Image-guided irreversible electroporation directed CAR T-cell delivery to solid tumors

图像引导不可逆电穿孔引导 CAR T 细胞递送至实体瘤

• 批准号: 10221646
• 负责人: Prasad S. Adusumilli
• 金额: $ 41.08万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221646
• 关键词: 3-Dimensional; Ablation; Address; Adoptive Transfer; Anatomy; Blood Vessels; C57BL/6 Mouse; Cell Death; Cell Therapy; Cell membrane; Cell physiology; Cells; Characteristics; Clinical; Clinical Trials; Coculture Techniques; Computer Simulation; Electroporation; Excision; Extracellular Matrix; Goals; Histology; Image; Immune; Immunofluorescence Immunologic; Immunosuppressive Agents; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; Inflammation; Inflammatory; Label; Lymphocyte; MSLN gene; Malignant Neoplasms; Malignant Pleural Effusion; Malignant Pleural Mesothelioma; Malignant mesothelioma; Maps; Mediating; Membrane; Metabolic; Modeling; Monitor; Mus; Operative Surgical Procedures; Patients; Pattern; Penetration; Physiologic pulse; Positron-Emission Tomography; Protocols documentation; Reproducibility; Research Design; Resected; Solid Neoplasm; Specimen; Surface Antigens; Surrogate Markers; T cell therapy; T-Lymphocyte; Techniques; Translating; Translations; Tumor-Infiltrating Lymphocytes; Tumor-infiltrating immune cells; Validation; Work; base; bioluminescence imaging; cancer cell; cancer therapy; cell killing; cellular targeting; chemokine; chimeric antigen receptor; chimeric antigen receptor T cells; clinical practice; clinically relevant; comparative; cytotoxic; cytotoxicity; design; electric field; experimental study; fluorodeoxyglucose positron emission tomography; genetically modified cells; image guided; in vivo; innovation; intravenous administration; mesothelin; metabolic imaging; mouse model; neoplasm immunotherapy; novel strategies; overexpression; success; targeted delivery; tool; tumor; tumor ablation; tumor microenvironment; tumor-immune system interactions; uptake

PROJECT ABSTRACT Tumor infiltrating lymphocytes (TILs) can significantly increase the success of immunotherapy. Solid tumors present barriers to TIL penetration and function. Strategies to increase TILs and their function are challenged by the inability to direct the delivery of T cells into solid tumors and a lack of techniques to predict their uptake. Through this proposal, we will develop the use of image-guided irreversible electroporation (IRE) as a novel approach to promote TILs in solid tumors. IRE is clinically used for the treatment of solid tumors by applying microsecond long electric pulses that kill cells through rapid permeabilization of the cell membrane. A unique feature of IRE is that, while killing cells, it spares the extracellular matrix and blood vessels within the treatment region, which is ideal for stimulating local inflammation and promoting TILs. We will investigate our proposed strategy in translational mouse models and in patients with malignant pleural mesothelioma (MPM), which is a solid tumor where a higher ratio of cytotoxic TIL to immunosuppressor cells has already been shown to increase patient survival. In our experiments, we will use chimeric antigen receptor (CAR) T cells that are genetically modified to recognize mesothelin, a cell surface antigen that is overexpressed on mesothelioma cancer cells. Based on prior work by the PIs' team, we rationalize that cell death from IRE: (i) can be imaged using intra-procedural FDG-PET; (ii) can generate an intratumoral chemokine gradient that promotes TILs; and (iii) therefore, FDG-PET guided IRE can serve as a surrogate to both map and drive tumor-targeted CAR T-cell delivery. Furthermore, our strategy of IRE-directed CAR T-cell infiltration can be repeated using electric pulse parameters designed to be cancer cell-targeted/TIL-sparing, thereby augmenting immunotherapeutic efficacy. The long-term goal of our work is to develop image-guided IRE as a platform for predictable, reproducible, and controllable delivery of CAR T cells and other cellular anticancer therapies to solid tumors. In Aim 1, we will create an integrated tumor immune microenvironment map (iTIMM) combining FDG-PET imaging, computer simulations, and quantitative multiplex immunofluorescence histology to develop a tool that will determine T- cell localization in solid tumors treated with IRE. iTIMM will then be used to develop a clinically relevant protocol that, in combination of intravenous administration of T cells, will be used to increase TILs. In Aim 2, we will use dual imaging with PET (cancer cells) and bioluminescence imaging (T cells) to validate IRE pulse parameters to selectively enrich and increase functional CAR T cells within MPM. In Aim 3, we will perform a pilot clinical trial using PET-guided transthoracic IRE of MPM prior to resection to validate the concept of iTIMM and its effect on TILs. The modulation of the tumor microenvironment with image-guided IRE to map and localize T-cell delivery, promoting CAR T cell function, and translational validation of iTIMM presents innovative rational combinations of clinically used techniques and is an ambitious attempt to address a key challenge to solid tumor immunotherapy.

项目摘要 肿瘤浸润淋巴细胞（TIL）可以显着提高免疫治疗的成功率。实体瘤 对 TIL 渗透和功能构成障碍。增加 TIL 的策略及其功能受到挑战 无法直接将 T 细胞输送到实体瘤中，并且缺乏预测其摄取的技术。 通过这个提案，我们将开发使用图像引导的不可逆电穿孔（IRE）作为一种新颖的方法 促进实体瘤中 TIL 的方法。 IRE在临床上用于实体瘤的治疗 微秒长的电脉冲通过细胞膜的快速透化来杀死细胞。一个独特的 IRE的特点是在杀死细胞的同时，在治疗过程中不伤害细胞外基质和血管 该区域非常适合刺激局部炎症和促进 TIL。我们将调查我们的提议 转化小鼠模型和恶性胸膜间皮瘤（MPM）患者的策略，这是一种 已证明细胞毒性 TIL 与免疫抑制细胞比例较高的实体瘤 提高患者的生存率。在我们的实验中，我们将使用嵌合抗原受体 (CAR) T 细胞 经过基因改造以识别间皮素，这是一种在间皮瘤上过度表达的细胞表面抗原 癌细胞。根据 PI 团队之前的工作，我们合理解释了 IRE 导致的细胞死亡：(i) 可以成像 使用术中 FDG-PET； (ii) 可以产生促进 TIL 的瘤内趋化因子梯度；和 (iii) 因此，FDG-PET 引导的 IRE 可以作为定位和驱动肿瘤靶向 CAR T 细胞的替代品 送货。此外，我们的 IRE 定向 CAR T 细胞浸润策略可以使用电脉冲重复 参数设计为癌细胞靶向/保留 TIL，从而增强免疫治疗功效。 我们工作的长期目标是开发图像引导 IRE 作为可预测、可重复和可预测的平台 CAR T 细胞和其他细胞抗癌疗法向实体瘤的可控递送。在目标 1 中，我们将 结合 FDG-PET 成像、计算机创建集成肿瘤免疫微环境图 (iTIMM) 模拟和定量多重免疫荧光组织学来开发一种工具来确定 T- IRE 治疗实体瘤中的细胞定位。然后，iTIMM 将用于开发临床相关的 结合静脉注射 T 细胞，将用于增加 TIL 的方案。在目标 2 中，我们 将使用 PET（癌细胞）和生物发光成像（T 细胞）双重成像来验证 IRE 脉冲 参数来选择性地富集和增加 MPM 内的功能性 CAR T 细胞。在目标 3 中，我们将执行 在切除前使用 PET 引导经胸 IRE 进行 MPM 的试点临床试验，以验证 iTIMM 的概念 及其对 TIL 的影响。使用图像引导 IRE 来调节肿瘤微环境以绘制和 iTIMM 的本地化 T 细胞递送、促进 CAR T 细胞功能和翻译验证呈现出创新 临床使用技术的合理组合，是解决关键挑战的雄心勃勃的尝试 实体瘤免疫治疗。