GENETICALLY ENGINEERED T CELLS FOR DIPG

用于 DIPG 的基因工程 T 细胞

基本信息

批准号：
10672286
负责人：
Giedre Krenciute
金额：
$ 50.36万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10672286
关键词：
Ablation Address Adoptive Transfer Adult Affect Antigens Antitumor Response Brain Neoplasms CAR T cell therapy CD276 gene CD28 gene CRISPR screen Cancer Etiology Cell Communication Cell Therapy Cell physiology Cell surface Cells Cellular biology Child Childhood Brain Neoplasm Clinical Clinical Research Data Development Diffuse intrinsic pontine glioma Disease model Endoplasmic Reticulum Engineering Evaluation Flow Cytometry Frequencies GRP78 gene Gene Deletion Genes Genetic Engineering Goals Golgi Apparatus Human Immune Immune response Immunocompetent Immunologic Deficiency Syndromes Immunooncology Immunotherapy In Vitro Intervention Intrinsic factor Knock-out Knowledge Learning Literature Macrophage Mediating Methods Modeling Molecular Molecular Target Mouse Strains Mus Myeloid Cells Outcome Pathway interactions Patients Phase Phenotype Population Proteins Publishing Role Safety Signal Pathway Solid Neoplasm Subgroup Surface T-Cell Activation T-Cell Receptor T-Lymphocyte TNFRSF5 gene Testing Translations Tumor Antigens Xenograft Model antigen-specific T cells cancer cell childhood cancer mortality chimeric antigen receptor chimeric antigen receptor T cells clinically relevant cytokine design diffuse midline glioma effective therapy effector T cell efficacy evaluation endoplasmic reticulum stress engineered T cells eosinophil exhaustion experience genome wide screen genome-wide glioma cell line glucose-regulated proteins human disease immune cell infiltrate improved improved outcome in vitro activity in vivo innovative technologies monocyte mouse model multidisciplinary neoplastic cell novel novel therapeutics pediatric patients pre-clinical prototype response safety testing tool tumor tumor heterogeneity tumor-immune system interactions

项目摘要

PROJECT ABSTRACT Genetically engineered T cells for DIPG The overall objective of this study is to engineer an effective and safe chimeric antigen receptor (CAR) T-cell therapy for diffuse intrinsic pontine glioma (DIPG), a subgroup of diffuse midline gliomas for which currently there is no cure. CAR T-cell therapy is an innovative technology based on adoptive transfer of antigen-specific T cells engineered to elicit a clinically effective and specific immune response against tumor cells. Early clinical studies in adult patients demonstrated the safety of CAR T-cell therapy for brain tumors, yet found only limited benefits. This lack of efficacy is most likely multifactorial and includes T-cell exhaustion, the immunosuppressive tumor microenvironment, and the paucity of brain tumor-specific antigens. Thus, we propose to develop CAR T cells targeting glucose-regulated protein 78 (GRP78), a novel DIPG-specific antigen. To further improve the effector function of GRP78-CAR T cells, we also propose to target a novel negative T-cell regulator called RASA2. We hypothesize that CAR T cells targeting GRP78 on the surface of tumor cells and lacking RASA2 can be developed as a safe, effective treatment for DIPGs and that evaluation of this intervention in DIPG mouse models that closely mimic human disease will identify T-cell extrinsic negative regulators of CAR T-cell function. Here we propose 3 interrelated Specific Aims to test this hypothesis. The rationale of each Aim is outlined below. In Aim 1, we will first target GRP78 and optimize the CAR design. To that end, we will generate CARs containing different co-stimulatory domains (41BB.ζ, MyD88.ζ, or MyD88.CD40.ζ) and compare their activity in vitro. Then we will evaluate the efficacy and safety of the active CAR(s) in vivo by using immunodeficient and immunocompetent mouse models of DIPG. A genome-wide screen in primary T cells has identified key regulators of T-cell activation after T-cell receptor stimulation. One of the identified genes, RASA2, has not been studied in T cells. In Aim 2, we will elucidate the mechanism by which the deletion of RASA2 enhances the effector function of CAR T cells and then delete the gene from GRP78-CAR T cells to enhance their function. Finally, in Aim 3, we will characterize the components of the DIPG microenvironment and examine the interactions between endogenous DIPG immune cells and CAR T–cell treatment. State-of-the-art methods will be used in all 3 Aims to study not only the function and in vivo fate of CAR T cells but also their antitumor activity and how they interact with DIPG-infiltrating immune cells (i.e., macrophages, eosinophils, monocytes). Our preliminary studies indicate that prototype GRP78-CAR T cells readily recognize and kill DIPG cells in vitro and have antitumor activity against solid tumors in vivo, highlighting that the developed models are well suited for the proposed Aims. To support the rapid translation of findings from this project to the clinical setting, we will use multiple DIPG mouse models that closely recapitulate human disease. Upon completion of this study, we will have defined the optimal GRP78-CAR design that safely eliminates DIPG tumors and persists long term in the DIPG immune microenvironment.

项目摘要用于 DIPG 的基因工程 T 细胞本研究的总体目标是设计一种有效且安全的嵌合抗原受体 (CAR) T 细胞弥漫性内质性脑桥胶质瘤（DIPG）的治疗，这是弥漫性中线胶质瘤的一个亚组，目前有治疗方法 CAR T 细胞疗法是一种基于抗原特异性 T 细胞过继转移的创新技术。旨在引发针对肿瘤细胞的临床有效和特异性免疫反应。在成年患者中证明了 CAR T 细胞疗法治疗脑肿瘤的安全性，但发现效果有限。这种功效的缺乏很可能是多因素造成的，包括 T 细胞耗竭、免疫抑制性肿瘤微环境和脑肿瘤特异性抗原的缺乏因此，我们建议开发 CAR T 细胞。靶向葡萄糖调节蛋白78（GRP78），一种新型DIPG特异性抗原，进一步改善效应子。为了研究 GRP78-CAR T 细胞的功能，我们还建议靶向一种名为 RASA2 的新型负 T 细胞调节因子。靶向肿瘤细胞表面 GRP78 且缺乏 RASA2 的 CAR T 细胞可以开发为一种安全、有效的 DIPG 治疗方法，并对 DIPG 小鼠的这种干预进行了评估密切模仿人类疾病的模型将识别 CAR T 细胞的 T 细胞外在负调节因子这里我们提出了 3 个相互关联的具体目标来检验这个假设，每个目标的基本原理是：在目标 1 中，我们将首先针对 GRP78 并优化 CAR 设计，为此，我们将生成。包含不同共刺激结构域（41BB.z、MyD88.z 或 MyD88.CD40.z）的 CAR 并比较它们然后我们将使用体内活性CAR来评估其有效性和安全性。免疫缺陷和免疫功能正常的 DIPG 小鼠模型已在原代 T 细胞中进行全基因组筛选。确定了 T 细胞受体刺激后 T 细胞激活的关键调节因子，其中之一是 RASA2，尚未在 T 细胞中进行研究。在目标 2 中，我们将阐明 RASA2 缺失的机制。增强CAR T细胞的效应功能，然后从GRP78-CAR T细胞中删除该基因以增强最后，在目标 3 中，我们将描述 DIPG 微环境的组成部分并进行检查。内源性 DIPG 免疫细胞与最先进的 CAR T 细胞治疗方法之间的相互作用。将用于所有 3 个目标，不仅研究 CAR T 细胞的功能和体内命运，还研究其抗肿瘤作用活性以及它们如何与 DIPG 浸润免疫细胞（即巨噬细胞、嗜酸性粒细胞、单核细胞）相互作用。我们的初步研究表明，原型 GRP78-CAR T 细胞很容易在体外识别并杀死 DIPG 细胞并对体内实体瘤具有抗肿瘤活性，这凸显了所开发的模型非常适合为了实现拟议的目标，为了支持该项目的研究结果快速转化为临床环境，我们将完成这项研究后，我们使用多个 DIPG 小鼠模型来密切再现人类疾病。将确定最佳的 GRP78-CAR 设计，可安全消除 DIPG 肿瘤并长期持续存在 DIPG 免疫微环境。