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.

这项建议的目标是深入了解控制Ys血统承诺的分子过程。 以及效应器命运的详细说明。Y的血统承诺和效应者命运的指定都发生了 在胸腺的发育过程中；然而，我们对控制发育线索的理解 这些命运决定仍未完成。越来越多的证据表明，它们受到 T细胞受体(TCR)信号强度的差异表现为对E盒DNA的分级抑制 由E蛋白拮抗剂Ids介导的结合蛋白(E蛋白)。然而，E蛋白的目标是 对这些命运决定至关重要的因素仍然没有明确的定义。目前也不清楚不同的Y5是否 与交替命运选择相关联的TCR复合体通过自主地传递 不同的强度或是否需要配基参与。在解决这些问题时，我们将利用一个 非常适合ySTCR转基因模型(KN6)，其已知的选择配体非经典的MHC-I NoleculeT-10/22，CanbemanipulatedtoaltertheresultantTCRsignal.InAimi，我们将：使用KN6tg 以及内源性T-10/22反应性Ys祖细胞，以确定T-10/22如何特异性消融T- 10/22配体影响Ys的谱系承诺、谱系选择和效应器功能。AIM2寻求 理解矛盾观察的基础，即IDs是VY2+和VY2+发展所必需的 VyS-t-Ys T细胞，但抑制Vyi 1+天然Ys T细胞的发育。我们会评估是否会 在缺乏Ids的情况下，Vyi.1+先天Ys T细胞的扩增是Vyl.l A/66.3的自主属性 TCR复合体或需要配基结合。AIMS解决了关键的悬而未决的问题 Y6血统承诺和效应者命运的指定是可分离的或同时发生的。要做到这一点， 我们将利用我们新描述的Ys谱系承诺标记CD7S诱导。全基因组ChlP- 对E蛋白靶标的SEQ也将在CD73标记的细胞上进行，以组装一个全球调控 定义承诺流程的网络。这些努力需要所有 这个项目的成员承诺，将揭示Ys T细胞如何发育的关键新见解 控制住了。 相关性(请参阅说明)： Y5 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，了解控制其发育和功能的分子过程 可以使它们的操控达到治疗的效果。此外，我们对分子效应器的研究 控制T血统承诺对其他发育过程也是至关重要的， 因为控制细胞的生长和分化是发育和转化中反复出现的主题。