Molecular Basis for gamma/delta T Lineage Specification

gamma/delta T 谱系规范的分子基础

• 批准号: 8608275
• 负责人: DAVID L. WIEST
• 金额: $ 186.17万
• 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 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8608275
• 关键词: Ablation; Address; Affect; Affinity; Bioinformatics; Cell Differentiation process; Cell Nucleus; ChIP-seq; Complex; Cues; Cutaneous; Cytokine Signaling; DNA Binding; DNA-Binding Proteins; Development; Developmental Process; E protein; Epithelial; Event; Family; Gene Expression; Generations; Genes; Genetic screening method; Genomics; Goals; Immune response; In Vitro; Individual; Inflammation; Investigation; Laboratories; Ligands; Link; Molecular; Molecular Analysis; Molecular Models; Network-based; Outcome; Pathway interactions; Play; Process; Production; Receptor Signaling; Research Personnel; Role; Signal Transduction; System; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Test Result; Therapeutic; Thymus Gland; Transgenic Mice; Transgenic Model; base; cell growth; cytokine; extracellular; genetic analysis; in vitro Model; in vivo; insight; killings; member; molecular modeling; notch protein; novel; novel marker; pathogen; progenitor; programs; protein function; skills; transcription factor; tumor

DESCRIPTION (provided by applicant): Both the commitment of progenitors to the ?? 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 ?? 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 ?? 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 ?? 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 ?? 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 ?? lineage commitment and effector fate from extracellular signals to the network of targets in the nucleus. RELEVANCE: ?? T cells are increasingly understood to play critical roles in immune responses to pathogens and tumors that are unique and thus not overlapping with those of ???lineage T cells. Therefore, a greater understanding of how the development of ?? T cells if controlled at the molecular level may enable the manipulation of their production or function for therapeutic benefit. Project 1: Influence of ligand on specification of gamma/delta fate and function Project Leader (PL): Wiest, D DESCRIPTION (provided by applicant): The goal of this proposal is to understand the molecular processes controlling ?? lineage commitment and specification of effector fate. Both events occur during development in the thymus; however, our understanding of the developmental cues controlling these fate decisions remains incomplete. Accumulating evidence suggests that both ?? 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. We will employ this approach to elucidate the E protein targets that are crucial for these fate decisions, as well as, the DNA-binding proteins and signaling cascades with which they cooperate. In doing so, we will exploit the ?? TCR transgenic (Tg) model, KN6, whose known selecting ligand, the non-classical MHC-I molecule T10d, can be manipulated to alter TCR signaling. In Aiml, we will employ KN6 Tg mice, and endogenous T10/22 reactive ?? progenitors, to determine how specific ablation of the T10/22 ligand affects ?? lineage commitment, repertoire selection, and effector function. Aim2 will exploit the network approach described above to determine how Id3 is able to promote development of V?2+ and V?3+ ?? T cells, but restrains the development of V?1.1+ innate ?? T cells. Aim3 will exploit our novel marker of ?? lineage commitment, CD73, to determine whether ?? lineage commitment and specification of effector fate are separable or occur simultaneously. The molecular processes defining lineage commitment will be elucidated through the comprehensive network being assembled in Project 4, and by genetic tests of the resulting molecular model. These efforts require the capabilities of all program members and promise to reveal critical new insights into how ?? T cell development is controlled. RELEVANCE: ?? T cells regulate inflammation, preserve epithelial barriers, and are particularly adept at killing cutaneous tumors. Accordingly, understanding the molecular processes controlling their development and function may enable their manipulation for therapeutic benefit. Moreover, our investigation of molecular effectors controlling T lineage commitment is also of fundamental importance for other developmental processes, since control of cell growth and differentiation is a recurring theme in development and transformation.

描述（由申请人提供）：祖细胞对？？在胸腺发育过程中，其效应子命运的谱系和特化发生;然而，我们对控制这些命运决定的发育线索的理解仍然不完整。越来越多的证据表明，这两个？谱系定型和效应子命运受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如何？产生不同强度的TCR信号，并了解E蛋白活性和合作DNA结合蛋白的改变在影响谱系和？效应子命运全面了解这样一个多方面的发育过程超出了任何单个实验室的范围，因为它需要多种实验方法来操纵命运决定线索并评估它们对命运的影响，无论是功能上的还是分子上的，无论是在体外还是体内。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来协助所有项目在分子上定义关键里程碑？通过组装围绕E蛋白靶点组装的全球调控网络来开发。总的来说，这些努力有望通过全面定义控制？？从细胞外信号到细胞核中靶点网络的谱系定型和效应子命运。 相关性：？？越来越多的人认识到T细胞在对病原体和肿瘤的免疫反应中起着关键作用，这些病原体和肿瘤是独特的，因此与T细胞的免疫反应不重叠。谱系T细胞。因此，更好地了解如何发展？？如果在分子水平上控制T细胞，则可以操纵它们的产生或功能以获得治疗益处。 项目1：配体对γ/δ命运和功能质量标准的影响 项目负责人（PL）：Wiest，D 描述（由申请人提供）：本提案的目标是了解控制？？谱系定型和效应子命运的特化。这两个事件都发生在胸腺的发育过程中;然而，我们对控制这些命运决定的发育线索的理解仍然不完整。越来越多的证据表明，这两个？谱系定型和效应子命运受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。我们将采用这种方法来阐明E蛋白的目标是至关重要的这些命运的决定，以及DNA结合蛋白和信号级联与他们合作。在此过程中，我们将利用？？TCR转基因（Tg）模型，KN 6，其已知的选择配体，非经典MHC-I分子T10 d，可以被操纵以改变TCR信号传导。在Aiml中，我们将使用KN 6 Tg小鼠，和内源性T10/22反应性？祖细胞，以确定T10/22配体的特异性消融如何影响？谱系定型、库选择和效应子功能。目标2将利用上述网络方法，以确定如何识别码3能够促进V？2+和V？3+？？T细胞，但抑制V？1.1+先天？？T细胞。目标3将利用我们的新标记？？谱系承诺，CD 73，以确定是否？谱系定型和效应子命运的特化是可分离的或同时发生的。定义谱系承诺的分子过程将通过在项目4中组装的综合网络以及由此产生的分子模型的遗传测试来阐明。这些努力需要所有计划成员的能力，并承诺揭示关键的新见解如何？T细胞的发育受到控制。 相关性：？？T细胞调节炎症，保护上皮屏障，特别擅长杀死皮肤肿瘤。因此，了解控制它们的发育和功能的分子过程可以使它们能够操纵以获得治疗益处。此外，我们对控制T细胞系定型的分子效应物的研究对于其他发育过程也具有根本的重要性，因为细胞生长和分化的控制是发育和转化中反复出现的主题。