Simplified Preparation of α_vβ₃ Integrin-Targeted Microbubbles Based on a Clinically Available Ultrasound Contrast Agent: Validation in a Tumor-Bearing Mouse Model

The usefulness of ultrasound molecular imaging with α_vβ₃ integrin-targeted microbubbles for detecting tumor angiogenesis has been demonstrated. Recently, we developed α_vβ₃ integrin-targeted microbubbles by modifying clinically available microbubbles (Sonazoid, Daiichi-Sankyo Pharmaceuticals, Tokyo, Japan) with a secreted glycoprotein (lactadherin). The aims of our present study were to simplify the preparation of lactadherin-bearing Sonazoid and to examine the diagnostic utility of lactadherin-bearing Sonazoid for α_vβ₃ integrin-expressing tumor vessels by using SK-OV-3-tumor-bearing mice. By incubating 1.2 × 10⁷ Sonazoid microbubbles with 1.0 µg lactadherin, the complicated washing and centrifugation steps during the microbubble preparation could be omitted with no significant reduction in labeling ratio of lactadherin-bearing Sonazoid. In addition, the number of Sonazoid microbubbles accumulated in the SK-OV-3 tumor was significantly increased by modifying Sonazoid with lactadherin. Our data suggest that the lactadherin-bearing Sonazoid is an easily prepared and potentially clinically translatable targeted microbubble for α_vβ₃ integrin-expressing vessels.