Project 2: Elucidating how TIGIT agonists regulate CNS autoimmunity exacerbated by PD-1 blockade

项目 2：阐明 TIGIT 激动剂如何调节因 PD-1 阻断而加剧的 CNS 自身免疫

• 批准号: 10643434
• 负责人: Arlene H. Sharpe
• 金额: $ 63.14万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643434
• 关键词: Affect; Agonist; Antibodies; Autoimmune Diseases; Autoimmunity; Binding; Blood; CD8-Positive T-Lymphocytes; CNS autoimmunity; Cell physiology; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Cytometry; Cytoplasmic Tail; Data; Defect; Effector Cell; Equilibrium; Experimental Autoimmune Encephalomyelitis; FOXP3 gene; Frequencies; Functional disorder; Genes; Genetic Transcription; Glioma; Goals; Heterogeneity; Immune; In Vitro; Inflammation; Interleukin-10; Knockout Mice; Malignant Glioma; Malignant Neoplasms; Mediating; Mediator; Modeling; Molecular; Monoclonal Antibodies; Mus; Oncogenic; PD-1 blockade; PD-1/PD-L1; PDL1 pathway; Pathogenicity; Pathway interactions; Peripheral; Play; Production; Program Research Project Grants; Regulatory T-Lymphocyte; Research Personnel; Role; Severities; Signal Transduction; Spleen; T cell receptor repertoire sequencing; T-Cell Activation; T-Lymphocyte; Testing; Tissues; Tumor Immunity; Work; anti-PD-1; anti-PD-L1; cancer immunotherapy; cell type; conditional knockout; cytotoxicity; design; effector T cell; genetic approach; high dimensionality; immune-related adverse events; insight; migration; multiple omics; multiple sclerosis patient; neoplastic cell; novel; novel therapeutic intervention; novel therapeutics; programmed cell death ligand 1; programmed cell death protein 1; programs; receptor; single-cell RNA sequencing; therapeutic target; tool; transcriptomics; tumor; tumor growth

The immunoinhibitory receptors PD-1 and TIGIT play critical roles in regulating tolerance and are key mediators of T cell dysfunction in cancer. We, along with our collaborators in this PPG, have shown that PD-1:PD-L1 interactions regulate peripheral T cell tolerance and TIGIT has T cell-intrinsic inhibitory effects that mediate regulatory T (Treg) cell function. Based on our new preliminary data showing PD-1 blockade increases the frequency of TIGIT+ Treg cells and data of Project 3 showing that TIGIT agonist reversed in vitro defects of Treg function in MS, we tested if TIGIT agonism could control pathogenic T cells and ameliorate autoimmunity exacerbated by PD-1 blockade using the EAE model. Remarkably, TIGIT agonist mAb diminished EAE severity in mice given anti-PD-1 blocking mAb to a greater extent than TIGIT agonism alone. Importantly, TIGIT agonism combined with PD-1 blockade did not impair efficacy of PD-1 blockade in controlling tumors, suggesting TIGIT as a potential target for treating immune-related adverse events (irAEs) associated with PD-1 blockade therapy. The goal of Project 2 is to determine how interactions between the TIGIT/CD155 and PD-1/PD-L1 pathways control CD4+ Treg and CD4+ FoxP3— T cells, and define the cellular and molecular circuits underlying these interactions. In Aim 1, we will test the hypothesis that cellular and molecular interactions between PD-1 and TIGIT control Treg and CD4+ FoxP3—T cells to regulate autoimmunity. We will use novel tools and approaches developed by PPG investigators, including TIGIT agonist antibodies, TIGIT and PD-1 conditional knockout (KO) mice that restrict deletion to specific T cell types, high-dimensional cytometry, single- cell RNA sequencing (scRNAseq), spatial transcriptomics and gene perturbation approaches to interrogate cellular and molecular mechanisms. In Aim 2, we will test the hypothesis that interactions between CD155 and PD-L1 on APC and tumors balance Treg and T effector cell functions to regulate tissue inflammation. We will evaluate functional interactions between CD155 and PD-L1, focusing on their cell-intrinsic functions in DC and tumor cells, and cell-extrinsic effects on Treg and CD4+ FoxP3— T cells, using CD155 and PD-L1 conditional KO mice that restrict deletion to DC and tumor cells lacking CD155 and/or PD-L1 or only their cytoplasmic domains, and similar approaches as in Aim 1. Our results will provide critical insights into unique and overlapping nodes by which PD-1/PD-L1 and TIGIT/CD155 interact to modulate Treg and CD4+FoxP3— cells to regulate T cell tolerance. Moreover, our results will contribute directly to Projects 1 and 3 by providing insight into bidirectional interactions between TIGIT and CD155 (Project 1) and functional differences in T cells from patients with MS or malignant gliomas based on PD-1/PD-L1 and TIGIT/CD155 expression and activity (Project 3). Understanding mechanisms of TIGIT/CD155 and PD-1/PD-L1 interactions should enable design of new therapeutic strategies for spontaneous autoimmune disorders and irAEs seen with cancer immunotherapies. We will benefit from Core A (admin), Core B (gene perturbation), and Core C (scNuc-Seq and Multi-omics).

免疫抑制受体 PD-1 和 TIGIT 在调节耐受性中发挥关键作用，并且是关键介质 我们与本 PPG 中的合作者一起证明了 PD-1:PD-L1。 相互作用调节外周 T 细胞耐受性，TIGIT 具有介导 T 细胞内在抑制作用 根据我们新的初步数据显示 PD-1 阻断可增强调节性 T (Treg) 细胞功能。 TIGIT+ Treg 细胞的频率和项目 3 的数据显示 TIGIT 激动剂逆转了 Treg 的体外缺陷 为了在多发性硬化症中发挥作用，我们测试了 TIGIT 激动是否可以控制致病性 T 细胞并改善自身免疫 使用 EAE 模型阻断 PD-1 会加剧这种情况 值得注意的是，TIGIT 激动剂 mAb 降低了 EAE 的严重程度。 与单独使用 TIGIT 激动剂相比，给予抗 PD-1 阻断性单克隆抗体的小鼠的效果更佳。 与 PD-1 阻断联合使用并不会损害 PD-1 阻断控制肿瘤的功效，表明 TIGIT 作为治疗与 PD-1 阻断疗法相关的免疫相关不良事件 (irAE) 的潜在目标。 项目 2 的目标是确定 TIGIT/CD155 和 PD-1/PD-L1 之间如何相互作用 控制通路 CD4+ Treg 和 CD4+ FoxP3— T 细胞，并定义细胞和分子回路 在目标 1 中，我们将检验细胞和分子相互作用的假设。 PD-1 和 TIGIT 控制 Treg 和 CD4+ FoxP3-T 细胞来调节自身免疫。 以及 PPG 研究人员开发的方法，包括 TIGIT 激动剂抗体、TIGIT 和 PD-1 限制删除特定 T 细胞类型的条件敲除 (KO) 小鼠、高维细胞计数、单 细胞 RNA 测序 (scRNAseq)、空间转录组学和基因扰动方法进行询问 在目标 2 中，我们将检验 CD155 和 CD155 之间相互作用的假设。 APC 和肿瘤上的 PD-L1 平衡 Treg 和 T 效应细胞功能以调节组织炎症。 评估 CD155 和 PD-L1 之间的功能相互作用，重点关注它们在 DC 和 使用 CD155 和 PD-L1 条件 KO 观察肿瘤细胞，以及对 Treg 和 CD4+ FoxP3-T 细胞的细胞外在影响 将缺失限制为缺乏 CD155 和/或 PD-L1 或仅缺乏细胞质结构域的 DC 和肿瘤细胞的小鼠， 以及与目标 1 中类似的方法。我们的结果将为独特和重叠节点提供重要见解 通过PD-1/PD-L1和TIGIT/CD155相互作用来调节Treg和CD4+FoxP3细胞来调节T细胞 此外，我们的结果将通过提供对双向的洞察直接为项目 1 和 3 做出贡献。 TIGIT 和 CD155 之间的相互作用（项目 1）以及 MS 或 MS 患者 T 细胞的功能差异 基于 PD-1/PD-L1 和 TIGIT/CD155 表达和活性的恶性胶质瘤（项目 3）。 TIGIT/CD155 和 PD-1/PD-L1 相互作用的机制应有助于设计新的治疗策略 对于癌症免疫疗法中出现的自发性自身免疫性疾病和 irAE，我们将从 Core 中受益。 A（管理）、核心 B（基因扰动）和核心 C（scNuc-Seq 和 Multi-omics）。