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: 10685628
• 负责人: Maria Ciofani
• 金额: $ 37.03万
• 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/10685628
• 关键词: Area; Binding Proteins; Cell Differentiation process; Cell Fate Control; Cells; ChIP-seq; Chromosome Mapping; Clonal Expansion; DNA-Binding Proteins; Development; 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; Molecular; Mus; Nature; Neonatal; Outcome; Perinatal; Play; Population; Protein Family; Proteins; Role; Signal Transduction; Specific qualifier value; Spleen; Stereotyping; System; T-Cell Development; T-Lymphocyte; TCF3 gene; Testing; Thymus Gland; Tissues; Transgenes; Transgenic Organisms; Untranslated RNA; V(D)J Recombination; Work; cytokine; experimental study; fetal; gene regulatory network; genome-wide; in vitro Model; insight; molecular imaging; mouse genetics; mouse model; permissiveness; 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.

项目总结/摘要 γδNKT（也称为NKγδT或Vδ6.3 T）谱系代表了一种先天性γδT细胞类型，其在哺乳动物中产生。 新生儿胸腺，随后在整个生命周期中维持在组织中，如胸腺、脾和 肝脏这些细胞与表达半恒定αβTCR的αβ谱系iNKT细胞密切相关， 它们一起构成了先天性样T淋巴细胞的主要群体。先天性类T细胞的一个主要功能 淋巴细胞的作用是调节其他免疫细胞的发育和功能，包括常规T细胞， 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。的 胸腺中γδNKT细胞的生成受DNA结合蛋白E盒家族的活性严格控制 （E蛋白），在T细胞发育中起着重要作用。Id蛋白抑制E蛋白的活性。 Id家族蛋白Id 3由与TCR信号强度成比例的TCR信号传导诱导，并且它是Id 3-E蛋白。 关键调节γδNKT细胞生成的蛋白质平衡。本方案项目完成的工作 已经揭示了TCR信号协调E蛋白结合到一个星座的全基因组重构， 增强子在γδT谱系特化、分化和扩增过程中的作用。然而，关键的E 控制单个γδ T亚系发育的蛋白质靶点仍然是知识上的一个主要空白。 我们与项目1一起确定了两个调节γδNKT发育的关键E蛋白靶点 细胞：编码定型的Vγ1.1Vδ6.3 TCR的基因组元件和一种转录因子， 在执行TCR启动的分化程序中发挥关键作用。我们现建议澄清 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。在目的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链的产生。在目标2中，我们将审查陈规定型观念的独特性质， Vγ1.1Vδ6.3 TCR和E蛋白如何协调由Vγ1.1Vδ6.3 TCR触发的基因表达变化 信号.目前的建议是建立在从方案项目得出的关键意见的基础上。拟议 实验将继续在TCR信号传导（项目1），小鼠 遗传学（项目2），小鼠和人类T细胞发育的体外建模（项目3），基因组学和 分子成像技术（项目4）。我们的具体目标不仅将导致更好地了解 γδNKT发展的机制，但也将提供独特的洞察一般机制， 哪些E蛋白控制细胞命运