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在确定公差方面起关键作用，并且是关键介体 T细胞功能障碍的癌症。我们与我们在PPG中的合作者一起表明PD-1：PD-L1 相互作用调节周围T细胞的耐受性和TIGIT具有介导的T细胞内抑制作用 调节t（Treg）细胞功能。根据我们的新初步数据，显示PD-1封锁增加了 Tigit+ Treg细胞的频率和项目3的数据，表明Tigit激动剂逆转了Treg的体外缺陷 在MS中的功能，我们测试了Tigit激动剂是否可以控制致病性T细胞并改善自身免疫性 使用EAE模型通过PD-1阻滞加剧。值得注意的是，Tigit激动剂mab减轻了EAE的严重性 在给予抗PD-1的小鼠中，仅比Tigit激动剂更大程度地阻断mAb。重要的是，蒂吉特激动剂 结合PD-1封锁并不会损害PD-1阻滞在控制肿瘤中的效率，这表明Tigit 作为治疗与PD-1封锁治疗相关的与免疫相关广告事件（IRAE）的潜在目标。 项目2的目标是确定Tigit/CD155和PD-1/PD-L1之间的相互作用 途径控制CD4+ Treg和CD4+ Foxp3 -T细胞，并定义细胞和分子电路 这些相互作用的基础。在AIM 1中，我们将测试细胞和分子相互作用的假设 在PD-1和Tigit Control Treg和CD4+ Foxp3之间，以调节自身免疫性。我们将使用新颖的工具 以及PPG研究人员开发的方法，包括Tigit激动剂抗体，Tigit和PD-1 条件敲除（KO）小鼠将缺失限制为特定的T细胞类型，高维细胞术，单个 细胞RNA测序（SCRNASEQ），空间转录组学和基因扰动方法 细胞和分子机制。在AIM 2中，我们将测试CD155和 APC和肿瘤上的PD-L1平衡Treg和T效应细胞功能以调节组织注射。我们将 评估CD155和PD-L1之间的功能相互作用，重点是其在DC和 使用CD155和PD-L1有条件KO 将缺失限制为DC和缺乏CD155和/或PD-L1或仅其细胞质结构域的小鼠， 和类似的方法与AIM 1一样。我们的结果将提供对独特和重叠节点的关键见解 PD-1/PD-L1和TIGIT/CD155相互作用以调节Treg和CD4+Foxp3-细胞调节T细胞 宽容。此外，我们的结果将通过提供双向的洞察力直接为项目1和3做出贡献 Tigit和CD155（项目1）之间的相互作用以及MS或MS患者的T细胞的功能差异 基于PD-1/PD-L1和Tigit/CD155表达和活性的恶性神经瘤（项目3）。理解 Tigit/CD155和PD-1/PD-L1相互作用的机制应启用新的治疗策略 对于具有癌症免疫疗法的赞助商 - 自动免疫性疾病和伊拉斯。我们将从核心中受益 A（Admin），Core B（基因扰动）和Core C（SCNUC-SEQ和多词）。