Selective targeting of adenovirus to alphavbeta3 integrins, VEGFR2 and Tie2 endothelial receptors by angio-adenobodies.
Tumor angiogenesis is a prominent mechanism, driving the development and progression of solid tumors and the formation of cancer cell metastasis. Newly formed tumor vessels represent an elective target for the activity and the delivery of cancer therapeutics. We targeted adenovirus (Ad5) to endothelial receptors which are up-regulated during the formation of new blood vessels, to enhance the efficiency of anticancer gene therapy applications. Bifunctional angio-adenobodies were constructed by the fusion of a single chain antibody directed against the adenoviral fiber knob, to different peptides recognizing the alpha(v)beta(3) integrins, VEGFR2 and Tie2 receptors on endothelial cells. The angio-adenobodies were coupled to the adenoviral vector, containing luciferase and GFP as reporter genes. In vitro data showed selective targeting of the Ad5 to the endothelial receptors both in mouse (H5V) and human cell lines (HUVEC). H5V cells, refractory to Ad5 infection, showed high level of luciferase expression when cells were infected with targeted virus. Viral transgene expression increased in HUVEC cells when cells were infected with Ad5 conjugated with angio-adenobody thereby demonstrating the affinity of the peptides for human endothelial cells also. In vivo data obtained from mice bearing a C26 colon carcinoma subcutaneously show viral transgene expression only in tumors infected with angio-adenobodies retargeted adenovirus. The results of the present study demonstrate that endothelial targeted angio-adenobodies represent a versatile tool to direct adenovirus from its native receptors to the integrins alpha(v)beta(3), VEGFR2 and Tie2 receptors that are fundamental in many angiogenesis related diseases such as cancer.
Haisma
Hidde J
HJ
Department of Pharmaceutical Gene Modulation, Groningen Research Institute for Pharmacy, University of Groningen, Ant Deusinglaan 1, 9713 AV Groningen, The Netherlands. h.j.haisma@rug.nl
Kamps
Gera K
GK
Bouma
Arend
A
Geel
Tessa M
TM
Rots
Marianne G
MG
Kariath
Anu
A
Bellu
Anna Rita
AR
D000256:Adenoviridae / Q000276:immunology / Q000472:pathogenicity*; D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000276:immunology; D002460:Cell Line; D045744:Cell Line, Tumor; D016503:Drug Delivery Systems / Q000379:methods*; D042783:Endothelial Cells / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000008:administration & dosage* / Q000276:immunology; D042787:Receptor, TIE-2 / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000008:administration & dosage*; D015854:Up-Regulation / Q000276:immunology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
D000970:Antineoplastic Agents; D039302:Integrin alphaVbeta3; D010455:Peptides; D057127:Single-Chain Antibodies; D042787:Receptor, TIE-2; D040301:Vascular Endothelial Growth Factor Receptor-2