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Integration of adeno-associated virus vectors in CD34+ human hematopoietic
progenitor cells after transduction
G Fisher-Adams, KK Wong , G Podsakoff, SJ Forman and S Chatterjee
Department of Hematology and Bone Marrow Transplantation, City of Hope
National Medical Center, Duarte CA 91010, USA.
Gene transfer vectors based on adeno-associated virus (AAV) appear
promising because of their high transduction frequencies regardless of cell
cycle status and ability to integrate into chromosomal DNA. We tested
AAV-mediated gene transfer into a panel of human bone marrow or umbilical
cord-derived CD34+ hematopoietic progenitor cells, using vectors encoding
several transgenes under the control of viral and cellular promoters. Gene
transfer was evaluated by (1) chromosomal integration of vector sequences
and (2) analysis of transgene expression. Southern hybridization and
fluorescence in situ hybridization analysis of transduced CD34 genomic DNA
showed the presence of integrated vector sequences in chromosomal DNA in a
portion of transduced cells and showed that integrated vector sequences
were replicated along with cellular DNA during mitosis. Transgene
expression in transduced CD34 cells in suspension cultures and in myeloid
colonies differentiating in vitro from transduced CD34 cells approximated
that predicted by the multiplicity of transduction. This was true in CD34
cells from different donors, regardless of the transgene or selective
pressure. Comparisons of CD34 cell transduction either before or after
cytokine stimulation showed similar gene transfer frequencies. Our findings
suggest that AAV transduction of CD34+ hematopoietic progenitor cells is
efficient, can lead to stable integration in a population of transduced
cells, and may therefore provide the basis for safe and efficient ex vivo
gene therapy of the hematopoietic system.
Volume 88,
Issue 2,
pp. 492-504,
07/15/1996
Copyright © 1996 by The American Society of Hematology
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