Improving CAR T cell therapy for brain tumors using immune competent models

使用免疫活性模型改善脑肿瘤的 CAR T 细胞疗法

基本信息

批准号：
10322192
负责人：
Dalia Haydar
金额：
$ 12.5万
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：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10322192
关键词：
Adoptive Transfer Adult Adverse effects American Antigen Targeting Antigens Biological Brain Neoplasms CAR T cell therapy CBLB gene CD276 gene CD28 gene CRISPR screen CRISPR/Cas technology Cell Cycle Cell surface Cells Cellular immunotherapy Childhood Childhood Brain Neoplasm Clinical Clinical Research Core Facility Disease Down-Regulation Engineering Generations Genes Genetic Glioma Goals Hematologic Neoplasms Heterogeneity Homing IL3RA gene Image Immune Immune response Immunocompetent Immunology Immunotherapy Impaired cognition Impairment In Vitro Inflammatory Intervention Life Mediating Mediator of activation protein Memory Modeling Modification Molecular Mus Mutate Operative Surgical Procedures Outcome Patients Penetration Performance Persons Phase Phenotype Play Positioning Attribute Pre-Clinical Model Primary Brain Neoplasms Radiation Receptor Signaling Research Research Personnel Resistance Role Safety Saint Jude Children&apos s Research Hospital Shapes Subgroup T-Cell Activation T-Cell Receptor T-Lymphocyte Techniques Testing Training Transmembrane Domain Tumor Antigens Tumor-associated macrophages Tumor-infiltrating immune cells Xenograft Model antigen-specific T cells antitumor effect base career chemotherapy chimeric antigen receptor chimeric antigen receptor T cells cytokine cytotoxicity design engineered T cells exhaustion extracellular genome wide screen human disease improved improved outcome in vitro activity in vivo innovative technologies interest macrophage motor impairment mouse model neoplastic cell novel prototype receptor research clinical testing success targeted treatment tool trafficking transcriptomics tumor tumor microenvironment tumor xenograft tumor-immune system interactions

项目摘要

PROJECT SUMMARY/ABSTRACT The overall objective of this study is to engineer a potent CAR T cell therapy for high grade glioma (HGG), a subgroup of brain tumors for which outcomes remain poor. 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 HGG patients demonstrated safety of CAR T cell therapy for brain tumors; yet, only limited benefits were observed. Lack of efficacy is most likely multifactorial and include heterogenous antigen expression, limited homing to and penetration of tumors, T cell exhaustion and limited persistence, as well as the immunosuppressive tumor microenvironment (TME). Therefore, the central hypothesis of this proposal is that optimized design and additional genetic modifications of CAR T cells will improve their antitumor effects, and that these interventions will be evaluated in immune competent mouse models that closely mimic human disease. Three interrelated research aims are proposed to test this hypothesis and the rationale of each is outlined below. First, CAR design has to be optimized and evaluated in immune competent glioma models. In Aim 1, I will generate CAR T cells containing different co- stimulatory domains (CD28.ζ, 41BB.ζ, CD28.mutζ, and 41BB.mutζ), and compare their activity in vitro and in vivo. Second, a genome wide screen in primary T cells has identified key regulators of T cell activation post T- cell receptor (TCR) stimulation. Identified genes belong to molecules that regulate cell cycle, proliferation, and downstream TCR signaling including SOCS1, RASA2, or CBLB. In Aim 2, I will therefore explore if CRISPR- Cas9 mediated silencing of Socs1, Rasa2, and/or Cblb enhances the effector functions of CAR T cells. The independent Aim 3 will then explore a dual CAR targeting approach in which I will not only target glioma cells but also immunosuppressive cells within the glioma microenvironment. These studies are focused on tumor associated macrophages (TAMs), since they are abundantly present in gliomas and play a critical role in shaping the TME. To support the feasibility of this project, I have adapted the well-established immune competent GL261 glioma model to study CAR T cell therapies targeting the relevant tumor antigen B7-H3, which is not only expressed by GL261 cells but also in a broad range of pediatric and adult brain tumors. In addition, my preliminary studies indicate that `prototype' B7- H3 CAR T-cells readily recognize and kill GL261 cells in vitro and have antitumor activity in vivo, highlighting that the developed model is well suited for the proposed aims of this project. State of the art technique will be used in all three Aims to not only study the function and in vivo fate of CAR T-cells, but also their antitumor activity, and how CAR T-cells interact with glioma- infiltrating immune cells. Completion of this study will define the most optimal CAR design that best controls HGG tumors and persists longer in the context of inflammatory brain tumors. Additionally, results will illustrate if targeting TAMs will overcome the suppressive effects of TME on B7-H3 CAR T cells.

项目摘要/摘要这项研究的总体目的是设计高级神经胶质瘤（HGG）的潜在汽车T细胞疗法，结局仍然很差的脑肿瘤亚组。汽车T细胞疗法是一种创新技术基于抗原特异性T细胞的自适应转移，该细胞设计为引起临床有效和特异性对肿瘤细胞的免疫反应。 HGG患者的早期临床研究表明汽车T细胞的安全性治疗脑肿瘤；但是，只有有限的好处。缺乏效率很可能是多因素并包括异源抗原表达，肿瘤的归巢和渗透，T细胞耗尽并且持久性有限，以及免疫抑制性肿瘤微环境（TME）。因此，该提案的中心假设是对汽车的优化设计和其他遗传修饰 T细胞将改善其抗肿瘤作用，并将这些干预措施评估为免疫胜任的小鼠模型，这些模型紧密模仿了人类疾病。提出了三个相互关联的研究目的为了检验这一假设，下面概述了每个假设。首先，必须优化汽车设计，并且在免疫胜任的神经胶质瘤模型中进行了评估。在AIM 1中，我将生成含有不同共同的汽车T细胞刺激结构域（CD28.ζ，41BB.ζ，CD28.MUTζ和41BB.MUTζ），并在体外和In比较其活性体内。其次，原代T细胞中的基因组宽筛网已经确定了T细胞激活的关键调节剂T- 细胞受体（TCR）刺激。确定的基因属于调节细胞周期，增殖和下游TCR信号传导，包括SOCS1，RASA2或CBLB。因此，在AIM 2中，我将探讨CRISPR- CAS9介导SOCS1，RASA2和/或CBLB的沉默增强了CAR T细胞的效应函数。这然后，独立AIM 3将探索双重车的靶向方法，我不仅将靶向神经胶质瘤细胞但也内神经胶质瘤微环境中的免疫抑制细胞。这些研究集中于肿瘤相关的巨噬细胞（TAM），因为它们在神经胶质瘤中大量存在，并且在塑造中起关键作用 TME。为了支持该项目的可行性，我改编了良好的免疫胜任GL261 胶质瘤模型研究针对相关肿瘤抗原B7-H3的汽车T细胞疗法，这不仅是由GL261细胞表达，但也处于各种儿科和成人脑肿瘤的范围内。另外，我的初步研究表明，“原型” B7-H3 CAR T细胞很容易识别并杀死GL261细胞的体外并在体内具有抗肿瘤活性，强调开发的模型非常适合提议的目的这个项目。最先进的技术将在所有三个目的中都用于研究功能和体内命运汽车T细胞的抗肿瘤活性，以及CAR T细胞与神经胶质瘤的相互作用细胞。这项研究的完成将定义最佳控制HGG肿瘤和最佳的汽车设计在炎症性脑肿瘤的背景下，持续时间更长。此外，结果将说明是否针对TAM 将克服TME对B7-H3 CAR T细胞的抑制作用。