Percutaneous transplantation of genetically-modified autologous fibroblasts in the rabbit femoral artery: A novel approach for cardiovascular gene therapy.
OBJECTIVE
Arterial cell and gene therapies are promising strategies for the treatment of cardiovascular diseases; however, the optimal cell type and delivery technique for such treatment remain to be determined. The aim of the present study was to design a new approach for arterial cell and gene therapy in which genetically modified autologous skin fibroblasts are percutaneously delivered in stented rabbit femoral arteries in vivo.


METHODS
Autologous skin fibroblasts underwent in vitro transfection with the cationic lipid FuGene and plasmids expressing the human form of the tissue inhibitor of metalloproteinase (hTIMP-1) or nls-LacZ reporter genes.


RESULTS
Transfection efficiency was about 50% and there were high levels of hTIMP-1 secretion up to 14 days after gene transfer. We demonstrated the feasibility of in vivo percutaneous transplantation of fluorescent fibroblasts in the rabbit femoral artery. Results were confirmed by scanning electron microscopy. In vivo local delivery of hTIMP-1-expressing fibroblasts in stented femoral arteries also resulted in high-levels of hTIMP-1 secretion ex vivo for 7 days. Fibroblast transplantation resulted in a modest increase in intimal hyperplasia at the target site, which was reversed with hTIMP-1-transfected fibroblasts.


CONCLUSIONS
Percutaneous transplantation of genetically modified autologous fibroblasts could be used as a cellular platform for locoregional secretion of therapeutic proteins to treat either specific arterial diseases or the diseased organ (eg, the heart) supplied by the target artery.


CONCLUSIONS
Cell and gene therapies are potential new treatments for cardiovascular diseases. We demonstrated that autologous fibroblasts could be easily harvested from a skin biopsy specimen, genetically modified in vitro with nonviral vectors, and percutaneously seeded in vivo in rabbit femoral arteries, leading to locoregional secretion of abundant amounts of recombinant proteins. This new approach has important advantages over alternative approaches that use endothelial cells, viral vectors, and intraoperative cell delivery. Clinical applications may include local treatment of atherosclerotic plaques or aneurysms and also treatment of the diseased organs supplied by the target artery (eg, ischemic or failing heart).
Mazighi
Mikael
M
Unité INSERM U698, hôpital Bichat, Paris, France.
Tchétché
Didier
D
Gouëffic
Yann
Y
San Juan
Aurélie
A
Louedec
Liliane
L
Hénin
Dominique
D
Michel
Jean-Baptiste
JB
Jacob
Marie-Paule
MP
Feldman
Laurent J
LJ
D000818:Animals; D001157:Arterial Occlusive Diseases / Q000473:pathology / Q000601:surgery*; D004247:DNA / Q000235:genetics; D004195:Disease Models, Animal; D005263:Femoral Artery / Q000601:surgery* / Q000648:ultrastructure; D005347:Fibroblasts / Q000201:enzymology / Q000637:transplantation* / Q000648:ultrastructure; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D010957:Plasmids; D011817:Rabbits; D012867:Skin / Q000166:cytology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D014162:Transfection; D014182:Transplantation, Autologous; D016896:Treatment Outcome
D019715:Tissue Inhibitor of Metalloproteinase-1; D004247:DNA