In vivo optical imaging of integrin αv-β3 in mice using multivalent or monovalent cRGD targeting vectors

Background: The cRGD peptide is a promising probe for early non-invasive detection of tumors. This study aimed to demonstrate how RAFT-c(-RGDfK-)4, a molecule allowing a tetrameric presentation of cRGD, improved cRGD-targeting potential using in vivo models of alpha or negative tumors. Results: We chose the human embryonic kidney cells HEK293(beta(3)) (high levels of alpha(v)beta(3)) or HEK293(beta(1)) (alpha(v)beta(3)-negative but expressing alpha(v) and beta(1)) engrafted subcutaneously (s.c.)in mice. Non-invasive in vivo optical imaging demonstrated that as compared to its monomeric cRGD analogue, Cy5-RAFT-c(-RGDfK-)(4) injected intravenously had higher uptake, prolonged retention and markedly enhanced contrast in HEK293(beta(3)) than in the HEK293(alpha(1)) tumors. Blocking studies further demonstrated the targeting specificity and competitive binding ability of the tetramer. Conclusion: In conclusion, we demonstrated that Cy5-RAFT-c(-RGDfK-)(4) was indeed binding to the alpha(v)beta(3) receptor and with an improved activity as compared to its monomeric analog, confirming the interest of using multivalent ligands. Intravenous injection of Cy5-RAFT-c(-RGDfK-)4 in this novel pair of HEK293(beta(3)) and HEK293(alpha(1)) tumors, provided tumor/skin ratio above 15. Such an important contrast plus the opportunity to use the HEK293(alpha(1)) negative control cell line are major assets for the community of researchers working on the design and amelioration of RGD-targeted vectors or on RGD-antagonists.