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

• 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
• 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

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：我们建议使用遗传学设计的T细胞在下面递送细胞毒性细胞因子 控制放射性诱导启动子。当一般设计的T细胞本地化在肿瘤床上时， 细胞因子的激活是通过放射疗法（RT）实现的，从而使肿瘤暴露于治疗作用 RT和细胞因子疗法的。这种“分子转换”策略是由我们的实验室开创的 重组腺病毒载体。在这个概念中，RT用可诱导的启动子激活嵌合基因 与细胞因子基因有关。此外，RT的直接抗肿瘤作用具有与 适当转移的T细胞的抗肿瘤作用。 假设：通过肿瘤浸润的T细胞外源控制细胞因子的产生的能力将会（i） 抵消肿瘤的免疫抑制作用，影响关键的抗肿瘤T细胞效应子功能，（ii） 最小化因细胞因子产生的局部激活而导致的全身毒性，（iii）潜力协同作用 在RT和自适应转移的T细胞之间，因此（IV）改善了RT的治疗比率。 具体目的：1）逆转录病毒载体中T细胞特异性诱导型启动子的克隆驱动表达 用于T细胞翻译的报告基因，并使用纵向肿瘤对RT进行基因诱导的体内分析 成像； 2）逆转录病毒载体中T细胞特异性诱导型启动子的克隆以驱动表达 T细胞中的治疗性细胞因子以及用射线诱导的鼠原发性和寡聚肿瘤的治疗 表达细胞因子的T细胞和局部RT。 我们的成像技术独特地使我们能够检查相同的肿瘤，以及肿瘤的相同区域， 每次RT剂量后的小时或几天，并测量EGFP表达的增加与之前的水平相比 RT。这将允许体内比较和验证在 目的1。此外，T细胞的抗肿瘤作用将被视为癌细胞和血管的破坏，从而可以看到 为我们提供机械数据。我们的提案中的其他尖端技术包括CT扫描图像 - 引导的小动物辐照剂，以毫米精度照射小鼠癌组织。 我们提出的RT，基因疗法和自适应T细胞疗法的组合用于局部晚期治疗 或转移性癌症是巨大的，因此有可能超越传统方法 在癌症治疗中。我们新范式的最初临床应用是对难治性固体的处理 肿瘤和寡聚酶具有适当转移的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 我们方法的翻译将导致治疗方法增加，并显着降低毒性 癌症患者。值得注意的是，我们的系统可以应用于需要激活的任何其他治疗基因 本地。