© 1995 American Heart Association, Inc.
Articles |
Endothelium-Specific In Vivo Gene Transfer
From the Molecular Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health (A.H.S., G.D., K.D.N., D.A.D.), Bethesda, Md, and the Division of Cardiovascular Pathology, Armed Forces Institute of Pathology (R.V.), Washington, DC.
Correspondence to David A. Dichek, MD, Gladstone Institute of Cardiovascular Disease, PO Box 419100, San Francisco CA 94141-9100. E-mail david_dichek@quickmail.ucsf.edu.
Abstract Targeted expression of genetic material within the
vascular endothelium is potentially a powerful tool for
the investigation of endothelial cell (EC) biology. We
developed, optimized, and characterized an efficient somatic transgenic
model of EC-specific gene transfer. Rat carotid arteries were infused
with adenovirus expressing a ß-galactosidase (ß-gal) gene. The
level and cell-type specificity of recombinant gene expression were
measured by assaying ß-gal activity in vessel extracts and by
counting transduced cells in histological sections.
Toxicity was evaluated by counting total ECs (3 days) and by measuring
neointimal formation (14 days). Effects of transduction on
the proliferation of vascular cells were measured with
bromodeoxyuridine and [3H]thymidine. Maximum recombinant
gene expression resulted from infusion of 1x1010 to
1x1011 plaque-forming units (pfu) per milliliter; 
35%
of luminal ECs were transduced. A high degree of EC specificity (90%
to 98% of total transduced cells) was maintained over this range of
virus concentrations. More highly concentrated virus resulted in loss
of ß-gal expression and a large decrease in luminal EC number (97%
decrease, P<.001). Gene transfer at 4x1010
pfu/mL was efficient, preserved EC integrity, and caused minimal
neointimal formation. After gene transfer, there were early
(3-day) increases in both EC and smooth muscle cell proliferation. At
14 days, only EC proliferation remained elevated (18% versus 1.4% in
vehicle-infused arteries, P=.005). This animal model permits
efficient highly EC-specific gene transfer. Vascular toxicity is
minimal, although the EC proliferative index is elevated. This model
will be useful in experiments that elucidate the biological role of EC
gene products and define pathways of EC gene regulation and signal
transduction in vivo.
Key Words: endothelium • gene transfer • adenovirus • carotid arteries
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