Ultrasound-Controlled Immunotherapy for Targeted Treatment of Solid Tumors

超声控制免疫疗法用于实体瘤的靶向治疗

• 批准号: 10399420
• 负责人: Justin Lee
• 金额: $ 5.18万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2023-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10399420
• 关键词: Address; Adverse effects; Animal Model; Antigen Targeting; Basic Science; Blood Vessels; Cell Therapy; Cell physiology; Cells; Cellular immunotherapy; Cessation of life; Clinical Trials; Communication; DNA; Data; Development; Diagnostic; Disease model; ERBB2 gene; Elements; Engineering; Enterobacteria phage P1 Cre recombinase; Environment; Exposure to; Feedback; Focused Ultrasound; Focused Ultrasound Therapy; Future; Gene Expression; Genes; Genetic; Goals; Heat-Shock Response; Heating; Hematologic Neoplasms; Immune; Immune system; Immunology; Immunomodulators; Immunotherapy; In Vitro; Interleukin-2; Life; Location; Malignant Neoplasms; Mammalian Cell; Mentors; Methods; Modeling; Mus; Oncology; Operative Surgical Procedures; Organism; Output; Performance; Peripheral; Pharmaceutical Preparations; Production; Proteins; Resolution; SKBR3; Safety; Signal Transduction; Solid Neoplasm; Stimulus; Structure of parenchyma of lung; Syndrome; Synthetic Genes; System; T cell therapy; T-Lymphocyte; Techniques; Technology; Temperature; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Toxic effect; Training; Trans-Activators; Transgenes; Translational Research; Tumor Antigens; Work; autocrine; base; cancer immunotherapy; cancer therapy; cell type; cellular engineering; chimeric antigen receptor T cells; clinical translation; cytokine; cytokine therapy; design; direct application; engineered T cells; experimental study; genetic element; improved; in vitro testing; in vivo; millimeter; mouse model; multidisciplinary; novel; operation; overexpression; programs; promoter; protein expression; recruit; remote control; spatiotemporal; success; synthetic biology; targeted treatment; therapeutic gene; therapeutic protein; time use; tool; tumor; ultrasound

Project Summary Advances in synthetic biology have enabled the development of increasing numbers of cell-based diagnostic and therapeutic tools. However, controlling these cells in vivo is difficult and current methods to communicate with engineered cells either suffer from poor spatiotemporal resolution or require invasive operations. In order to improve safety and efficacy of cell-based therapies, robust methods to control gene expression in vivo are required. Temperature is a unique communication signal as it can be modulated with millimeter precision deep within tissues non-invasively using focused ultrasound (FUS). In addition, cells have already evolved the ability to sense changes in temperature through heat shock promoters (HSPs). These genetic elements are ubiquitous across organisms, providing a platform to develop genetic circuits that will activate upon FUS heating. Combining the spatial control offered by FUS with cellular engineering to confer thermal sensitivity will allow us to create thermally responsive cells that will activate in specific locations following FUS treatment. One rapidly growing field that could benefit from spatially controlled gene expression is cell-based cancer immunotherapy. Immunotherapy has recently emerged as a promising new class of cancer therapies with transformative results in hematological malignancies. However, engineered cell-based immunotherapies such as CAR T-cells and immunomodulatory agents such as cytokines must overcome significant challenges before becoming more widely applicable for solid tumors. In recent clinical trials, CAR T-cells have attacked healthy tissues if their targeted antigen is not tumor limited, resulting in massive peripheral toxicity and death. Systemic cytokine therapy can also cause life-threatening adverse effects such as vascular leak syndrome. Both types of immunotherapy could benefit from spatially controlled therapeutic expression. This project’s overall goal is to develop HSP-driven circuits in primary T-cells that will allow thermal stimuli delivered by FUS to active therapeutic genes expression. Initial experiments will focus on developing T-cells in vitro that will respond to bulk heating by transiently releasing cytokine to boost CAR performance in an autocrine fashion or activating permanent CAR expression specifically in the tumor. We will accomplish this by developing HSP driven circuits that feature drug induced transactivators or Cre Recombinase. After validating these circuits in vitro, we will test their performance upon FUS treatment in vivo using a murine SKBR3 tumor model and an anti-HER2 CAR. This model will allow us to develop and demonstrate the performance of robust, FUS-activated primary T-cells with two distinct payload outputs. This proposed approach represents a novel combination of cellular engineering and therapeutic ultrasound to spatiotemporally control cell-based immunotherapies with direct application to solid tumor treatment. This technology also has substantial potential for further development and adaption to other cell types which may facilitate both basic science and translational research.

项目概要 合成生物学的进步使得越来越多的基于细胞的诊断方法得以发展 和治疗工具。然而，在体内控制这些细胞很困难，目前的通讯方法 工程细胞要么时空分辨率差，要么需要侵入性操作。为了 为了提高细胞疗法的安全性和有效性，控制体内基因表达的稳健方法是 必需的。温度是一种独特的通信信号，因为它可以进行毫米级精度深度调制 使用聚焦超声 (FUS) 在组织内进行非侵入性检测。此外，细胞已经进化出这种能力 通过热休克促进剂（HSP）感知温度变化。这些遗传因素无处不在 跨生物体，提供一个开发遗传电路的平台，该电路将在 FUS 加热时激活。组合 FUS 提供的空间控制与细胞工程赋予热敏感性将使我们能够创造 FUS 治疗后，热响应细胞将在特定位置激活。 细胞癌症是一个快速发展的领域，可以从空间控制的基因表达中受益 免疫疗法。免疫疗法最近已成为一种有前景的新型癌症疗法， 血液系统恶性肿瘤的变革性结果。然而，基于细胞的工程免疫疗法，例如 因为 CAR T 细胞和细胞因子等免疫调节剂必须克服重大挑战 越来越广泛地应用于实体瘤。在最近的临床试验中，CAR T细胞攻击健康 如果其靶向抗原不限于肿瘤，则会在组织中产生大量外周毒性和死亡。系统性 细胞因子治疗还可引起危及生命的不良反应，例如血管渗漏综合征。两种类型的 免疫疗法可以受益于空间控制的治疗表达。 该项目的总体目标是在原代 T 细胞中开发 HSP 驱动电路，以允许热刺激 由 FUS 传递至活性治疗基因表达。初步实验将集中于开发 T 细胞 体外，将通过瞬时释放细胞因子来响应大量加热，以提高自分泌中的 CAR 性能 时尚或激活肿瘤中的永久 CAR 表达。我们将通过开发来实现这一目标 HSP 驱动电路具有药物诱导的反式激活因子或 Cre 重组酶。验证这些电路后 在体外，我们将使用鼠 SKBR3 肿瘤模型和 抗HER2 CAR。该模型将使我们能够开发并展示强大的、FUS 激活的性能 具有两个不同有效负载输出的初级 T 细胞。这种提出的方​​法代表了一种新颖的组合 细胞工程和治疗超声来时空控制基于细胞的免疫疗法 直接应用于实体瘤治疗。该技术还具有进一步发展的巨大潜力 以及对其他细胞类型的适应，这可能会促进基础科学和转化研究。