Phenotypic, Functional and Transcriptional Heterogeneity in T Cell Exhaustion

T 细胞耗竭中的表型、功能和转录异质性

基本信息

批准号：
10428867
负责人：
WEIGUO CUI
金额：
$ 2.85万
依托单位：
VERSITI WISCONSIN, INC.
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-16 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10428867
关键词：
ATAC-seq Acetylation Adoptive Transfer Affect Antigens Antiviral Agents Binding Blocking Antibodies CD4 Positive T Lymphocytes CD8-Positive T-Lymphocytes Cell Differentiation process Cells ChIP-seq Chromatin Chronic Computer Analysis Cytokine Signaling Development Effector Cell Enhancers Epigenetic Process Flow Cytometry Genetic Genetic Transcription Goals Heterogeneity Individual Infection Inflammatory Interferons Interleukin-10 Knowledge Lymphocytic choriomeningitis virus Malignant Neoplasms Mediating Modeling Molecular Genetics Names PD-1 blockade Pathway interactions Phenotype Population Process RNA Interference Regulator Genes Research Signal Transduction Site Solid Surface T cell differentiation T-Lymphocyte T-Lymphocyte Subsets Testing Therapeutic Time Viral Virus Virus Diseases acute infection antiviral immunity base cancer immunotherapy chronic infection cytokine design effector T cell exhaust exhaustion extracellular improved insight progenitor programmed cell death protein 1 receptor response self-renewal single-cell RNA sequencing stem cells success synergism

项目摘要

ABSTRACT T cell exhaustion is a differentiation state that is marked by the loss of effector function and increased expression of inhibitory receptors such as PD-1. Although virus-specific CD8 T cells are commonly considered as a homogeneous population that gradually become exhausted over time, recent research has clearly demonstrated that a CXCR5hi TCF-1hi subset is serving as a self-renewing progenitor population that can give rise to a more terminally exhausted CXCR5lo TCF-1lo subset. To better dissect the heterogeneity of “exhausted” CD8 T cells, the lab applied single cell RNA-seq (scRNA-seq) to the chronic LCMV infection and identified three major subsets of virus-specific CD8 T cells that are phenotypically, functionally and transcriptionally distinct. Not only had the lab validated the existence of these subsets of T cells experimentally by flow cytometry, more advanced computational analyses were also performed to further predict their core transcriptional networks and developmental trajectories. Collectively, the findings reveal that a TCF-1hi progenitor subset can give rise to either a truly exhausted PD-1hi subset or a newly identified functional effector population that is named CX3CR1hi subset. This discovery laid a solid framework that allows testing of how extracellular signals and intrinsic genetic circuits regulate the formation and function of these three subsets of CD8 T cells. More importantly, it provides unprecedented opportunities to explore the possibility of generating more functional CX3CR1hi cells from TCF-1hi progenitors to overcome T cell exhaustion. This conceptual breakthrough is obviously applicable to control over chronic viral infection as well as cancer. Intriguingly, the preliminary study has also demonstrated that CD4 helper T cells, possibly through producing IL-21, are critical for TCF-1hi  CX3CR1hi transition. This led to the hypothesis that inflammatory cytokines (such as CD4- derived IL-21) modulate the cellular, functional and transcriptional diversity of virus-specific CD8 T cells during chronic LCMV infection. Blocking antibodies, RNA interference and genetic deletion models will be used to further dissect how inflammatory cytokines and transcriptional networks regulate heterogeneity in T cell exhaustion. Furthermore, the lab proposes to redirect CD8 T cell differentiation away from “exhaustion” by providing additional “CD4 help”, either alone or in combination with PD-1 blockade. The lab will test if providing IL-21 producing CD4 T cells through adoptive transfer could drive TCF-1hi progenitor cell differentiation into functional CX3CR1hi effector cells. Overall, knowledge gained from this research will provide mechanistic insights into how to redirect the formation of functional effector T cells and simultaneously limit T cell exhaustion for improved viral control over chronic infection.

抽象的 T细胞耗尽是一种区分状态，其标志着效应函数的丧失和增加通常认为抑制性受体（例如PD-1）。就像随着时间的流逝逐渐疲惫的同质人群一样，最近的研究显然已经证明CXCR5HI TCF-1HI子集是一个自我更新人群的服务上升到更耗尽的CXCR5LO TCF-1LO子集。 CD8 T细胞，实验室应用了单细胞RNA-Seq（SCRNA-Seq）慢性LCMV感染并鉴定病毒特异性CD8 T细胞的三个主要子集，在功能和转录上是表型的不同的。还进行了细胞仪，更先进的计算分析以进一步预测其核心转录网络和发展轨迹。祖细胞子集可能会引起耗尽的PD-1HI子集或新的iDenticational效应器名为CX3CR1HI子集的人口。细胞外信号和内在遗传回路调节地层，并关闭子集 CD8 T细胞。来自TCF-1HI祖细胞的功能性CX3CR1HI细胞克服T细胞的耗竭突破显然适用于控制慢性病毒感染和癌症初步研究还表明，尽管产生IL-21，CD4辅助T细胞可能是至关重要的对于TCF-1HICX3CR1HI过渡。在病毒特异性CD8 T细胞中的细胞，功能和转录多样性范围内慢性LCMV感染。进一步剖析炎症细胞因子和转录网络如何调节t细胞的异质性精疲力尽。单独或与PD-1封锁结合使用其他“ CD4帮助”。产生CD4的IL-21通过收养转移告诉TCF-1HI祖细胞细胞细胞分化可能功能性CX3CR1HI效应细胞总体上从研究中获得的知识提供了机械深入了解如何重定向功能性T细胞的形成和同时限制T细胞提高对慢性感染的病毒控制的呼气。