Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein.
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 alpha(V)beta(3/5) 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.
Krom
Y D
YD
Department of Human Genetics, Leiden University Medical Center, Leiden, The Netherlands. Y.D.Krom@lumc.nl
Gras
J C E
JC
Frants
R R
RR
Havekes
L M
LM
van Berkel
T J
TJ
Biessen
E A L
EA
van Dijk
K Willems
KW
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000096:biosynthesis* / Q000235:genetics; D016023:Integrins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D019030:Receptors, Vitronectin / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D039302:Integrin alphaVbeta3; D016023:Integrins; D009842:Oligopeptides; D019030:Receptors, Vitronectin; D011993:Recombinant Fusion Proteins; C064660:integrin alphaVbeta5; C047981:arginyl-glycyl-aspartic acid