Programming multi-pronged immune response to glioblastoma with IL-13Ra2/TGF-b CAR-T cell therapy.

使用 IL-13Ra2/TGF-b CAR-T 细胞疗法对胶质母细胞瘤进行多管齐下的免疫反应。

• 批准号: 10586951
• 负责人: Yvonne Yu-Hsuan Chen
• 金额: $ 61.3万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586951
• 关键词: Alpha Interleukin 2 Receptor; Antigens; Autopsy; Biopsy; Bone Marrow; Brain; C57BL/6 Mouse; CAR T cell therapy; Cell physiology; Cells; Cessation of life; Clinic; Clinical; Clinical Treatment; Cytometry; Data; Dominant-Negative Mutation; Engineering; Environment; Epitope spreading; Evaluation; Feedback; Flow Cytometry; Genetic Heterogeneity; Glioblastoma; Glioma; Heterogeneity; Human; IL13RA1 gene; Immune; Immune response; Immune system; Immunity; Immunocompetent; Immunofluorescence Immunologic; Immunohistochemistry; Immunosuppression; Immunosuppressive Agents; Immunotherapy; Infiltration; Inflammatory; Intravenous; Libraries; Lymphocyte; Malignant neoplasm of brain; Medical; Mesenchymal; Modeling; Monitor; Mus; Myelogenous; Myeloid Cells; Myeloid-derived suppressor cells; Neoplasm Metastasis; Newly Diagnosed; Patients; Phase I Clinical Trials; Phenotype; Play; Primary Brain Neoplasms; Proliferating; Reporting; Resected; Route; Safety; Signal Transduction; Specificity; Spleen; Stimulant; Surface Antigens; Survival Rate; T-Lymphocyte; Testing; Time; Toxic effect; Transforming Growth Factor Beta 2; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Tumor Promotion; Tumor-Infiltrating Lymphocytes; Tumor-associated macrophages; Tumor-infiltrating immune cells; Work; Xenograft Model; Xenograft procedure; beta Chain Antigen T Cell Receptor; chimeric antigen receptor; chimeric antigen receptor T cells; clinical translation; cytokine; effective therapy; engineered T cells; immunoregulation; improved; in vivo; mouse model; neoplastic cell; novel; pre-clinical; preservation; pressure; recruit; response; single-cell RNA sequencing; targeted treatment; translational potential; treatment group; tumor; tumor growth; tumor microenvironment; tumor progression

ABSTRACT Glioblastoma multiforme (GBM) is the most common type of primary brain tumor, with a five-year survival rate of only 5.5%. Chimeric antigen receptor (CAR)-T cell therapy has shown safety but limited efficacy in the treatment of patients with GBM to date. GBM is characterized by dramatic antigen heterogeneity, thus immunotherapy targeting any single antigen is unlikely to achieve complete and durable response. In addition, GBM cells and surrounding tumor stroma overproduce transforming growth factor beta (TGF-β), which not only promotes tumor growth and metastasis, but also actively modulate the immune response by suppressing T-cell function and recruiting suppressive myeloid cells. Our analysis of patient GBM biopsies further confirmed that tumor-infiltrating myeloid cells (TIMs) express both TGF-β and TGF-β receptors, indicating the presence of an immunosuppressive feedback loop in the GBM tumor microenvironment (TME). Here, we aim to develop a multi- pronged immunotherapy featuring bispecific CAR-T cells that can not only attack GBM directly, but also modify the TME to overcome immunosuppression and induce multiclonal immune responses against GBM. Our group has developed a bispecific CAR that can directly target GBM cells through recognition of IL- 13Rα2, a clinically validated GBM-associated surface antigen, while simultaneously converting TGF-β into a stimulant for the engineered T cells. We have demonstrated that bispecific IL-13Rα2/TGF-β CAR-T cells are superior to single-input IL-13Rα2 CAR-T cells in both human GBM xenograft and immunocompetent mouse models. Here, we aim to demonstrate the safety and efficacy of IL-13Rα2/TGF-β CAR-T cells for clinical translation. In addition to demonstrating clearance of IL-13Rα2+ GBM, we will explore the potential for IL- 13Rα2/TGF-β CAR-T cells to induce endogenous immune response against IL-13Rα2– tumor cells using mouse models of heterogenous GBM. We will perform in-depth analyses by flow cytometry, multiplexed immuno- fluorescence, cytometry by time of flight (CyTOF), and single-cell RNA sequencing to understand the impact of bispecific CAR-T cells on the immune composition and function in the GBM TME. We will also perform T-cell receptor beta (TCRβ) sequencing on tumor-infiltrating lymphocytes to quantify the potential for epitope spreading. We will perform rigorous safety evaluations in immunocompetent mouse models, applying worst-case scenario pressure tests to explore the toxicity profile of IL-13Rα2/TGF-β CAR-T cells. Finally, we will verify efficacy of IL-13Rα2/TGF-β CAR-T cell therapy in heterogeneous human gliomas ex vivo & in vivo, using both fresh patient GBM biopsies containing TIMs and GliomaPDOX models—i.e., mice bearing patient-derived GBM tumors that have never been passaged ex vivo and retain the genetic heterogeneity seen in human patients. Successful completion of this project will generate a comprehensive set of preclinical data in support of a phase- 1 clinical trial to treat GBM patients with this novel, multi-pronged immunotherapy option.

抽象的 胶质母细胞瘤多形（GBM）是最常见的原发性脑肿瘤类型，生存率为五年 仅5.5％。嵌合抗原受体（CAR）-T细胞疗法已显示出安全性，但效率有限 迄今为止GBM患者的治疗。 GBM的特征是戏剧性的抗原异质性，因此 靶向任何单一抗原的免疫疗法不可能实现完整耐用的反应。此外， GBM细胞和周围的肿瘤基质过量转化生长因子β（TGF-β），不仅 促进肿瘤的生长和转移，但也通过抑制T细胞积极调节免疫反应 功能和募集抑制性髓样细胞。我们对患者GBM活检的分析进一步证实了 肿瘤浸润的髓样细胞（TIMS）同时表达TGF-β和TGF-β受体，表明存在 GBM肿瘤微环境（TME）中的免疫抑制反馈回路。在这里，我们旨在开发一个多 支撑的免疫疗法具有双特异性CAR-T细胞，不仅可以直接攻击GBM，还可以修改 TME克服免疫抑制并诱导针对GBM的多隆免疫反应。 我们的小组已经开发了双特异性汽车，该汽车可以通过识别IL-直接靶向GBM细胞 13Rα2，一种经过临床验证的GBM相关的表面抗原，同时将TGF-β转化为A 工程T细胞的刺激性。我们已经证明了双特异性IL-13Rα2/TGF-βCAR-T细胞是 在人类GBM特征和免疫功能的小鼠中，都优于单输入IL-13Rα2CAR-T细胞 型号。在这里，我们旨在证明IL-13Rα2/TGF-βCar-T细胞对临床的安全性和效率 翻译。除了证明IL-13Rα2+ GBM的清除率外，我们还将探索IL-的潜力 13Rα2/TGF-βCAR-T细胞使用小鼠对IL-13Rα2-肿瘤细胞诱导内源性免疫响应 异源GBM的模型。我们将通过流式细胞仪，多重免疫 - 进行深入分析 荧光，按飞行时间（cytof）进行的细胞仪和单细胞RNA测序以了解的影响 双特异性CAR-T细胞在GBM TME中的免疫组成和功能上。我们还将执行T细胞 受体β（TCRβ）在肿瘤浸润淋巴细胞上进行测序，以量化表位扩散的潜力。 我们将在免疫能力的小鼠模型中进行严格的安全评估，应用最差的案例 方案压力测试以探索IL-13Rα2/TGF-βCAR-T细胞的毒性特征。最后，我们将验证 IL-13Rα2/TGF-βCAR-T细胞疗法在异质人神经胶质瘤的体内和体内的功效 新鲜的患者GBM活检，其中包含TIMS和Gliomapdox模型 - I.E。，带有患者衍生GBM的小鼠 从未被体内传播并保留人类患者中遗传异质性的肿瘤。 该项目的成功完成将产生一组全面的临床前数据，以支持阶段 1临床试验，以这种新型的多管齐下免疫疗法选择治疗GBM患者。