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: 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
• 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

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 自身免疫调节因子（Aire）基因，一种在胸腺髓质上皮中表达的关键转录调节因子 细胞（mTECs），已被证明是至关重要的中枢耐受诱导组织特异性抗原（TSA） 在mTEC中的表达。有趣的是，Aire也存在于胸腺外表达Aire的细胞（eTAC）中， 次级淋巴器官，如脾和淋巴结。我们之前发现eTAC 造血抗原呈递细胞（APC），由两种相似的细胞类型组成：CCR 7 + Aire表达细胞 迁移性树突状细胞（AmDCs）和共表达Aire和视黄酸受体的Aire-高群体， 相关孤儿受体γt（ROR γt），我们称之为Janus细胞（JC）。在功能上，eTAC能够 在自身反应性T细胞上强制缺失和无反应性，并且eTAC中的自身抗原表达足以 防止自身免疫。eTAC（JC和ETAC）之间的转录、基因组和功能对称性是不确定的。 AmDCs）和mTECs可能识别出由Aire驱动的可能影响自我表征的核心程序 和耐受性。然而，eTAC的血统关系， Aire在这些人群中的作用，以及胸腺外Aire和eTAC亚群如何有助于免疫 体内平衡仍然是未知。该提议将检验Aire诱导耐受原性的假设。 胸腺外Aire和eTAC对于增强外周免疫是重要的， 宽容本提案的目的1将定义eTAC亚群、其抗原 加工和呈递功能，以及它们的迁移能力。目标2将定义细胞内在功能 Aire在eTAC中的转录和染色质水平。目标3将研究 胸腺外Aire和eTAC在维持正常免疫稳态中的作用。该提案将使用 多种方法，包括单细胞多组学、流式细胞术和功能性方法，例如离体 共培养和利用新的遗传小鼠模型体内监测自身免疫。通过表征 eTAC亚群和研究胸腺外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，明确外周耐受的基本机制。此外，委员会认为， 了解这些致耐受性群体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。该研究项目和 奖学金培训将在一个顶级资助的研究机构，加州大学， 旧金山弗朗西斯科，在詹姆斯·加德纳博士和马克·安德森博士的实验室。加德纳医生 在研究外周Aire/eTAC和遗传小鼠模型的生成方面具有专业知识。安德森博士是一位 在艾尔生物学、胸腺选择和免疫耐受方面的世界专家，以及一位备受尊敬的导师和领导者 在免疫学领域。这些导师和机构将提供一个丰富的培训环境， 这种研究和发展的专业技能，必要的职业生涯中的学术研究。