3-D Super-Resolution Ultrasound Imaging for Monitoring Early Changes in Breast Cancer after Treatment with a Vascular-Disrupting Agent

The purpose of this research project was to evaluate the use of 3-dimensional (3-D) super-resolution ultrasound (SR-US) imaging to assess any early changes in breast cancer after treatment with a vascular-disrupting agent (VDA). A Vevo 3100 ultrasound system (FUJIFILM VisualSonics Inc) equipped with an MX 201 transducer was used for image acquisition. A total of
$2.5\times 10^{7}$
microbubbles (MBs) were injected into the tail vein of anesthetized breast cancer-bearing mice using repeat bolus injections every 5 min. A total of 10 stacks of ultrasound images were collected as the transducer was mechanically moved across the tumor at 0.6 mm intervals yielding a 6-mm thick volume. At each tumor location, a stack contained
$1\ \times\ 10^{4}$
frames of ultrasound data that were acquired at 463 frames/sec and stored as in-phase/quadrature (IQ) format. After motion correction, each temporal stack of ultrasound images was processed separately for clutter signal removal, which was followed by MB localization and enumeration before generation of the final SR-US image. After reconstruction of the 3-D SR-US volume dataset, the tumor microvasculature was enhanced using a multiscale vessel enhancement filter. Vessels from the resultant microvascular network were then segmented using an adaptive thresholding method. Finally, mean microvascular density (MVD) measurements from each tumor volume were computed as a summarizing statistic. While no differences were found between baseline SR-US image-derived measures of MVD (
$p=0.76$
), these same measurements were significantly lower at 24
$h$
after VDA treatment (
$p < 0.001$
). Overall, 3-D SR-US imaging detected early tumor changes following treatment with a vascular-targeted drug.