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Fibronectin improves transduction of reconstituting hematopoietic stem
cells by retroviral vectors: evidence of direct viral binding to
chymotryptic carboxy-terminal fragments
T Moritz, P Dutt, X Xiao, D Carstanjen, T Vik, H Hanenberg and DA Williams
Herman B Wells Center for Pediatric Research, Riley Hospital for Children,
Indiana University School of Medicine, Indianapolis 46202- 5225, USA.
Efficient transduction of reconstituting hematopoletic stem cells (HSC) is
currently only possible by cocultivation of target cells directly on
producer cell lines, a method not applicable to human gene therapy
protocols. Our laboratory has previously shown adhesion of primitive
hematopoletic stem and progenitor cells to the carboxy-terminal 30/35- kD
fragment of the extracellular matrix molecule fibronectin (FN 30/35)
(Nature 352:438, 1991) and increased transduction of human hematopoietic
progenitor cells via retroviral vectors while adherent to this fragment (J
Clin Invest 93:1451, 1994). Here we report that (1) transduction of
reconstituting murine HSC assayed 12 months after infection with retrovirus
supernatant on FN 30/35 is as effective as cocultivation directly on
producer cells; (2) recombinant retrovirus particles directly adhere to FN
30/35 in a quantitative and dose- dependent fashion; and (3) increased
transduction efficiency on FN 30/ 35 does not appear to be associated with
increased cell proliferation or activation of protein phosphorylation
typically induced by integrin- fibronectin interactions. Therefore, we
speculate that supernatant infection of HSC on FN 30/35 leads to
colocalization of retrovirus particles and target cells on FN 30/35
molecule with a large increase in local virus titer presented to the cell.
These findings have direct and important implications for the modification
of current human gene therapy protocols.
Volume 88,
Issue 3,
pp. 855-862,
08/01/1996
Copyright © 1996 by The American Society of Hematology
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