Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein

• Published in: Biochemical and biophysical research communications Vol. 338; no. 2; pp. 847 - 854
• Main Authors: Krom, Y.D., Gras, J.C.E., Frants, R.R., Havekes, L.M., van Berkel, T.J., Biessen, E.A.L., van Dijk, K. Willems
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.12.2005
• Subjects: Adenoviridae - genetics; Adenovirus; Animals; CHO Cells; Cricetinae; Cricetulus; Cyclic RGD peptide; Endothelial cells; Genetic Vectors - genetics; Humans; Integrin alphaVbeta3 - biosynthesis; Integrin alphaVbeta3 - genetics; Integrins - biosynthesis; Integrins - genetics; Male; Mice; Mice, Inbred C57BL; Muscle, Smooth, Vascular - metabolism; Myocytes, Smooth Muscle - metabolism; Oligopeptides - genetics; Oligopeptides - metabolism; Protein Engineering - methods; Receptors, Vitronectin - biosynthesis; Receptors, Vitronectin - genetics; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Targeting; Transfection - methods; Vascular smooth muscle cells; Cyclic RGD peptide; Targeting; Adenovirus; Endothelial cells; Vascular smooth muscle cells
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2005.10.073

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 Vβ 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.