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Glycoprotein Ibalpha promoter drives megakaryocytic lineage-restricted expression after hematopoietic stem cell transduction using a self-inactivating lentiviral vector.
Stem Cells. 2007 Jun; 25(6):1571-7.SC

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 with an integrative vector such as a lentiviral vector. Glycoprotein (Gp) IIb 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, bone marrow, and peripheral-blood mobilized CD34(+) cells were transduced with a human immunodeficiency virus-derived self-inactivating lentiviral vector encoding the green fluorescent protein (GFP) under the transcriptional control of GpIbalpha, GpIIb, or EF1alpha gene regulatory sequences. Both GpIbalpha and GpIIb promoters restricted GFP expression (analyzed by flow cytometry and immunoelectron microscopy) in MK cells among the maturing progeny of transduced cells. However, only the GpIbalpha promoter was strictly MK-specific, whereas 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 fragment of the human GpIbalpha 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. Disclosure of potential conflicts of interest is found at the end of this article.

Authors+Show Affiliations

Institut Cochin, Department of Hematology, Hôpital de Port-Royal, 123 Bd de Port-Royal, Paris 75014, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17379771

Citation

Lavenu-Bombled, Cécile, et al. "Glycoprotein Ibalpha Promoter Drives Megakaryocytic Lineage-restricted Expression After Hematopoietic Stem Cell Transduction Using a Self-inactivating Lentiviral Vector." Stem Cells (Dayton, Ohio), vol. 25, no. 6, 2007, pp. 1571-7.
Lavenu-Bombled C, Izac B, Legrand F, et al. Glycoprotein Ibalpha promoter drives megakaryocytic lineage-restricted expression after hematopoietic stem cell transduction using a self-inactivating lentiviral vector. Stem Cells. 2007;25(6):1571-7.
Lavenu-Bombled, C., Izac, B., Legrand, F., Cambot, M., Vigier, A., Massé, J. M., & Dubart-Kupperschmitt, A. (2007). Glycoprotein Ibalpha promoter drives megakaryocytic lineage-restricted expression after hematopoietic stem cell transduction using a self-inactivating lentiviral vector. Stem Cells (Dayton, Ohio), 25(6), 1571-7.
Lavenu-Bombled C, et al. Glycoprotein Ibalpha Promoter Drives Megakaryocytic Lineage-restricted Expression After Hematopoietic Stem Cell Transduction Using a Self-inactivating Lentiviral Vector. Stem Cells. 2007;25(6):1571-7. PubMed PMID: 17379771.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Glycoprotein Ibalpha promoter drives megakaryocytic lineage-restricted expression after hematopoietic stem cell transduction using a self-inactivating lentiviral vector. AU - Lavenu-Bombled,Cécile, AU - Izac,Brigitte, AU - Legrand,Faézeh, AU - Cambot,Marie, AU - Vigier,Agathe, AU - Massé,Jean-Marc, AU - Dubart-Kupperschmitt,Anne, Y1 - 2007/03/22/ PY - 2007/3/24/pubmed PY - 2007/8/19/medline PY - 2007/3/24/entrez SP - 1571 EP - 7 JF - Stem cells (Dayton, Ohio) JO - Stem Cells VL - 25 IS - 6 N2 - 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 with an integrative vector such as a lentiviral vector. Glycoprotein (Gp) IIb 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, bone marrow, and peripheral-blood mobilized CD34(+) cells were transduced with a human immunodeficiency virus-derived self-inactivating lentiviral vector encoding the green fluorescent protein (GFP) under the transcriptional control of GpIbalpha, GpIIb, or EF1alpha gene regulatory sequences. Both GpIbalpha and GpIIb promoters restricted GFP expression (analyzed by flow cytometry and immunoelectron microscopy) in MK cells among the maturing progeny of transduced cells. However, only the GpIbalpha promoter was strictly MK-specific, whereas 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 fragment of the human GpIbalpha 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. Disclosure of potential conflicts of interest is found at the end of this article. SN - 1066-5099 UR - https://www.unboundmedicine.com/medline/citation/17379771/Glycoprotein_Ibalpha_promoter_drives_megakaryocytic_lineage_restricted_expression_after_hematopoietic_stem_cell_transduction_using_a_self_inactivating_lentiviral_vector_ L2 - https://doi.org/10.1634/stemcells.2006-0321 DB - PRIME DP - Unbound Medicine ER -