Molecular Basis for gamma/delta T Lineage Specification

描述（由申请方提供）：在胸腺发育期间，祖细胞向γδ谱系的定型及其效应物命运的特化均发生;然而，我们对控制这些命运决定的发育线索的理解仍不完整。越来越多的证据表明，γδ谱系定型和效应子命运都受到T细胞受体（TCR）信号强度差异的影响。TCR信号强度的差异通过诱导E蛋白DNA结合的拮抗剂Id 3影响命运。虽然E蛋白显然起着核心作用，但它们对发育结果的影响几乎肯定是由控制其表达和功能的额外转录因子和细胞外信号调节的。因此，这些命运决定太复杂，无法通过专注于一个基因或途径来理解，因此需要Murre开创的全面的，基于网络的方法。该计划的总体目标是确定不同强度的γδ TCR信号是如何产生的，并了解E蛋白活性和协同DNA结合蛋白的变化在影响谱系和γδ效应子命运中的作用。获得这样一个多方面的发展过程的全面理解是超出了任何单个实验室的范围，因为它需要设施与众多的实验方法来操纵命运决定线索和评估其对命运的影响，功能和分子，在体外和体内。组成该计划的调查人员拥有必要的独特但互补的技能。Wiest实验室（项目1）已经产生了体内和体外模型，其中发育命运可以通过改变TCR信号强度来操纵。Zuniga-Pflucker博士（项目3）已经建立了优雅的体外和体内系统，其中可以操纵Notch和细胞因子输入，以评估对效应子命运的影响。庄博士（项目2）和Murre博士（项目4）是E蛋白及其Id家族拮抗剂的遗传和分子分析专家。最后，Murre博士（基因组学核心）将利用他新颖的生物信息学方法，通过组装围绕E蛋白靶点组装的全球调控网络，协助所有项目在分子上定义γδ发育的关键里程碑。总的来说，这些努力有望通过全面定义控制γδ谱系定型和效应物命运的过程来推动该领域的发展，这些过程从细胞外信号到细胞核中的靶点网络。

DESCRIPTION (provided by applicant): Both the commitment of progenitors to the γδ lineage and specification of their effector fates occurs during development in the thymus; however, our understanding of the developmental cues controlling these fate decisions remains incomplete. Accumulating evidence suggests that both γδ lineage commitment and effector fate are influenced by differences in T cell receptor (TCR) signal strength. The differences in TCR signal strength influence fate by inducing Id3, an antagonist of E protein DNA binding. While E proteins clearly play a central role, their influence on developmental outcomes is almost certainly modulated by additional transcription factors and the extracellular signals that control their expression and function. Accordingly, these fate decisions are too complex to be understood by focusing on one gene or pathway and so require the comprehensive, network-based approach pioneered by Murre. The overall goal of this program is to determine how γδ TCR signals of varying intensities are generated and understand the role that the resultant alterations in E protein activity and cooperating DNA-binding proteins play in influencing lineage and γδ effector fate. Gaining a comprehensive understanding of such a multifaceted developmental process is beyond the scope of any individual laboratory, as it requires facility with numerous experimental approaches to manipulate fate-determining cues and assess their effect on fate, both functionally and molecularly, in vitro and in vivo. The investigators comprising this program possess the necessary distinct, yet complementary, skills to do so. The Wiest lab (Project 1) has generated in vivo and in vitro models in which developmental fates can be manipulated by altering TCR signal intensity. Dr. Zuniga-Pflucker (Project 3) has established elegant in vitro and in vivo systems in which Notch and cytokine input can be manipulated to assess the impact on effector fate. Drs. Zhuang (Project 2) and Murre (Project 4) are experts in the genetic and molecular analysis of E proteins and their Id family antagonists. Finally, Dr. Murre (Genomics Core) will utilize his novel bioinformatic approach to assist all projects in molecularly defining critical milestones in γδ development by assembling global regulatory networks assembled around E protein targets. Collectively, these efforts promise to move the field forward by comprehensively defining the processes controlling γδ lineage commitment and effector fate from extracellular signals to the network of targets in the nucleus.