Genetic screens for key transcriptional regulators of antiviral T cell immunity

抗病毒 T 细胞免疫关键转录调节因子的基因筛选

基本信息

批准号：
8810644
负责人：
Shane P Crotty
金额：
$ 212.71万
依托单位：
LA JOLLA INSTITUTE FOR IMMUNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8810644
关键词：
Acute Antibody Response Antiviral Agents B-Lymphocytes Biology CD4 Positive T Lymphocytes CD8B1 gene Cell physiology Cells Cellular biology Chromatin Cytotoxic T-Lymphocytes Dermal Effector Cell Equilibrium Future Gene Expression Generations Genes Genetic Screening Genetic Transcription Goals Health Human Humoral Immunities Immune response Immunity Immunology Infection Intervention Knockout Mice Knowledge Link Lymphocyte Lymphocyte Biology Malignant Neoplasms Medical Memory Metabolic Mus Nucleosomes Nutrient Oxygen Pathway interactions Perception Population Process Provider Regulation Research Role Staging System T cell differentiation T cell response T memory cell T-Cell Activation T-Lymphocyte Testing Time Tissues Vaccine Design Vaccines Virus Diseases adaptive immunity base cost effective cytotoxic in vivo pathogen programs receptor transcription factor vaccine development vaccinology vector

项目摘要

DESCRIPTION (provided by applicant): Our projects propose a highly integrated approach to revealing the biology of regulation and differentiation of Tfh CD4 T cells, Th1 CD4 T cells, CTL CD8 T cells, and memory CD8 T cells, making use of high throughput genetic screens of T cell response in mice to multiple viral infections. 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 are central regulators of these differentiation processes. The identification of key transcription factors (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, and the large majority of analyses of TF networks have been correlative or computational. The generation of double and triple knockout mice is excessively time consuming. Furthermore, the compelling FANTOM study highlights the importance of moderate changes in TF expression for most cellular differentiation processes, not complete on-off switches. That has always been a clear limitation of knockout mice, and it continues to bias our perception of lymphocyte differentiation and function. Current experimental approaches are insufficient to dramatically advance our understanding of CD4 and CD8 T cell differentiation and function, and lymphocyte biology in general, due to conceptual, time, and monetary limitations. Therefore, our approach to this serious problem has been focused on generating an experimental approach whereby we can modulate and test 100 genes in parallel for their roles in antiviral T cell responses in vivo, using a new shRNAmir vector based approach. We have established this system, and are now able to perform genetic screens, in vivo, in primary CD4 or CDS T cells, probing differentiation and function. 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?

描述（由申请人提供）：我们的项目提出了一种高度集成的方法，利用高通量遗传技术来揭示 Tfh CD4 T 细胞、Th1 CD4 T 细胞、CTL CD8 T 细胞和记忆 CD8 T 细胞的调节和分化生物学。筛选小鼠 T 细胞对多种病毒感染的反应。 T 细胞分化为各种效应细胞以及分化为记忆细胞的能力是针对病原体和癌症的适应性免疫的重要组成部分。转录因子是这些分化过程的核心调节因子。调节 CD4 和 CD8 T 细胞分化不同途径的关键转录因子 (TF) 的鉴定对于了解这些细胞的生物学至关重要。然而，非常清楚的是，TF 并不是孤立地发挥作用，许多 TF 可能是 T 细胞分化途径的重要诱导剂或抑制剂。研究 TF 网络生物学的最大挑战是，在受控条件下一次对 1 个以上因子进行实验操作通常不太可行，特别是在体内原代细胞中。因此，几十年来，每次关注 1 个基因一直是实验的必需，并且 TF 网络的绝大多数分析都是相关的或计算的。双重和三重敲除小鼠的产生非常耗时。此外，引人注目的 FANTOM 研究强调了 TF 表达的适度变化对于大多数细胞分化过程的重要性，而不是完全的开关。这一直是基因敲除小鼠的明显局限性，并且它继续影响我们对淋巴细胞分化和功能的看法。由于概念、时间和资金的限制，目前的实验方法不足以显着增进我们对 CD4 和 CD8 T 细胞分化和功能以及淋巴细胞生物学的理解。因此，我们解决这个严重问题的方法一直集中在生成一种实验方法，通过这种方法，我们可以使用基于 shRNAmir 载体的新方法并行调节和测试 100 个基因在体内抗病毒 T 细胞反应中的作用。我们已经建立了这个系统，现在能够在原代 CD4 或 CDS T 细胞中进行体内遗传筛选，探测分化和功能。这三个项目积极追求对抗病毒 CD4 和 CD8 T 细胞的理解，其主题是：哪些转录因子调节这些细胞以及它们是如何调节这些细胞的？