Circulation, Vol 85, 1110-1117, Copyright © 1992 by American Heart Association
MY Flugelman, MT Jaklitsch, KD Newman, W Casscells, GL Bratthauer and DA Dichek
BACKGROUND. Gene transfer into the arterial wall may provide a novel
therapeutic strategy for the treatment of coronary artery restenosis.
Previously described methods for gene transfer into the arterial wall
require total vessel occlusion for 30 minutes. We sought to develop a
protocol for gene transfer within a more clinically relevant time frame.
METHODS AND RESULTS. We used a perforated balloon (Wolinsky) catheter to
inject retroviral vector-containing virions into rabbit aortas in vivo. The
virions were injected within 1 minute. Aortas were removed 5-14 days after
injection and analyzed for evidence of gene transfer. In initial studies,
nine rabbits were injected with a vector expressing the beta-galactosidase
gene, and nine rabbits were injected with either
non-beta-galactosidase-containing vectors or with a vehicle control.
Histochemical staining of aortic tissues revealed blue (positive) cells in
eight of nine experimental rabbits and six of nine controls. Because of the
lack of specificity of the beta-galactosidase detection system, we adopted
a polymerase chain reaction-based protocol in which oligonucleotide primers
were used to amplify specific vector- related sequences from aortic tissue
extracts. The polymerase chain reaction protocol, calibrated with standards
containing known numbers of transduced cells, revealed low amounts of
vector-related sequences in six of 12 vector-injected rabbits and in one of
13 controls (p less than 0.03). Comparison with standards indicated that
fewer than 100 transduced cells were present in a 2-cm length of the
injected aortic tissue. CONCLUSIONS. Although in vivo gene transfer through
an infusion balloon catheter can be accomplished within 1 minute, the
therapeutic use of this protocol is limited by the small number of cells
that are transduced.
ARTICLES
Low level in vivo gene transfer into the arterial wall through a perforated balloon catheter
Molecular Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
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