SEARCH:   Advanced

Prepublished online as a Blood First Edition Paper on July 5, 2002; DOI 10.1182/blood-2002-02-0599. This Article Full Text (PDF) All Versions of this Article: 2002-02-0599v1 100/4/1257    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Nicolini, F. E Articles by Eaves, C. J Search for Related Content PubMed PubMed Citation Articles by Nicolini, F. E Articles by Eaves, C. J Social Bookmarking                   What's this? Submitted February 27, 2002 Accepted March 29, 2002 Expression Of A Human ß-globin Transgene In Erythroid Cells Derived From Retrovirally Transduced Transplantable Human Fetal Liver And Cord Blood Cells Franck E Nicolini, Suzan Imren, Il-Hoan Oh, R. Keith Humphries, Philippe Leboulch, Mary E Fabry, Ronald L Nagel, and Connie J Eaves* The Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, Canada Harvard-MIT Division of Health Sciences and Technology, Massachussets Institute of Technology, Cambridge, MA, USA; Genetix Pharmaceuticals, Cambridge, MA, USA Albert Einstein College of Medicine, Bronx, NY, USA * Corresponding author; email: ceaves{at}bccancer.bc.ca. Transfer of therapeutic genes to human hematopoietic stem cells (HSCs) using complex vectors at clinically relevant efficiencies remains a major challenge. Recently we described a stable retroviral vector that sustains long-term expression of GFP and a human ß-globin gene in the erythroid progeny of transduced murine HSCs. We now report the efficient transduction of primitive human CD34+ fetal liver or cord blood cells with this vector and expression of the ß-globin transgene in the erythroid progeny of these human cells for at least 2 months. After growth factor pre-stimulation and then a 2-3 day exposure to the virus, 35-55% GFP+ progeny were seen in assays of transduced colony-forming cells, primitive erythroid precursors that generate large numbers of glycophorin A-positive cells in 3-week suspension cultures, and 6-week long-term culture-initiating cells. In immunodeficient mice injected with unselected infected cells, 5-15% of the human cells regenerated in the marrow (including the erythroid cells) were GFP+ 3 and 6 weeks post-transplant. Importantly, the numbers of GFP+ human lymphoid and either granulopoietic or erythroid cells in individual mice 6 weeks post-transplant were significantly correlated, indicative of the initial transduction of human multi-potent cells with in vivo repopulating activity. Expression of the transduced ß-globin gene in human cells obtained directly from the mice or after their differentiation into erythroid cells in vitro was demonstrated by reverse transcriptase-PCR using specific primers. These experiments represent a significant step towards the realization of a gene therapy approach for human ß-globin gene disorders. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: G. F. Atweh, J. DeSimone, Y. Saunthararajah, H. Fathallah, R. S. Weinberg, R. L. Nagel, M. E. Fabry, and R. J. Adams Hemoglobinopathies Hematology, January 1, 2003; 2003(1): 14 - 39. [Abstract] [Full Text] [PDF]

	Copyright © 2002 by American Society of Hematology         Online ISSN: 1528-0020