Recombinant adeno-associated virus vector for gene transfer to the transplanted rat heart.
Efficient durable viral vector transduction of the transplanted heart remains elusive. This study assesses the potential of recombinant adeno-associated virus (rAAV) mediated gene delivery to the transplanted rat heart. rAAV serotype 1, 2 and 5 vectors encoding the green fluorescent protein (GFP) gene (1 x 10(11) viral particles/ml) were diluted in cold University of Wisconsin solution and circulated through the coronary vasculature of the donor organs for 30 min before syngeneic rat heterotopic heart transplantation was performed. Study 1: animals (n = 5 each serotype) were killed at 21 days post-transplant to evaluate the efficiency of GFP transduction using RT-PCR and expression by fluorescence microscopy. Study 2: using rAAV-1, animals (n = 5 each group) were killed at 7, 21 and 84 days to evaluate the durability of GFP expression. The maximum cardiac GFP expression at 21 days was observed in rAAV-1. GFP expression by rAAV-1 was detectable at 7 days, improved at 21 days, and was still evident at 84 days. This study demonstrates cardiac rAAV gene transduction with a cold perfusion preservation system of the donor heart. These data show that AAV-1 is superior to AAV-2 and AAV-5 for this purpose and that durable expression is achievable.
Schirmer
Johannes M
JM
William J. von Liebig Transplant Center, Mayo Clinic, Rochester, MN 55905, USA.
Miyagi
Naoto
N
Rao
Vinay P
VP
Ricci
Davide
D
Federspiel
Mark J
MJ
Kotin
Robert M
RM
Russell
Stephen J
SJ
McGregor
Christopher G A
CG
D000256:Adenoviridae; D000818:Animals; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins; D016027:Heart Transplantation; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014161:Transduction, Genetic / Q000379:methods*
D049452:Green Fluorescent Proteins