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

• 批准号: 10311054
• 负责人: WEIGUO CUI
• 金额: $ 41.25万
• 依托单位: VERSITI WISCONSIN, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-16 至 2022-08-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10311054
• 关键词: ATAC-seq; Acetylation; Adoptive Transfer; Affect; Antigens; 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; 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; gene regulatory network; improved; insight; progenitor; programmed cell death protein 1; receptor; response; self-renewal; single-cell RNA sequencing; stem cells; success; synergism

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。