Proj. 3: Immunosuppressive circuits in T cells and other immune cells in GBM patients enrolled in clinical trials

项目。

基本信息

批准号：
10684029
负责人：
Kai W Wucherpfennig
金额：
$ 35.2万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-03 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10684029
关键词：
Antibodies Blocking Antibodies C Type Lectin Receptors CD8-Positive T-Lymphocytes CD8B1 gene Cell Line Cell physiology Cell surface Cell-Mediated Cytolysis Cells Cellular Immunity Clinical Trials Clonal Expansion Clone Cells Collaborations Color Combined Modality Therapy Data Diagnosis Dinoprostone Effector Cell Flow Cytometry Gene Expression Gene Expression Profile Genes Glioblastoma Glioma Goals Human Immune Immunocompetent Immunodeficient Mouse Immunohistochemistry Immunosuppression Immunotherapy Interferon Type II KLRB1 gene Label Length Ligands Malignant Neoplasms Mediating Messenger RNA Molecular Monoclonal Antibodies Mus Myeloid Cells Natural Killer Cells Operative Surgical Procedures Pathway interactions Patients Peptide Vaccines Peptide/MHC Complex Phase Population Proteins Relapse Role Sampling Signal Transduction Spatial Distribution T cell infiltration T-Cell Activation T-Cell Receptor T-Lymphocyte Therapeutic Tumor Immunity WFDC2 gene Work anti-PD1 antibodies brain tissue cancer infiltrating T cells cytokine cytotoxic cytotoxicity effector T cell exhaust experimental study genetic approach humanized mouse implantation interest mouse model neoantigen vaccine neoplastic cell overexpression participant enrollment population based programmed cell death ligand 1 programmed cell death protein 1 programs receptor receptor expression response single-cell RNA sequencing transcriptome sequencing tumor

项目摘要

Abstract T cells are central effector cells of protective anti-tumor immunity, but little is currently known about these important immune cells in human GBM. We have generated single-cell RNA-seq data on tumor-infiltrating T cell populations from GBM patients at initial diagnosis or relapse. We used these full-length RNA-seq data to identify clonally expanded T cell populations based on their TCRα and β chain sequences and then examined which genes were overexpressed by such expanded T cells. This analysis highlighted the KLRB1 gene which encodes the CD161 receptor that was previously shown to inhibit NK cell-mediated cytotoxicity. The CD161 ligand, CLEC2D, is expressed at the cell surface of human GBM cells. We therefore hypothesize that the CD161 – CLEC2D pathway inhibits the anti-tumor function of both CD8 and CD4 effector T cell populations in GBM. Preliminary data show that inactivation of the KLRB1 gene in primary human T cells greatly enhances their effector function in a humanized mouse model of GBM. Our preliminary data also demonstrate that several other inhibitory receptors are expressed by substantial populations of GBM-infiltrating T cells, including CD96 and two prostaglandin E2 receptors (EP2 and EP4). Aim 1 will focus on the analysis of tumor-infiltrating T cells in GBM patients enrolled in the phase 1b NeoVax plus PD-1 antibody trial described in Project 1. These studies will primarily focus on the expression of inhibitory receptors by T cells and their ligands by tumor cells and myeloid cells. Expression of inhibitory receptors and their ligands will be examined by 16-color spectral flow cytometry and single-cell RNA-seq (in collaboration with Cores 1 and 2), with an emphasis on CD161, PD-1, CD96 and prostaglandin E2 receptors. We will investigate paired tumor samples from the same patient obtained at initial surgery and relapse in order to determine how expression of these inhibitory receptors and their ligands changes following immunotherapy with NeoVax plus PD-1 antibody. In collaboration with Project 1, we will also examine the spatial distribution of T cells that express CD161 and other inhibitory receptors. Aim 2 will investigate the therapeutic significance of the CD161 – CLEC2D pathway. We will first use a genetic approach to study this inhibitory receptor by inactivating the KLRB1 gene in primary T cells. Blocking mAbs specific for human CD161 will also be used to examine the therapeutic potential of these findings. We will also examine combination therapies (collaboration with Projects 2, 4 and Core 3) involving the inhibitory receptors identified by single-cell RNA-seq in human GBM infiltrating T cells, with a particular focus on CD161, PD-1, CD96 and the prostaglandin E2 receptors. These studies will significantly advance our understanding of T cell function in GBM and characterize important inhibitory receptor – ligand interactions that constrain effector T cell function.

抽象的 T 细胞是保护性抗肿瘤免疫的中心效应细胞，但目前对这些细胞知之甚少我们已经生成了肿瘤浸润 T 细胞的单细胞 RNA 测序数据。我们使用这些全长 RNA-seq 数据来识别 GBM 患者的初始诊断或复发人群。根据 TCRα 和 β 链序列克隆扩增 T 细胞群，然后检查哪些该分析强调了编码 KLRB1 的基因。 CD161 受体先前已被证明可以抑制 NK 细胞介导的细胞毒性。 CLEC2D 在人类 GBM 细胞的细胞表面表达，因此我们捕获了 CD161 –。 CLEC2D 通路抑制 CD8 和 CD4 效应 T 细胞群的抗肿瘤功能初步数据表明，原代人类 T 细胞中 KLRB1 基因的失活大大增强了 GBM。我们的初步数据还表明，它们在 GBM 人源化小鼠模型中的效应功能。其他抑制性受体由大量 GBM 浸润 T 细胞表达，包括 CD96 和两种前列腺素 E2 受体（EP2 和 EP4）。目标 1 将重点分析肿瘤浸润 T 细胞。在参加项目 1 中描述的 1b 期 NeoVax 加 PD-1 抗体试验的 GBM 患者中。这些研究主要关注 T 细胞抑制性受体的表达及其肿瘤细胞和骨髓细胞的配体抑制性受体及其配体的表达将通过 16 色光谱流式细胞术进行检查。和单细胞 RNA-seq（与 Cores 1 和 2 合作），重点关注 CD161、PD-1、CD96 和我们将研究最初从同一患者获得的配对肿瘤样本。手术和复发以确定这些抑制性受体及其配体的表达如何变化在使用 NeoVax 加 PD-1 抗体进行免疫治疗后，我们还将与项目 1 合作进行检查。目标 2 将研究表达 CD161 和其他抑制性受体的 T 细胞的空间分布。 CD161 – CLEC2D 通路的治疗意义我们将首先使用遗传学方法来研究这一点。通过灭活原代 T 细胞中的 KLRB1 基因来抑制人 CD161 特异性阻断单克隆抗体。还将用于检查这些发现的治疗潜力我们还将检查组合。涉及单细胞识别的抑制性受体的疗法（与项目 2、4 和核心 3 合作）人 GBM 浸润 T 细胞中的 RNA 测序，特别关注 CD161、PD-1、CD96 和前列腺素这些研究将显着增进我们对 GBM 和 T 细胞功能的理解。表征限制效应 T 细胞功能的重要抑制性受体 - 配体相互作用。