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: 10627822
• 负责人: Ashish Ranjan
• 金额: $ 7.2万
• 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 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10627822
• 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; Cytoplasm; Data; Disease; Elements; Focused Ultrasound; Focused Ultrasound Therapy; Gene Delivery; Generations; Genetic Engineering; Genetically Engineered Mouse; Goals; Heating; Human; Hyperthermia; Immune; Immune Evasion; Immune Tolerance; Immune checkpoint inhibitor; Immunity; Immunologic Surveillance; Immunosuppression; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; Injections; Interferon Type I; Investigation; Leucocytic infiltrate; Liposomes; Magnetic Resonance; Malignant Neoplasms; Mechanical Stress; Mechanics; Mediating; Melanoma Cell; Metastatic Melanoma; Methodology; Methods; Modeling; Molecular; Mus; Mutation; Natural Immunity; Neoplasm Metastasis; Oral; Pathway interactions; Patients; Phagocytes; Plasmids; Production; Prostate; Proteins; Radiation therapy; Refractory; Role; Signal Pathway; Signal Transduction; Solid Neoplasm; Survival Rate; Systemic Therapy; T cell infiltration; T-Lymphocyte; T-cell inflamed; Temperature; Testing; Therapeutic; Tissues; Toxic effect; Transfection; Translating; Translations; Tumor Antigens; Tumor Immunity; Up-Regulation; Variant; adaptive immunity; anti-PD-L1 therapy; biomaterial compatibility; calreticulin; cancer infiltrating T cells; checkpoint modulation; chemokine; chemotherapy; clinically relevant; combinatorial; cytokine; cytotoxic; dosage; efficacy evaluation; image guided; immune checkpoint; immunogenic; immunosuppressed; improved; in vivo; inhibitor; innovation; melanoma; nanoparticle; neoplastic cell; novel; phase I trial; prevent; programmed cell death ligand 1; programmed cell death protein 1; response; success; survival outcome; thermal stress; tumor; tumor microenvironment; ultrasound

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形式 并且中位生存期通常小于4年。为了提高响应率，靶向CTL-4的抗体， PD1和PDL1已获得突出性，在转移性情况下可以达到约50％的响应率。这是 有希望，但很大一部分患者由于没有 渗透T细胞，以及钝性的异常和刺激信号通路的存在 检查点蛋白质（例如CD47，PDL1）。为了促进发炎的黑色素瘤微环境，最近的研究 表明图像引导的聚焦超声（FUS）可以）介导精确的机械扰动和 肿瘤温度的升高以诱导肿瘤抗原释放和ii）上调钙网蛋白（CRT），一种蛋白 这是激活局部和全身反免疫的关键。但是，确切的机制以及如何 对这种恶性黑色素瘤临床治疗的方法翻译得很糟糕。 该项目的目的是将超声引导的FUS与新型CRT-NP（脂质体质量化）相结合 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 鼠类黑色素瘤表明，与CRT-NP/FUS（CFU）合并局部治疗会增强 CRT并调节先天（CD47）和自适应检查点蛋白（PDL1）；所有这些都显着增强了 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 是CRT/CD47/PDL1轴的CFU目标优化将提供强大的免疫启动和 对恶性黑色素瘤的全身免疫力产生。 为了检验我们的假设，我们将在B16中进行机械剖析并理解CFUS介导的免疫启动 原位和基因工程的小鼠黑色素瘤模型（AIM 1＆2），将这种方法转化为临床 使用自发口服黑色素瘤的客户拥有的狗在试验中使用（Aims 3）。具体来说，我们将评估 CFUS治疗序列，FUS暴露和CRT激活机制的影响 黑色素瘤并翻译此信息以提高鼠和犬肿瘤中检查点阻塞的功效 型号。 我们预计，该项目中当地CFU的成功优化将使肿瘤免疫 - 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张，并改善 免疫疗法功效与癌症复杂性无关。如果成功，此方法将提供有希望的 可显着治疗黑色素瘤和其他类型的实体瘤（例如乳房，前列腺）的新途径 克服当前的免疫疗法障碍。