Genetic Approaches to Optimize CAR T cells for Glioblastoma Therapy

优化 CAR T 细胞用于胶质母细胞瘤治疗的基因方法

• 批准号: 9790997
• 负责人: Irina V Balyasnikova
• 金额: $ 53.1万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9790997
• 关键词: Address; Adoptive Transfer; Animal Model; Animals; Antibodies; Antigens; Antitumor Response; Blood - brain barrier anatomy; Brain; Brain Neoplasms; CAR T cell therapy; CCL2 gene; Cells; Clinical Research; Complement; Data; Development; Engineering; Environment; Expression Profiling; Failure; Flow Cytometry; Frequencies; Future; Genetic Engineering; Glioblastoma; Glioma; Goals; Grant; Hematologic Neoplasms; Human; Image; Immune; Immunosuppressive Agents; Immunotherapy; In Vitro; Individual; Infiltration; Injections; Interleukin-15; Membrane; Modeling; Monitor; Mus; Normal tissue morphology; Outcome; Patients; Population; Primary Brain Neoplasms; Production; Proliferating; Receptor Cell; Regulatory T-Lymphocyte; Research; Site; Solid Neoplasm; Survival Analysis; Survival Rate; T cell therapy; T-Lymphocyte; Therapeutic; Time; Tissues; Transgenic Model; Transgenic Organisms; Treatment Failure; Tropism; Tumor Antigens; Tumor Burden; Tumor Escape; Variant; Xenograft Model; Xenograft procedure; base; chemokine; chemokine receptor; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine; cytotoxic; cytotoxicity; engineered T cells; genetic approach; improved; in vivo; novel therapeutics; overexpression; pre-clinical; prevent; receptor; response; tool; trafficking; tumor; tumor microenvironment; virtual

Glioblastoma (GBM), the most frequently occurring and aggressive primary brain tumor, remains virtually incurable. Thus, there is an urgent need to develop new therapies. Genetically modified T cells expressing chimeric antigen receptors (CARs) have the potential to serve as a unique cytotoxic tool to specifically target GBM. CAR T cell therapy has been successful for hematological malignancies, but multiple challenges posed by the brain tumor environment require a multifaceted approach for CAR T cells to succeed for GBM. To study this, we have developed a single-chain variable fragment (scFv) specific for IL13Rα2, a GBM-associated tumor antigen, and have generated an IL13Rα2-CAR. IL13Rα2-CAR T cells only recognize IL13Rα2-positive glioma cells and had anti-glioma activity in preclinical xenograft and immune-competent animal models. However, tumors eventually recurred, paralleling the situation in humans. Major causes of treatment failure include (i) the inability of CAR T cells to persist within an immunosuppressive tumor environment, (ii) antigen-loss variants when a single antigen is targeted, and (iii) the inability of CAR T cells to efficiently traffic to tumor sites due to a mismatch between chemokines produced by the tumor and chemokine receptors expressed by CAR T cells. In mechanistic studies, we have demonstrated limited IL13Rα2-CAR T cell persistence and the development of antigen-loss variants. In addition, we showed in xenograft models that transgenic expression of IL15 in CAR T cells enhances their persistence and anti-glioma activity. However, these xenograft studies are limited; the goal of this R01 is to perform mechanistic studies in immune-competent animal models and evaluate genetic approaches to enhance the anti-glioma activity of IL13Rα2-CAR T cells. Thus, we now hypothesize that IL13Rα2-CAR T cells can be further genetically engineered to optimize their anti-GBM activity by enhancing their persistence, targeting multiple tumor antigens, and improving their trafficking to tumor sites. Aim 1 investigates whether IL15-expressing CAR T cells can resist the immunosuppressive tumor environment in syngeneic GBM models. Aim 2 optimizes CAR T cells to target both IL13Rα2 and EphA2, two glioma-associated antigens. Aim 3 investigates if trafficking of CAR T cells to GBMs can be improved by the transgenic expression of CCR2, a chemokine receptor that recognizes CCL2, a chemokine produced by GBMs. At the conclusion of the grant, we will have addressed three major hurdles of CAR T cell therapy for GBM. While we will use our data to justify the development of a future clinical study utilizing optimized IL13Rα2-CAR T cells for patients with GBMs; our modified approach to T cell therapy should be applicable to a broad range of solid tumors.

胶质母细胞瘤 (GBM) 是最常发生且最具侵袭性的原发性脑肿瘤，实际上仍然存在 因此，迫切需要开发表达基因修饰的 T 细胞。 嵌合抗原受体（CAR）有潜力作为一种独特的细胞毒性工具来特异性靶向 GBM CAR T 细胞疗法在治疗血液恶性肿瘤方面取得了成功，但也带来了多重挑战。 脑肿瘤环境需要对 CAR T 细胞采取多方面的方法才能成功进行 GBM 研究。 为此，我们开发了一种针对 IL13Rα2（一种 GBM 相关肿瘤）特异的单链可变片段 (scFv) 抗原，并产生了仅识别 IL13Rα2 阳性神经胶质瘤的 IL13Rα2-CAR T 细胞。 细胞并在临床前异种移植和免疫活性动物模型中具有抗神经胶质瘤活性。 肿瘤最终复发，与人类的情况相似，治疗失败的主要原因包括（i） CAR T 细胞无法在免疫抑制肿瘤环境中持续存在，(ii) 抗原丢失变异 当针对单一抗原时，以及（iii）由于 肿瘤产生的趋化因子与 CAR T 细胞表达的趋化因子受体之间不匹配。 通过机制研究，我们证明了有限的 IL13Rα2-CAR T 细胞持久性和 此外，我们在异种移植模型中展示了 CAR T 中 IL15 的转基因表达。 然而，这些异种移植研究的目标是有限的。 该 R01 的目的是在具有免疫能力的动物模型中进行机制研究并评估遗传 因此，我们现在发现了增强 IL13Rα2-CAR T 细胞的抗神经胶质瘤活性的方法。 IL13Rα2-CAR T 细胞可以通过进一步基因工程来优化其抗 GBM 活性 增强它们的持久性，靶向多种肿瘤抗原，并改善它们向肿瘤的运输 目标 1 研究表达 IL15 的 CAR T 细胞是否可以抵抗免疫抑制肿瘤。 Aim 2 优化 CAR T 细胞以同时靶向 IL13Rα2 和 EphA2 这两种细胞。 目标 3 研究是否可以通过以下方法改善 CAR T 细胞向 GBM 的运输。 CCR2 的转基因表达，CCR2 是一种趋化因子受体，可识别 CCL2（GBM 产生的趋化因子）。 在资助结束时，我们将解决 GBM CAR T 细胞疗法的三个主要障碍。 我们将利用我们的数据来证明利用优化的 IL13Rα2-CAR T 细胞开展未来临床研究的合理性 对于 GBM 患者；我们改进的 T 细胞治疗方法应适用于广泛的实体瘤 肿瘤。