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.

项目概要 T 辅助细胞 (Th) 是表达 CD4 的 T 淋巴细胞，可分化为多个亚类以支持不同的功能 免疫反应。在 Th 亚群分化选项的多样性中，产生 IL-17A 的炎症 Th17 细胞因其相对较高水平的固有可塑性而脱颖而出。事实上，这个子集 通常在针对细菌和真菌的粘膜免疫中发挥作用，可以很容易地采用其他 T 的特征 当环境条件发生变化时的辅助子集。虽然此功能在 清除感染、Th17 细胞功能失调与许多自身免疫性疾病有关 疾病，包括炎症性肠病、多发性硬化症和类风湿性关节炎。而且， 炎症性疾病背景下表现出的 Th17 细胞可塑性往往呈现出类似 Th1 的特征，例如 IFNg 或 T-bet 的表达，也与病理增加有关。在我们目前资助的研究中， 我们确定了 JunB-T-bet 轴作为控制 Th17 细胞谱系稳定性的调控网络的中心节点 与可塑性。虽然有几个调节因子影响 Th17 细胞身份和转录程序 灵活性已被确定，但对三维基因组结构的调节剂知之甚少 这在细胞类型和状态之间有所不同，并且是基因调控的关键决定因素。因此，我们假设 谱系特异性因子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，我们建议综合评估染色质构型的动态变化 以及体内控制 Th17 细胞效应器转换的顺式基因组元件。为此，在目标 1 中，我们将 应用基因命运图谱小鼠模型、整体染色质构象测定和表观基因组分析 表征小鼠模型中 Th17 细胞可塑性期间染色质环动态的工具 实验性自身免疫性脑脊髓炎（EAE）。我们将确定循环对 Th17 细胞的贡献 通过评估染色质组织蛋白和谱系调节的作用来进行塑料转化 转录因子。在目标 2 中，我们计划确定控制 Th17 细胞可塑性的顺式调控网络。 为此，我们将应用 EAE 中 Th 细胞的 scATACseq 分析来识别可塑性相关的顺式元件。我们 将使用新颖的体内高通量报告测定法验证候选物的活性和功能， CRISPR/Cas9 表观基因组筛选。这将确定新的监管网络、监管机构和基因目标 对 Th17 细胞效应可塑性至关重要。总而言之，拟议的工作将填补知识方面的重要空白 关于控制 Th17 细胞稳定性与可塑性的分子机制。