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 DIPG的基因工程T细胞这项研究的总体目的是设计有效且安全的嵌合抗原受体（CAR）T细胞弥漫性内在蓬托胶质瘤（DIPG）的治疗，这是当前在那里的弥漫性中线胶质瘤的亚组无法治愈。 CAR T细胞疗法是一种基于抗原特异性T细胞自适应转移的创新技术设计以引起针对肿瘤细胞的临床有效和特异性免疫反应。早期临床研究在成年患者中，汽车T细胞治疗对脑肿瘤的安全性，但发现有限的益处。缺乏效率很可能是多因素的，包括T细胞疲劳，免疫抑制肿瘤微环境和脑肿瘤特异性抗原的缺乏。那，我们建议开发汽车T细胞靶向葡萄糖调节的蛋白78（GRP78），一种新型DIPG特异性抗原。进一步改善效应器 GRP78-CAR T细胞的功能，我们还建议针对一种称为RASA2的新型负T细胞调节剂。我们假设靶向GRP78在肿瘤细胞表面且缺乏RASA2的CAR T细胞可以是作为DIPGS的安全有效治疗而开发的，对DIPG小鼠中这种干预的评估紧密模仿人类疾病的模型将识别CAR T细胞的T细胞外部负调节剂功能。在这里，我们提出了3个相互关联的特定目的来检验这一假设。每个目标的理由是下面概述了。在AIM 1中，我们将首先针对GRP78并优化汽车设计。为此，我们将生成包含不同共刺激域（41bb.ζ，myD88.ζ或myd88.cd40.ζ）的汽车，并比较它们体外活性。然后，我们将通过使用体内评估活性汽车的效率和安全性 DIPG的免疫缺陷和免疫能力的小鼠模型。原代T细胞中的全基因组筛查具有鉴定出T细胞受体刺激后T细胞激活的关键调节剂。已鉴定的基因RASA2，尚未在T细胞中研究。在AIM 2中，我们将阐明删除RASA2的机制增强CAR T细胞的效应子功能，然后从GRP78-CAR T细胞中删除该基因以增强他们的功能。最后，在AIM 3中，我们将表征DIPG微环境和检查的组成部分内源性DIPG免疫细胞与CAR T –细胞处理之间的相互作用。最先进的方法将在所有三个目标中都使用，不仅研究CAR T细胞的功能和体内命运，还研究其抗tuumoror 活性及其与浸入浸润的免疫细胞（即巨噬细胞，嗜酸性粒细胞，单核细胞）的相互作用。我们的初步研究表明，原型GRP78-CAR T细胞在体外容易识别并杀死DIPG细胞并具有对体内实体瘤的抗肿瘤活性，强调了开发的模型非常适合对于拟议的目的。为了支持从该项目到临床环境的发现的快速翻译，我们将使用多种仔细概括人类疾病的小鼠模型。完成这项研究后，我们将定义最佳的GRP78卡车设计，该设计可安全地消除DIPG肿瘤并长期持续 DIPG免疫微环境。