Circulation, Vol 89, 785-792, Copyright © 1994 by American Heart Association
DW Losordo, JG Pickering, S Takeshita, G Leclerc, D Gal, L Weir, M Kearney, J Jekanowski and JM Isner
BACKGROUND: The development of molecular strategies for the treatment of
restenosis has been hindered by low efficiencies of in vivo arterial
transfection. Expression of intracellular marker proteins is generally
evident in < 1% of vascular smooth muscle cells after in vivo arterial
transfection. Efforts to improve the efficiency of in vivo gene transfer
have been further impeded by the use of transgenes encoding for
intracellular marker proteins, necessitating tissue removal and limiting
survey for expression to one point in time. METHODS AND RESULTS: To study
gene expression on a serial basis in vivo and determine the relation
between a secreted gene product and transfection efficiency after in vivo
arterial gene transfer, a method for performing and serially monitoring
gene expression in vivo was developed using the central artery of the
rabbit ear. Liposome-mediated transfection of plasmid DNA containing the
gene for human growth hormone (hGH) was successfully performed in 18 of 23
arteries. Serum hGH levels measured 5 days after transfection ranged from
0.1 to 3.8 ng/mL (mean, 0.97 ng/mL); in contrast, serum drawn from the
control arteries demonstrated no evidence of hGH production. Serial
measurement of hGH from transfected arteries demonstrated maximum hGH
secretion 5 days after transfection and no detectable hormone after 20
days. Despite these levels of secreted gene product documented in vivo,
immunohistochemical staining of sections taken from the rabbit ear artery
at necropsy disclosed only rare cells in which there was evidence of
successful transfection. CONCLUSIONS: These experiments demonstrate a
useful method of performing serial in vivo analyses of gene expression
after vascular transfection and that anatomic analyses of transfection
efficiency may underestimate the potential magnitude of expression in the
case of a secreted gene product. These findings have implications for the
clinical application of somatic gene therapy because low-efficiency
transfection with a gene encoding for a secreted protein may achieve
therapeutic effects not realized by transfection with genes encoding for
proteins that remain intracellular.
ARTICLES
Use of the rabbit ear artery to serially assess foreign protein secretion after site-specific arterial gene transfer in vivo. Evidence that anatomic identification of successful gene transfer may underestimate the potential magnitude of transgene expression
Department of Medicine (Cardiology), St Elizabeth's Hospital, Tufts University School of Medicine, Boston, Mass. 02135.
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