Project 3 - Zuniga

摘要--项目3--祖尼加： 组织驻留记忆(Trm)CD8+T细胞是记忆细胞的一个独特子集，它为 针对黏膜屏障和非屏障组织的微生物的前线防御。尽管Trm现在是 公认在宿主抵御急性感染中的关键作用、功能和分子调控 在持续性感染期间，很大程度上是未被探索的。这种知识差距从根本上说是重要的。 鉴于感染主要持续在慢性感染后的非淋巴组织中。作为坚持不懈的 艾滋病毒、丙型肝炎病毒和乙肝病毒等感染在世界范围内造成了巨大的疾病负担， 了解非淋巴组织中的免疫调节将有助于制定新的策略来控制Trm 慢性感染和肿瘤。 我们提出了创新的方法来阐明具体促进Trm耗竭的新机制， 效应器功能的渐进性丧失，导致组织中持续存在病原体并增强敏感性 继发性感染。这些研究可能会重新定义目前CD8+T细胞分化、命运和 在持续病毒感染过程中的作用，因为目前对T细胞耗竭的了解几乎完全是 基于对循环淋巴细胞群体的研究。具体地说，我们将：(1)破译独特的分子 尖端单细胞RNA-SEQ和小细胞数对耗竭的Trm的调节和异质性 表观遗传学方法与新的综合计算分析相结合。新的调节剂 这些研究中确定的耗尽的Trm将用高通量的在体功能shRNA进行测试 放映。(2)阐明T-box转录因子、T-bet和eome的作用机制。 转录抑制因子Ezh2在急性和慢性感染中控制Trm分化的不同作用。 (3)研究非典型性转化生长因子β信号成分如何调控TRM的发育和耗竭。我们的 研究将利用其他项目和核心实验室中代表的数据和专业知识来定义 Trm分化、滞留和衰竭的功能和分子决定因素，提供了 有效操纵CD8+T细胞亚群对抗慢性传染病的基础 以及肿瘤浸润性淋巴细胞耗尽的癌症。由于LCMV是该病毒的原型成员 我们的工作也将有助于解释出血热引起的T细胞抑制 阿雷纳病毒。此外，我们预计将从以下方面收集关于TRM监管的基本知识 我们的研究将对其他免疫相关疾病具有广泛的意义，在这些疾病中，Trm已被证明是 发挥致病作用，如过敏和自身免疫。

SUMMARY-PROJECT 3-ZUNIGA: Tissue-resident memory (Trm) CD8+ T cells are a distinct subset of memory cells that provide an essential frontline of defense against microbes at mucosal barriers and non-barrier tissues. Although Trm are now recognized to play critical roles in host defense against acute infection, their function and molecular regulation during persistent infections have been largely unexplored. This knowledge gap is fundamentally important given that infection persists primarily in non-lymphoid tissues following chronic infection. As persistent infections such as HIV, HCV and HBV are responsible for tremendous disease burden worldwide, understanding immune regulation in non-lymphoid tissues will inform novel strategies to harness Trm against chronic infections and tumors. We propose innovative approaches to elucidate new mechanisms that specifically promote Trm exhaustion, the progressive loss of effector functions, leading to persistent pathogen in tissues and enhancing susceptibility to secondary infections. These studies will likely redefine current models of CD8+ T cell differentiation, fate and function during persistent viral infection as the current understanding of T cell exhaustion is almost exclusively based on studying circulating lymphocyte populations. Specifically, we will: (1) Decipher the unique molecular regulation and heterogeneity of exhausted Trm using cutting-edge single-cell RNA-seq and small-cell-number epigenetic methodologies combined with novel integrative computational analyses. Novel regulators of exhausted Trm identified in these studies will be tested with high-throughput functional in vivo shRNA screening. (2) Elucidate the mechanisms by which T-box transcription factors, T-bet and Eomes, and the transcriptional repressor Ezh2 differentially operate to control Trm differentiation in acute and chronic infection. (3) Investigate how non-canonical TGFβ signaling components regulate Trm development and exhaustion. Our studies will leverage the data and expertise represented in the other Project and Core laboratories to define the functional and molecular determinants of Trm differentiation, retention, and exhaustion, providing the foundation with which to effectively manipulate this CD8+ T cell subset to combat chronic infectious diseases and cancer where tumor infiltrating lymphocytes undergo exhaustion. As LCMV is a prototypic member of the family arenaviridae, our work will also help explain T cell suppression caused by hemorrhagic fever arenaviruses. Moreover, we anticipate that the basic knowledge on Trm regulation that will be gathered from our studies will have broad implications for other immune-related diseases in which Trm have been shown to play a pathogenic role, such as allergy and autoimmunity.