Influence of ligand on specification of gamma/delta fate and function

配体对 γ/δ 命运和功能规范的影响

• 批准号: 8608276
• 负责人: DAVID L. WIEST
• 金额: $ 32.44万
• 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/8608276
• 关键词: Ablation; Address; Adopted; Adoption; Affect; Affinity; Attenuated; Automobile Driving; Binding; Boxing; Cell Differentiation process; Cell Lineage; Cells; Commit; Complex; Cues; Cutaneous; DNA-Binding Proteins; Development; Developmental Process; E protein; Epithelial; Goals; In Vitro; Inflammation; Instruction; Investigation; Ligands; Link; Major Histocompatibility Complex; Mediating; Modeling; Molecular; Mus; Outcome; Pathway interactions; Peripheral; Process; Receptor Signaling; Repression; Role; Shapes; Signal Pathway; Signal Transduction; Specific qualifier value; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Technology; Testing; Therapeutic; Thymus Gland; Transgenic Mice; Transgenic Model; Transgenic Organisms; Zinc Fingers; base; cell growth; genome-wide; in vivo; insight; killings; member; novel; nuclease; progenitor; programs; response; thymocyte; transcriptome sequencing; tumor

The goal of this proposal is to gain insight into the molecular processes controlling yS lineage commitment and specification of effector fate. Both yS lineage commitment and specification of effector fate occur during development in the thymus; however, our understanding of the developmental cues controlling these fate decisions remains incomplete. Accumulating evidence suggests that they are governed by differences in T cell receptor (TCR) signal strength that manifest through graded repression of E box DNA binding proteins (E proteins) mediated by the E protein antagonist, IdS. Nevertheless, the E protein targets that are crucial for these fate decisions remain poorly defined. It is also unclear whether the different Y5 TCR complexes linked to alternate fate choices promote them by autonomously transducing signals of differing intensities or if they require ligand-engagement. In addressing these questions, we will exploit an ideally suited ySTCR transgenic model (KN6) whose known selecting ligand, the non-classical MHC-I nnoleculeT-10/22,CanbemanipulatedtoaltertheresultantTCRsignal.InAimi,wewill:employKN6tg mice as well as endogenous T-10/22 reactive yS progenitors to determine how specific ablation of the T- 10/22 ligand affects yS lineage commitment, repertoire selection, and effector function. Aim2 seeks to understand the basis for the paradoxical observation that IdS is required for the development of VY2+ and VyS-t- yS T cells, but restrains the development of Vyi.1+ innate yS T cells. We will assess whether the expansion of Vyi.1+ innate yS T cells in the absence of IdS is an autonomous attribute of the Vyl.l A/66.3 TCR complex or requires ligand-engagement. AimS addresses the critical unresolved question of whether y6 lineage commitment and specification of effector fate are separable or occur simultaneously. To do so, we will utilize our newly described marker of yS lineage commitment, CD7S induction. Genome wide ChlP- Seq on E protein targets will also be performed on CD73-marked cells to assemble a global regulatory network defining the commitment process. These efforts, which require the combined capabilities of all of the members of this program, promise to reveal critical new insights into how yS T cell development is controlled. RELEVANCE (See instructions): Y5 T cells regulate inflammation, preserve epithelial barriers, and are particularty 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.

该提案的目标是深入了解控制 yS 谱系承诺的分子过程 和效应器命运的规范。 yS谱系承诺和效应子命运的规范都发生 在胸腺发育期间；然而，我们对控制发育线索的理解 这些命运决定仍然不完整。越来越多的证据表明它们受 T 细胞受体 (TCR) 信号强度的差异通过 E 盒 DNA 的分级抑制表现出来 由 E 蛋白拮抗剂 IdS 介导的结合蛋白（E 蛋白）。尽管如此，E蛋白的目标是 对这些命运决定至关重要的因素仍然不明确。还不清楚是否不同的Y5 与替代命运选择相关的 TCR 复合物通过自主转导以下信号来促进它们： 不同的强度或是否需要配体接合。在解决这些问题时，我们将利用 非常适合 ySTCR 转基因模型 (KN6)，其已知的选择配体是非经典 MHC-I nnoleculeT-10/22，可以操纵以改变TCR信号。在Aimi中，我们将：使用KN6tg 小鼠以及内源性 T-10/22 反应性 yS 祖细胞，以确定 T-10/22 反应性 yS 祖细胞的特异性消融如何 10/22 配体影响 yS 谱系定型、库选择和效应子功能。 Aim2 旨在 了解 IdS 是 VY2+ 发展所必需的这一矛盾观察的基础 VyS-t- yS T 细胞，但抑制 Vyi.1+ 先天 yS T 细胞的发育。我们将评估是否 在 IdS 不存在的情况下 Vyi.1+ 先天 yS T 细胞的扩增是 Vyl.1 A/66.3 的自主属性 TCR 复合物或需要配体接合。 AimS 解决了一个尚未解决的关键问题： y6 谱系承诺和效应子命运的指定是分开的或同时发生的。为此， 我们将利用我们新描述的 yS 谱系定型标记，CD7S 诱导。全基因组 ChlP- E 蛋白靶点的测序也将在 CD73 标记的细胞上进行，以组装全球监管 网络定义承诺过程。这些努力需要所有各方的综合能力 该计划的成员承诺揭示有关 yS T 细胞发育如何的重要新见解 受控。 相关性（参见说明）： Y5 T 细胞调节炎症、保护上皮屏障，并且特别擅长杀死皮肤细胞 肿瘤。因此，了解控制其发育和功能的分子过程 可能使他们能够进行治疗以获得治疗效果。此外，我们对分子效应器的研究 控制 T 谱系承诺对于其他发育过程也至关重要， 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。