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

• 批准号: 10536557
• 负责人: Jiaxi Wang
• 金额: $ 3.86万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 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/10536557
• 关键词: ATAC-seq; 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 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; Proteins; RNA; Regulator Genes; Regulatory T-Lymphocyte; Reporter; Research; Research Project Grants; Retinoic Acid Receptor; Role; San Francisco; Self Tolerance; Serum; Skin; Spleen; Surface; System; T-Cell Proliferation; T-Lymphocyte; Testing; Thymic epithelial cell; Thymus Gland; Time; 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

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.

项目摘要/摘要 自身免疫调节剂（AIRE）基因，一种在髓质胸腺上皮表达的键转录调节剂 通过诱导组织特异性抗原（TSA），细胞（MTEC）已被证明对中央耐受至关重要 MTEC中的表达。有趣的是，在表达外激体AIRE的细胞（ETAC）中也发现了AIRE 继发性淋巴机构，例如脾脏和淋巴结。我们以前发现ETAC是 造血抗原细胞（APC），由两种类似的细胞组成：CCR7+表达AIRE 迁移的树突状细胞（AMDC）和AIRE-HIGH人群共表达AIRE和维黄酸受体 - 我们称为Janus细胞（JCS）的相关孤儿接收器γT（RORγT）。在功能上，ETAC能够 在自反应性T细胞上执行删除和不含糊不清，并且在ETAC中的自我抗原表达足以使 防止自身免疫性。 ETAC之间的转录，基因组和功能对称性（JCS和 AMDCS）和MTEC可能会识别由AIRE驱动的核心计划，可能会影响自我代表 和跨免疫发展范围的耐受性。但是，ETAC的血统关系，什么 Aire在这些人群中扮演角色，以及外激发AIRE和ETAC子集有助于免疫 稳态仍然未知。该提案将检验以下假设，即AIRE被诱导了耐受性 ETAC中的表型以及外激发AIRE和ETAC对于强迫外周免疫很重要 宽容。该提案的目标1将定义ETACS子集之间的谱系关系，其抗原 处理和表现功能及其迁徙能力。 AIM 2将定义细胞中的功能 在转录和染色质水平的ETAC中AIR的AIR。 AIM 3将调查 外肌aire和ETAC保持正常的免疫稳态。该建议将使用 多种方法在内 利用新型遗传小鼠模型的自身免疫的共培养和体内监测。通过表征 ETAC子集并调查外激发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围耐受性机制。此外， 了解这些耐受性种群的生物学可能对一系列临床具有重要意义 从自身免疫到肿瘤免疫学到母亲易耐受性的应用。这个研究项目和 奖学金培训将在加利福尼亚大学一家资金资助的研究机构进行 旧金山（UCSF），詹姆斯·加德纳（James Gardner）博士和马克·安德森（Mark Anderson）博士的实验室。加德纳博士有 研究周围AIRE/ETAC和遗传小鼠模型的产生方面的专业知识。安德森博士是 世界AIR生物学，胸腺选择和免疫耐受性的世界专家，以及备受尊敬的精神和领导者 在免疫学领域。这些导师和机构将为完成提供丰富的培训环境 研究和发展学术研究所需的专业技能。