The overall objective of this project is to develop gutless adenovirus vectors for use in experimental animals, to determine the best protocol for prolonged expression following delivery in vivo, and to use these vectors to express low density lipoprotein receptor (LDLR) and very low density lipoprotein receptor (VLDLR) in animal models of familial hypercholesterolemia (FH).
The specific aims are (i) We will examine the effect of long-term expression of VLDLR and LDLR in transgenic mice with an LDLR -/- background using an inducible binary transactivation system. This system was developed by investigators in Project 3. Different levels of expression of the transgene can be accomplished by graded subphysiological doses of an exogenous compound RU486. (ii) We will develop gutless adenovirus vectors expressing reporter genes (e.g. alpha1- antitrypsin), VLDLR and LDLR which will be tested in mice and rhesus monkeys. (iii) We will develop a protocol for the repeated administration of gutless vectors to mice and rhesus monkeys. Transient immunosuppression protocols and vectors of different serotypes will be tested. (iv) We will test and compare the gutless vectors expressing mouse LDLR and VLDLR in LDLR -/- mice for their efficacy in reversing the hypercholesterolemia and their general health effects, and the extent of aortic atherosclerosis. (v) We will examine the effect of hepatic transfer of the rhesus LDLR and VLDLR genes in heterozygous and possibly homozygous LDLR-deficient rhesus monkeys. This project interacts closely with Project 2 for the gutless vector development, and Project 3 for the regulated expression system. It will be supported by the scientific Cores A (Primate Core), B (Vector Production Core) and C (Pathology Core).

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Specialized Center (P50)
Project #
3P50HL059314-05S1
Application #
6565063
Study Section
Project Start
2001-09-01
Project End
2003-08-31
Budget Start
Budget End
Support Year
5
Fiscal Year
2002
Total Cost
$126,320
Indirect Cost
Name
Baylor College of Medicine
Department
Type
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
Oka, K; Mullins, C E; Kushwaha, R S et al. (2015) Gene therapy for rhesus monkeys heterozygous for LDL receptor deficiency by balloon catheter hepatic delivery of helper-dependent adenoviral vector. Gene Ther 22:87-95
Palmer, Donna J; Ng, Philip (2011) Rescue, amplification, and large-scale production of helper-dependent adenoviral vectors. Cold Spring Harb Protoc 2011:857-66
Palmer, Donna J; Ng, Philip (2011) Characterization of helper-dependent adenoviral vectors. Cold Spring Harb Protoc 2011:867-70
Stephen, Sam Laurel; Montini, Eugenio; Sivanandam, Vijayshankar Ganesh et al. (2010) Chromosomal integration of adenoviral vector DNA in vivo. J Virol 84:9987-94
Sakai, Keiko; Tiebel, Oliver; Ljungberg, M Cecilia et al. (2009) A neuronal VLDLR variant lacking the third complement-type repeat exhibits high capacity binding of apoE containing lipoproteins. Brain Res 1276:11-21
Brunetti-Pierri, Nicola; Stapleton, Gary E; Law, Mark et al. (2009) Efficient, long-term hepatic gene transfer using clinically relevant HDAd doses by balloon occlusion catheter delivery in nonhuman primates. Mol Ther 17:327-33
Palmer, Donna J; Ng, Philip (2008) Methods for the production of helper-dependent adenoviral vectors. Methods Mol Biol 433:33-53
Palmer, Donna J; Ng, Philip (2008) Methods for the production of first generation adenoviral vectors. Methods Mol Biol 433:55-78
Brunetti-Pierri, Nicola; Stapleton, Gary E; Palmer, Donna J et al. (2007) Pseudo-hydrodynamic delivery of helper-dependent adenoviral vectors into non-human primates for liver-directed gene therapy. Mol Ther 15:732-40
Oka, K; Belalcazar, L M; Dieker, C et al. (2007) Sustained phenotypic correction in a mouse model of hypoalphalipoproteinemia with a helper-dependent adenovirus vector. Gene Ther 14:191-202

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