Regulatory mechanisms governing Th17 cell effector identity and plasticity

控制 Th17 细胞效应子身份和可塑性的调控机制

• 批准号: 10211809
• 负责人: Maria Ciofani
• 金额: $ 38.21万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10211809
• 关键词: 3-Dimensional; Adopted; Adoption; Anti-Inflammatory Agents; Architecture; Autoimmune Diseases; Autoimmunity; Automobile Driving; Bacteria; Biological Assay; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Lineage; Cell physiology; Cells; Characteristics; Chromatin; Chromatin Loop; Disease; Effector Cell; Elements; Environment; Exhibits; Experimental Autoimmune Encephalomyelitis; Funding; Gene Expression Regulation; Gene Targeting; Genetic; Genetic Transcription; Genome; Genomics; Helper-Inducer T-Lymphocyte; Immune response; Immunity; Infection; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Interleukin-17; Intestines; Knowledge; Maps; Mediator of activation protein; Molecular; Molecular Conformation; Mucosal Immunity; Mucous Membrane; Multiple Sclerosis; Pathogenicity; Pathology; Play; Proteins; Psoriatic Arthritis; Regulation; Regulator Genes; Regulatory Element; Regulatory T-Lymphocyte; Reporter; Reporting; Rheumatoid Arthritis; Role; Series; Site; Spinal Cord; Structure; Surface; T cell differentiation; T-Lymphocyte; T-Lymphocyte Subsets; Therapeutic Intervention; Work; autoimmune inflammation; cell type; cytokine; effector T cell; epigenomics; flexibility; fungus; in vivo; mouse model; novel; preservation; programs; tool; trait; transcription factor

PROJECT SUMMARY T helper (Th) cells are CD4-expressing T lymphocytes that diversify into several subclasses to support distinct immune responses. Among the diversity of Th subset differentiation options, IL-17A-producing inflammatory Th17 cells stand out as unique by virtue of their relatively high level of inherent plasticity. Indeed, this subset that normally functions in mucosal immunity against bacteria and fungi can easily adopt features of other T helper subsets when environmental conditions change. While this feature can be advantageous during the clearance of an infection, dysregulated Th17 cell function has been implicated in numerous autoimmune conditions, including Inflammatory Bowel Disease, Multiple Sclerosis, and Rheumatoid Arthritis. Moreover, Th17 cell plasticity exhibited in the context of inflammatory disease tends to take on Th1-like traits, such as expression of IFNg or T-bet, that are also associated with increased pathology. In our current funded studies, we identified a JunB-T-bet axis as a central node in the regulatory network governing Th17 cell lineage stability versus plasticity. While several regulators influencing the transcriptional program of Th17 cell identity and flexibility have been identified, less is known about the regulators of the three-dimensional genome architecture that differs between cell types and states and is a critical determinant of gene regulation. Thus, we hypothesize that regulation of chromatin confirmation by lineage-specific factors is key to the control of CD4 T cell effector conversions. Here, we propose to comprehensively evaluate the dynamic changes in chromatin configuration and the cis genomic elements that govern Th17 cell effector conversions in vivo. To this end, in Aim 1, we will apply genetic fate-mapping mouse models, global chromatin conformation assays, and epigenomic profiling tools to characterize the chromatin looping dynamics during Th17 cell plasticity in a mouse model of experimental autoimmune encephalomyelitis (EAE). We will determine the contribution of looping to Th17 cell plastic conversion by assessing the role of both chromatin organizing proteins and lineage-regulating transcription factors. In Aim 2, we plan to determine the cis regulatory network that governs Th17 cell plasticity. For this, we will apply scATACseq profiling of Th cells in EAE to identify plasticity-associated cis elements. We will validate candidates for activity and function using novel in vivo high throughput reporter assays and CRISPR/Cas9 epigenomic screens. This will identify new regulatory networks, regulators, and gene targets critical to Th17 cell effector plasticity. Taken together, the proposed work will fill an important gap in knowledge concerning the molecular mechanism governing stability versus plasticity of Th17 cells.

项目摘要 T助手（Th）细胞是表达CD4的T淋巴细胞，将多样化成几个子类别以支持不同 免疫反应。在子集分化选项的多样性中，产生IL-17A的炎症 Th17细胞由于其相对较高的固有可塑性而脱颖而出。确实，这个子集 通常在粘膜免疫中对细菌和真菌的功能很容易采用其他T的特征 当环境条件发生变化时，助手子集。虽然此功能在 感染的清除，TH17细胞功能失调与许多自身免疫有关 疾病，包括炎症性肠病，多发性硬化症和类风湿关节炎。而且， 在炎症性疾病的背景下表现出的Th17细胞可塑性倾向于采用Th1样性状，例如 IFNG或T-BET的表达也与病理增加有关。在我们目前的资助研究中， 我们将一个junb-t-bet轴确定为控制TH17细胞谱系稳定性的监管网络中的中央节点 与可塑性相对于可塑性。而几个监管机构影响了Th17细胞身份的转录程序和 已经确定了灵活性，对三维基因组结构的调节剂的了解少 细胞类型和状态之间的不同，是基因调节的关键决定因素。因此，我们假设 通过谱系特异性因素对染色质确认的调节是控制CD4 T细胞效应子的关键 转换。在这里，我们建议全面评估染色质配置的动态变化 以及在体内控制Th17细胞效应子转化的顺式基因组元素。为此，在AIM 1中，我们将 应用遗传命运映射小鼠模型，全局染色质构象测定和表观基因组分析 在Th17细胞可塑性期间，在鼠标模型中表征染色质循环动力学的工具 实验性自身免疫性脑脊髓炎（EAE）。我们将确定循环对Th17单元的贡献 通过评估染色质组织蛋白和谱系调节的作用，通过评估塑料转化 转录因子。在AIM 2中，我们计划确定控制TH17细胞可塑性的顺式调节网络。 为此，我们将应用EAE中Th细胞的ScatacSeq分析以鉴定与可塑性相关的CIS元素。我们 将使用新型的体内高通量记者分析和 CRISPR/CAS9表观基因组筛选。这将确定新的监管网络，调节器和基因目标 对Th17细胞效应的可塑性至关重要。综上所述，拟议的工作将填补知识的重要空白 关于控制稳定性与Th17细胞可塑性的分子机制。