Selective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display.

2.50
Hdl Handle:
http://hdl.handle.net/10541/90040
Title:
Selective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display.
Authors:
Nicklin, Stuart A; White, Steve J; Watkins, Sarah J; Hawkins, Robert E; Baker, Andrew H
Abstract:
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.
Affiliation:
Bristol Heart Institute, University of Bristol, UK.
Citation:
Selective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display. 2000, 102 (2):231-7 Circulation
Journal:
Circulation
Issue Date:
11-Jul-2000
URI:
http://hdl.handle.net/10541/90040
PubMed ID:
10889136
Type:
Article
Language:
en
ISSN:
1524-4539
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorNicklin, Stuart Aen
dc.contributor.authorWhite, Steve Jen
dc.contributor.authorWatkins, Sarah Jen
dc.contributor.authorHawkins, Robert Een
dc.contributor.authorBaker, Andrew Hen
dc.date.accessioned2010-01-19T17:07:46Z-
dc.date.available2010-01-19T17:07:46Z-
dc.date.issued2000-07-11-
dc.identifier.citationSelective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display. 2000, 102 (2):231-7 Circulationen
dc.identifier.issn1524-4539-
dc.identifier.pmid10889136-
dc.identifier.urihttp://hdl.handle.net/10541/90040-
dc.description.abstractBACKGROUND: 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.en
dc.language.isoenen
dc.subject.meshAdenoviridae-
dc.subject.meshAdenoviridae Infections-
dc.subject.meshCells, Cultured-
dc.subject.meshCloning, Molecular-
dc.subject.meshEndothelium, Vascular-
dc.subject.meshGene Transfer Techniques-
dc.subject.meshHela Cells-
dc.subject.meshHumans-
dc.subject.meshLiver-
dc.subject.meshMuscle, Smooth, Vascular-
dc.subject.meshPeptide Fragments-
dc.subject.meshPeptide Library-
dc.subject.meshPolylysine-
dc.subject.meshUmbilical Veins-
dc.titleSelective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display.en
dc.typeArticleen
dc.contributor.departmentBristol Heart Institute, University of Bristol, UK.en
dc.identifier.journalCirculationen

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