Project 3 - Differential contribution of thymic APCs to central tolerance during the perinatal to adult transition

项目 3 - 胸腺 APC 在围产期到成人过渡期间对中枢耐受的不同贡献

基本信息

批准号：
10022939
负责人：
Lauren Ilyse Richie EHRLICH
金额：
$ 51.96万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10022939
关键词：
Activities of Daily Living Adolescent Adult Affinity Age Agonist Antigen Presentation Antigen-Presenting Cells Antigens Autoantigens Autoimmune Process Autoimmunity Bioinformatics Biological Assay Biometry Cell Compartmentation Cell Differentiation process Cells Cellularity Characteristics Conflict (Psychology)Data Dendritic Cells Environment Gene Expression Gene Expression Profiling Generations Genetic Models Genetic Transcription Goals Growth Hematopoietic Human Impairment Life Location Maps Mediating Molecular Molecular Profiling Mus Organizational Change Output Perinatal Phenotype Play Process Property Regulatory T-Lymphocyte Reporting Role Self Tolerance Slice Stromal Cells T cell response T-Lymphocyte Testing Therapeutic Thymic epithelial cell Thymocyte Selection Thymus Gland Tissue imaging Tissues autoreactive T cell autoreactivity central tolerance experimental study multiplexed imaging pathogen perinatal period programs response single-cell RNA sequencing thymocyte two photon microscopy

项目摘要

ABSTRACT Project 3 The perinatal thymus plays a unique role in promoting central tolerance. Central tolerance results from negative selection of autoreactive conventional T cells (Tconv) and differentiation of regulatory T cells (Treg). Notably, Tregs selected during the perinatal period are more autoreactive and are uniquely required for life- long protection against autoimmunity. Perinatal Tconv cells are also more autoreactive and have unique molecular and functional properties compared with their juvenile/adult counterparts. Both negative selection and Treg induction occur when thymocytes encounter auto-antigens presented by medullary thymic epithelial cells (mTECs) or hematopoietic antigen presenting cells (HAPCs), including dendritic cells (DCs). TECs and thymic HAPCs undergo profound changes during the perinatal to juvenile transition. The rationale for RP3 is that thymic HAPCs are distinct in cellular composition and molecular profiles in the perinatal period, when thymic selection is critical for differentiation of the first wave of Tconv cells and establishment of self-tolerance. We will test the hypothesis that unique properties of mTECs and HAPCs in the perinatal period create an altered environment for central tolerance induction and are responsible for selection of Tconv and Treg cells with increased self-reactivity and age-specific functional properties. In Aim 1, we will use single cell transcriptional profiling (scRNA-seq) and multiplex imaging to identify the cellular, transcriptional, and organizational changes that occur in mouse and human HAPCs over the perinatal to juvenile transition which could impact central tolerance. In Aim 2, we will use phenotypic analyses, transcriptional profiling, genetic models, and functional assays to identify the APCs responsible for selecting tissue-protective Treg in the perinatal period. We will also determine whether CCR7-mediated cross-talk with TECs alters the DC compartment in perinatal mice, with downstream consequences for Treg selection. In Aim 3, we will determine if negative selection is impaired in perinates in response to antigens across a range of affinities. Live-cell 2- photon microscopy will be used to determine if distinct APCs induce negative selection in the perinatal period. RP3 has multiple points of intersection with all Projects and Cores. We will use scRNA-seq to identify altered molecular signatures of HAPCs in mice and humans (with Cores B and C) during the perinatal to juvenile transition that could impact central tolerance. Parallel data from RP1 and RP2 will provide a comprehensive map of changes in TECs and thymic stromal cells over the perinatal to juvenile transtition that could to alter central tolerance. We will also collaborate with the Manley lab (RP2) for MiCasa analysis to reveal organizational changes in the thymic environment during the perinatal to juvenile transition. Results generated in RP3 are integral to achieving the overall Program goals of elucidating mechanisms by which thymic stromal properties alter Treg and Tconv cell differentiation and selection in the perinatal period, with implications for devising rational therapeutic strategies to safely enhance T cell output.

摘要项目 3 围产期胸腺在促进中枢耐受方面发挥着独特的作用。中心公差的结果是自身反应性常规 T 细胞 (Tconv) 的阴性选择和调节性 T 细胞 (Treg) 的分化。值得注意的是，在围产期选择的 Tregs 具有更强的自身反应性，并且是生命所独特需要的。长期防止自身免疫。围产期 Tconv 细胞也更具自身反应性，并且具有独特的与幼年/成年对应物相比的分子和功能特性。均为负选择当胸腺细胞遇到髓质胸腺上皮呈现的自身抗原时，就会发生 Treg 诱导细胞（mTEC）或造血抗原呈递细胞（HAPC），包括树突状细胞（DC）。 TEC 和胸腺 HAPC 在从围产期到青少年的过渡过程中经历了深刻的变化。 RP3 的基本原理是胸腺 HAPC 在围产期的细胞组成和分子谱上是不同的，当胸腺选择对于第一波 Tconv 细胞的分化和自我耐受的建立至关重要。我们将检验以下假设：mTEC 和 HAPC 在围产期的独特特性创造了一种改变中枢耐受诱导的环境，并负责 Tconv 和 Treg 细胞的选择具有增强的自身反应性和特定年龄的功能特性。在目标 1 中，我们将使用单细胞转录分析 (scRNA-seq) 和多重成像来识别细胞、转录和小鼠和人类 HAPC 在从围产期到青少年的过渡过程中发生的组织变化可能会影响中枢耐受性。在目标 2 中，我们将使用表型分析、转录谱分析、遗传分析模型和功能测定来识别负责选择组织保护性 Treg 的 APC 围产期。我们还将确定 CCR7 介导的与 TEC 的串扰是否会改变 DC 围产期小鼠的隔室，对 Treg 选择具有下游影响。在目标 3 中，我们将确定如果围产期动物对一系列亲和力抗原的反应中的负选择受损。活细胞2- 光子显微镜将用于确定不同的 APC 是否会在围产期诱导负选择。 RP3 与所有项目和核心有多个交叉点。我们将使用 scRNA-seq 来识别在围产期改变小鼠和人类（核心 B 和 C）HAPC 的分子特征可能影响中枢耐受性的青少年过渡。来自 RP1 和 RP2 的并行数据将提供围产期到幼年期 TEC 和胸腺基质细胞变化的综合图可以改变中枢耐受性。我们还将与 Manley 实验室 (RP2) 合作进行 MiCasa 分析，以揭示围产期到青少年过渡期间胸腺环境的组织变化。结果 RP3 中产生的结果对于实现总体计划目标（阐明机制）是不可或缺的。胸腺基质特性改变围产期 Treg 和 Tconv 细胞的分化和选择，对设计合理的治疗策略以安全地增强 T 细胞输出的影响。