Sa. Nicklin et al., Selective targeting of gene transfer to vascular endothelial cells by use of peptides isolated by phage display, CIRCULATION, 102(2), 2000, pp. 231-237
Citations number
33
Categorie Soggetti
Cardiovascular & Respiratory Systems","Cardiovascular & Hematology Research
Background-Gene transfer to vascular cells is a highly inefficient and nons
elective process, defined by the lack of specific cell-surface receptors fo
r 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 quie
scent human umbilical vein endothelial cells (HUVECs) with reduced or negli
gible binding to nonendothelial cells, including vascular smooth muscle cel
ls and hepatocytes. By direct biopanning on HUVECs and a second approach in
volving preclearing steps before panning on HUVECs, we isolated and sequenc
ed 140 individual phages and identified 59 peptides, We selected 7 candidat
es for further investigation by secondary screening of homogeneous phages o
n a panel of cell types. Using adenovirus-mediated gene transfer as a model
gene delivery system, we cloned the peptide SIGYPLP and the positive contr
ol peptide KKKKKKK upstream of the S11e single-chain Fv (" adenobody") dire
cted against the knob domain of the adenovirus to create fusion proteins. A
denovirus-mediated gene transfer via fiber-dependent infection was blocked
with S11e, whereas inclusion of the KKKKKKK peptide retargeted gene transfe
r. The peptide SIGYPLP, however, retargeted gene delivery specifically to e
ndothelial 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 pe
ptides isolated via phage display to target gene delivery specifically and
efficiently to HUVECs and highlights their use for retargeting both viral a
nd nonviral gene transfer to vascular endothelial cells for future clinical
applications.