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First published online August 25, 2005
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2004-0364v1
24/1/199    most recent
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Submitted on December 21, 2004
Accepted on June 14, 2005

Original Article

Promoter Dependence of Transgene Expression by Lentivirus-Transduced Human Blood-Derived Endothelial Progenitor Cells

JIA WEI LIU 1, GILLES PERNOD 2, SYLVIE DUNOYER-GEINDRE 1, RICHARD J. FISH 1, HONG YANG 1, HENRI BOUNAMEAUX 1, EGBERT K.O. KRUITHOF 1*

1 Division of Angiology and Haemostasis, University Hospital, Geneva, Switzerland
2 Unité d'Hémostase, DBPC, GREPI EA 2938, CHU Grenoble, France

* To whom correspondence should be addressed. E-mail: Egbert.Kruithof{at}hcuge.ch.


  Abstract

Peripheral blood-derived endothelial progenitor cells (EPC) have considerable potential for the autologous therapy of vascular lesions or ischemic tissues. By introducing stable genetic modifications into these cells this potential might be further enhanced. We investigated to what extent transgene expression can be controlled by using different transgene promoters. This was investigated in early or late outgrowth human EPC obtained by culturing blood mononuclear cells for one or four weeks on type 1 collagen in medium containing endothelial growth supplements. A large fraction of these cells were stably transduced using lentiviral vectors for expression of the enhanced green fluorescent protein (EGFP). Transgene expression in vitro or in vivo after injection into nude mice, was highest when under the control of the CMV promoter, intermediate with the EF1{alpha} promoter and lowest with the PGK promoter. When blood mononuclear cells were cultured for one week, in the absence of endothelial growth supplements, CMV promoter-driven expression of EGFP was two orders of magnitude lower than in similarly transduced EPC. Our results show that lentiviral vectors are useful tools for the stable introduction of exogenous genes into EPC and for their expression at desired levels using the appropriate gene promoter.

Key Words. lentiviral vector, endothelial, gene therapy




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