Defining the Role of Aire in eTACs and its Contribution to Peripheral Immune Tolerance

定义 Aire 在 eTAC 中的作用及其对外周免疫耐受的贡献

基本信息

批准号：
10660969
负责人：
Jiaxi Wang
金额：
$ 3.98万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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：
2022-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660969
关键词：
ATAC-seq Acceleration Adaptive Immune System Antigen Presentation Antigen Presentation Pathway Antigen-Presenting Cells Antigens Autoantibodies Autoantigens Autoimmune Autoimmune Diseases Autoimmunity B-Lymphocytes Bioinformatics Biology CD4 Positive T Lymphocytes CD80 gene CD86 gene California Cell Count Cell physiology Cells Chromatin Coculture Techniques Complex Data Data Set Dendritic Cells Development Diabetes Mellitus Diphtheria Toxin Education Environment Experimental Autoimmune Encephalomyelitis Fellowship Flow Cytometry Funding G6PC2 gene Generations Genes Genetic Genetic Transcription Genomics Helper-Inducer T-Lymphocyte Hematopoietic Histology Homeostasis Host Defense Human Immune Immune Tolerance Immune system Immunology Incubated Institution Insulin Interleukin-10 Knock-out Knockout Mice Laboratories MHC Class II Genes Maintenance Measures Mentors Methods Monitor Mus Mutation Organ Organism Orphan Paint Pancreas Peptides Peripheral Phenotype Physiologic pulse Play Polyglandular Autoimmune Syndrome Type I Population Population Biology Proliferating Proteins RNA Reaction Regulator Genes Regulatory T-Lymphocyte Reporter Research Research Project Grants Retinoic Acid Receptor Role San Francisco Self Tolerance Serum Skin Sorting Spleen Surface System T-Cell Proliferation T-Lymphocyte Testing Thymic epithelial cell Thymus Gland Tissues Training Transgenic Mice Tumor Immunity Universities Venus Vertebrates Work aged anergy autoreactive T cell career cell type central tolerance clinical application fetal fluorophore in vitro testing in vivo monitoring lymph nodes migration mouse model multiple omics novel overexpression pathogen peripheral tolerance prevent programs promoter receptor research and development response secondary lymphoid organ single-cell RNA sequencing skills tool uptake

项目摘要

PROJECT SUMMARY/ABSTRACT The autoimmune regulator (Aire) gene, a key transcriptional regulator expressed in medullary thymic epithelial cells (mTECs), has been shown to be crucial for central tolerance by inducing tissue specific antigen (TSA) expression in mTECs. Interestingly, Aire is also found in extrathymic Aire-expressing cells (eTACs) in the secondary lymphoid organs such as the spleen and lymph nodes. We previously found that eTACs are hematopoietic antigen-presenting cells (APCs) and consist of two similar cell types: CCR7+ Aire-expressing migratory dendritic cells (AmDCs) and an Aire-high population co-expressing Aire and retinoic acid receptor– related orphan receptor γt (ROR γt) that we termed Janus cells (JCs). Functionally, eTACs are capable of enforcing deletion and anergy on self-reactive T cells, and self-antigen expression in eTACs is sufficient to prevent autoimmunity. The transcriptional, genomic, and functional symmetry between eTACs (both JCs and AmDCs) and mTECs potentially identifies a core program driven by Aire that may influence self-representation and tolerance across the spectrum of immune development. However, the lineage relationship of eTACs, what role Aire plays in these populations, and how extrathymic Aire and eTAC subsets contribute to immune homeostasis are still unknown. This proposal will test the hypothesis that Aire is inducing a tolerogenic phenotype in eTACs and that extrathymic Aire and eTACs are important for enforcing peripheral immune tolerance. Aim 1 of this proposal will define the lineage relationship between eTACs subsets, their antigen processing and presentation functions, and their migratory abilities. Aim 2 will define the cell-intrinsic functions of Aire in eTACs at both the transcriptional and chromatin level. Aim 3 will investigate the contribution of extrathymic Aire and eTACs in maintaining normal immune homeostasis. This proposal will be carried out using a variety of methods including single cell multiomics, flow cytometry, and functional approaches such as ex vivo co-cultures and in vivo monitoring of autoimmunity utilizing novel genetic mouse models. By characterizing eTAC subsets and investigating the functional roles of extrathymic Aire and eTACs, this work will help further elucidate the function of Aire and define basic peripheral tolerance mechanisms. Furthermore, understanding the biology of these tolerogenic populations may have significance for a range of clinical applications from autoimmunity to tumor immunity to maternal-fetal tolerance. This research project and fellowship training will be conducted at a top-funded research institution, the University of California, San Francisco (UCSF), in the laboratories of Dr. James Gardner and Dr. Mark Anderson. Dr. Gardner has expertise in studying peripheral Aire/eTACs and the generation of genetic mouse models. Dr. Anderson is a world expert on Aire biology, thymic selection, and immune tolerance, and a highly respected mentor and leader in the field of immunology. These mentors and institution will provide a rich training environment for completion of this research and development of professional skills necessary for a career in academic research.

项目概要/摘要自身免疫调节因子 (Aire) 基因，在髓质胸腺上皮中表达的关键转录调节因子细胞（mTEC）已被证明通过诱导组织特异性抗原（TSA）对于中枢耐受至关重要在 mTEC 中隐含表达，Aire 也存在于胸腺外表达 Aire 的细胞 (eTAC) 中。我们之前发现 eTAC 是次要淋巴器官，例如脾脏和淋巴结。造血抗原呈递细胞 (APC)，由两种相似的细胞类型组成：CCR7+ Aire 表达迁移树突状细胞（AmDC）和共同表达 Aire 和视黄酸受体的 Aire 高细胞群 - 相关的孤儿受体 γt (ROR γt)，我们称为 Janus 细胞 (JC) 在功能上，eTAC 具有以下功能。对自身反应性 T 细胞实施删除和无反应性，并且 eTAC 中自身抗原的表达足以 eTAC（JC 和 JC）之间的转录、基因组和功能对称性。 AmDC）和 mTEC 可能确定由 Aire 驱动的可能影响自我表现的核心计划然而，eTAC 的谱系关系是什么？ Aire 在这些人群中发挥的作用，以及胸腺外 Aire 和 eTAC 亚群如何促进免疫体内平衡仍然未知，该提案将检验艾尔正在诱导耐受性的假设。 eTAC 中的表型以及胸腺外 Aire 和 eTAC 对于增强外周免疫非常重要该提案的目标 1 将定义 eTAC 子集及其抗原之间的谱系关系。目标 2 将定义细胞固有功能。 Aire 在 eTAC 中在转录和染色质水平上的贡献。胸腺外的 Aire 和 eTACs 在维持正常的免疫稳态中将被使用。多种方法，包括单细胞多组学、流式细胞术和离体等功能方法利用遗传小鼠模型对自身免疫进行共培养和体内监测。 eTAC 子集并研究胸腺外 Aire 和 eTAC 的功能作用，这项工作将有助于进一步阐明 Aire 的功能并定义基本的外周耐受机制。了解这些耐受性人群的生物学可能对一系列临床治疗具有重要意义从自身免疫到肿瘤免疫再到母胎耐受的应用。奖学金培训将在顶级资助的研究机构加州大学进行，旧金山（加州大学旧金山分校），在詹姆斯·加德纳博士和马克·加德纳博士的实验室中。 Anderson 博士在研究外周 Aire/eTAC 和基因小鼠模型的生成方面拥有丰富的专业知识。艾尔生物学、胸腺选择和免疫耐受方面的世界专家，也是一位备受尊敬的导师和领导者这些导师和机构将为免疫学领域的完成提供丰富的培训环境。从事学术研究职业所需的专业技能的研究和发展。