Adenovirus-Mediated Gene Transfer Into Rat Cardiac Allografts

Abstract

With the ultimate goal of modulating the host immune response in organ transplantation, gene therapy studies have demonstrated that direct plasmid DNA injection into transplanted myocardium can result in detectable levels of transgene expression. However, the restricted distribution and low level of transgene expression evident in these studies have limited its application. Recently, replication-defective adenovirus vectors have been shown to be an efficient gene-transfer vehicle in vivo whose infection does not require target-cell proliferation. In the present study, adenovirus vectors encoding reporter genes were delivered into transplanted hearts by either direct injection into the myocardium or perfusion via aorta of the donor hearts. The efficacy and stability of the transgene expression by perfusion and by direct injection were examined and compared. Using the adenovirus vector encoding the firefly luciferase gene, we found that a higher level of transgene expression was achieved by direct injection, but that more evenly distributed transgene expression was observed in hearts perfused with viral vector. These results were further confirmed by 5-bromo-4-chloro-3-indolyl-beta-d-galactoside histochemical staining of another adenoviral vector encoding beta-galactosidase. The transgene expression was not stable and decreased within 1 month with either delivery method. Nevertheless, these results indicate that adenovirus-mediated gene transfer can result in short-term expression of the gene throughout the heart and may be useful as a gene vector in organ transplantation.

Lipid-mediated gene transfer of viral IL-10 prolongs vascularized cardiac allograft survival by inhibiting donor-specific cellular and humoral immune responses

“…[31][32][33][34] The higher efficiency of viral vectors at delivering genes in vivo than nonviral vectors is illustrated by a direct comparison in which luciferase transgene activity was 10-fold higher in rat cardiac grafts transduced with an adenoviral vector compared with a plasmid vector. 35 None the less, even with viral vectors, investigators have documented inefficient gene transfer by allograft perfusion, with most studies reporting less than 1% of the cells expressing the transgene. 30,33 Although appealing because they can transduce a wide variety of cell types relatively efficiently, adenoviral vectors induce an exuberant immune response and can be especially problematic in human gene therapy applications, in that many individuals have pre-existing immunity to adenovirus.…”

Efficient and Durable Gene Transfer to Transplanted Heart Using Adeno-associated Virus 9 Vector

“…Gene therapy offers a potential approach to modify the donor organ to modulate the effects of ischemic-reperfusion injury and acute or chronic immune rejection after organ transplantation. Various strategies including direct injection into the myocardium, (1,2) bolus injection into the coronary vasculature (3,4) and in situ warm perfusion, (5) have been used to deliver genes to the transplanted heart. We have developed a cold ex vivo perfusion gene delivery system using adenoviral vectors in both rat (6) and pig (7) heart transplantation models.…”

Interleukin-10 produced by recombinant adenovirus prolongs survival of cardiac allografts in rats

“…The results obtained with AdlacZ are in agreement with those of other groups where recombinant adenovirus were injected into cardiac muscle. [12][13][14] Comparison of intramyocardial injection with intracoronary delivery of adenoviruses in pigs showed a higher transduction efficiency for the former method. 15 Despite the fact that inclusion of compounds which regulate vascular tone and permeability in the adenovirus perfusion solution have been shown greatly to increase transduction efficiency, 16 been followed by heart transplantation, and therefore it is difficult to conclude whether it is applicable in the transplantation setting.…”

“…14,15 This transient expression may be due to immune responses against adenoviruses or the transgene itself (such as ␤-galactosidase). 17,18 As opposed to gene therapy for genetic deficiencies, expression of transgenes in transplantation can be short term.…”