Linking the behavior of individual host cells to their transcriptional signatures

将单个宿主细胞的行为与其转录特征联系起来

• 批准号: 8711564
• 负责人: LALITA RAMAKRISHNAN
• 金额: $ 77.22万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8711564
• 关键词: Bacteria; Behavior; Cell Aggregation; Cells; Communicable Diseases; Disease; Elements; Extreme drug resistant tuberculosis; Gene Expression; Genetic Techniques; Genus Mycobacterium; Granuloma; Immune; Individual; Infection; Inflammatory; Intervention; Kinetics; Laboratories; Larva; Link; Modeling; Molecular; Molecular Profiling; Monitor; Movement; Mycobacterium marinum; Pathogenesis; Peripheral; Phagocytes; Phagocytosis; Plant Roots; Preventive Intervention; Process; Recruitment Activity; Reporter; Site; Structure; Techniques; Therapeutic Intervention; Time; Tissues; Tuberculosis; Vaccines; Work; Zebrafish; abstracting; cell motility; fighting; immunological intervention; innovation; insight; laser capture microdissection; novel therapeutic intervention; resistant strain; tool

DESCRIPTION Abstract: Linking the behavior of individual host cells to their transcriptional signatures in tuberculosis Tuberculosis is an infectious disease of enormous proportions with no effective vaccine and the emergence of extensively drug resistant strains. Major holes in our understanding of its pathogenesis present roadblocks to new therapeutic and preventive interventions. My laboratory has developed zebrafish infection by Mycobacterium marinum as a powerful surrogate model for exploring tuberculosis pathogenesis. The transparency of zebrafish larvae allows real-time examination of cellular movements and behaviors in unprecedented detail. We have made surprising discoveries about central aspects of tuberculosis pathogenesis, in particular regarding the organized aggregate of immune cells called the granuloma. This hallmark structure was long thought to be a key host protective element. However, detailed kinetic monitoring of granuloma formation revealed that granulomas are converted by mycobacteria into tools for expansion and dissemination. Phagocytes are recruited to the nascent granuloma, where they phagocytose the contents of dying infected cells. A subset of newly infected cells leaves the granuloma to disseminate infection to new foci. Here I propose to systematically link the behaviors of immune cells to their expression profiles to gain insight into the molecular blueprint of the host processes that promote pathogenesis. I propose to use laser capture microdissection techniques combined with multiple fluorescent bacterial and host reporters to isolate cells engaged in distinct movements, i.e. migrating towards newly infecting bacteria, returning to tissues from peripheral infection sites, migrating to forming granulomas, and departing granulomas. The gene expression profiles of these cells will then be deciphered using single cell gene expression techniques. We will then perturb these molecular signatures with genetic techniques to determine their impact on infection. These studies should yield a broad mechanistic understanding of basic processes such as cell migration and aggregation that are at the root of all inflammatory diseases and suggest new host-directed intervention strategies to fight them.

描述 摘要： 结核病中单个宿主细胞的行为与其转录特征的联系结核病是一种比例巨大的传染病，没有有效的疫苗，并出现了广泛的耐药菌株。我们对其发病机制的理解存在重大漏洞，阻碍了新的治疗和预防干预措施。我的实验室已经开发了斑马鱼感染海洋分枝杆菌作为一个强大的替代模型，探索结核病的发病机制。斑马鱼幼体的透明性允许以前所未有的细节实时检查细胞运动和行为。我们在结核病发病机制的核心方面取得了令人惊讶的发现，特别是关于称为肉芽肿的免疫细胞的有组织聚集。这种标志性结构长期以来被认为是宿主的关键保护元件。然而，肉芽肿形成的详细动力学监测显示，肉芽肿被分枝杆菌转化为扩张和传播的工具。吞噬细胞被募集到新生的肉芽肿中，在那里它们吞噬垂死的感染细胞的内容物。一部分新感染的细胞离开肉芽肿，将感染扩散到新的病灶。在这里，我建议系统地将免疫细胞的行为与其表达谱联系起来，以深入了解促进发病机制的宿主过程的分子蓝图。我建议使用激光捕获显微切割技术结合多种荧光细菌和宿主报告分离细胞从事不同的运动，即向新感染的细菌迁移，从外周感染部位返回组织，迁移形成肉芽肿，并离开肉芽肿。这些细胞的基因表达谱将使用单细胞基因表达技术来破译。然后，我们将用遗传技术干扰这些分子特征，以确定它们对感染的影响。这些研究应该产生一个广泛的机制的基本过程，如细胞迁移和聚集的理解，在所有炎症性疾病的根源，并提出新的主机定向干预策略，以打击他们。