PMID- 35039461 OWN - NLM STAT- MEDLINE DCOM- 20220316 LR - 20221009 IS - 2051-1426 (Electronic) IS - 2051-1426 (Linking) VI - 10 IP - 1 DP - 2022 Jan TI - Combination of ultrasound-based mechanical disruption of tumor with immune checkpoint blockade modifies tumor microenvironment and augments systemic antitumor immunity. LID - 10.1136/jitc-2021-003717 [doi] LID - e003717 AB - BACKGROUND: Despite multimodal adjuvant management with radiotherapy, chemotherapy and hormonal therapies, most surgically resected primary breast cancers relapse or metastasize. A potential solution to late and distant recurrence is to augment systemic antitumor immunity, in part by appropriately presenting tumor antigens, but also by modulating the immunosuppressive tumor microenvironment (TME). We previously validated this concept in models of murine carcinoma treated with a novel predominately microcavitating version of high-intensity focused ultrasound (HIFU), mechanical high-intensity focused ultrasound (M-HIFU). Here we elucidated the mechanisms of enhanced antitumor immunity by M-HIFU over conventional thermal high-intensity focused ultrasound (T-HIFU) and investigated the potential of the combinatorial strategy with an immune checkpoint inhibitor, anti-PD-L1 antibody. METHODS: The antitumor efficacy of treatments was investigated in syngeneic murine breast cancer models using triple-negative (E0771) or human ErbB-2 (HER2) expressing (MM3MG-HER2) tumors in C57BL/6 or BALB/c mice, respectively. Induction of systemic antitumor immunity by the treatments was tested using bilateral tumor implantation models. Flow cytometry, immunohistochemistry, and single-cell RNA sequencing were performed to elucidate detailed effects of HIFU treatments or combination treatment on TME, including the activation status of CD8 T cells and polarization of tumor-associated macrophages (TAMs). RESULTS: More potent systemic antitumor immunity and tumor growth suppression were induced by M-HIFU compared with T-HIFU. Molecular characterization of the TME after M-HIFU by single-cell RNA sequencing demonstrated repolarization of TAM to the immunostimulatory M1 subtype compared with TME post-T-HIFU. Concurrent anti-PD-L1 antibody administration or depletion of CD4(+) T cells containing a population of regulatory T cells markedly increased T cell-mediated antitumor immunity and tumor growth suppression at distant, untreated tumor sites in M-HIFU treated mice compared with M-HIFU monotherapy. CD8 T and natural killer cells played major roles as effector cells in the combination treatment. CONCLUSIONS: Physical disruption of the TME by M-HIFU repolarizes TAM, enhances T-cell infiltration, and, when combined with anti-PD-L1 antibody, mediates superior systemic antitumor immune responses and distant tumor growth suppression. These findings suggest M-HIFU combined with anti-PD-L1 may be useful in reducing late recurrence or metastasis when applied to primary tumors. CI - (c) Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. FAU - Abe, Shinya AU - Abe S AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. AD - Department of Surgical Oncology, Faculty of Medicine, The University of Tokyo Graduate School of Medicine, Bunkyo-ku, Tokyo, Japan. FAU - Nagata, Hiroshi AU - Nagata H AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. AD - Department of Surgical Oncology, Faculty of Medicine, The University of Tokyo Graduate School of Medicine, Bunkyo-ku, Tokyo, Japan. FAU - Crosby, Erika J AU - Crosby EJ AUID- ORCID: 0000-0002-4872-6711 AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Inoue, Yoshiyuki AU - Inoue Y AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. AD - Department of Surgery, Jichi Medical University, Shimotsuke, Tochigi, Japan. FAU - Kaneko, Kensuke AU - Kaneko K AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. AD - Department of Surgical Oncology, Faculty of Medicine, The University of Tokyo Graduate School of Medicine, Bunkyo-ku, Tokyo, Japan. FAU - Liu, Cong-Xiao AU - Liu CX AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Yang, Xiao AU - Yang X AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Wang, Tao AU - Wang T AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Acharya, Chaitanya R AU - Acharya CR AUID- ORCID: 0000-0001-7149-1749 AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Agarwal, Pankaj AU - Agarwal P AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Snyder, Joshua AU - Snyder J AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Gwin, William AU - Gwin W AD - Department of Medicine, University of Washington, Seattle, Washington, USA. FAU - Morse, Michael A AU - Morse MA AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. AD - Department of Medicine, Duke University Medical Center, Durham, North Carolina, USA. FAU - Zhong, Pei AU - Zhong P AD - Department of Mechanical Engineering and Materials Science, Duke University, Durham, North Carolina, USA. FAU - Lyerly, Herbert Kim AU - Lyerly HK AUID- ORCID: 0000-0002-0063-4770 AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA. FAU - Osada, Takuya AU - Osada T AUID- ORCID: 0000-0003-1424-5001 AD - Department of Surgery, Duke University Medical Center, Durham, North Carolina, USA osada001@duke.edu. LA - eng GR - P30 CA014236/CA/NCI NIH HHS/United States GR - T32 CA009111/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - England TA - J Immunother Cancer JT - Journal for immunotherapy of cancer JID - 101620585 RN - 0 (Immune Checkpoint Inhibitors) SB - IM MH - Animals MH - Cell Line, Tumor MH - Combined Modality Therapy/*methods MH - Female MH - Humans MH - Immune Checkpoint Inhibitors/pharmacology/*therapeutic use MH - Immunotherapy/*methods MH - Mice MH - Neoplasms/*diagnostic imaging/*drug therapy MH - Tumor Microenvironment MH - Ultrasonography/*methods PMC - PMC8765068 OTO - NOTNLM OT - breast neoplasms OT - immunotherapy OT - macrophages OT - translational medical research OT - tumor microenvironment COIS- Competing interests: None declared. EDAT- 2022/01/19 06:00 MHDA- 2022/03/17 06:00 PMCR- 2022/01/17 CRDT- 2022/01/18 05:46 PHST- 2021/12/13 00:00 [accepted] PHST- 2022/01/18 05:46 [entrez] PHST- 2022/01/19 06:00 [pubmed] PHST- 2022/03/17 06:00 [medline] PHST- 2022/01/17 00:00 [pmc-release] AID - jitc-2021-003717 [pii] AID - 10.1136/jitc-2021-003717 [doi] PST - ppublish SO - J Immunother Cancer. 2022 Jan;10(1):e003717. doi: 10.1136/jitc-2021-003717.