The overall goal of this project is the development of new adenovirus- based gene transfer vectors that are better suited than those that are currently available for human gene transfer therapy of cystic fibrosis and other genetic diseases.
The specific aims of this development include: 1) helper-dependent adenoviral vectors that accommodate a high molecular weight DNA insert; 2) site-specific chromosomal integration (safe harbor) by legitimate recombination; 3) long-term tissue-specific expression; and 4) control of expression by small molecular weight inducers (drugs). These objectives, if achieved, will overcome three major disadvantages of the adenovirus system: lack of efficient nuclear integration, limited insert packaging capacity, and control of expression in particular cell types. An adenovirus amplicon plasmid with a minimum cis-element for packaging of adenovirus genome will be constructed. This vector has the potential for creating an adenovirus-based vector with capacity of an insert of up to 37 kb. Such a vector will remove size limitations for inclusion of cis- regulatory elements and will also open a new possibility to add a feature of efficient nuclear targeting by homologous recombination. The gene targeting efficiency of adenoviral vectors will be studied in mouse and human chromosomal sites. Attempts to target in vivo will be performed in mice and baboons. We propose to introduce a drug-responsive promoter, Cypla-1, into adenoviral vectors and examine the inducibility in vivo. If successful, these advances could lead to a long-term expression capable of drug-mediated regulation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL051754-02
Application #
3759203
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Baylor College of Medicine
Department
Type
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
Sakuma, Tsutomu; Gu, Xiu; Wang, Zheng et al. (2006) Stimulation of alveolar epithelial fluid clearance in human lungs by exogenous epinephrine. Crit Care Med 34:676-81
Toietta, Gabriele; Mane, Viraj P; Norona, Wilma S et al. (2005) Lifelong elimination of hyperbilirubinemia in the Gunn rat with a single injection of helper-dependent adenoviral vector. Proc Natl Acad Sci U S A 102:3930-5
Pastore, Lucio; Belalcazar, L Maria; Oka, Kazuhiro et al. (2004) Helper-dependent adenoviral vector-mediated long-term expression of human apolipoprotein A-I reduces atherosclerosis in apo E-deficient mice. Gene 327:153-60
Morral, Nuria; O'Neal, Wanda K; Rice, Karen et al. (2002) Lethal toxicity, severe endothelial injury, and a threshold effect with high doses of an adenoviral vector in baboons. Hum Gene Ther 13:143-54
Toietta, Gabriele; Pastore, Lucio; Cerullo, Vincenzo et al. (2002) Generation of helper-dependent adenoviral vectors by homologous recombination. Mol Ther 5:204-10
O'Neal, W K; Zhou, H; Morral, N et al. (2000) Toxicity associated with repeated administration of first-generation adenovirus vectors does not occur with a helper-dependent vector. Mol Med 6:179-95
Morral, N; O'Neal, W; Rice, K et al. (1999) Administration of helper-dependent adenoviral vectors and sequential delivery of different vector serotype for long-term liver-directed gene transfer in baboons. Proc Natl Acad Sci U S A 96:12816-21
Lee, B; Dennis, J A; Healy, P J et al. (1999) Hepatocyte gene therapy in a large animal: a neonatal bovine model of citrullinemia. Proc Natl Acad Sci U S A 96:3981-6
O'Neal, W K; Zhou, H; Morral, N et al. (1998) Toxicological comparison of E2a-deleted and first-generation adenoviral vectors expressing alpha1-antitrypsin after systemic delivery. Hum Gene Ther 9:1587-98
Morral, N; Parks, R J; Zhou, H et al. (1998) High doses of a helper-dependent adenoviral vector yield supraphysiological levels of alpha1-antitrypsin with negligible toxicity. Hum Gene Ther 9:2709-16

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