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Volume 271, Number 52, Issue of December 27, 1996 pp. 33647-33653
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Targeting DNA to Cells with Basic Fibroblast Growth Factor (FGF2)

(Received for publication, August 13, 1996, and in revised form, October 3, 1996)

Barbara A. Sosnowski , Ana Maria Gonzalez , Lois A. Chandler , Ying J. Buechler , Glenn F. Pierce and Andrew Baird

From PRIZM Pharmaceuticals, San Diego, California 92121

Ligand-mediated targeting of DNA was validated by condensing a plasmid DNA encoding the beta -galactosidase (beta -gal) gene with a basic fibroblast growth factor (FGF2) that was first chemically conjugated to polylysine (K). The conditions that gave optimal binding of this FGF2 to DNA also generated the highest level of beta -gal expression when added to FGF2 target cells like COS-1, 3T3, baby hamster kidney (BHK), or endothelial cells. This beta -gal activity increased in a time- and dose-dependent manner and was dependent on the inclusion of FGF2 in the complex. FGF receptor specificity was demonstrated by competition of the complex with FGF2 and heparin, and by the failure of cytochrome c or histone H1 to mimic the gene-targeting effects of FGF2. The expression of beta -gal was also endosome dependent because chloroquine increased beta -gal expression 8-fold and endosome disruptive peptides increased expression of beta -gal 26-fold. Taken together these findings establish that DNA can be introduced into cells through the high affinity FGF receptor complex, and while its efficiency will require significant enhancements to achieve sustained and elevated transgene expression, the possibility that the technique could be used to deliver DNAs encoding cytotoxic molecules is discussed.


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