In vivo gene transfer and expression in normal uninjured blood vessels using replication-deficient recombinant adenovirus vectors.

Abstract

Replication-deficient recombinant adenovirus vectors do not require target cell replication for transfer and expression of exogenous genes and thus may be useful for in vivo gene therapy in the endothelium. To evaluate the feasibility of adenovirus-mediated gene transfer in vivo in normal intact blood vessels, adenovirus vectors containing the Escherichia coli lacZ gene or a human alpha 1-antitrypsin (alpha 1AT) cDNA were injected in vivo into the lumen of an occluded vessel segment of sheep jugular vein and/or carotid artery. After 15 minutes of incubation, circulation was restored; the vessels were harvested 1-28 days later and evaluated for gene transfer and expression. Three days after in vivo exposure to the lacZ adenovirus vector, the endothelium of jugular veins and carotid arteries expressed beta-galactosidase. Exposure of jugular veins and carotid arteries in vivo to the alpha 1AT adenovirus vector resulted in the expression of alpha 1AT mRNA transcripts detected by Northern analysis and in the synthesis and secretion of alpha 1AT detected by ex vivo [35S]methionine labeling. Expression with the adenovirus vectors was efficient and easily detectable 1-14 days after injection, with maximum expression at 7 days. Expression was no longer evident at 28 days. Thus, adenovirus vectors are capable of transferring exogenous genes to the endothelium of normal arteries and veins with expression for at least 2 weeks, suggesting that these vectors have the potential for a variety of cardiovascular experimental and clinical applications.

Tissue-specific gene therapy directed to tumor angiogenesis

“…[8][9][10] Lemarchand et al 11,12 examined the feasibility and stability of gene expression in the ligated region of sheep carotid arteries and veins. One of the limitations of these studies was the local application, which is invasive and clinically impractical especially in tumor metastasis.…”

Pulmonary inflammation associated with repeated, prenatal exposure to an E1, E3-deleted adenoviral vector in sheep

“…[1][2][3][4][5][6] Gene expression is transient and appears to be limited by host inflammatory and immune responses to adenoviral proteins. 2,4,[7][8][9][10] Adenovirus-mediated gene transfer and expression are more effective when the ability of the recipient's immune system to respond to vector administration is suppressed.…”

Adeno-associated virus (AAV)-7 and -8 poorly transduce vascular endothelial cells and are sensitive to proteasomal degradation

“…Most preclinical studies have utilized adenoviral vectors. At sufficiently high doses, adenoviral gene delivery to intact blood vessels, 4 atherosclerotic lesions 5 and vein grafts 6 can be achieved. However, this is not without the complications of toxicity and immunogenicity and mediates a purely transient transgene expression when first-generation vectors are utilized.…”