Ameliorating off-target toxicities of CAR T cells by engineering NOT gates

通过设计 NOT 门改善 CAR T 细胞的脱靶毒性

基本信息

批准号：
10657356
负责人：
WENDELL A LIM
金额：
$ 44.29万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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/10657356
关键词：
Affinity Antigen Targeting Antigens B-Lymphocytes Bioinformatics Brain Cell Death Induction Cell physiology Cells Clinical Trials Cross Reactions Discrimination Engineering Epidermal Growth Factor Receptor Goals Immunocompetent Immunotherapeutic agent In Vitro Lung Malignant Neoplasms Modeling Mus Neuroblastoma Neurons Normal tissue morphology Organ Pediatric Neoplasm Preclinical Testing Production Publishing Safety Series Signal Transduction Solid Solid Neoplasm T-Cell Activation T-Cell Proliferation T-Cell Receptor T-Lymphocyte Technology Testing Therapeutic Time Tissues Toxic effect Transcription Repressor Treatment-related toxicity Tumor Antigens Tumor Tissue Work Xenograft Model antigen detection cancer cell chimeric antigen receptor T cells cross reactivity design engineered T cells high risk improved in vitro Assay in vivo in vivo evaluation mesothelin mouse model paracrine preclinical study prevent prototype recoverin protein success synthetic biology tool tumor

项目摘要

Project Summary/Abstract Engineered therapeutic T cells have shown transformative success in treating B cell cancers but applying this approach to solid tumors has proven far more difficult. There do not appear to be absolutely tumor-specific single antigen targets for solid cancers, and thus, CAR T cells that attack most tumor-associated antigens have led to toxic cross-reaction with normal organs that also express the antigen. If we are to successfully and safely treat solid tumors with CAR T cells, it will be essential to mitigate toxic cross-reaction with normal tissues. To prevent off-tumor toxicity of therapeutic T cells, we propose to engineer multi-receptor T cell circuits that can recognize a tumor based on a multi-antigen profile. In this proposal, we focus specifically on engineering NOT gate circuits -- circuits that can override and inhibit CAR T cell activation and killing upon detecting an antigen that is uniquely indicative of a cross-reactive normal tissue (i.e., antigen is absent in the tumor). Our recently published bioinformatic analysis shows that there are numerous tissue-specific antigens that could be used as signals to induce T cell inactivation in common cross-reactive tissues like the brain and lung. Nonetheless, there is currently a lack of robust NOT-gate circuits demonstrated to work well in tumor models. Thus, we will develop and test new NOT circuits that can inactivate a CAR T cell in an antigen-induced manner. Our specific aims are: Aim 1. Engineer, prototype and optimize new NOT gate circuits that use diverse mechanisms to block therapeutic T cell activation in antigen-induced manner Aim 1.1. T cell NOT gates using transcriptional repressors of CAR expression. Aim 1.2. T cell NOT gates that inhibit T cell proliferation by antigen-induction of cell death effectors. Aim 1.3. T cell NOT gates that locally induce production of secreted immunosuppressive factors (paracrine) Aim 2. Applying NOT gate circuits to prevent anti-GD2 CAR T cross-reaction with brain/CNS tissue. Aim 2.1. in vitro prototyping of NOT gate circuit targeting the brain antigen MOG to turn off anti-GD2 CAR Aim 2.2. Test if brain NOT gates block CNS toxicity of anti-GD2 CAR T cells in vivo, while maintaining efficacy against murine neuroblastoma xenograft models (GD2+). Aim 2.3. Test in vivo safety & efficacy of NOT gate CAR T cells in an immunocompetent model of neuroblastoma. This work should provide important general capabilities for engineering CAR T cells that selectively turn themselves OFF when they are in the wrong, cross-reactive tissue. These are much needed tools that are currently missing in the toolbox for T cell engineering, but which will be critical for engineering T cells that safely treat solid cancers.

项目摘要/摘要设计的治疗性T细胞在治疗B细胞癌中表现出了变革性的成功，但应用了实体瘤的方法已被证明要困难得多。似乎没有绝对的肿瘤特异性单曲固体癌症的抗原靶标，因此，攻击大多数肿瘤相关抗原的CAR T细胞已导致与也表达抗原的正常器官的有毒交叉反应。如果我们要成功，安全地对待带有CAR T细胞的实体瘤，减轻与正常组织的有毒交叉反应至关重要。为了防止治疗性T细胞的肿瘤毒性，我们提议设计可以进行多受体T细胞电路识别基于多抗原特征的肿瘤。在此提案中，我们专门关注工程而不是栅极电路 - 可以覆盖并抑制CAR T细胞激活并在检测抗原时杀死的电路这是唯一表明交叉反应性正常组织的（即肿瘤中不存在抗原）。我们最近发表的生物信息学分析表明，有许多组织特异性抗原可以用作在常见的交叉反应性组织（如大脑和肺）中诱导T细胞失活的信号。尽管如此，那里目前，缺乏在肿瘤模型中表现出良好效果的强大的非门电路。因此，我们将发展并测试新的不是可以以抗原引起的方式使汽车T细胞失活的电路。我们的具体目的是： AIM 1。工程师，原型和优化新的Not Gate电路，使用各种机制阻止治疗性T细胞以抗原诱导的方式激活目标1.1。 T细胞不是使用CAR表达的转录阻遏物的门。目标1.2。 T细胞不是通过细胞死亡效应子抗原诱导抑制T细胞增殖的门。目标1.3。 T细胞不是局部诱导分泌免疫抑制因子产生的门（旁分泌） AIM 2。不应用栅极电路，以防止抗GD2 CAR T与脑/CNS组织交叉反应。目标2.1。靶向脑抗原mog的非门电路的体外原型制作以关闭抗GD2汽车目标2.2。在维持的同时，测试大脑不盖在体内抗CNS抗GD2 CAR T细胞的毒性针对鼠神经母细胞瘤异种移植模型（GD2+）的功效。目标2.3。在免疫能力模型中测试非栅极T细胞的体内安全性和功效成神经细胞瘤。这项工作应为工程汽车T单元提供重要的一般能力，以选择性地转动当他们处于错误的交叉反应组织中时。这些是急需的工具目前在T细胞工程工具箱中缺少缺少治疗固体癌症。