Regulatory T cell lineage stability controlled by Foxp3 CNS2

Foxp3 CNS2 控制的调节性 T 细胞谱系稳定性

• 批准号: 9197261
• 负责人: Ye Zheng
• 金额: $ 48.5万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9197261
• 关键词: Acute; Address; Autoimmune Diseases; Autoimmunity; Biology; Cell Lineage; Cell Therapy; Cells; Chronic; Collaborations; CpG Islands; Defect; Development; Drug Targeting; Elements; Exhibits; Experimental Autoimmune Encephalomyelitis; FOXP3 gene; Generations; Genetic; Genomic approach; Genomics; Goals; High-Throughput Nucleotide Sequencing; Homeostasis; Human; Immune System Diseases; Immune system; In Vitro; Individual; Inflammation; Inflammatory; Insulin-Dependent Diabetes Mellitus; Knock-out; Knockout Mice; Lead; Light; Link; LoxP-flanked allele; Lymphocyte Function; Lymphoproliferative Disorders; Maintenance; Methylation; Modeling; Molecular; Mucous Membrane; Multiple Sclerosis; Mus; Names; Organ Transplantation; Outcome; Output; Pathway interactions; Play; Population; Population Dynamics; Regulatory T-Lymphocyte; Role; Safety; Signal Transduction; Specificity; Symbiosis; System; T-Lymphocyte; T-Lymphocyte Subsets; Therapeutic; Thymus Gland; Transgenic Mice; Transplant Recipients; Universities; Untranslated RNA; Washington; base; comparative genomics; epigenetic marker; experience; immunopathology; in vivo; insight; knock-down; loss of function mutation; novel; novel therapeutics; prevent; public health relevance; small hairpin RNA; tool; transcription factor

DESCRIPTION (provided by applicant): Regulatory T cell (Treg) plays a critical role in maintaining immune system homeostasis and preventing autoimmunity and immunopathology. Compromised Treg function is linked to multiple autoimmune diseases. The development and maintenance of Treg cell lineage is dependent on transcription factor Foxp3, as loss of function mutations lead to severe lymphoproliferative disease in mice and humans. Thus, understanding the mechanisms that govern Foxp3 induction and stability may lead to novel therapies for autoimmune disease. Using a comparative genomic approach, Dr. Zheng and colleagues identified three evolutionarily conserved intronic cis-elements at the Foxp3 locus (conserved non-coding sequences (CNS) 1, 2 and 3). To elucidate the role of these CNS regions in Treg lineage development in vivo, they proceeded to knockout individual Foxp3 CNS regions in mice. Each of the three CNS knockout mice showed different defects in their Treg population, indicating the unique roles these CNS regions play in regulating Foxp3 expression. CNS2 is unique as it is located in the only CpG island at the Foxp3 locus. The methylation status of CNS2 is one of the most prominent epigenetic markers distinguishing Tregs from conventional T cells. Induction of Foxp3 is not affected by CNS2 deletion, but stabilization of Foxp3 expression is compromised. The mechanism underlying Treg lineage stability is not only a central question in the Treg biology field, but also has practical implications in Treg cell based therapy for autoimmune disease and organ transplant patients. The CNS2 KO mice provide a unique opportunity to explore the molecular and cellular mechanisms of Treg lineage stability in vitro and in vivo. Dr. Zheng's group will study the role of CNS2 in long-term maintenance of Treg stability in vivo, and determine if ex-Tregs are a major driver of autoimmunity (Aim 1). Additionally, they will address how Treg lineage stability is maintained under inflammation, a critical checkpoint for development of autoimmunity (Aim 2). They will dissect the molecular pathways that are involved in safeguarding Tregs under inflammatory conditions, which can potentially lead to drug targets for stabilizing Tregs for therapeutic purpose. Furthermore, Dr. Zheng's group will employ state-of-the-art mouse genetic tools to study the lineage stability of gut commensal induced iTregs (Aim 3). Since Treg lineage stability is directly related to the safety of Treg cell- based therapy for a variety of immune diseases, Dr. Zheng's study on CNS2 will provide insight on an important regulatory circuitry safeguarding Treg identity.

描述（由申请人提供）：调节性T细胞（Treg）在维持免疫系统稳态和预防自身免疫和免疫病理学中起关键作用。Treg功能受损与多种自身免疫性疾病有关。Treg细胞谱系的发育和维持依赖于转录因子Foxp3，因为功能缺失突变导致小鼠和人类严重的淋巴组织增生性疾病。因此，了解Foxp3诱导和稳定性的机制可能会导致自身免疫性疾病的新疗法。使用比较基因组方法，Zheng博士及其同事在Foxp3基因座上鉴定了三个进化上保守的内含子顺式元件（保守的非编码序列（CNS）1，2和3）。为了阐明这些CNS区域在体内Treg谱系发育中的作用，他们继续敲除小鼠中的单个Foxp3 CNS区域。三个CNS敲除小鼠中的每一个在其Treg群体中显示出不同的缺陷，表明这些CNS区域在调节Foxp3表达中发挥着独特的作用。CNS2是独特的，因为它位于Foxp3基因座上唯一的CpG岛上。CNS2的甲基化状态是区分TcB与常规T细胞的最突出的表观遗传标记之一。Foxp3的诱导不受CNS2缺失的影响，但Foxp3表达的稳定性受到损害。Treg谱系稳定性的机制不仅是Treg生物学领域的中心问题，而且在自身免疫性疾病和器官移植患者的基于Treg细胞的治疗中具有实际意义。CNS2 KO小鼠提供了一个独特的机会来探索Treg谱系在体外和体内稳定性的分子和细胞机制。Zheng博士的研究小组将研究CNS2在体内长期维持Treg稳定性中的作用，并确定前胸腺肽是否是自身免疫的主要驱动因素（Aim 1）。此外，他们将解决如何在炎症下维持Treg谱系稳定性，这是自身免疫发展的关键检查点（目的2）。他们将剖析参与在炎症条件下保护TdR的分子途径，这可能导致稳定TdR用于治疗目的的药物靶点。此外，Zheng博士的研究小组将采用最先进的小鼠遗传工具来研究肠道菌群诱导的iTbR的谱系稳定性（目的3）。由于Treg谱系的稳定性直接关系到基于Treg细胞的治疗对各种免疫疾病的安全性，Zheng博士对CNS2的研究将提供对保护Treg身份的重要调控电路的见解。