Stem Cells
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

First published online March 22, 2007
This Article
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
2006-0321v1
25/6/1571    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Reprints/Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lavenu-Bombled, C.
Right arrow Articles by Dubart-Kupperschmitt, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lavenu-Bombled, C.
Right arrow Articles by Dubart-Kupperschmitt, A.
Submitted on May 26, 2006
Accepted on March 12, 2007

Technology Development

GpIb{alpha} promoter drives megakaryocytic lineage- restricted expression after hematopoietic stem cell transduction using a SIN lentiviral vector

Cécile Lavenu-Bombled 1, Brigitte Izac 2, Faézeh Legrand 3, Marie Cambot 2, Agathe Vigier 2, Jean-Marc Massé 4, Anne Dubart-Kupperschmitt 2*

1 Institut Cochin, Département d'Hématologie, Paris, France; Inserm U567, Paris, France; CNRS, UMR 8104, Paris, France; Université Paris Descartes, Paris, France; Service d' Hématologie biologique, Hôpital Henri Mondor, Créteil, France
2 Institut Cochin, Département d'Hématologie, Paris, France; Inserm U567, Paris, France; CNRS, UMR 8104, Paris, France; Université Paris Descartes, Paris, France
3 Institut Cochin, Département d'Hématologie, Paris, France; Inserm U567, Paris, France; CNRS, UMR 8104, Paris, France; Université Paris Descartes, Paris, France; Service d'Hématologie, Hôpital Minjoz, Besançon, France
4 Institut Cochin, Plateforme de microscopie électronique, Paris, France; Inserm U567, Paris, France; CNRS, UMR 8104, Paris, France; Université Paris Descartes, Paris, France

* To whom correspondence should be addressed. E-mail: dubart{at}cochin.inserm.fr.


  Abstract

Megakaryocytic (MK) lineage is an attractive target for cell/gene therapy approaches, aiming at correcting platelet protein deficiencies. However, MK cells are short-lived cells and their permanent modification requires modification of hematopoietic stem cells (HSC) with an integrative vector such as a lentiviral vector. Glycoprotein IIb (GpIIb) promoter, the most studied among the MK regulatory sequences, is also active in stem cells. To strictly limit transgene expression to the MK lineage after transduction of human CD34+ hematopoietic cells with a lentiviral vector, we looked for a promoter activated later during MK differentiation.

Human cord blood (CB), bone marrow (BM) and peripheral blood mobilized (PBM) CD34+ cells were transduced with a HIV-derived SIN lentiviral vector encoding the green fluorescent protein (GFP) under the transcriptional control of GpIb{alpha}, GpIIb or EF1{alpha} gene regulatory sequences. Both GpIb{alpha} and GpIIb promoters restricted GFP expression (analyzed by flow-cytometry and immuno-electron microscopy) in MK cells among the maturing progeny of transduced cells. However, only the GpIb{alpha} promoter was strictly MK specific while GpIIb promoter was leaky in immature progenitor cells not yet engaged in MK cell lineage differentiation.

We thus demonstrate the pertinence of using a 328 base pair (bp) fragment of the human GpIb{alpha} gene regulatory sequence, in the context of a lentiviral vector, to tightly restrict transgene expression to the MK lineage after transduction of human CD34+ hematopoietic cells.


This article has been cited by other articles:


Home page
 JEMHome page
H. Nishikii, K. Eto, N. Tamura, K. Hattori, B. Heissig, T. Kanaji, A. Sawaguchi, S. Goto, J. Ware, and H. Nakauchi
Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
J. Exp. Med., August 4, 2008; 205(8): 1917 - 1927.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
STEM CELLS THE ONCOLOGIST CME ALPHAMED PRESS JOURNALS
http://www.peprotech.com/
Copyright © 2007 by AlphaMed Press.