Molecular and genomic control of innate γδ T cell development

先天γδ T 细胞发育的分子和基因组控制

• 批准号: 10462548
• 负责人: Yuan Zhuang
• 金额: $ 37.69万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462548
• 关键词: Area; Binding Proteins; Cell Differentiation process; Cells; ChIP-seq; Clonal Expansion; DNA-Binding Proteins; Development; E protein; EGR2 gene; Elements; Embryo; Enhancers; Epigenetic Process; Equilibrium; Event; Family; Funding; Gene Expression; Generations; Genes; Genetic Recombination; Genetic Transcription; Genomics; Human; IL4 gene; Imaging technology; Immune; Individual; Inhibitor of Differentiation Proteins; Interferon Type II; Knowledge; Lead; Life; Ligands; Liver; Maps; Molecular; Mus; Nature; Neonatal; Outcome; Perinatal; Play; Population; Protein Family; Proteins; Role; Signal Transduction; Spleen; Stereotyping; System; T-Cell Development; T-Lymphocyte; TCF3 gene; Testing; Thymus Gland; Tissues; Transgenes; Transgenic Organisms; Untranslated RNA; V(D)J Recombination; Work; base; cytokine; experimental study; fetal; gene regulatory network; genome-wide; in vitro Model; insight; molecular imaging; mouse genetics; mouse model; postnatal; programs; protein function; recruit; stem cells; transcription factor; transgene expression; γδ T cells

PROJECT SUMMARY/ABSTRACT The γδNKT (also known as NKγδT or Vδ6.3 T) lineage represents an innate type of γδT cells that is generated in the neonatal thymus and subsequently maintained throughout life in tissues such as the thymus, spleen, and liver. These cells are closely related to αβ lineage iNKT cells, which express a semi-invariant αβTCR, and together they constitute the main populations of innate-like T lymphocytes. A major function of innate-like T lymphocytes is to modulate the development and function of other immune cells including conventional T lymphocytes. γδNKT cells use a stereotyped Vγ1.1Vδ6.3 TCR; however, their ligand(s) remains unknown. The generation of γδNKT cells in the thymus is tightly controlled by the activity of E box family of DNA-binding proteins (E proteins), which play profound roles in T cell development. The activity of E proteins is inhibited by Id proteins. The Id family protein, Id3, is induced by TCR signaling in proportion to TCR signal strength and it is the Id3-E protein balance that critically regulates the generation of γδNKT cells. Work completed by this Program Project has revealed that TCR signals orchestrate genome-wide remodeling of E protein binding to a constellation of enhancers during γδT lineage specification, differentiation, and expansion. However, the identity of the critical E protein targets that control the development of individual γδ T sublineages remains a major gap in knowledge. We, together with Project 1, have identified two critical E protein targets that regulate the development of γδNKT cells: the genomic elements encoding the stereotyped Vγ1.1Vδ6.3 TCR and a transcription factor that plays a crucial role in executing the differentiation program initiated by the TCR. We now propose to elucidate the mechanism by which E-proteins regulate γδNKT development and restrict it to neonatal life. We will do so through the following aims. In aim 1, we will investigate E-protein function in regulating the recombination events responsible for generation of the Vδ6.3 TCR chain. In aim 2, we will examine the unique nature of the stereotyped Vγ1.1Vδ6.3 TCR and how E-proteins coordinate the changes in gene expression triggered by Vγ1.1Vδ6.3 TCR signals. The current proposal is built upon key observations derived from the Program Project. The proposed experiments will continue to draw the complementary expertise in the areas of TCR signaling (Project 1), mouse genetics (Project 2), in vitro modeling of mouse and human T cell development (Project 3), and genomics and molecular imaging technologies (Project 4). Our specific aims will not only lead to a better understanding of the mechanism of γδNKT development but will also provide unique insight into the general mechanisms through which E proteins control cell fate.

项目概要/摘要 γδNKT（也称为 NKγδT 或 Vδ6.3 T）谱系代表 γδT 细胞的先天类型，产生于 新生儿胸腺，随后终生维持在胸腺、脾脏和 肝。这些细胞与 αβ 谱系 iNKT 细胞密切相关，后者表达半不变的 αβTCR，并且 它们共同构成了先天样 T 淋巴细胞的主要群体。先天类T的主要功能 淋巴细胞的作用是调节其他免疫细胞（包括常规 T 细胞）的发育和功能 淋巴细胞。 γδNKT 细胞使用定型的 Vγ1.1Vδ6.3 TCR；然而，它们的配体仍然未知。这 胸腺中 γδNKT 细胞的生成受到 DNA 结合蛋白 E box 家族活性的严格控制 （E 蛋白），在 T 细胞发育中发挥着深远的作用。 E蛋白的活性被Id蛋白抑制。 Id 家族蛋白 Id3 由 TCR 信号传导诱导，与 TCR 信号强度成比例，它是 Id3-E 关键调节 γδNKT 细胞生成的蛋白质平衡。本计划项目完成的工作 研究表明，TCR 信号协调 E 蛋白与一系列蛋白结合的全基因组重塑。 γδT 谱系规范、分化和扩展过程中的增强子。然而，关键 E 的身份 控制个体 γδ T 亚系发育的蛋白质靶点仍然是一个主要的知识空白。 我们与项目 1 一起确定了调节 γδNKT 发育的两个关键 E 蛋白靶点 细胞：编码定型 Vγ1.1Vδ6.3 TCR 的基因组元件和发挥作用的转录因子 在执行 TCR 发起的分化计划中发挥着至关重要的作用。我们现在建议阐明 E蛋白调节γδNKT发育并将其限制在新生儿生命中的机制。我们将通过 以下目标。在目标 1 中，我们将研究 E 蛋白在调节重组事件中的功能 负责 Vδ6.3 TCR 链的生成。在目标 2 中，我们将研究刻板印象的独特性质 Vγ1.1Vδ6.3 TCR 以及 E 蛋白如何协调 Vγ1.1Vδ6.3 TCR 触发的基因表达变化 信号。当前的提案是建立在从该计划项目中得出的关键观察结果的基础上的。拟议的 实验将继续汲取 TCR 信号传导（项目 1）、小鼠 遗传学（项目 2）、小鼠和人类 T 细胞发育的体外模型（项目 3）以及基因组学和 分子成像技术（项目 4）。我们的具体目标不仅会让人更好地理解 γδNKT 发育的机制，但也将通过以下方式提供对一般机制的独特见解 其中E蛋白控制细胞命运。