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(Circulation. 2000;102:231.)
© 2000 American Heart Association, Inc.

Basic Science Reports

Selective Targeting of Gene Transfer to Vascular Endothelial Cells by Use of Peptides Isolated by Phage Display

Stuart A. Nicklin, PhD; Steve J. White, PhD; Sarah J. Watkins, PhD; Robert E. Hawkins, MD; Andrew H. Baker, PhD

From the Bristol Heart Institute, University of Bristol (S.A.N., S.J.W., A.H.B.), and the Paterson Institute for Cancer Research, University of Manchester (S.J.W., R.E.H.), UK. Drs Nicklin and Baker are now at the Department of Medicine and Therapeutics, University of Glasgow, Western Infirmary, UK. The first 2 authors contributed equally to this article.

Correspondence to Dr Andrew H. Baker, Department of Medicine and Therapeutics, University of Glasgow, Western Infirmary, Glasgow G11 6NT, UK. E-mail ab11f{at}clinmed.gla.ac.uk

Background—Gene transfer to vascular cells is a highly inefficient and nonselective process, defined by the lack of specific cell-surface receptors for both nonviral and viral gene delivery vectors.

Methods and Results—We used filamentous phage display to isolate a panel of peptides that have the ability to bind selectively and efficiently to quiescent human umbilical vein endothelial cells (HUVECs) with reduced or negligible binding to nonendothelial cells, including vascular smooth muscle cells and hepatocytes. By direct biopanning on HUVECs and a second approach involving preclearing steps before panning on HUVECs, we isolated and sequenced 140 individual phages and identified 59 peptides. We selected 7 candidates for further investigation by secondary screening of homogeneous phages on a panel of cell types. Using adenovirus-mediated gene transfer as a model gene delivery system, we cloned the peptide SIGYPLP and the positive control peptide KKKKKKK upstream of the S11e single-chain Fv ("adenobody") directed against the knob domain of the adenovirus to create fusion proteins. Adenovirus-mediated gene transfer via fiber-dependent infection was blocked with S11e, whereas inclusion of the KKKKKKK peptide retargeted gene transfer. The peptide SIGYPLP, however, retargeted gene delivery specifically to endothelial cells with a significantly enhanced efficiency over nontargeted adenovirus and without transduction of nontarget cells.

Conclusions—Our study demonstrates the feasibility of using small, novel peptides isolated via phage display to target gene delivery specifically and efficiently to HUVECs and highlights their use for retargeting both viral and nonviral gene transfer to vascular endothelial cells for future clinical applications.

Key Words: bacteriophages • peptides • gene therapy • viruses • cells

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