Define the role of REV-ERB in colonic RORgt+ regulatory T cells

定义 REV-ERB 在结肠 RORgt 调节性 T 细胞中的作用

• 批准号: 10753360
• 负责人: Ye Zheng
• 金额: $ 28.5万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-11 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10753360
• 关键词: Agonist; Antibodies; Autoimmune Diseases; Autoimmunity; Binding; Biological Assay; CTLA4 gene; Cell Lineage; Cell physiology; Cells; Colitis; Colon; Colonic inflammation; Data; Development; Equilibrium; FOXP3 gene; Family; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Homeostasis; Human; Immune system; Immunosuppressive Agents; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Insulin-Dependent Diabetes Mellitus; Interleukin-10; Knock-out; Knockout Mice; Lamina Propria; Lead; Link; Lymphoproliferative Disorders; Maintenance; Maps; Mediating; Molecular; Multiple Sclerosis; Mus; Nuclear Hormone Receptors; Nuclear Receptors; Orphan; Outcome; Outcome Study; Play; Population; Predisposition; Receptor Inhibition; Regulatory T-Lymphocyte; Repression; Research; Role; Running; Side; Spleen; T-Lymphocyte; Testing; Tissues; Transcription Coactivator; Transcription Factor 3; Transcription Repressor; Tretinoin; Work; anti-tumor immune response; conditional knockout; experimental study; genome-wide; immunopathology; in vivo; in vivo Model; insight; loss of function mutation; member; mouse model; murine colitis; new therapeutic target; next generation sequencing; prevent; protective effect; receptor; receptor function; therapeutic development; therapeutic target; transcription factor; transcriptome sequencing

PROJECT SUMMARY Regulatory T cells (Treg) play a crucial role in keeping the immune system in balance and preventing autoimmune disease. Defective Treg function leads to autoimmune diseases, while intra-tumor Tregs can block effective anti-tumor immune responses. Tregs can undergo further specialization in adaptation to their tissue microenvironment. Recent studies showed that a unique RORgt (retinoic acid–related orphan receptor gamma t) expressing Treg cell population could be induced in the colon lamina propria. These RORgt+ Tregs play an important role in the suppression of colon inflammation. In Treg cells, REV-ERBa is highly expressed in RORgt+ colonic Tregs, but not in RORgt- colonic Tregs or spleen Tregs. Mice with Treg-specific deletion of REV-ERB were more susceptible to TNBS-induced colitis with decreased RORgt+ Treg cells and increased Th1/Th17 cell- mediated inflammation in the colon. RNA-sequencing experiment showed that the expression of a set of genes related to Treg function, including CTLA-4, IL-10, and c-Maf, were reduced in REV-ERB deficient colonic Tregs compared to WT controls. To study the mechanism of gene regulation by REV-ERB in Tregs, Dr. Zheng’s lab performed Cut & Run assays with WT RORgt+ iTregs using Foxp3, REV-ERBa, and RORgt antibodies to map their binding peaks genome-wide. The results showed a significant overlap of the binding peaks of these 3 transcription factors, suggesting coordination in gene regulation among Foxp3, REV-ERB, and RORgt in Treg cells. The overall objective of the proposed study is to define the role of REV-ERB in Treg function. This goal will be accomplished by dissecting the molecular mechanism of REV-ERB function in RORgt+ Treg cells (Aim 1), and characterizing the role of REV-ERB in Treg in mouse models of IBD (Aim 2). The outcomes of the proposed study are expected to reveal the functional and mechanistic role of REV-ERB in RORgt+ Tregs. Such results are expected to further advance our understanding of how the identity and function of RORgt+ Tregs are established and maintained. Additionally, this study will provide key insights into how transcriptional repressor such as REV- ERB serves as key regulator and tune the function of transcriptional activators in Treg cells. Finally, the outcomes of this study could provide evidence supporting the development of therapeutics targeting REV-ERB for IBD treatment.

项目概要 调节性T细胞（Treg）在保持免疫系统平衡和预防疾病方面发挥着至关重要的作用。 自身免疫性疾病。 Treg 功能缺陷会导致自身免疫性疾病，而肿瘤内的 Tregs 可以阻断 有效的抗肿瘤免疫反应。 Tregs 可以进一步专业化以适应其组织 微环境。最近的研究表明，一个独特的 RORgt（视黄酸相关孤儿受体 γ t) 可以在结肠固有层中诱导表达 Treg 细胞群。这些 RORgt+ Tregs 发挥着 对抑制结肠炎症具有重要作用。在Treg细胞中，REV-ERBa在RORgt+中高表达 结肠 Tregs，但不在 RORgt-结肠 Tregs 或脾 Tregs 中。 REV-ERB 特异性 Treg 缺失小鼠 随着 RORgt+ Treg 细胞减少和 Th1/Th17 细胞增加，更容易受到 TNBS 诱导的结肠炎的影响 介导结肠炎症。 RNA测序实验表明一组基因的表达 与 Treg 功能相关的 Treg 功能，包括 CTLA-4、IL-10 和 c-Maf，在 REV-ERB 缺陷的结肠 Tregs 中减少 与 WT 对照相比。郑博士实验室研究REV-ERB对Tregs基因的调控机制 使用 Foxp3、REV-ERBa 和 RORgt 抗体对 WT RORgt+ iTreg 进行 Cut & Run 分析以进行图谱分析 它们的结合峰在全基因组范围内。结果显示这 3 个结合峰显着重叠 转录因子，表明 Treg 中 Foxp3、REV-ERB 和 RORgt 之间的基因调控协调 细胞。本研究的总体目标是明确 REV-ERB 在 Treg 功能中的作用。这个目标将 通过剖析 RORgt+ Treg 细胞中 REV-ERB 功能的分子机制来完成（目标 1）， 并描述 IBD 小鼠模型中 REV-ERB 在 Treg 中的作用（目标 2）。拟议的结果 研究有望揭示 REV-ERB 在 RORgt+ Tregs 中的功能和机制作用。这样的结果是 有望进一步加深我们对 RORgt+ Tregs 的身份和功能如何建立的理解 并维护。此外，这项研究还将提供有关 REV- 等转录抑制因子如何发挥重要作用的重要见解。 ERB 是关键调节因子，调节 Treg 细胞中转录激活因子的功能。最后，结果 这项研究的成果可以为支持针对 IBD 的 REV-ERB 疗法的开发提供证据 治疗。