Administrative Core

项目摘要/摘要 行政核心的主要作用是协调4个项目地点之间的研究活动和 基因组学核心。生物学研究已经远远超出了对单一基因或途径的分析。 因此，这个项目试图使用全基因组分析来洞察T细胞的差异 细胞受体(Tcr)信号强度引导胸腺祖细胞采用γδ命运，以及它们可能如何 影响效应器命运规范。为了做到这一点，我们使用了最初集中在细胞上的基因组分析 E盒DNA结合蛋白(E蛋白)的靶点，调节αβ和 γδT细胞。我们在上一个资助周期中对E蛋白结合位点的全基因组分析揭示了一些 关键的E蛋白靶点和协同转录因子对采用γδT细胞命运至关重要。我们 现在寻求通过基因组学核心来利用这些发现来评估这些E蛋白的功能 我们关注全基因组E蛋白网络中的靶点，并确定它们如何影响γδ谱系的命运。 该计划整合了γδT细胞开发和E蛋白功能方面的四位领导者的努力。项目1将 探索编排γδ谱系中的E蛋白靶标TCF7和长非编码RNA(LncRNA)Gm15417 承诺和采用产生IL-17的效应器命运。项目2将评估E蛋白的作用 TCRVδ元件在控制非编码转录中的调控元件 NKγδT细胞和转录因子EGR2，在协调NKγδT细胞的发育过程中起着关键作用。我们的节目 还发现特定的E蛋白家族成员具有特定的、非冗余的支持作用 γδT细胞的发展和项目3将阐明其特定作用的机制基础。项目3有 还开发了一种基于多能干细胞的人类γδT细胞开发模型，将使我们能够 在动物模型中评估程序发现的人类相关性。最后，项目4将评估角色 E蛋白控制的lncRNA，ThymoD，在调节T谱系承诺中的作用以及它的缺失如何导致 致力于αβ命运的细胞中的γδ命运潜能的熄灭。具体地说，Project 4还将使用成像 为了可视化胸腺激素对γδT细胞发育过程中染色体环路动力学的影响，这是 基因表达的关键调节器。行政核心将通过以下方式协调这些活动：1)重新- 建立管理体制；2)促进试剂的分配；3)协调科学 项目地点之间的交流和学员对基因组核心的访问，以及必要时的其他计划 实验室。总而言之，我们的计划承诺不仅揭示对分子控制的新见解 γδT细胞开发，但也将装备一批新的科学家，掌握应用集成湿凳的技能 以及生物信息学方法来解决生物学中的重要问题。

PROJECT SUMMARY/ABSTRACT The primary role of the Administrative Core is to coordinate research activities among the 4 Project Sites and Genomics Core. Biological research has moved well beyond the analysis of single genes or pathways. Accordingly, this program seeks to use genome-wide analysis to gain insight into the way that differences in T cell receptor (TCR) signal strength direct thymic progenitors to adopt the γδ fate, as well as how they might influence effector fate specification. To do so, we have employed genomic analysis focused initially on the cellular targets of E box DNA-binding proteins (E proteins), which regulate critical checkpoints in development of αβ and γδ T cells. Our genome-wide analysis of E protein binding sites in the last funding cycle revealed a number of critical E protein targets and cooperating transcription factors that are critical in adoption of the γδ T cell fate. We now seek to leverage those findings through the Genomics Core to assess the function of those E protein targets within our genome-wide E protein focused network and determine how they influence γδ lineage fate. The program integrates the efforts of four leaders in γδ T cell development and E protein function. Project 1 will explore the E protein targets, Tcf7 and long noncoding RNA (lncRNA) Gm15417, in orchestrating γδ lineage commitment and adoption of the IL-17 producing effector fate. Project 2 will assess the role of E protein regulated elements in controlling noncoding transcription from the TCRVδ element employed by innate-like NKγδT cells and the TF Egr2, which plays a critical role in orchestrating NKγδ T cell development. Our program has also discovered that particular E protein family members have specific, non-redundant roles in supporting γδ T cell development and Project 3 will elucidate the mechanistic basis for their specific roles. Project 3 has also developed a pluripotent stem cell (PSC)-based human γδ T cell development model that will enable us to assess the human relevance of program findings made in animal models. Finally, Project 4 will evaluate the role of an E protein controlled lncRNA, ThymoD, in regulating T lineage commitment and how its loss results in the extinguishing of αβ fate potential in cells committed to the γδ fate. Specifically, Project 4 will also employ imaging to visualize the effect of ThymoD on the kinetics of chromosome looping during γδ T cell development, which is a critical regulator of gene expression. The Administrative Core will coordinate these activities by: 1) re- establishing a management structure; 2) facilitating the distribution of reagents; and 3) coordinating scientific interchanges between project sites and trainee visits to the Genomics Core, and as necessary other Program Laboratories. Collectively, our program promises not only to reveal novel insights into the molecular control of γδ T cell development, but will also equip a new cadre of scientists with the skills to apply integrated wet bench and bioinformatic approaches to important questions in biology.