(Circulation. 1996;93:301-309.)
© 1996 American Heart Association, Inc.
Articles |
Enhanced In Vivo Antithrombotic Effects of Endothelial Cells Expressing Recombinant Plasminogen Activators Transduced With Retroviral Vectors
From the Division of Hematology and Oncology, Department of Medicine, and Yerkes Regional Primate Research Center, Emory University School of Medicine, Atlanta, Ga; and the Molecular Hematology Branch (D.A.D.), National Heart, Lung, and Blood Institute, Bethesda, Md.
Correspondence to Laurence A. Harker, MD, Division of Hematology and Oncology, Emory University School of Medicine, PO Drawer AR, Atlanta, GA 30322.
Background The effects of regulating endothelial cell (EC) plasminogen activator production on thrombus accumulation in vivo are incompletely understood. By overexpressing plasminogen activators in ECs via gene transfer, the hypothesis was tested that increased levels of plasminogen activators inhibit the accumulation of thrombus in vivo.
Methods and Results Cultured baboon ECs transduced with human
cDNAs for wild-type tissue plasminogen
activator (TPA) or for
glycosylphosphatidylinositol-anchored urokinase-type
plasminogen activator (a-UPA) were seeded onto
collagen-coated segments of vascular graft (collagen segments)
and exposed overnight to flow using an in vitro perfusion circuit. The
antigenic levels of TPA and UPA each increased 10-fold in the media
perfusing the corresponding transduced ECs compared with untransduced
ECs (P
.05 in both cases). In baboons the antithrombotic
effects of TPA-transduced or a-UPA–transduced ECs were measured
as 111In-platelet deposition and
125I-fibrin accumulation on collagen segments bearing
sparsely attached ECs (transduced versus untransduced) interposed in
exteriorized arteriovenous femoral shunts. Platelet-rich
thrombus formed on the collagen segments with fibrin-rich thrombus
propagated distally. The presence of TPA-transduced or
a-UPA–transduced ECs on collagen segments at a density of 25 000
ECs/cm2 decreased 111In-platelet
deposition and 125I-fibrin accumulation on collagen
surfaces compared with untransduced ECs present at equivalent
density (P<.05 for platelet deposition with
TPA-transduced ECs and P<.05 for platelet deposition on
the propagated tail, as well as fibrin accumulation on the graft with
a-UPA–transduced ECs). The systemic levels of
fibrinopeptide A, thrombin-antithrombin complex,
D-dimer, and both local and systemic levels of TPA and UPA
were not increased by transduced ECs compared with untransduced ECs.
The focal antithrombotic effects of transduced ECs appear to be due to
local enhancement of thrombolysis.
Conclusions ECs transduced with recombinant TPA and a-UPA enhance local antithrombotic activity in vivo. This strategy of attaching transduced ECs overexpressing plasminogen activators may be therapeutically useful by preventing thrombo-occlusive failure of implanted cardiovascular devices or mechanically denuded vessels.
Key Words: plasminogen activators • thrombosis • endothelium
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