Gene delivery to in situ veins: differential effects of adenovirus and adeno-associated viral vectors.
OBJECTIVE
Gene transfer offers the potential to modify vein graft biology at the time of surgical implantation. Efficiency of gene delivery, stability of expression, and host responses are critical parameters for candidate vectors. We compared the effects of intraluminal exposure with adenovirus (AD) and adeno-associated virus (AAV) vectors on transgene expression and monocyte adhesion (MA) in treated vein segments.


METHODS
Adult New Zealand white rabbits (N = 51) were anesthetized, and the jugular veins were cannulated bilaterally. Veins were gently distended with either vector (2.10(8) to 1.10(10) infective particles/mL) or vehicle (control) for 30 minutes, after which venous flow was restored. AD and AAV vectors encoding for the marker genes beta-galactosidase (LacZ) and green fluorescent protein (GFP) were used. Vessels were explanted 2 to 40 days postinfection for analysis of gene expression (X-gal staining, reverse transcriptase-polymerase chain reaction), MA, and immunohistochemistry. Ex vivo adhesion assays used (51)Cr-labeled THP-1 cells. Statistical significance was tested by using analysis of variance with a P value less than.05.


RESULTS
All animals survived, and all treated veins were patent at sacrifice. Intraluminal exposure to AD at a titer of 1.10(9) resulted in near complete transduction of the endothelium at 2 days, with no detectable expression by day 14. At an equal titer of infectious particles, transgene expression was markedly less for AAV at 2 to 7 days, but improved at 2 weeks and persisted to 40 days. MA was significantly increased 2 days after AD exposure (2.7-fold vs control, *P <.002); AAV treatment had no discernible effect on MA.


CONCLUSIONS
AD-mediated gene transfer to vein segments resulted in robust, transient gene expression that disappeared after 2 weeks. In comparison, AAV-mediated gene delivery was less efficient, but resulted in delayed onset, persistent expression beyond 30 days. AD exposure induced an early increase in MA to the vein surface that was not seen with AAV treatment. Current generations of both AD and AAV vectors have significant, albeit different, limitations for vascular gene therapy.
Eslami
M H
MH
Division of Vascular Surgery, Brigham and Women's Hospital, Boston, MA 02115, USA.
Gangadharan
S P
SP
Sui
X
X
Rhynhart
K K
KK
Snyder
R O
RO
Conte
M S
MS
D000256:Adenoviridae / Q000235:genetics*; D000704:Analysis of Variance; D000818:Animals; D002448:Cell Adhesion / Q000235:genetics; D002860:Chromium Radioisotopes; D002863:Chromogenic Compounds; D000229:Dependovirus / Q000235:genetics*; D005697:Galactosides; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D005819:Genetic Markers / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D007211:Indoles; D007763:Lac Operon / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D009000:Monocytes / Q000502:physiology; D014677:Pharmaceutical Vehicles; D016133:Polymerase Chain Reaction; D011817:Rabbits; D019275:Radiopharmaceuticals; D012039:Regional Blood Flow / Q000502:physiology; D012501:Saphenous Vein / Q000502:physiology* / Q000637:transplantation; D019076:Transgenes / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D002860:Chromium Radioisotopes; D002863:Chromogenic Compounds; D005697:Galactosides; D005819:Genetic Markers; D007202:Indicators and Reagents; D007211:Indoles; D008164:Luminescent Proteins; D014677:Pharmaceutical Vehicles; D019275:Radiopharmaceuticals; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside