Sustained gene expression in retrovirally transduced, engrafting human hematopoietic stem cells and their lympho-myeloid progeny

Inefficient retroviral-mediated gene transfer to human hematopoietic stem cells (HSC) and insufficient gene expression in progeny cells derived from transduced HSC are two major problems associated with HSC-based gene therapy. In this study we evaluated the ability of a murine stem cell virus (MSCV)...

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Bibliographic Details
Published in:Blood 1998-07, Vol.92 (1), p.83-92
Main Authors: LINZHAO CHENG, CHANGCHUN DU, LAVAU, C, CHEN, S, JIE TONG, CHEN, B. P, SCOLLAY, R, HAWLEY, R. G, HILL, B
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Biotechnology
Fundamental and applied biological sciences. Psychology
Gene Expression
Gene Transfer Techniques
Genes, Reporter
Genetic engineering
Genetic technics
Genetic Vectors
Graft Survival
Hematopoiesis - physiology
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells - cytology
Hematopoietic Stem Cells - physiology
Humans
Leukocytes - cytology
Leukocytes - physiology
Methods. Procedures. Technologies
Mice
Mice, SCID
Receptors, Nerve Growth Factor - genetics
Retroviridae
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary:Inefficient retroviral-mediated gene transfer to human hematopoietic stem cells (HSC) and insufficient gene expression in progeny cells derived from transduced HSC are two major problems associated with HSC-based gene therapy. In this study we evaluated the ability of a murine stem cell virus (MSCV)-based retroviral vector carrying the low-affinity human nerve growth factor receptor (NGFR) gene as reporter to maintain gene expression in transduced human hematopoietic cells. CD34(+) cells lacking lineage differentiation markers (CD34(+)Lin-) isolated from human bone marrow and mobilized peripheral blood were transduced using an optimized clinically applicable protocol. Under the conditions used, greater than 75% of the CD34(+) cell population retained the Lin- phenotype after 4 days in culture and at least 30% of these expressed a high level of NGFR (NGFR+) as assessed by fluorescence-activated cell sorter analysis. When these CD34(+)Lin-NGFR+ cells sorted 2 days posttransduction were assayed in vitro in clonogenic and long-term stromal cultures, sustained reporter expression was observed in differentiated erythroid and myeloid cells derived from transduced progenitors, and in differentiated B-lineage cells after 6 weeks. Moreover, when these transduced CD34(+)Lin-NGFR+ cells were used to repopulate human bone grafts implanted in severe combined immunodeficient mice, MSCV-directed NGFR expression could be detected on 37% +/- 6% (n = 5) of the donor-type human cells recovered 9 weeks postinjection. These findings suggest potential utility of the MSCV retroviral vector in the development of effective therapies involving gene-modified HSC.
ISSN:0006-4971
1528-0020
DOI:10.1182/blood.v92.1.83.413k09_83_92