scholarly journals Chromosomal Integration of Adenoviral Vector DNA In Vivo

2010 ◽  
Vol 84 (19) ◽  
pp. 9987-9994 ◽  
Author(s):  
Sam Laurel Stephen ◽  
Eugenio Montini ◽  
Vijayshankar Ganesh Sivanandam ◽  
Muhseen Al-Dhalimy ◽  
Hans A. Kestler ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Adenoviral Vector ◽  
High Capacity ◽  
Median Frequency ◽  
Chromosomal Dna ◽  
Genomic Locus ◽  
Vector Integration ◽  
Fumarylacetoacetate Hydrolase ◽  
Vector Dna

ABSTRACT So far there has been no report of any clinical or preclinical evidence for chromosomal vector integration following adenovirus (Ad) vector-mediated gene transfer in vivo. We used liver gene transfer with high-capacity Ad vectors in the FAHΔexon5 mouse model to analyze homologous and heterologous recombination events between vector and chromosomal DNA. Intravenous injection of Ad vectors either expressing a fumarylacetoacetate hydrolase (FAH) cDNA or carrying part of the FAH genomic locus resulted in liver nodules of FAH-expressing hepatocytes, demonstrating chromosomal vector integration. Analysis of junctions between vector and chromosomal DNA following heterologous recombination indicated integration of the vector genome through its termini. Heterologous recombination occurred with a median frequency of 6.72 × 10−5 per transduced hepatocyte, while homologous recombination occurred more rarely with a median frequency of 3.88 × 10−7. This study has established quantitative and qualitative data on recombination of adenoviral vector DNA with genomic DNA in vivo, contributing to a risk-benefit assessment of the biosafety of Ad vector-mediated gene transfer.

scholarly journals In vivo adenoviral vector-mediated gene transfer into balloon-injured rat carotid arteries.

1993 ◽  
Vol 73 (5) ◽  
pp. 797-807 ◽  
Author(s):  
S W Lee ◽  
B C Trapnell ◽  
J J Rade ◽  
R Virmani ◽  
D A Dichek
Keyword(s):  
Gene Transfer ◽  
Adenoviral Vector ◽  
Carotid Arteries

scholarly journals Assessing the potential for AAV vector genotoxicity in a murine model

Blood ◽  
2011 ◽  
Vol 117 (12) ◽  
pp. 3311-3319 ◽  
Author(s):  
Hojun Li ◽  
Nirav Malani ◽  
Shari R. Hamilton ◽  
Alexander Schlachterman ◽  
Giulio Bussadori ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Normal Tissue ◽  
Expression Patterns ◽  
Tumor Formation ◽  
High Dose ◽  
Adjacent Normal Tissue ◽  
Aav Vector ◽  
Vector Integration ◽  
Integration Sites ◽  
Vector Dna

AbstractGene transfer using adeno-associated virus (AAV) vectors has great potential for treating human disease. Recently, questions have arisen about the safety of AAV vectors, specifically, whether integration of vector DNA in transduced cell genomes promotes tumor formation. This study addresses these questions with high-dose liver-directed AAV-mediated gene transfer in the adult mouse as a model (80 AAV-injected mice and 52 controls). After 18 months of follow-up, AAV-injected mice did not show a significantly higher rate of hepatocellular carcinoma compared with controls. Tumors in mice treated with AAV vectors did not have significantly different amounts of vector DNA compared with adjacent normal tissue. A novel high-throughput method for identifying AAV vector integration sites was developed and used to clone 1029 integrants. Integration patterns in tumor tissue and adjacent normal tissue were similar to each other, showing preferences for active genes, cytosine-phosphate-guanosine islands, and guanosine/cysteine-rich regions. Gene expression data showed that genes near integration sites did not show significant changes in expression patterns compared with genes more distal to integration sites. No integration events were identified as causing increased oncogene expression. Thus, we did not find evidence that AAV vectors cause insertional activation of oncogenes and subsequent tumor formation.

High-Capacity Adenoviral Vector-Mediated Reduction of Huntingtin Aggregate Load In Vitro and In Vivo

2007 ◽  
Vol 18 (4) ◽  
pp. 303-311 ◽  
Author(s):  
Bin Huang ◽  
Johannes Schiefer ◽  
Christian Sass ◽  
G. Bernhard Landwehrmeyer ◽  
Christoph M. Kosinski ◽  
...  
Keyword(s):  
Adenoviral Vector ◽  
High Capacity ◽  
Mediated Reduction

scholarly journals Variables Affecting In Vivo Performance of High-Capacity Adenovirus Vectors

2002 ◽  
Vol 76 (4) ◽  
pp. 1600-1609 ◽  
Author(s):  
Gudrun Schiedner ◽  
Sabine Hertel ◽  
Marion Johnston ◽  
Volker Biermann ◽  
Volker Dries ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Transfer ◽  
High Capacity ◽  
Fold Increase ◽  
Expression Cassette ◽  
Matrix Attachment Region ◽  
Hepatic Expression ◽  
Vector Genome

ABSTRACT In high-capacity adenovirus (HC-Ad) vectors the size and/or composition of the vector genome influences vector stability during production and the expression profile following gene transfer. Typically, an HC-Ad vector will contain both a gene or an expression cassette and stuffer DNA that is required to balance the final vector genome to a size of between 27 and 36 kb. To gain an improved understanding of factors that may influence gene expression from HC-Ad vectors, we have generated a series of vectors that carry different combinations of human alpha-1 antitrypsin (hAAT) expression constructs and stuffer DNAs. Expression in vitro did not predict in vivo performance: all vectors expressed hAAT at similar levels when tested in cell culture. Hepatic expression was evaluated following in vivo gene transfer in C57BL/6J mice. hAAT levels obtained from genomic DNA were significantly higher than levels achieved with small cDNA expression cassettes. Expression was independent of the orientation and only marginally influenced by the location of the expression cassette within the vector genome. The use of lambda stuffer DNA resulted in low-level but stable expression for at least 3 months when higher doses were applied. A potential matrix attachment region element was identified within the hAAT gene and caused a 10-fold increase in expression when introduced in an HC-Ad vector genome carrying a phosphoglycerate kinase (pgk) hAAT cDNA construct. We also illustrate the influence of the promoter on anti-hAAT antibody formation in C57BL/6J mice: a human cytomegalovirus but not a pgk promoter resulted in an anti-hAAT antibody response. Thus, the overall design of HC-Ad vectors may significantly influence amounts and duration of gene expression at different levels.

scholarly journals 851. Molecular Determination of High-Capacity Helper Dependent Adenoviral Vector Genomes In Vitro and In Vivo

2006 ◽  
Vol 13 ◽  
pp. S328
Author(s):  
Mariana Puntel ◽  
James F. Curtin ◽  
Jeffrey M. Zirger ◽  
A.K.M. Muhammad ◽  
Xiong Weidong ◽  
...  
Keyword(s):  
Adenoviral Vector ◽  
High Capacity ◽  
Molecular Determination
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