Defining the Effector T cell KLF2 Transcriptome

定义效应 T 细胞 KLF2 转录组

• 批准号: 10391460
• 负责人: Geoffrey S. Kansas
• 金额: $ 24万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-12 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10391460
• 关键词: ATAC-seq; B-Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Compartmentation; Cell physiology; Cells; Chromatin; Data; Defect; Development; Disease; Down-Regulation; Effector Cell; Enterobacteria phage P1 Cre recombinase; Epigenetic Process; Failure; Family; Gene Expression; Generations; Genes; Genetic Transcription; Helper-Inducer T-Lymphocyte; Homing; Immune response; Immune system; Immunologics; Impairment; Infection; Infectious Agent; Inflammatory; Leukocytes; Lymphocytic choriomeningitis virus; Mature T-Lymphocyte; Metabolism; Modeling; Mutant Strains Mice; Outcome; Peripheral; Physiological; Play; Research Personnel; Role; T cell response; T-Lymphocyte; Time; Transcriptional Activation; adaptive immune response; arm; cell type; effector T cell; member; migration; mouse model; novel; pathogen; programs; receptor; response; trafficking; transcription factor; transcriptome; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Transcriptional programs which control the activation, differentiation and migration of T and B lymphocytes have been studied for decades, yet remain incompletely understood. “Master” transcription factors which drive the differentiation of different classes of CD4 helper cells and CD8 effector cells have been well characterized, but functions of other transcription factors which regulate the outcome of an immune response are less well defined. KLF2, a member of the Kruppel-like factor family, has diverse roles in the immune system, and in particular plays a critical role in controlling migration of T (and B) cells via its control of expression of key homing receptors which determine selective trafficking capabilities. Other functions of KLF2 remain mostly undefined. A striking and unusual feature of KLF2 is its rapid loss of expression in T, B and multiple other cell types following cellular activation. Despite this feature of KLF2 being known for almost two decades, the physiologic importance of this loss of KLF2 remains poorly defined, despite efforts by several investigators. We have generated a novel, powerful, and sophisticated mouse model which for the first time makes possible interrogation of the functions of KLF2 downregulation. In this model, loss of KLF2 expression is completely and permanently blocked in any cell type which expresses or which once expressed cre recombinase. We present extensive data showing that this novel mouse model has a normal peripheral T cell compartment prior to immunologic challenge, yet induction of both CD4 and CD8 T cell effectors in response to infection with LCMV is sharply impaired. Using this powerful and novel mouse model, we propose to identify the transcriptional and epigenetic basis underlying defects in the generation of effector T cells associated with failure to turn off KLF2 expression. In Aim 1, we will identify aspects of T cell effector development which require loss of KLF2 expression, focusing on possible defects in defective or dysregulated activation, proliferation, or survival, and alterations in cellular metabolism or traffic. In Aim 2, we will use RNAseq and ATACseq to identify how the transcriptome and regulome of effector CD4 and CD8 T cells changes as a result of failure to extinguish KLF2 expression. These studies will greatly advance our understanding of how KLF2 controls the T cell arm of the adaptive immune response.

项目摘要/摘要 控制T和B淋巴细胞激活、分化和迁移的转录程序 已经研究了几十年，但仍然不完全清楚。“大师”转录因子 驱使不同类别的CD4辅助细胞和CD8效应细胞分化良好 具有调节免疫反应结果的其他转录因子的特征和功能 定义不那么明确。KLF2是Kruppel样因子家族的成员之一，在免疫中具有不同的作用 系统，特别是在控制T(和B)细胞迁移方面起着关键作用。 决定选择性转运能力的关键归巢受体的表达。的其他功能 KLF2大部分仍未定义。KLF2的一个显著和不同寻常的特征是它在T、B 以及细胞激活后的多种其他类型的细胞。尽管KLF2的这一特性几乎为人所知 二十年来，这种KLF2缺失的生理重要性仍然没有得到很好的定义，尽管 几名调查人员。我们已经生成了一种新颖、强大和复杂的鼠标模型，用于 第一次使询问KLF2下调的功能成为可能。在这个模型中，KLF2的损失 在任何表达或一旦表达的细胞类型中，表达都被完全和永久地阻止 Cre重组酶。我们提供的大量数据表明，这种新的小鼠模型具有正常的外周T 免疫攻击前的细胞隔间，但同时诱导CD4和CD8T细胞效应器 对LCMV感染的反应急剧减弱。使用这个强大而新颖的鼠标模型，我们 建议确定效应器T产生缺陷的转录和表观遗传学基础 与未能关闭KLF2表达相关的细胞。在目标1中，我们将确定T细胞效应器的各个方面 需要丢失KLF2表达的开发，重点关注缺陷或 失调的激活、增殖或存活，以及细胞代谢或运输的改变。在目标2中，我们 将使用RNAseq和ATACseq来确定效应器CD4和CD8T的转录和调节组如何 KLF2表达不能被抑制，导致细胞发生变化。这些研究将极大地促进我们的 了解KLF2如何控制获得性免疫反应的T细胞臂。