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

基本信息

批准号：
10426457
负责人：
Michael Lawrence Dougan
金额：
$ 61.22万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
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/10426457
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 Enrollment 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 Pulmonary Inflammation Randomized Regulation Regulatory T-Lymphocyte Reporting Role Signal Transduction Small Intestines Specimen Stromal Cells Surface Suspensions T memory cell T-Cell Receptor T-Lymphocyte TNF gene Techniques Technology Therapeutic Therapeutic Agents Time Tissue Embedding Tissues anti-PD1 antibodies base 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 pathogen programmed cell death protein 1 receptor receptor function response single cell analysis single-cell RNA sequencing therapeutic target tool tumor-immune system interactions

项目摘要

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

抽象的肠粘膜具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维持共生微生物群和膳食蛋白质的调节稳态。记忆 T 细胞因其快速反应的能力而对于粘膜表面的免疫稳态至关重要 CTLA-4 和 PD-1 受体抑制 T 细胞功能，已成为主要的病原体。然而，增强癌症患者 T 细胞介导的免疫力的治疗靶点。抑制性受体经常诱发炎症不良事件，而结肠炎是最常见和最常见的炎症不良事件之一。我们最近报道了由检查点抑制（CPI）引起的严重炎症不良事件。 CPI 结肠炎患者和健康受试者粘膜免疫细胞的单细胞分析。 CD8 T 细胞群的急剧积累，具有高度的增殖和细胞毒性状态我们使用 CD8 T 细胞的 T 细胞受体 (TCR) 序列来研究所有研究的 CPI 结肠炎患者。这些克隆扩增T细胞的起源，我们发现很大一部分与结肠炎相关。因此，细胞毒性 CD8 T 细胞与组织驻留记忆 CD8 T 细胞具有相同的 TCR 序列。最近，CTLA-4 和 PD-1 抑制受体将组织驻留记忆 T 细胞 (Trm) 保留在检查，并且这些抑制信号的丢失可以诱导 Trm 的大规模克隆扩张，这是一个主要的抗体阻断 CTLA-4 和 PD-1 受体是 CPI 结肠炎发展的一个步骤。因此，免疫学正常的粘膜可以深入了解这些受体的生理功能在目标 1 中，我们将研究 CTLA-4 和 PD-1 受体如何调节 Trm 细胞的功能，我们将在人类和小鼠模型中研究炎症途径的靶向是否可能恢复。通过执行深入的单细胞治疗，T 细胞从高度增殖、细胞毒性状态恢复到 Trm 状态对参加单独资助的随机 2 期临床的 CPI 结肠炎患者的结肠活检进行分析评估 TNFα 阻断与皮质类固醇的试验我们的单细胞分析表明上调。 CPI 结肠炎中的 TNF 基因表达特征和回顾性临床数据表明 TNFα 阻断是我们还将通过评估小鼠模型中的影响来研究这一假设。 CTLA-4 和 PD-1 抗体对肠道粘膜中 CD8 T 细胞的激活和细胞毒性状态的影响。目标 2。我们将研究肠道中 T 细胞群与免疫细胞、上皮细胞和基质细胞的空间相互作用使用 CODEX 技术对粘膜进行高度多重免疫荧光分析这种高维成像方法提供了使用 DNA 条形码单克隆抗体组进行切片的机会。研究健康肠道粘膜和 CPI 结肠炎中的细胞-细胞相互作用，包括来自以下组织的活检标本目标 1 中描述的临床试验。因此，该项目解决了粘膜免疫学这一基本问题。 1