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.

项目总结/摘要 控制T和B淋巴细胞活化、分化和迁移的转录程序 已经研究了几十年，但仍然没有完全理解。“主”转录因子， 驱动不同类别的CD 4辅助细胞和CD 8效应细胞的分化已经很好地 特征性的，但是调节免疫应答结果的其他转录因子的功能 不太明确。KLF 2是Kruppel样因子家族的成员，在免疫系统中具有多种作用。 系统，特别是在通过其控制T（和B）细胞的迁移中起关键作用。 决定选择性运输能力的关键归巢受体的表达。的其他功能 KLF 2大部分尚未定义。KLF 2的一个显著的和不寻常的特征是其在T、B中表达的快速丧失 和多种其它细胞类型。尽管KLF 2的这一功能几乎 20年来，KLF 2丢失的生理重要性仍然不清楚，尽管 几个调查员。我们已经产生了一种新颖、强大和复杂的小鼠模型， 第一次使KLF 2下调的功能的询问成为可能。在这个模型中，KLF 2的损失 在表达或曾经表达的任何细胞类型中， cre重组酶。我们提出了大量的数据表明，这种新的小鼠模型具有正常的外周T细胞， 细胞区室免疫攻击前，但诱导CD 4和CD 8 T细胞效应， 对LCMV感染的反应严重受损。使用这种强大而新颖的小鼠模型，我们 建议确定转录和表观遗传基础的缺陷产生的效应T 与未能关闭KLF 2表达相关的细胞。在目标1中，我们将鉴定T细胞效应物的方面， 需要KLF 2表达缺失的发展，重点关注缺陷或 失调的活化、增殖或存活，以及细胞代谢或运输的改变。在目标2中， 将使用RNAseq和ATACseq来鉴定效应CD 4和CD 8 T细胞的转录组和调节组如何表达， 细胞的变化是由于未能消除KLF 2表达。这些研究将大大促进我们的 了解KLF 2如何控制适应性免疫反应的T细胞臂。