Therapeutic use of T cells engineered to produce radiation-inducible cytokines

经改造可产生辐射诱导细胞因子的 T 细胞的治疗用途

基本信息

批准号：
9810289
负责人：
RALPH R WEICHSELBAUM
金额：
$ 17.62万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9810289
关键词：
Adenovirus Vector Affect Animals Area Automobile Driving Beds Blood flow CD8-Positive T-Lymphocytes Cancer Patient Cells Cloning Cytokine Activation Cytokine Gene Data Disease Disseminated Malignant Neoplasm Dose Elements Enzyme-Linked Immunosorbent Assay Flow Cytometry Genetic Engineering Hour Image Imaging Techniques Imaging technology Immune Immunosuppression Interferon Type II Label Laboratories Link Locally Advanced Malignant Neoplasm MC38 Malignant Neoplasms Measures Modality Modeling Molecular Mus Primary Neoplasm Process Assessment Production Property Radiation Radiation therapy Radiation-Sensitizing Agents Radio Recombinants Refractory Reporter Genes Retroviral Vector Roentgen Rays Schedule Solid Neoplasm Spleen Subcutaneous Injections System T cell therapy T-Lymphocyte TNF gene Technology Testing Therapeutic Therapeutic Effect Therapeutic Uses Time Tissues Toxic effect Transduction Gene Transgenes Transgenic Organisms Translations Tumor-infiltrating immune cells Validation X-Ray Computed Tomography antitumor effect cancer cell cancer imaging cancer therapy cellular transduction chimeric gene clinical application cytokine cytokine therapy cytotoxic design effector T cell engineered T cells experience experimental study gene induction gene therapy image guided improved in vivo in vivo evaluation irradiation mouse model neoplastic cell promoter radiosensitizing serial imaging synergism systemic toxicity therapeutic cloning therapeutic cytokines therapeutic evaluation therapeutic gene tumor vector

项目摘要

Project Summary: We propose to use genetically-engineered T cells to deliver cytotoxic cytokines under the control of a radio-inducible promoter. When the genetically engineered T cells localize to the tumor bed, activation of the cytokine is achieved by radiotherapy (RT), thereby exposing the tumor to the therapeutic effects of both RT and cytokine therapy. This “molecular switch” strategy was pioneered by our laboratory, employing recombinant adenoviral vectors. In this concept, RT activates a chimeric gene with a radio-inducible promoter linked to a cytokine gene. Furthermore, the direct anti-tumor effects of RT have the potential to synergize with the anti-tumor effects of adoptively-transferred T cells. HYPOTHESIS: The ability to exogenously control the production of cytokines by tumor-infiltrating T cells will (i) counteract the immunosuppressive effects of the tumor, affecting a critical anti-tumor T cell effector function, (ii) minimize systemic toxicity due to controlled local activation of cytokine production, (iii) potentiate the synergy between RT and adoptive transferred T cells, and therefore (iv) improve the therapeutic ratio of RT. Specific Aims: 1) Cloning of T cell-specific radio-inducible promoters in retroviral vectors driving expression of a reporter gene for transduction of T cells, and in vivo analysis of gene induction by RT using longitudinal tumor imaging; 2) Cloning of T cell-specific radio-inducible promoters in retroviral vectors to drive expression of therapeutic cytokines in T cells and treatment of murine primary and oligometastatic tumors with radio-inducible cytokines-expressing T cells plus local RT. Our imaging technology uniquely allows us to examine the same tumor, and an identical area of the tumor, hours or days after each RT dose, and measure the increase in EGFP expression compared to levels before RT. This will allow in vivo comparison and validation of radio-induction of T cell-specific promoters identified in Aim 1. Furthermore, anti-tumor effects of T cells will be visible as destruction of cancer cells and vessels, thereby providing us with mechanistic data. Other cutting-edge technologies in our proposal include a CT-scan image- guided small animal irradiator for irradiation of mouse cancer tissues with millimetric precision. Our proposed combination of RT, gene therapy and adoptive T-cell therapy for treatment of locally advanced or metastatic cancer is dramatically new, and therefore has the potential to move beyond traditional approaches in the treatment of cancer. The initial clinical application of our new paradigm is the treatment of refractory solid tumors and oligometastases with the combined effects of adoptively-transferred T cells carrying radio-inducible cytokine cargo and RT, since RT has been shown to be an immunostimulatory modality; therefore, it is likely that translation of our approach would result in increased cures and significantly lower toxicities for cancer patients. Of note, our system could be applied to any other therapeutic gene that needs to be activated locally.

项目摘要：我们建议使用基因工程T细胞在以下条件下传递细胞毒性细胞因子：当基因工程 T 细胞定位于肿瘤床时，放射诱导启动子的控制通过放射治疗 (RT) 激活细胞因子，从而使肿瘤受到治疗效果这种“分子开关”策略是我们实验室首创的，采用了 RT 和细胞因子疗法。在这个概念中，RT激活具有放射诱导启动子的嵌合基因。此外，RT 的直接抗肿瘤作用有可能与细胞因子基因产生协同作用。过继转移的 T 细胞的抗肿瘤作用。假设：肿瘤浸润 T 细胞外源控制细胞因子产生的能力将 (i) 抵消肿瘤的免疫抑制作用，影响关键的抗肿瘤 T 细胞效应功能，(ii) 通过控制细胞因子产生的局部激活，最大限度地减少全身毒性，(iii) 增强协同作用 RT 和过继转移 T 细胞之间的差异，因此 (iv) 提高 RT 的治疗率。具体目标： 1) 在逆转录病毒载体中克隆 T 细胞特异性放射诱导启动子，驱动 T 细胞表达用于 T 细胞转导的报告基因，以及使用纵向肿瘤通过 RT 进行基因诱导的体内分析成像；2) 在逆转录病毒载体中克隆 T 细胞特异性放射诱导启动子以驱动 T 细胞中的治疗性细胞因子以及放射诱导治疗小鼠原发性和寡转移性肿瘤表达细胞因子的 T 细胞加局部 RT。我们独特的成像技术使我们能够检查相同的肿瘤以及肿瘤的相同区域，每次 RT 剂量后数小时或数天，并测量 EGFP 表达与之前水平相比的增加 RT. 这将允许对 T 细胞特异性启动子的放射诱导进行体内比较和验证。目标 1. 此外，T 细胞的抗肿瘤作用将通过破坏癌细胞和血管而显现出来，从而我们提案中的其他尖端技术包括 CT 扫描图像。引导小动物照射器，用于以毫米级精度照射小鼠癌组织。我们建议结合放疗、基因疗法和过继性 T 细胞疗法来治疗局部晚期或转移性癌症是全新的，因此有可能超越传统方法我们的新范例的最初临床应用是难治性固体的治疗。肿瘤和寡转移与携带放射诱导的过继转移 T 细胞的综合作用细胞因子货物和 RT，因为 RT 已被证明是一种免疫刺激方式，因此很可能；我们的方法的转化将增加治愈率并显着降低毒性值得注意的是，我们的系统可以应用于任何其他需要激活的治疗基因。本地。