Direct in vivo gene transfer into the coronary and peripheral vasculatures of the intact dog.

Abstract

Gene therapy approaches have been suggested for the treatment of cardiovascular disease. Recently, direct transfer of the gene encoding beta-galactosidase into peripheral arteries of the pig has been demonstrated. To determine whether this approach is applicable to other arterial beds and to other species, we first evaluated the use of beta-galactosidase as a marker protein in the canine model. We demonstrate that variable but substantial endogenous beta-galactosidase-like activity is induced by manipulation of canine peripheral arteries, which precludes the use of this marker protein in evaluating the efficiency of gene transfer in this model. A marker gene encoding firefly luciferase was then evaluated, and background luciferase activity was found to be low in the dog even after arterial manipulation. Using the luciferase gene, we then demonstrated lipid-mediated gene transfer directly into both coronary and peripheral arteries of the intact dog. These results indicate the feasibility of in vivo gene transfer into coronary arteries and demonstrate the use of the luciferase marker protein in quantifying recombinant protein expression following gene transfer in canine models. This simple and effective method for direct in vivo gene transfer into coronary and peripheral arteries may be applicable to the localized production of therapeutically important proteins for the treatment of cardiovascular diseases.

Recombinant platelet-derived growth factor B gene expression in porcine arteries induce intimal hyperplasia in vivo.

“…5 A, B, C), and were not seen in E. coli f-galactosidase transduced arteries (Fig. 5 D) 7 d after direct gene transfer. In PDGF transfected vessels, the PDGF immunoreactive staining was mostly not associated with macrophages (Fig.…”

“…It is often difficult to determine whether a specific gene product induces intimal hyperplasia directly or indirectly through other gene products synthesized in response to injury. Recently, it has become possible to express recombinant genes in vivo by direct gene transfer utilizing retroviral vectors or DNA liposome complexes (6)(7)(8)(9). Direct gene transfer permits the delivery of a specific recombinant gene into vascular cells at specific sites in vivo and affords the opportunity to determine the effects of a specific gene product in the arterial wall ( 10).…”

Transgene delivery of plasmid DNA to smooth muscle cells and macrophages from a biostable polymer-coated stent

“…The luciferase expression plasmid (pCl) was kindly supplied by CS Lim and JL Swain and encodes the firefly luciferase gene. 30 All plasmid vectors express the transgene under the transcriptional control of the CMV immediate early promoter-enhancer. The 3.0 Â 12 mm stainless-steel stents (Guidant Corporation) were coated with a polymer prepared and applied by Surface Solutions Labs, Inc. (Carlisle, MA, USA).…”

Increased gene expression after liposome-mediated arterial gene transfer associated with intimal smooth muscle cell proliferation. In vitro and in vivo findings in a rabbit model of vascular injury.

“…Beginning with the report by Nabel et al (I), work from several laboratories (2)(3)(4)(5) has convincingly demonstrated the feasibility of using catheter delivery systems to transfer recombinant genes to normal and/or atherosclerotic arteries of experimental animals. All of these previous demonstrations, however, have been compromised by low transfection efficiencies regardless of the vector used.…”