Project 2 - Chang

项目2-张

• 批准号: 10214457
• 负责人: John T Chang
• 金额: $ 45.25万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-17 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214457
• 关键词: Acute; Body Surface; CD8-Positive T-Lymphocytes; Cell Differentiation process; Cells; Collaborations; Computer Analysis; Data Set; Effector Cell; Exhibits; Exposure to; FURIN gene; Face; Failure; Gene Expression; Gene Expression Profiling; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Goals; Growth; Heterogeneity; Host Defense; Immune; Immune response; Immunity; Immunology; Individual; Infection; Knock-out; Lymphocyte; Malignant Neoplasms; Measurement; Memory; Modeling; Molecular; Mucous Membrane; Nature; Outcome; Population; Principal Investigator; Process; Proteins; RNA; RNA interference screen; Regulation; Sentinel; Signal Transduction; Site; Specific qualifier value; T cell differentiation; T memory cell; T-Lymphocyte; Testing; Therapeutic Intervention; Time; Tissues; Vaccine Design; Validation; Work; acute infection; base; chronic infection; cost effective; design; exhaust; exhaustion; first responder; gene discovery; genome-wide; in vivo; innovation; insight; intestinal epithelium; microbial; new therapeutic target; phenotypic biomarker; single-cell RNA sequencing; small hairpin RNA; transcriptome; transcriptomics

SUMMARY-PROJECT 2-CHANG: During an immune response to microbial infection, a naive CD8+ T lymphocyte can give rise to terminal effector cells that provide acute host defense and functionally distinct subsets of memory cells that provide durable immunity. Generation of one such subset, tissue-resident memory cells (Trm), is now recognized to be essential for protection at mucosal and body surfaces. Exhaustion, the failure of immune cells to achieve or maintain optimal function, is an alternative outcome that occurs in the setting of chronic infections and cancer. Prior studies have advanced our understanding of how CD8+ T cells differentiate into Trm or exhausted cells, but the field faces several important challenges that this Project seeks to overcome. We propose to take an unbiased, `phenotypic marker-agnostic' approach exploiting an innovative sequencing approach that enables simultaneous measurement of proteins and the transcriptome in the same single cells to overcome the inherent limitations of prior studies that have performed transcriptional profiling at the bulk population level. We will utilize an in vivo shRNA-based approach enabling us to reiteratively evaluate the function of 50 genes simultaneously, thereby overcoming the slow pace of discovery resulting from traditional genetic knockout models that evaluate one gene at a time. Specifically, we will use these approaches to achieve the following goals: (1) Investigate the molecular heterogeneity and functional importance of Trm cells within and among tissues; (2) Elucidate the ontogeny of Trm cells by reconstructing the transcriptomic roadmap from the naive state through progressive states of differentiation, within and across tissue sites; (3) Identify early, functionally important molecular determinants of exhausted circulating and tissue-resident CD8+ T cells. A major strength of our proposal is the integrated and synergistic design that exploits the strength of the individual Project and Core Principal Investigators, generating datasets that inform multiple Projects. We envision that insights resulting from our studies may have important implications for the rational design of vaccines and for therapeutic interventions that reverse the exhausted state.

总结-项目2-变更： 在对微生物感染的免疫应答过程中，初始CD 8 + T淋巴细胞可以产生终末效应子。 提供急性宿主防御的细胞和提供持久免疫的功能不同的记忆细胞亚群。 免疫力一个这样的子集，组织驻留存储器细胞（Trm）的生成现在被认为是可实现的。 对粘膜和身体表面的保护至关重要。衰竭，免疫细胞无法实现或 维持最佳功能，是在慢性感染和癌症的情况下发生的替代结果。 先前的研究已经推进了我们对CD 8 + T细胞如何分化为Trm或衰竭细胞的理解， 但该领域面临着本项目寻求克服的几个重要挑战。我们建议采取 无偏见的，“表型标记不可知”的方法， 创新的测序方法， 在相同的单细胞中同时测量蛋白质和转录组 克服 先前在群体水平上进行转录谱分析的研究的固有局限性。我们 将利用一种基于shRNA的体内方法，使我们能够粘附地评估50个基因的功能， 同时，从而克服了传统基因敲除导致的缓慢发现速度 一次评估一个基因的模型。具体来说，我们将使用这些方法来实现以下目标 目的：（1）研究Trm细胞在细胞内和细胞间的分子异质性和功能重要性 （2）通过从原始组织中重建转录组路线图来阐明Trm细胞的个体发育 通过组织部位内和跨组织部位的渐进分化状态来表达状态;（3）识别早期、功能性 耗尽的循环和组织驻留的CD 8 + T细胞的重要分子决定因素。一个主要优势 我们的建议是综合和协同设计，利用个别项目的优势， 核心主要研究者，生成为多个项目提供信息的数据集。我们设想， 我们的研究结果可能对疫苗的合理设计和 治疗干预，以扭转疲惫的状态。