Title
Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein
Author
Krom, Y.D.
Gras, J.C.E.
Frants, R.R.
Havekes, L.M.
van Berkel, T.J.
Biessen, E.A.L.
van Dijk, K.W.
TNO Kwaliteit van Leven
Publication year
2005
Abstract
Vascular smooth muscle (VSMC) and endothelial cells (EC) are particularly resistant to infection by type 5 adenovirus (Ad) vectors. To overcome this limitation and target Ad vectors to ubiquitously expressed α Vβ3/5 integrins, we have generated a linker protein consisting of the extracellular domain of the coxsackie adenovirus receptor (CAR) connected via avidin to a biotinylated cyclic (c) RGD peptide. After optimization of CAR to cRGD and to Ad coupling, infection of mouse heart endothelial cells (H5V) could be augmented significantly, as demonstrated by 600-fold increased transgene expression levels. In EOMAs, a hemangioendothelioma-derived cell line, the fraction of infected cells was enhanced 4- to 6-fold. Furthermore, the fraction of infected primary mouse VSMC was increased from virtually 0% to 25%. Finally, in human umbilical vein endothelial cells, the number of GFP positive cells was enhanced from 2% to 75%. In conclusion, CAR-cRGD is a versatile and highly efficient construct to target Ad vectors to both transformed and primary VSMC and EC. © 2005 Elsevier Inc. All rights reserved. Chemicals / CAS: arginylglycylaspartic acid, 99896-85-2; arginyl-glycyl-aspartic acid, 99896-85-2; Integrin alphaVbeta3; integrin alphaVbeta5; Integrins; Oligopeptides; Receptors, Vitronectin; Recombinant Fusion Proteins
Subject
Biomedical Research
Adenovirus
Cyclic RGD peptide
Endothelial cells
Targeting
Vascular smooth muscle cells
adenovirus vector
arginylglycylaspartic acid
avidin
coxsackie virus and adenovirus receptor
cyclopeptide
green fluorescent protein
integrin
animal cell
animal tissue
biotinylation
cell fractionation
cell line
endothelium cell
gene expression
gene targeting
heart
hemangioendothelioma
human cell
infection
mouse
nonhuman
priority journal
protein targeting
transgene
umbilical vein
vascular smooth muscle
Adenoviridae
Animals
CHO Cells
Cricetinae
Cricetulus
Genetic Vectors
Humans
Integrin alphaVbeta3
Integrins
Male
Mice
Mice, Inbred C57BL
Muscle, Smooth, Vascular
Myocytes, Smooth Muscle
Oligopeptides
Protein Engineering
Receptors, Vitronectin
Recombinant Fusion Proteins
Transfection
Animalia
Coxsackievirus
To reference this document use:
http://resolver.tudelft.nl/uuid:9a7f79b6-49b4-404e-bfc9-11a2ed873b17
DOI
https://doi.org/10.1016/j.bbrc.2005.10.073
TNO identifier
239031
ISSN
0006-291X
Source
Biochemical and Biophysical Research Communications, 338 (2), 847-854
Document type
article