Signal transduction and gene induction in lymphocytes

淋巴细胞中的信号转导和基因诱导

• 批准号: 10459427
• 负责人: Patrick Hogan
• 金额: $ 54万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10459427
• 关键词: ATAC-seq; Adoptive Transfer; Antibodies; Antigens; Antitumor Response; Biological; Biological Models; CD19 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; CRISPR/Cas technology; CTLA4 gene; Calcineurin; Calcium; Cancer Patient; Cell Nucleus; Cell Surface Receptors; Cell model; Cells; ChIP-seq; Characteristics; Chronic; Combined Modality Therapy; Complement; Data; Engineering; Family; Family member; Feedback; Fluorescence-Activated Cell Sorting; Functional disorder; Funding; Gene Expression Profile; Genetic Transcription; Grant; Head and Neck Squamous Cell Carcinoma; Human; Immune; Immune system; Individual; Kinetics; Lead; Lymphocyte; Malignant Neoplasms; Mediating; Modeling; Molecular; Mus; Mutation; Paper; Phenotype; Physiological; Production; Property; Role; Signal Transduction; Solid Neoplasm; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Transcription Factor AP-1; Transgenic Organisms; Tumor Antigens; Tumor-infiltrating immune cells; Up-Regulation; Ursidae Family; Variant; Viral Antigens; Viral Cancer; Virus Diseases; anergy; anti-CTLA4; anti-PD-1; cancer immunotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine; exhaust; exhaustion; gene induction; immune checkpoint blockade; improved; in vivo; mouse model; neoplastic cell; programmed cell death protein 1; programs; receptor; receptor expression; response; single-cell RNA sequencing; small hairpin RNA; targeted treatment; transcription factor; transcriptome sequencing; tumor

ABSTRACT In the previous project period of this grant, we investigated the transcriptional mechanisms of CD8+ T cell “exhaustion”, a hyporesponsive state observed under conditions of sustained antigen stimulation in cancer and chronic viral infections. Exhausted T cells show decreased proliferation and cytokine production, and upregu- late inhibitory cell surface receptors including CTLA4, PD-1, LAG3 and TIM3. Antibodies to these receptors reverse T cell exhaustion, and their administration to cancer patients forms the basis for “immune checkpoint blockade”, a strategy that has recently been remarkably successful in cancer immunotherapy. Combinations of antibodies to inhibitory receptors show greater efficacy than administration of individual antibodies alone, consistent with the fact that exhausted T cells typically express several inhibitory receptors. Thus understanding the molecular mechanisms that lead to inhibitory receptor expression and T cell “exhaustion/ dysfunction” would complement and enhance the effects of therapies that target combinations of individual inhibitory receptors. We previously showed that the transcriptional programs of anergy and exhaustion are initiated by the transcription factor NFAT, acting in the absence of its partner AP-1 (Fos-Jun). In the course of these studies, we developed an engineered NFAT, CA-RIT-NFAT1, which induces the characteristic features of exhaustion when transduced into CD8+ T cells. To understand the biological implications of this hyporesponsive program, we used mouse models of anti-tumor responses that involving adoptive transfer of tumor-reactive or unreactive TCR- transgenic T cells or T cells bearing chimeric antigen receptors (CAR-T cells). Using these models, we identified Nr4a transcription factors, and other families of transcription factors, as “exhaustion-related” target of NFAT. We further showed that tumor-infiltrating CD8+ T cells lacking all three Nr4a family members displayed a gene expression profile similar to that of activated T cells and rejected tumors more efficiently than control CD8+ T cells. In this application we will test the hypothesis that mouse and human CD4+ and CD8+ tumor-infiltrating T cells (TILs) are functionally silenced by a cell-intrinsic transcriptional program mediated, at least in part, by NFAT and Nr4a transcription factors, and other transcription factors induced by NFAT. In Aim 1, we will define mechanistically the cell-intrinsic roles of Nr4a transcription factors in CD8+ T cell exhaustion. We will ask how Nr4a deletion in TILs overcomes exhaustion, enhances the effector phenotype and promotes tumor regression. In Aim 2, we will examine the roles of other pertinent transcription factors in CD8+ T cell exhaustion, and define the kinetics with which the exhaustion program unfolds. In Aim 3, we will examine the roles of NFAT and Nr4a in primary human T cells bearing exhaustion markers or transduced with CA-RIT-NFAT1, and in TILs isolated from human tumors. Our proposed studies will contribute to a broad mechanistic understanding of the transcriptional mechanisms operating in tumor-infiltrating immune cells, and may spark improved immunotherapies for cancer patients.

摘要 在本基金的前期研究中，我们研究了CD 8 + T细胞的转录机制， “衰竭”，在癌症中持续抗原刺激的条件下观察到的低应答状态， 慢性病毒感染耗尽的T细胞显示增殖和细胞因子产生减少，并且上调细胞因子水平。 晚期抑制性细胞表面受体包括CTLA 4、PD-1、LAG 3和TIM 3。这些受体的抗体 逆转T细胞耗竭，并将其施用于癌症患者，形成了“免疫检查点”的基础 阻断”，这是一种最近在癌症免疫治疗中非常成功的策略。的组合 抑制性受体的抗体显示出比单独施用单个抗体更大的功效， 这与耗尽的T细胞通常表达几种抑制性受体的事实一致。从而理解 导致抑制性受体表达和T细胞“衰竭/功能障碍”的分子机制将 补充和增强靶向单个抑制性受体的组合的疗法的效果。 我们先前表明，无反应性和耗竭的转录程序是由转录因子启动的。 转录因子NFAT，在没有其伴侣AP-1（Fos-Jun）的情况下起作用。在这些研究中，我们 开发了一种工程NFAT，CA-RIT-NFAT 1，它在以下情况下会诱导疲惫的特征： 转导入CD 8 + T细胞。为了理解这种低反应程序的生物学意义，我们使用了 涉及肿瘤反应性或非反应性TCR的过继转移的抗肿瘤应答的小鼠模型， 转基因T细胞或携带嵌合抗原受体的T细胞（CAR-T细胞）。利用这些模型，我们发现 Nr 4a转录因子和其他转录因子家族作为NFAT的“耗竭相关”靶标。我们 进一步表明，缺乏所有三个Nr 4a家族成员的肿瘤浸润性CD 8 + T细胞显示出一种基因， 表达谱与活化T细胞相似，并且比对照CD 8 + T细胞更有效地排斥肿瘤。 在本申请中，我们将检验小鼠和人的CD 4+和CD 8+肿瘤浸润性T细胞（TIL） 在功能上被至少部分由NFAT和Nr 4a介导的细胞内在转录程序沉默 转录因子和由NFAT诱导的其他转录因子。在目标1中，我们将机械地定义 Nr 4a转录因子在CD 8 + T细胞耗竭中的细胞内在作用。我们将询问TIL中的Nr 4a缺失是如何 克服衰竭，增强效应表型，促进肿瘤消退。在目标2中，我们将 检查其他相关转录因子在CD 8 + T细胞衰竭中的作用，并定义动力学 这是耗尽程序所展现的。在目的3中，我们将研究NFAT和Nr 4a在原代人中的作用。 携带耗竭标志物或用CA-RIT-NFAT 1转导的T细胞，以及从人肿瘤分离的TIL。 我们提出的研究将有助于对转录机制的广泛理解 在肿瘤浸润免疫细胞中起作用，并可能为癌症患者带来更好的免疫治疗。