Transcription factor regulation of CD4 and CD8 T cell effector and memory differentiation and function

CD4 和 CD8 T 细胞效应及记忆分化和功能的转录因子调节

• 批准号: 10488579
• 负责人: Matthew Eugene Pipkin
• 金额: $ 227.56万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10488579
• 关键词: Antibody Response; B-Lymphocytes; Biology; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; CHD7 gene; Cells; Cellular biology; Chromatin; Complex; Consumption; Custom; Cytotoxic T-Lymphocytes; Data; Development; Effector Cell; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic Epistasis; Genetic Screening; Genetic Transcription; Helper-Inducer T-Lymphocyte; Heterogeneity; Histone Deacetylase; Human; IRF4 gene; Immunity; Intervention; Knockout Mice; Knowledge; Libraries; Link; Malignant Neoplasms; Medical; Memory; Molecular; Nature; NuRD complex; Paper; Pathway interactions; Population; Process; Provider; RUNX3 gene; Regulation; Regulator Genes; Role; System; T cell differentiation; T cell response; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Time; Tissues; Vaccine Design; Vaccines; Virus Diseases; Work; adaptive immunity; base; cost effective; cytotoxic CD8 T cells; effector T cell; epigenetic regulation; experience; experimental study; in vivo; insight; memory CD4 T lymphocyte; neoplasm immunotherapy; pathogen; programs; synergism; technology development; transcription factor; tumor; vector

ABSTRACT (Overall Component) The three Projects vigorously pursue an understanding of antiviral CD4 and CD8 T cells, linked by the theme: what transcription factors regulate these cells and how do they do so? T cell differentiation into various effector cells, and the capacity to differentiate into memory cells, are important parts of adaptive immunity to pathogens and cancers. Transcription factors (TFs) are central regulators of these differentiation processes. The identification of key TFs regulating different pathways of CD4 and CD8 T cell differentiation have been central to understanding the biology of these cells. However, it is abundantly clear that TFs do not act in isolation and many TFs may be important inducers or repressors of a T cell differentiation pathway. The biggest challenge to studying TF network biology is that experimental manipulation of more than 1 factor at a time under controlled conditions has not been generally feasible, particularly in primary cells in vivo. Therefore, the focus on 1 gene at a time has been an experimental necessity for decades; the generation of double and triple knockout mice is excessively time consuming. Therefore, our approach as a group to this serious problem has been focused on experimental approaches whereby we can modulate and test many genes in parallel for their roles in antiviral T cell responses in vivo, using custom shRNAmir vector-based approaches. We have established these systems, and are able to perform genetic screens, in vivo, in primary CD4 or CD8 T cells, probing differentiation and function. Our projects propose a highly integrated approach to revealing the biology of regulation and differentiation of T follicular helper (Tfh) CD4 T cells, memory CD4 T cells, CTL CD8 T cells, circulating memory CD8 T cells, and tissue-resident CD8 and CD4 T cells (Trm). The three PIs have worked intensively together over the past 5 years to develop important insights into TF biology in T cells. The interactions are not simply via shared cores, they involve extensive shared experiments and ideas regarding TFs, CRs, epigenetics, and gene regulation networks. This is most clearly seen in our 2017 Nature paper on Trm and Runx3. That work is connected to our earlier work together predicting TF regulators of gene expression in T cells, and our most recent paper together, defining the dominant pioneer factor role of Runx3 in the earliest steps of effector CD8 T cell differentiation and defining how Runx3 interacts with other TFs that we have studied, including T-bet, Blimp-1, IRF4, TCF1, and Id2. Our collective experience is that our screens identify multiple critical factors, more than can be comprehensively studied in the context of a single project. Therefore, the equally important challenge—and where our Program’s synergy stands out—is narrowing down the candidate molecules to particular ‘high value’ TFs and CRs that have a major influence on T cell programming. Our three Projects acquire such data in the orthogonal screens and epistasis experiments proposed by each Project, which allow us to converge quickly on important targets to study in detail.

抽象的 （总体组件） 这三个项目大力追求对抗病毒CD4和CD8 T细胞的理解，该细胞由主题联系在一起： 哪些转录因子调节了这些细胞？如何进行这些细胞？ T细胞分化为各种效应子 细胞以及分化为记忆细胞的能力，是对病原体自适应免疫学的重要部分 和癌症。转录因子（TF）是这些分化过程的中心调节因子。这 识别调查CD4和CD8 T细胞分化不同途径的关键TF已成为中心 了解这些细胞的生物学。但是，很明显，TFS不孤立地起作用，并且 许多TF可能是T细胞分化途径的重要诱导剂或复制品。最大的挑战 研究TF网络生物学是在受控的一次以上的实验操作。 条件通常是不可行的，尤其是在体内原代细胞中。因此，重点是1基因 几十年来，一次是一个实验性的。双重淘汰小鼠的产生是 过于耗时。因此，我们作为这个严重问题的小组的方法一直集中在 实验方法，我们可以在其中调节和测试许多基因在抗病毒t中的作用 使用基于自定义的Shrnamir矢量方法在体内进行细胞反应。我们已经建立了这些 系统，并且能够在一级CD4或CD8 T细胞中进行遗传筛选，体内，探测 分化和功能。我们的项目提出了一种高度综合的方法来揭示 T卵泡辅助器（TFH）CD4 T细胞，记忆CD4 T细胞，CTL CD8 T细胞的调节和分化 循环记忆CD8 T细胞以及组织居民CD8和CD4 T细胞（TRM）。这三个PI工作了 在过去的五年中，密集地共同开发了对T细胞中TF生物学的重要见解。 互动不仅是通过共享内核，还涉及广泛的共享实验和关于 TF，CRS，表观遗传学和基因调节网络。这是我们2017年自然论文中最清楚地看到的 trm和runx3。这项工作与我们早期的工作相关，以预测基因的TF调节剂 在T细胞中的表达以及我们最近的论文中的表达，定义了Runx3在 最早的效应子CD8 T细胞分化的步骤，并定义Runx3与我们的其他TF相互作用的最早步骤 已经研究了，包括T-BET，Blimp-1，IRF4，TCF1和ID2。我们的集体经验是我们的屏幕 确定多个关键因素，而不是在一个项目的背景下全面研究。 因此，同样重要的挑战（以及我们计划的协同作用脱颖而出的地方）正在缩小 对T细胞产生重大影响的特定“高价值” TF和CR的候选分子 编程。我们的三个项目在正交屏幕和Epistasis实验中获取此类数据 每个项目提出的提议，这使我们能够快速汇聚到重要目标以详细研究。