Molecular Pathways Regulating Tissue-resident Memory T cells in the Gut

调节肠道组织驻留记忆 T 细胞的分子途径

• 批准号: 10579333
• 负责人: Michael Lawrence Dougan
• 金额: $ 59.52万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579333
• 关键词: Address; Adrenal Cortex Hormones; Advanced Malignant Neoplasm; Adverse event; Affect; Antibodies; Antibody Therapy; Autoimmune Diseases; Back; Bar Codes; Biopsy; Biopsy Specimen; Blocking Antibodies; CD8-Positive T-Lymphocytes; CTLA4 blockade; CTLA4 gene; Cancer Patient; Cell Communication; Cell physiology; Cells; Cellular Immunity; Clinical; Clinical Data; Clinical Trials; Clonal Expansion; Colitis; Colon; Colonic inflammation; Combined Modality Therapy; Cytotoxic T-Lymphocytes; DNA; Data; Development; Diagnosis; Diagnostic; Dietary Proteins; Epithelial Cells; FOXP3 gene; Fc Receptor; Formalin; Funding; Gene Expression Profile; Goals; Gut Mucosa; Homeostasis; Human; Imaging Techniques; Immune; Immune checkpoint inhibitor; Immunity; Immunofluorescence Immunologic; Immunologics; Immunology; Individual; Inflammatory; Intestinal Mucosa; Invaded; Lead; Lung; Memory; Molecular; Monoclonal Antibodies; Mucous Membrane; Mus; Myeloid Cells; Paraffin Embedding; Pathogenesis; Pathway interactions; Patients; Phase II Clinical Trials; Physiological; Population; Proliferating; Pulmonary Inflammation; Randomized; Receptor Inhibition; Regulation; Regulatory T-Lymphocyte; Reporting; Role; Signal Transduction; Small Intestines; Specimen; Stromal Cells; Surface; Suspensions; T-Cell Receptor; T-Lymphocyte; TNF gene; Techniques; Technology; Therapeutic; Therapeutic Agents; Time; Tissue Embedding; Tissues; anti-PD1 antibodies; checkpoint inhibition; chemokine; chemokine receptor; commensal microbes; cytokine; cytotoxic; cytotoxic CD8 T cells; enteritis; high dimensionality; human model; imaging approach; immune checkpoint blockade; immunoregulation; insight; intestinal homeostasis; mouse model; participant enrollment; pathogen; programmed cell death protein 1; receptor; receptor function; response; single cell analysis; single-cell RNA sequencing; therapeutic target; tissue resident memory T cell; tool; tumor-immune system interactions

Abstract The intestinal mucosa has a rich immune microenvironment capable of responding rapidly to invading pathogens while maintaining a regulated homeostatic state to commensal microbiota and dietary proteins. Tissue-resident memory T cells are critical for immune homeostasis at mucosal surfaces based on their ability to rapidly respond to invading pathogens. The CTLA-4 and PD-1 receptors inhibit T cell function and have become major therapeutic targets for boosting T cell-mediated immunity in cancer patients. However, targeting of these inhibitory receptors frequently induces inflammatory adverse events, and colitis is one of the most common and severe inflammatory adverse events induced by checkpoint inhibition (CPI). We recently reported an in-depth single cell analysis of immune cells in the mucosa of CPI colitis patients and healthy subjects. This analysis demonstrated dramatic accumulation of CD8 T cell populations with highly proliferative and cytotoxic states in all studied CPI colitis patients. We used the T cell receptor (TCR) sequences of CD8 T cells to investigate the origin of these clonally expanded T cells. Interestingly, we discovered that a large fraction of colitis-associated cytotoxic CD8 T cells had the same TCR sequences as tissue-resident memory CD8 T cells. We therefore hypothesize that the CTLA-4 and PD-1 inhibitory receptors hold tissue-resident memory T cells (Trm) in check, and that loss of these inhibitory signals can induce massive clonal expansion of Trm, a major step in the development of CPI colitis. Antibody blockade of CTLA-4 and PD-1 receptors in individuals with immunologically normal mucosa thus provides insight into the physiological function of these receptors in humans. In Aim 1, we will investigate how the CTLA-4 and PD-1 receptors regulate the function of Trm cells, both in humans and murine models. We will investigate whether targeting of inflammatory pathways may revert T cells from highly proliferative, cytotoxic states back into a Trm state by performing an in-depth single cell analyses on colon biopsies from CPI colitis patients enrolled in a separately funded randomized phase 2 clinical trial that evaluates TNFα blockade versus corticosteroids. Our single cell analysis demonstrated upregulated TNF gene expression signatures in CPI colitis, and retrospective clinical data indicate that TNFα blockade is efficacious in CPI colitis. We will also investigate this hypothesis in murine models by evaluating the impact of CTLA-4 and PD-1 antibodies on the activation and cytotoxic states of CD8 T cells in the gut mucosa. In Aim 2 we will study the spatial interactions of T cell populations with immune, epithelial and stromal cells in the intestinal mucosa using the CODEX technology that enables highly multiplexed immunofluorescence analysis of tissue sections using panels of DNA-barcoded mAbs. This high-dimensional imaging approach affords an opportunity to study the cell – cell interactions in the healthy gut mucosa and in CPI colitis, including biopsy specimens from the clinical trial described in Aim 1. This project thus addresses a fundamental question is mucosal immunology. 1

摘要 肠粘膜有丰富的免疫微环境，能够对入侵的病原体做出快速反应。 同时保持一种调节的动态平衡状态，以共生微生物区系和饮食蛋白质。纸巾驻留 基于记忆T细胞的快速反应能力，它们对粘膜表面的免疫稳态至关重要 入侵的病原体。CTLA-4和PD-1受体抑制T细胞功能，已成为主要的 提高癌症患者T细胞免疫功能的治疗靶点。然而，针对这些人 抑制性受体经常诱发炎性不良事件，结肠炎是最常见的和 检查点抑制(CPI)引起的严重炎症性不良事件。我们最近报道了一项深入的 CPI结肠炎患者和健康人粘膜免疫细胞的单细胞分析。这一分析 显示CD8 T细胞群在高度增殖和细胞毒状态下显著积累 所有研究对象均为CPI结肠炎患者。我们使用CD8T细胞的T细胞受体(TCR)序列来研究 这些克隆扩增的T细胞的起源。有趣的是，我们发现很大一部分与结肠炎相关的 细胞毒性CD8T细胞具有与组织驻留记忆CD8T细胞相同的TCR序列。因此，我们 假设CTLA-4和PD-1抑制受体持有组织驻留的记忆T细胞(Trm) 这些抑制信号的丢失可以诱导Trm的大规模克隆性扩张，Trm是一种主要的 参与CPI结肠炎的发展。慢性阻塞性肺疾病患者CTLA-4和PD-1受体的抗体阻断 因此，免疫正常的粘膜可以深入了解这些受体的生理功能。 人类。在目标1中，我们将研究CTLA-4和PD-1受体如何调节Trm细胞的功能， 在人类和小鼠模型中都是如此。我们将调查针对炎症途径的靶向是否可能恢复 T细胞从高度增殖、细胞毒状态恢复到Trm状态通过进行深入的单个细胞 一项单独资助的随机二期临床研究中CPI结肠炎患者的结肠活检分析 评估肿瘤坏死因子α阻滞剂与皮质类固醇的试验。我们的单细胞分析显示 肿瘤坏死因子基因在CPI结肠炎中的表达特征，回顾临床资料表明，肿瘤坏死因子α阻断是 治疗CPI结肠炎疗效确切。我们还将在小鼠模型中通过评估 CTLA-4和PD-1抗体对肠黏膜CD8 T细胞活化和细胞毒状态的影响在AIM 2 我们将研究T细胞群与肠道免疫细胞、上皮细胞和基质细胞的空间相互作用 使用Codex技术对组织进行高度多元化的免疫荧光分析 使用DNA条形码mAb板的切片。这种高维成像方法提供了一个机会 目的：研究健康肠粘膜和CPI结肠炎的细胞-细胞相互作用，包括来自 目标1中描述的临床试验。因此，该项目解决了一个基本问题，即粘膜免疫学。 1