Inverse targeting of retroviral vectors: Selective gene transfer in a mixed population of hematopoietic and nonhematopoietic cells

Adele K. Fielding, Marielle Maurice, Frances J. Morling, F. L. Cosset, Stephen J Russell

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

We previously reported that retroviral vectors displaying epidermal growth factor (EGF) as part of a chimeric envelope glycoprotein are sequestered upon binding to EGF receptor (EGFR)-positive target cells, leading to loss of infectivity. In the current study, we have displayed stem cell factor (SCF) on β-galactosidase-transducing ecotropic and amphotropic retroviral vector particles as a factor Xa protease-cleavable N-terminal extension of the envelope glycoprotein. Viral incorporation of the SCF chimeric envelopes was demonstrated by immunoblotting of pelleted virions and their specific attachment to Kit receptors was demonstrated by flow cytometry. Gene transfer studies showed that when SCF was displayed on an amphotropic envelope, the infectivity of the SCF-displaying vectors was selectively inhibited on Kit-expressing cells, but could be restored by adding soluble SCF to block the Kit receptors or by cleaving the displayed SCF domain from the vector particles with factor Xa protease. The host range properties of EGF-displaying and SCF-displaying vectors were then compared in cell mixing experiments. When EGFR-positive cancer cells and Kit-positive hematopoietic cells were mixed and exposed to the different engineered vector particles, the cancer cells were selectively transduced by the SCF-displaying vector and the hematopoietic cells were selectively transduced by the EGF- displaying vector. Retroviral display of polypeptide growth factors can therefore provide the basis for a novel inverse targeting strategy with potential use for selective transduction of hematopoietic or nonhematopoietic cells (eg, cancer cells) in a mixed cell population.

Original languageEnglish (US)
Pages (from-to)1802-1809
Number of pages8
JournalBlood
Volume91
Issue number5
StatePublished - Mar 1 1998
Externally publishedYes

Fingerprint

Gene transfer
Stem Cell Factor
Population
Genes
Cells
Epidermal Growth Factor
Factor Xa
Epidermal Growth Factor Receptor
Glycoproteins
Peptide Hydrolases
Galactosidases
Flow cytometry
Neoplasms
Host Specificity
Intercellular Signaling Peptides and Proteins
Immunoblotting
Virion
Display devices
Flow Cytometry
Peptides

ASJC Scopus subject areas

  • Hematology

Cite this

Inverse targeting of retroviral vectors : Selective gene transfer in a mixed population of hematopoietic and nonhematopoietic cells. / Fielding, Adele K.; Maurice, Marielle; Morling, Frances J.; Cosset, F. L.; Russell, Stephen J.

In: Blood, Vol. 91, No. 5, 01.03.1998, p. 1802-1809.

Research output: Contribution to journalArticle

Fielding, Adele K. ; Maurice, Marielle ; Morling, Frances J. ; Cosset, F. L. ; Russell, Stephen J. / Inverse targeting of retroviral vectors : Selective gene transfer in a mixed population of hematopoietic and nonhematopoietic cells. In: Blood. 1998 ; Vol. 91, No. 5. pp. 1802-1809.
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