Adenovirus vectors based on a new helper dependent system are being developed for use in gene therapy. Because cloning capacity of the vectors is greater than 35 kb and all viral genes are deleted, these vectors are well suited for gene therapy, and will be used to produce vectors carrying genes for therapy of cardiovascular disease. Both """"""""first generation"""""""" E1, E3 deleted viruses and helper dependent vectors will be used to identify and study gene regulatory elements that can improve duration of expression while retaining ability to express transgenes at high levels. A disadvantage of adenovirus vectors for some applications is the failure of the viral DNA to integrate a high frequency. To achieve integration of transgenes into mammalian DNA, the possibility of using components of a eukaryotic transposable element will be explored. The eukaryotic transposable element, mariner, has been shown to encode a transposase that can function in the absence of host cell factors and therefore should be capable of inducing transposition of sequences flanked by mariner terminal repeats when both are introduced into mammalian cells. This transpoase will be expressed in Ad vectors under the control of a T7 promoter with T7 RNA polymerase provided by a second vector or by cells transformed with a T7 pol expression cassette. Initially, enzymatic activity of the transposase expressed in vector infected cells will be studied in cell free systems. To detect transposition in transduced cells, appropriate vectors carrying minimariner elements and selectable markers encoding drug resistance will be constructed, used in combination with vectors expressing transposase to transduce cells, and transformed cells selected for drug resistance will be established and analyzed. Eventually experiments will be done in transgenic mice expressing the T7 pol to obtain transposition in vivo.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Specialized Center (P50)
Project #
5P50HL059314-03
Application #
6258929
Study Section
Project Start
1999-09-01
Project End
2000-08-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
3
Fiscal Year
1999
Total Cost
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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