Adenovirus-Mediated Gene Transfer to Lower Limb Artery of Patients with Chronic Critical Leg Ischemia

Abstract

Arterial gene transfer offers a promising new approach for the treatment of vascular disorders. However, no data are available about the gene transfer efficiency in human arteries in vivo. The aim of this study was to evaluate the safety and feasibility of catheter-mediated adenoviral gene transfer in human peripheral arteries. Ten patients (8 females, 2 males, mean age 80 +/- 8 years) suffering from chronic critical leg ischemia with a prior decision for amputation were recruited in the study. Gene transfer was performed in eight patients in conjunction with a conventional percutaneous transluminal angioplasty, using a perfusion coil balloon catheter. Two patients served as controls. Increasing concentrations of replication-deficient adenoviruses (titers from 1 x 10(8) to 4 x 10(10) PFU) containing a nuclear-targeted beta-galactosidase marker gene were administered into the arteries over 10 min via the catheter. Amputations were performed 20 to 51 hr after the procedures and gene transfer efficiency was evaluated in the transduced arteries using X-Gal staining for beta-galactosidase activity. Beta-galactosidase gene transfer was well tolerated and no adverse tissue responses or systemic complications were observed in any of the patients. Gene transfer was successful in six of the eight patients. Gene transfer efficiency varied between 0.04 and 5.0% of all arterial cells. Transgene expression was detected in smooth muscle cells, endothelial cells, and macrophages and in tunica adventitia. However, transgene activity was not evenly distributed in the arterial wall and no transgene activity was found beneath advanced atherosclerotic lesions. The safety and feasibility of in vivo gene transfer by adenoviral vectors to human peripheral arteries were established. Although improvements are still required in gene transfer efficiency, these findings suggest that adenoviruses can be used to deliver therapeutically active genes into human arteries.

Efficient adventitial gene delivery to rabbit carotid artery with cationic polymer–plasmid complexes

“…8,10,22 In human atherosclerotic arteries, 5% transfection efficiency via the intravascular route has been recently achieved with adenoviruses. 23 We show here that the transfection efficiency with fractured dendrimers and PEI is clearly higher than that achieved previously with DOTMA/ DOPE liposomes. 10 Thus, it is conceivable that gene transfer with cationic polymer-plasmid complexes will become competitive with adenoviruses for in vivo gene delivery applications.…”

“…6,8,23 Furthermore, human lesions are frequently rich in connective tissue and contain only a limited number of transfectable cells in the intimal part of the lesion. To circumvent these problems adventitial gene delivery can be performed with a silastic or biodegradable collar, 13 biodegradable gel, 5 or direct injection into adventitia.…”

Eight-year safety follow-up of coronary artery disease patients after local intracoronary VEGF gene transfer

“…4,9 Adenoviruses containing a CMV-hVEGF 165 expression cassette were produced under good clinical practice GMP in 293 cells, as described. 4 They were analyzed for the absence of replication-competent viruses by means of a cytopathic effect assay on A549 cells, and viral preparations were examined to be free of any microbiological or endotoxin contamination (o20 EU/dose, Whittaker).…”

“…The therapeutic effect of non-viral gene transfer is long lasting compared with that of adenoviral gene transfer. 9,7,11,13 In addition, they can also cause fever and moderate increases in CRP. It is also possible that elevated CRP and fever after non-viral gene transfer might also be related to the invasive procedure itself.…”

Baculovirus-mediated periadventitial gene transfer to rabbit carotid artery

“…10 Virus preparations were analyzed for lipopolysaccharide and bacteriological contaminants. 34 Preparation of recombinant adenoviruses Nuclear targeted LacZ encoding adenoviruses (pCMVnlsLacZ Ad5) were constructed and prepared as described earlier. 7 Virus preparations were analyzed for replicationcompetent viruses, lipopolysaccharide and bacteriological contaminants as described.…”