Multiply attenuated, self-inactivating lentiviral vectors efficiently transduce human coronary artery cells in vitro and rat arteries in vivo.

Details

Serval ID
serval:BIB_E3BD4E5335A1
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Multiply attenuated, self-inactivating lentiviral vectors efficiently transduce human coronary artery cells in vitro and rat arteries in vivo.
Journal
Journal of Molecular and Cellular Cardiology
Author(s)
Céfaï D., Simeoni E., Ludunge K.M., Driscoll R., von Segesser L.K., Kappenberger L., Vassalli G.
ISSN
0022-2828 (Print)
ISSN-L
0022-2828
Publication state
Published
Issued date
02/2005
Volume
38
Number
2
Pages
333-344
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
Endothelial cells (ECs) in normal vessels are poorly transducible by retroviral vectors, which require cell division for gene transduction. Among retroviruses, lentiviruses have the unique ability to integrate their genome into the chromatin of nondividing cells. Here we show that multiply attenuated, self-inactivating, lentiviral vectors transduce both proliferating and growth-arrested human umbilical vein ECs (HUVECs), human coronary artery ECs (HCAECs), and human coronary artery smooth muscle cells (HCASMCs), with high efficacy. Lentiviral vectors containing the enhanced green fluorescence protein (EGFP) transgene driven by either the cytomegalovirus or the elongation factor-1alpha promoter, but not the phosphoglycerate kinase promoter, directed high-level EGFP expression in endothelial and smooth muscle cells. The endothelium-specific Tie2 promoter also directed transgene expression in ECs. Re-insertion of cis-acting sequences from pol of human immunodeficiency virus type 1 (HIV-1) into the vectors improved transgene expression. A lentiviral vector containing the vascular endothelial growth factor transgene promoted EC proliferation and sprouting in vitro. In vivo gene transfer was studied by lumenal infusion of vector containing solutions into rat carotid arteries. Lentivirus-mediated EGFP gene transfer was observed in approximately 5% of ECs. Lentiviral vectors containing the LacZ transgene achieved detectable beta-galactosidase activity in rat arteries, albeit at a lower level compared with adenoviral vectors. This difference was mainly due to the lower concentration of lentiviral vector preparations. Lentivirus-mediated gene transfer was associated with minimal neointimal hyperplasia and scant inflammatory cell infiltrates in the media and adventitia. These observations indicate that lentiviral vectors may be useful for genetic modifications of vascular cells in vitro and in vivo.
Keywords
Adenoviridae/genetics, Animals, Carotid Arteries/cytology, Carotid Arteries/metabolism, Cell Proliferation, Cells, Cultured, Coronary Vessels/cytology, Coronary Vessels/metabolism, Endothelial Cells/cytology, Endothelial Cells/metabolism, Genetic Vectors/genetics, Humans, Inflammation/genetics, Inflammation/virology, Lentivirus/genetics, Lentivirus/pathogenicity, Male, Myocytes, Smooth Muscle/metabolism, Rats, Rats, Sprague-Dawley, Time Factors, Transduction, Genetic/instrumentation, Transduction, Genetic/methods, Vascular Endothelial Growth Factor A/genetics, Vascular Endothelial Growth Factor A/metabolism, Virus Inactivation
Pubmed
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15/02/2008 11:29
Last modification date
20/08/2019 16:07
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