Phase Changing Ultrasound Contrast Agents for Deep Tissue Imaging of Cellular Immunotherapies - PROJECT SUMMARY
Adoptive cellular immunotherapies are an established therapeutic weapon in the fight against cancer. However,
there is a large variability in anti-tumor responses and toxicity between patients, due primarily to differences in
immune cell trafficking to tumor and lymph tissues. This has urged the development of surveillance tools that
can monitor adoptive immune cell tissue migration in vivo to inform treatment regimens, empower interpretation
of therapeutic outcomes, and aid in the early detection of adverse events. To address this technologic need, our
goal is to develop nano-scale contrast agents that enable continuous and high-fidelity ultrasound (US) imaging
of immune cells in deep tissues, employing tumoricidal chimeric antigen receptor macrophages (CAR-M) as an
exemplary cellular model. Fundamental to this strategy is our development of phase-changing peptide
nanoemulsions (NPep) that are rapidly internalized and persist within macrophages for multiple days and can
generate echogenic bubble imaging nuclei on-demand without compromising cell viability. Our recent studies
show these capabilities allow NPeps to provide real-time and long-term monitoring of macrophages in tissues
using diagnostic B-mode and Doppler US imaging. Additional studies demonstrate NPeps provide a distinctive
Doppler ‘twinkling’ feature that permits unparalleled spatiotemporal resolution of contrast-enhanced features
from the tissue background to improve imaging resolution and identification of cellular locale. Bringing together
a multi-disciplinary team of materials scientists, ultrasound imaging experts, and immunologists, our objective is
to rationally tune NPep design to improve Doppler twinkling in cells (Aim 1), controllably modulate macrophage
behavior in situ via US release of immunostimulants loaded into the NPep carrier (Aim 2), and demonstrate non-
invasive, real-time, high contrast, and continuous imaging of NPep-labeled CAR-M intratumoral migration and
persistence in vivo (Aim 3). To achieve this, in aim 1 we perform multiplexed imaging of NPep formulations with
varying emulsion size and surface tension, two parameters linked to particle US response, to establish design
rules that can be used to improve the consistency, power, and duration of NPep Doppler twinkling. Aim 2 will
evaluate changes in macrophage viability, phenotype, and phagocytic function after uptake of NPep contrast
agents, as well as demonstrate our ability to productively modulate anti-tumor behavior via US-actuated
intracellular release of a loaded PPARγ agonist demonstrated to stimulate tumoricidal responses. In aim 3, we
evaluate the potential of this tool to provide real-time and long-term monitoring of adoptive CAR-M infiltration into
xenograft neuroblastoma tumors, as an exemplary in vivo model, using standard diagnostic US imaging
modalities. Collectively, this work will provide the foundation for a clinically relevant tool that can be broadly
adopted by the oncology community for real-time deep tissue imaging of adoptive cellular immunotherapies.
