Abstract 576: Neoadjuvant immunotherapy improves efficacy of image-guided thermal ablation to generate curative responses in a murine breast cancer model
Authors: Silvestrini Matthew T. , Ingham Elizabeth S. , Mahakian Lisa M. , Kheirolomoom Azadeh , Liu Yu , Fite Brett Z. , Tam Sarah M. , Tucci Samantha , Watson Katherine D. , Wong Andrew W. , Monjazeb Arta M. , Hubbard Neil E. , Murphy William J. , Borowsky Alexander D. , Ferrara Katherine W. 
Published: July 2017

Abstract
Magnetic resonance-guided focused ultrasound (MRgFUS) facilitates local tumor control via thermal ablation, however, the anti-tumor immune effects induced are weak and unable to consistently generate robust objective responses in distant lesions. Here, we set out to optimize a therapeutic approach for employing immunotherapy with thermal ablation for systemic cancer treatment. We assessed the efficacy of implementing MRgFUS ablation with blockade of the PD-1/PD-L1 axis (anti-PD-1) and activation of TLR9 (CpG oligonucleotide) under various protocols and in multiple models of murine cancer. Anti-PD-1 (200 µg, i.p., days 21 & 28) and CpG (100 µg, i.t., days 21, 24 and 28) were administered coincidentally with MRgFUS ablation (3 MHz central frequency, circular pattern with R=2 mm, 1 revolution per second, 65ºC for 1 min, days 21 and 28) over the course of a week in bilateral syngeneic neu deletion line (NDL), 4T-1 and B16 tumor bearing mice. Additionally, we evaluated the administration of immunotherapy prior to a course of thermal ablation (i.e., “primed ablation”), where anti-PD-1 (as above on days 21, 28 & 35), CpG (as above on days 21, 24, 28, 31, 38 and 45) and MRgFUS ablation (as above on days 31, 38 and 45) were administered in bilateral NDL tumor-bearing mice. Primed ablation generated a robust anti-tumor immune response in distant lesions two weeks after the start of treatment, where a threefold increase in tumor infiltrating leukocytes (reaching 40% CD8+ and 20% CD4+ T-cells) was observed. This led to a complete response in 80% of treated mice within 70 days after treatment commenced. This effect was also observed in animals with high tumor burden and when thermal ablation was performed sequentially at multiple independent sites; 80% of untreated lesions were eradicated at 50 days after the start of treatment. However, therapeutic efficacy was limited when thermal ablation was performed coincident with the first dose of immunotherapy; this protocol was not curative in any murine model. To elucidate the mechanism for this effect, we employed tumor histology and positron emission tomography immediately after MRgFUS ablation. We found that thermal ablation induced stromal inflammation, and the loss of cell-cell adhesion and local vascular integrity, which impacted the intratumoral transport of small molecules and proteins for 48 hours post treatment. These data suggest that tumor debulking using image-guided thermal therapy can be successfully incorporated within a curative protocol in which immunotherapy is initiated before ablation.
Citation Format: Matthew T. Silvestrini, Elizabeth S. Ingham, Lisa M. Mahakian, Azadeh Kheirolomoom, Yu Liu, Brett Z. Fite, Sarah M. Tam, Samantha Tucci, Katherine D. Watson, Andrew W. Wong, Arta M. Monjazeb, Neil E. Hubbard, William J. Murphy, Alexander D. Borowsky, Katherine W. Ferrara. Neoadjuvant immunotherapy improves efficacy of image-guided thermal ablation to generate curative responses in a murine breast cancer model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 576. doi:10.1158/1538-7445.AM2017-576