Investigating TOX2 as a novel regulator of T cell memory differentiation

研究 TOX2 作为 T 细胞记忆分化的新型调节剂

• 批准号: 9759532
• 负责人: Sierra McDonald
• 金额: $ 4.5万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759532
• 关键词: Antigen Targeting; Antigens; Automobile Driving; Binding; Binding Proteins; Binding Sites; Bioinformatics; CAR T cell therapy; CD19 Antigens; CD19 gene; Cancer Patient; Candidate Disease Gene; Cell physiology; Cells; Characteristics; Chromatin; Chronic Lymphocytic Leukemia; Clinic; Clinical; Collaborations; DNA; DNA Binding; DNA-Binding Proteins; Development; Disease remission; Effectiveness; Effector Cell; Engineering; Family; Frequencies; Gene Expression; Genes; Genetic Transcription; Genome; Goals; Growth; HMG Domain; HMG-Box; HMGB Proteins; High Mobility Group Proteins; Human; Immune; Immune system; Immunotherapy; In Vitro; Infusion procedures; Lymphocyte; Major Groove; Malignant Neoplasms; Measures; Memory; Messenger RNA; Molecular; Molecular Genetics; Mus; Natural Killer Cells; Patient-Focused Outcomes; Patients; Pattern; Phenotype; Proteins; Quantitative Reverse Transcriptase PCR; Receptor Cell; Role; SLEB2 gene; Sampling; Spleen; T cell differentiation; T memory cell; T-Lymphocyte; T-bet protein; Testing; Time; Tonsil; Tumor Immunity; Up-Regulation; Xenograft procedure; anticancer research; base; cancer cell; cancer immunotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; effector T cell; engineered T cells; experimental study; fighting; functional improvement; improved; in vivo; insight; knock-down; leukemia; member; novel; overexpression; patient response; personalized cancer therapy; potential biomarker; predicting response; programs; promoter; receptor; responders and non-responders; response; small hairpin RNA; transcription factor; transcriptome sequencing; treatment response; tumor

PROJECT ABSTRACT Recent scientific advancements have made immunotherapy a promising option for many cancer patients. Immunotherapy can bolster the ability of the immune system to recognize and destroy cancer cells. However, not all patients respond to immunotherapy, thus a better understanding of the mechanisms that underlie immune cell effector function is required to improve response rates. In collaboration with Dr. Carl June's group, our lab recently studied a CLL patient who had gone into complete remission following CAR-T therapy. We found that an overwhelming majority of the CAR+ cells had a disruption in the TET2 gene, and experimental knockdown of TET2 resulted in enhanced proliferative capacity and anti-tumor activity of CAR T cells in vitro. Furthermore, T cells with the TET2 knockdown displayed increased expression of TOX2, a member of the HMG-box family of DNA-binding proteins. TOX2 is an immune-specific transcription factor expressed in the human spleen and tonsils. Interestingly, the only known function of TOX2 is that it positively regulates the transcription factor T-BET during the development of natural killer cells, by repressing the inhibitory PD-1 protein. Owing to the increase in TOX2 mRNA levels in TET2 knockdown CAR T cells, in addition to its ability to downregulate a key inhibitory receptor, I hypothesize that TOX2 is a positive regulator of T cell effector function due to its ability to upregulate T-BET. To test this hypothesis, I will pursue three aims. Aim 1 is to manipulate TOX2 expression in human T cells to determine whether it improves T cell differentiation and effector function. Aim 2 is to determine the pattern of TOX2 binding to the genome, with the goal of identifying its transcriptional targets, at both the mRNA and protein levels. Very little is known about how HMG-box proteins bind chromatin in vivo. Thus, this aim will also provide a more fundamental understanding of the chromatin functions of HMG proteins. Aim 3 is to determine levels of TOX2 mRNA in CLL patients who have received CAR T therapy, to determine whether higher TOX2 levels are predictive of better response to CAR T therapy. This study could elevate TOX2 as a novel target for improving CAR T cell function. An understanding of the molecular mechanisms could make it possible to activate TOX2, ultimately improving patient responses to immunotherapy.

项目摘要 对于许多癌症患者来说，最近的科学进步使免疫疗法成为有前途的选择。 免疫疗法可以增强免疫系统识别和破坏癌细胞的能力。然而， 并非所有患者都对免疫疗法做出反应，因此可以更好地理解基于的机制 需要免疫细胞效应子功能以提高反应率。与Carl June博士的小组合作 我们的实验室最近研究了一名CLL患者，该患者在CAR-T治疗后已完全缓解。我们 发现绝大多数CAR+细胞在TET2基因中都有破坏，并且实验 TET2的敲低导致CAR T细胞体外的增殖能力和抗肿瘤活性增强。 此外，具有TET2敲低的T细胞显示出毒物的表达增加，TOX2是一个成员 DNA结合蛋白的HMG盒家族。 TOX2是一种免疫特异性转录因子，在 人脾脏和扁桃体。有趣的是，托克斯2的唯一已知功能是，它积极调节 通过抑制抑制性PD-1，在天然杀伤细胞的发展过程中T-BET的转录因子T-BET 蛋白质。由于Tet2敲低CAR T细胞中TEX2 mRNA水平的升高，除了其能力 为了下调关键的抑制受体，我假设TOX2是T细胞效应器的正调节剂 功能由于其上调T-bet的能力。为了检验这一假设，我将追求三个目标。目标1是 操纵人T细胞中的TOX2表达，以确定其是否改善T细胞分化和 效应子功能。 AIM 2是确定与基因组结合的TOX2的模式，目的是识别 它的转录靶标在mRNA和蛋白质水平上均具有。关于HMG-box的了解很少 蛋白质在体内结合染色质。因此，这个目标还将提供对 HMG蛋白的染色质功能。 AIM 3是确定患有CLL患者的TOX2 mRNA水平 接受的汽车治疗，以确定较高的TOX2水平是否可以预测对汽车T的更好反应 治疗。这项研究可以将TOX2提升为改善CAR T细胞功能的新目标。一种理解 分子机制可以使激活TOX2成为可能，最终改善患者反应 进行免疫疗法。