cytoplasm into the nucleus. DNA movement in the cytoplasm, DNA accumulation in the nucleus, and the probable dissociation of the complex in the cytoplasm will be measured using biochemical methods and digital imaging fluorescence microscopy.
Specific Aim 3 is to synthesize DNA binding ligands that facilitate transport of the DNA through the cytoplasm into the nucleus. The SV-40 nuclear localization sequence on a DNA binding polypeptide will be tested as a component of the DNA complex.
Specific Aim 4 is to demonstrate that the DNA complexes, constructed on the basis of in vitro experiments, are effective for gene delivery in vivo.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL050422-03
Application #
3737150
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Type
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
Waugh, J M; Li-Hawkins, J; Yuksel, E et al. (2000) Thrombomodulin overexpression to limit neointima formation. Circulation 102:332-7
Waugh, J M; Yuksel, E; Li, J et al. (1999) Local overexpression of thrombomodulin for in vivo prevention of arterial thrombosis in a rabbit model. Circ Res 84:84-92
Waugh, J M; Kattash, M; Li, J et al. (1999) Gene therapy to promote thromboresistance: local overexpression of tissue plasminogen activator to prevent arterial thrombosis in an in vivo rabbit model. Proc Natl Acad Sci U S A 96:1065-70
Rojas-Martinez, A; Wyde, P R; Montgomery, C A et al. (1998) Distribution, persistency, toxicity, and lack of replication of an E1A-deficient adenoviral vector after intracardiac delivery in the cotton rat. Cancer Gene Ther 5:365-70
Faith, R E; Montgomery, C A; Durfee, W J et al. (1997) The cotton rat in biomedical research. Lab Anim Sci 47:337-45
Gingras, M C; Arevalo, P; Aguilar-Cordova, E (1996) Potential salmon sperm origin of the E3 region insert of the adenovirus 5 dl309 mutant. Cancer Gene Ther 3:151-4
Sparrow, J T; Monera, O D (1996) Improvements to the TMSBr method of peptide resin deprotection and cleavage: application to large peptides. Pept Res 9:218-22
Gottschalk, S; Sparrow, J T; Hauer, J et al. (1996) A novel DNA-peptide complex for efficient gene transfer and expression in mammalian cells. Gene Ther 3:448-57
Guo, Z S; Wang, L H; Eisensmith, R C et al. (1996) Evaluation of promoter strength for hepatic gene expression in vivo following adenovirus-mediated gene transfer. Gene Ther 3:802-10
Gottschalk, S; Tweten, R K; Smith, L C et al. (1995) Efficient gene delivery and expression in mammalian cells using DNA coupled with perfringolysin O. Gene Ther 2:498-503

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