Novel focused ultrasound enhanced calreticulin-nanoparticle for immune primed melanoma immunotherapy

用于免疫引发黑色素瘤免疫治疗的新型聚焦超声增强钙网蛋白纳米颗粒

• 批准号: 10177969
• 负责人: Ashish Ranjan
• 金额: $ 32.96万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-04 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177969
• 关键词: Ablation; Adverse effects; Antibodies; Antigen Presentation; Antitumor Response; B-Lymphocytes; Breast; CD47 gene; CT26; Canis familiaris; Cells; Client; Clinical; Clinical Treatment; Clonal Expansion; Colon; Data; Disease; Elements; Focused Ultrasound; Focused Ultrasound Therapy; Gene Delivery; Generations; Genetic Engineering; Genetically Engineered Mouse; Goals; Heating; Human; Hyperthermia; Immune; Immune Tolerance; Immune checkpoint inhibitor; Immunity; Immunologic Surveillance; Immunosuppression; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; Injections; Interferon Type I; Investigation; Leukocytes; Liposomes; Magnetic Resonance; Malignant Neoplasms; Mechanical Stress; Mechanics; Mediating; Melanoma Cell; Metastatic Melanoma; Metastatic to; Methodology; Methods; Modeling; Molecular; Mus; Mutation; Natural Immunity; Neoplasm Metastasis; Oral; Pathway interactions; Patients; Phagocytes; Plasmids; Production; Prostate; Proteins; Refractory; Role; Signal Pathway; Signal Transduction; Solid Neoplasm; Survival Rate; Systemic Therapy; T-Lymphocyte; T-cell inflamed; Temperature; Testing; Therapeutic; Tissues; Toxic effect; Translating; Translations; Tumor Antigens; Tumor Immunity; Tumor-infiltrating immune cells; Ultrasonography; Up-Regulation; Variant; adaptive immunity; anti-PD-L1 therapy; base; biomaterial compatibility; calreticulin; chemokine; chemoradiation; clinically relevant; combinatorial; cytokine; cytotoxic; dosage; efficacy evaluation; image guided; immune checkpoint; immunogenic; immunosuppressed; improved; in vivo; inhibitor/antagonist; innovation; melanoma; nanoparticle; neoplastic cell; novel; phase I trial; pointed protein; prevent; programmed cell death ligand 1; programmed cell death protein 1; response; success; survival outcome; thermal stress; tumor; tumor microenvironment

Summary Malignant melanoma in advanced stages is a highly lethal form of cancer, refractory to chemo- and radio-therapy, and the median survival is typically less than 4years. To improve response rates, antibodies that target CTL-4, PD1, and PDL1 has gained prominence, and can achieve a response rate of ~50% in metastatic cases. This is promising but a large proportion of patients still do not respond to immune therapies due to the absence of infiltrating T-cells, and presence of aberrant and suppresive signaling pathways that blunt the expression of checkpoint proteins (e.g. CD47, PDL1). To promote inflamed melanoma microenvironment, recent studies indicate that image-guided focused ultrasound (FUS) can i) mediate precise mechanical perturbation and elevation of tumor temperature to induce tumor antigen release and ii) upregulate calreticulin (CRT), a protein that is key to the activation of local and systemic anti-immunity. However, the exact mechanisms and how to translate this approach for clinical treatment of malignant melanoma is poorly understood. The goal of this project is to combine ultrasound guided FUS with novel CRT-NP, a liposome-plasmid nanoparticle agent that transfects melanoma cells to induce expression of CRT. Our in vitro and in vivo data in murine melanoma suggest that combined local treatment with CRT-NP/FUS (CFUS) enhances expression of CRT and modulates innate (CD47) and adaptive checkpoint proteins (PDL1); all of which significantly enhance the local and systemic immune priming and anti-tumor responses. Based on this premise, our central hypothesis is that CFUS targeted optimization of the CRT/CD47/PDL1 axis will provide powerful immune priming and generation of systemic immunity against malignant melanoma. To test our hypothesis, we will mechanistically dissect and understand CFUS-mediated immune priming in a B16 orthotopic and genetically-engineered mouse melanoma model (Aim 1 & 2) and translate this approach to clinical use in trials using client-owned dogs with spontaneous oral melanoma (Aims 3). Specifically, we will evaluate the impact of the CFUS treatment sequence, FUS exposures, and CRT-activation mechanisms in murine melanoma and translate this information to improve efficacy of checkpoint blockage in murine and canine tumor models. We expect that the successful optimization of local CFUS in this project will liberate tumors from their immune- suppressive state, achieve consistent and predictable clonal expansion of cytotoxic immune cells, and improve immunotherapy efficacy independent of cancer complexity. If successful, this method will provide a promising new avenue for treating melanoma and other types of solid tumor (e.g., breast, prostate) by significantly overcoming current immunotherapy barriers.

概括 晚期恶性黑色素瘤是一种高度致命的癌症，对化疗和放疗无效， 中位生存期通常小于 4 年。为了提高缓解率，针对 CTL-4 的抗体， PD1 和 PDL1 已受到重视，在转移病例中可以达到约 50% 的缓解率。这是 有希望，但由于缺乏免疫疗法，很大一部分患者仍然对免疫疗法没有反应 浸润 T 细胞，以及异常和抑制性信号通路的存在会削弱 T 细胞的表达 检查点蛋白（例如 CD47、PDL1）。为了促进发炎的黑色素瘤微环境，最近的研究 表明图像引导聚焦超声（FUS）可以 i）介导精确的机械扰动和 升高肿瘤温度以诱导肿瘤抗原释放，以及 ii) 上调钙网蛋白 (CRT)（一种蛋白质） 这是激活局部和全身抗免疫的关键。然而，确切的机制以及如何 这种方法对于临床治疗恶性黑色素瘤的了解还很少。 该项目的目标是将超声引导的 FUS 与新型 CRT-NP（一种脂质体质粒）结合起来 转染黑色素瘤细胞以诱导 CRT 表达的纳米颗粒剂。我们的体外和体内数据 小鼠黑色素瘤表明，与 CRT-NP/FUS (CFUS) 联合局部治疗可增强 CRT 并调节先天 (CD47) 和适应性检查点蛋白 (PDL1)；所有这些都显着增强 局部和全身免疫启动和抗肿瘤反应。基于这个前提，我们的中心假设 CFUS 对 CRT/CD47/PDL1 轴的靶向优化将提供强大的免疫启动和 产生针对恶性黑色素瘤的全身免疫力。 为了检验我们的假设，我们将机械地剖析和理解 B16 中 CFUS 介导的免疫启动 原位和基因工程小鼠黑色素瘤模型（目标 1 和 2）并将这种方法转化为临床 在使用患有自发性口腔黑色素瘤的客户拥有的狗进行的试验中使用（目标 3）。具体来说，我们将评估 CFUS 治疗顺序、FUS 暴露和 CRT 激活机制对小鼠的影响 黑色素瘤并转化此信息以提高小鼠和犬肿瘤检查点阻断的功效 模型。 我们期望该项目中局部 CFUS 的成功优化将把肿瘤从免疫中解放出来。 抑制状态，实现细胞毒性免疫细胞的一致且可预测的克隆扩增，并改善 免疫疗法的疗效与癌症的复杂性无关。如果成功的话，该方法将提供一个有前途的 治疗黑色素瘤和其他类型实体瘤（例如乳腺癌、前列腺癌）的新途径 克服当前免疫治疗障碍。