Molecular basis of γδ T lineage specification

γδ T 谱系规范的分子基础

• 批准号: 10226992
• 负责人: DAVID L. WIEST
• 金额: $ 200.43万
• 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 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10226992
• 关键词: Address; Adoption; Affect; Architecture; Binding; Binding Proteins; Binding Sites; California; Cells; Chromatin Loop; Chromosomes; Complex; DNA-Binding Proteins; Development; Disease; E protein; Event; Exhibits; Family; Family member; Fox Chase Cancer Center; Funding; Gene Expression; Generations; Genetic Transcription; Genomics; Health; Host Defense; Human; Immune response; In Vitro; Inhibitor of Differentiation Proteins; Interleukin-17; Knowledge; Link; Mediating; Modeling; Molecular; Mus; Pathologic; Play; Pluripotent Stem Cells; Principal Investigator; Production; Public Health; Receptor Signaling; Regulation; Regulatory Element; Repression; Resources; Role; Signal Transduction; Specific qualifier value; Specificity; Stereotyping; Study models; System; Systems Development; T-Cell Development; T-Cell Receptor; T-Lymphocyte; TCF3 gene; Testing; Therapeutic; Time; Transcript; Universities; Untranslated RNA; base; genome-wide; human pluripotent stem cell; imaging approach; immunopathology; insight; member; novel; pathogen; professor; progenitor; programs; response; skills; stem cell model; stem cells; tumor; γδ T cells

PROJECT SUMMARY/ABSTRACT Despite the growing appreciation for the critical role played by γδ T cells in host defense and immunopathology, the molecular events controlling their development and effector function remain poorly understood. Our program seeks to fill this gap in knowledge using genome-wide approaches that ultimately focus on key regulatory nodes. During the last funding cycle, we provided compelling evidence that the commitment of T cell progenitors to the αβ and γδ T cell fates depends on differences in T cell receptor (TCR) signal strength. These signaling differences regulate fate by proportional induction of Id3, which causes graded repression of the function of E box DNA binding proteins (E proteins). Using genome-wide approaches to define changes in E protein-DNA binding, we generated a number of novel insights into the control of γδ T lineage specification, which can be distilled into 3 themes: Theme 1) E protein specificity – E protein family members exhibit distinct responses to the TCR signals of differing intensity/duration that specify fate and play distinct roles in supporting γδ T cell development (Proj1/3/4); Theme 2) Non-coding transcription - E protein binding is extensively associated long non-coding RNAs (lncRNA), such as ThymoD (all Projects), Importantly, Project 4 has developed novel imaging approaches to study lncRNA function, by visualizing lncRNA promotion of chromosome looping in real time in live cells; and Theme 3) Human γδ development - Project 3 developed a novel human pluripotent stem cell (PSC) based system for studying human γδ T cell development, which revealed a requirement for HEB. In our renewal application, we will explore these themes by integrating the complementary skills of the four project leaders. The Zúñiga-Pflücker lab, together with Michele Anderson, will focus on the specific roles of E protein family members, and their responsiveness to Id-mediated repression, in orchestrating lineage commitment in mouse (theme 1) and human (theme 3) differentiation models. Their observations will inform the efforts of the Murre lab to understand the molecular basis by which E protein family members control ThymoD expression, a lncRNA (theme 2), which orchestrates both the onset of αβ lineage commitment and the ultimate loss of αβ fate potential upon γδ lineage commitment. The efforts of both the Zúñiga-Pflücker and Murre labs will enable the Wiest lab (with Dietmar Kappes) to understand the role of E protein family members and non-coding transcription (theme 2) in specifying the IL-17 producing effector fate through E protein binding sites near the Tcf7 and Zbtb7b loci. Finally, all Projects will inform efforts of Zhuang to understand how E protein family members (theme 1) and non-coding transcription (theme 2) control the generation and function of the stereotyped γδ TCR complexes that drive NKγδT cell development. All projects will continue to rely on the genomic expertise of Core B to assess the implications of destroying E protein binding sites on the genomic architecture, and then link those changes to cell fate. The collective efforts of these four projects promise to provide great insight into the role of E proteins in controlling both γδ development and function, which is of critical importance to human health and disease.

项目概要/摘要 尽管人们越来越认识到 γδ T 细胞在宿主防御和免疫病理学中发挥的关键作用， 我们的程序对控制其发育和效应器功能的分子事件仍知之甚少。 寻求利用最终关注关键调控节点的全基因组方法来填补这一知识空白。 在上一个资助周期中，我们提供了令人信服的证据，证明 T 细胞祖细胞对 αβ 和 γδ T 细胞的命运取决于 T 细胞受体 (TCR) 信号强度的差异。 差异通过 Id3 的比例诱导来调节命运，从而导致 E 功能的分级抑制 盒 DNA 结合蛋白（E 蛋白）使用全基因组方法来定义 E 蛋白-DNA 的变化。 结合，我们对 γδ T 谱系规范的控制产生了许多新的见解，这可以是 提炼为 3 个主题： 主题 1) E 蛋白特异性 – E 蛋白家族成员表现出不同的反应 不同强度/持续时间的 TCR 信号指定命运并在支持 γδ T 细胞中发挥不同的作用 开发（Proj1/3/4）；主题 2）非编码转录 - E 蛋白结合通常与长相关 非编码 RNA (lncRNA)，例如 ThymoD（所有项目），重要的是，项目 4 开发了新颖的成像技术 研究 lncRNA 功能的方法，通过实时可视化 lncRNA 对染色体环的促进 活细胞；以及主题 3）人类 γδ 发育 - 项目 3 开发出一种新型人类多能干细胞 基于 PSC 的系统用于研究人类 γδ T 细胞发育，这揭示了我们对 HEB 的需求。 续订申请，我们将通过整合四个项目的互补技能来探索这些主题 Zúñiga-Pflücker 实验室将与 Michele Anderson 一起重点研究 E 蛋白的具体作用。 家庭成员，以及他们对身份介导的压抑的反应，在协调血统承诺方面 小鼠（主题 1）和人类（主题 3）分化模型的观察将为研究人员的努力提供信息。 Murre 实验室旨在了解 E 蛋白家族成员控制 ThymoD 表达的分子基础， lncRNA（主题2），协调αβ谱系承诺的开始和αβ命运的最终丧失 Zúñiga-Pflücker 和 Murre 实验室的努力将使 γδ 谱系承诺成为可能。 Wiest 实验室（与 Dietmar Kappes 合作）了解 E 蛋白家族成员和非编码转录的作用 （主题 2）通过 Tcf7 和 Zbtb7b 附近的 E 蛋白结合位点指定产生效应子命运的 IL-17 最后，所有项目都将告知 Zhuang 为了解 E 蛋白家族成员（主题 1）和 非编码转录（主题 2）控制定型 γδ TCR 复合物的生成和功能 驱动 NKγδT 细胞发育的所有项目将继续依赖 Core B 的基因组专业知识进行评估。 破坏 E 蛋白结合位点对基因组结构的影响，然后将这些变化联系起来 这四个项目的共同努力有望为 E 蛋白的作用提供深入的见解。 控制γδ的发育和功能，这对人类健康和疾病至关重要。