Fibronectin improves transduction of reconstituting hematopoietic stem cells by retroviral vectors: evidence of direct viral binding to chymotryptic carboxy-terminal fragments.

Abstract

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.

• Authors: T Moritz, P Dutt, X Xiao, D Carstanjen, T Vik, H Hanenberg, D A Williams
• Journal: Blood (Blood) Vol. 88 Issue 3 Pg. 855-62 (Aug 01 1996) ISSN: 0006-4971 [Print] United States
• PMID: 8704241 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Fibronectins
• Peptide Fragments
• Recombinant Proteins
• Chymotrypsin
• Adenosine Deaminase

Topics

• 3T3 Cells
• Adenosine Deaminase (genetics, metabolism)
• Animals
• Cell Cycle
• Cells, Cultured
• Chymotrypsin (metabolism)
• Fibronectins (metabolism, pharmacology)
• Genetic Vectors (genetics, metabolism)
• Hematopoietic Stem Cell Transplantation (methods)
• Hematopoietic Stem Cells (drug effects, metabolism, virology)
• Mice
• Peptide Fragments (metabolism)
• Recombinant Proteins (metabolism)
• Retroviridae (genetics, metabolism)
• Transfection (drug effects)