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Sustained Gene Expression in Retrovirally Transduced, Engrafting
Human Hematopoietic Stem Cells and Their Lympho-Myeloid Progeny
Linzhao Cheng,
Changchun Du,
Catherine Lavau,
Shirley Chen,
Jie Tong,
Benjamin P. Chen,
Roland Scollay,
Robert G. Hawley, and
Beth Hill
From SyStemix, Inc, Palo Alto, CA; the Oncology Gene Therapy Program,
The Toronto Hospital, Toronto; and the Department of Medical
Biophysics, University of Toronto, Toronto, Ontario, Canada.
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+LinNGFR+ 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+LinNGFR+ 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.
Blood, Vol. 92 No. 1 (July 1), 1998:
pp. 83-92
© 1998 by The American Society of Hematology.
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