Sensitive noninvasive monitoring of tumor perfusion during antiangiogenic therapy by intermittent bolus-contrast power Doppler sonography

Abstract

Intermittent bolus-contrast power Doppler ultrasound was used for noninvasive, quantitative monitoring of tumor perfusion during antiangiogenic therapy. Subcutaneous heterotransplants of human squamous cell carcinoma cells in nude mice were treated with a blocking antibody to vascular endothelial growth factor receptor 2 (DC101) and repeatedly examined at weekly intervals. Using replenishment kinetics of microbubbles (Levovist) tumor vascularization, including capillary blood flow, was clearly visualized by this dynamic ultrasound method allowing the determination of a comprehensive functional status of tumor vascularization (blood volume, blood flow, perfusion, and mean blood velocity) in all examined tumors. DC101 treatment decreased tumor blood flow (-64%) and volume (-73%) compared with untreated controls (+409% and +185%, respectively). Regression of functional vessel parameters was observed early well before reduction of tumor size. The treatment-related amount of reduction in tumor volume was directly correlated for the initial tumor blood flow before start of therapy and the perfusion calculated at the preceding examination. The vessel density (immunofluorescence staining with CD31 antibody at different time points) showed an excellent correlation with the calculated relative blood volume (k = 0.84, P < 0.01), thereby validating intermittent sonography as a useful monitoring method. We conclude that intermittent sonography is a promising tool for comprehensive monitoring of antiangiogenic or proangiogenic therapies, especially during early stages of treatment, thus yielding information regarding a prospective evaluation of therapy effects beyond the follow up of tumor size.