Genetic and metabolic regulation of macrophage activation at steady state

巨噬细胞稳态激活的遗传和代谢调控

• 批准号: 9978893
• 负责人: Celia E Shiau
• 金额: $ 38.29万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978893
• 关键词: Address; Binding; Biochemical; Biological; Biological Assay; Biological Models; Candidate Disease Gene; Cells; Chemicals; Chronic; Collaborations; Complex; Development; Diabetes Mellitus; Engineering; Equilibrium; Foundations; Funding; Gene Silencing; Genes; Genetic; Genetic Screening; Glucose; Goals; Health; Immune signaling; Immune system; Inflammation; Inflammatory; Innate Immune System; Knowledge; Lipids; Logic; Macrophage Activation; Malignant Neoplasms; Metabolic; Metabolic Pathway; Molecular; Obesity; Pathway interactions; Process; Property; Proteomics; Public Health; Regulation; Reporter; Research; Series; Shapes; Signal Pathway; Signal Transduction; Technology; Work; Zebrafish; design; disorder prevention; genetic analysis; genetic manipulation; high throughput screening; human disease; human model; immune activation; in vivo; in vivo imaging; innovation; macrophage; metabolomics; mouse model; mutant; novel; null mutation; prevent; receptor; screening; stem; transcriptomics

Project Summary Understanding how a cell is switched off and maintains its quiescence is fundamentally as important as how it is activated. The long-term goal of the proposed research is to determine how cell-intrinsic processes control and modulate activation states of macrophages in vivo. Because dysregulation and unprovoked activation of the immune system cause a host of human diseases associated with inappropriate inflammation, deciphering the molecular networks regulating immune activation normally is critical for addressing these health challenges. This will lead to new knowledge and technologies needed to harness the properties of macrophages for disease prevention and treatment. We are leveraging the unique advantages of the highly tractable vertebrate model system, Danio rerio, for exquisite genetic manipulations, high throughput screening, and in vivo imaging to dissect the complex relationship between intrinsic metabolic signaling and macrophage activation. The proposal encompasses a series of projects that collectively define essential negative regulators and their functions for keeping the innate immune system in check to maintain a normal equilibrium in macrophages. The starting basis of our projects stems from emerging evidence that metabolic and immune signaling pathways intersect to shape immune activation in macrophages, and a discovery of a null mutation in an intracellular NOD-like receptor (NLR) in zebrafish. A gene inactivation in this novel NLR, nlrc3l, causes unprovoked macrophage activation possibly due to metabolic dysregulation. The proposal seeks to define the network of molecular interactions of nlrc3l to understand this very important mechanism that keeps macrophages in check under normal biological conditions. We are taking a highly integrated approach at multiple levels-- using differential transcriptomics, proteomics, and metabolomics to inform candidate genes and pathways that constitute possible interactors and effectors of nlrc3l, and validating interactions using genetic mutants and biochemical studies. The proposal will also use the power of a forward genetic screen to discover additional genes akin to nlrc3l that prevent macrophage activation at steady state that act in the same or completely new pathways. We designed an innovative assay for the screen to assess macrophage activation using a live-cell reporter for an activation marker irg1. Finally, the proposal will examine the influence of lipid and glucose metabolic pathways on macrophage activation in zebrafish using genetic analyses and chemical screening. This work will benefit from collaboration with an expert group in extending our findings to mouse and human models. Taken together, these projects provide the important foundation for understanding the genetic and metabolic basis of how the innate immune system is kept in check, and will impact the direction of my lab far beyond the 5 years of MIRA funding.

项目概要 了解细胞如何关闭并保持其静止状态从根本上来说与了解细胞如何关闭并保持其静止状态同样重要。 被激活。拟议研究的长期目标是确定细胞内在过程如何控制 并调节体内巨噬细胞的激活状态。因为调节失调和无端激活 免疫系统会导致许多与不适当的炎症相关的人类疾病，破译 通常调节免疫激活的分子网络对于解决这些健康问题至关重要 挑战。这将带来利用其特性所需的新知识和技术 巨噬细胞用于疾病预防和治疗。我们正在利用高度的独特优势 易驯化的脊椎动物模型系统，Danio rerio，用于精致的基因操作，高通量筛选， 和体内成像来剖析内在代谢信号与巨噬细胞之间的复杂关系 激活。 该提案包含一系列项目，这些项目共同定义了基本的负面监管机构及其 具有控制先天免疫系统以维持巨噬细胞正常平衡的功能。 我们项目的起始基础源于新出现的证据，即代谢和免疫信号传导 途径交叉形成巨噬细胞中的免疫激活，并且发现了一个无效突变 斑马鱼细胞内 NOD 样受体 (NLR)。这种新型 NLR（nlrc3l）中的基因失活会导致 巨噬细胞无端激活可能是由于代谢失调所致。该提案旨在定义 nlrc3l 的分子相互作用网络，以了解这一非常重要的机制 巨噬细胞在正常生物条件下受到检查。我们正在采取高度集成的方法 多层次——使用差异转录组学、蛋白质组学和代谢组学来告知候选基因 以及构成 nlrc3l 可能的相互作用子和效应子的途径，并使用验证相互作用 基因突变和生化研究。该提案还将利用正向基因筛选的力量 发现与 nlrc3l 类似的其他基因，这些基因可防止巨噬细胞在稳定状态下激活，其作用相同 或全新的途径。我们设计了一种创新的筛选方法来评估巨噬细胞 使用活细胞报告基因激活激活标记 irg1。最后，该提案将考察影响 使用遗传分析和方法研究斑马鱼巨噬细胞激活的脂质和葡萄糖代谢途径 化学筛选。这项工作将受益于与专家组的合作，将我们的发现扩展到 小鼠和人体模型。总的来说，这些项目为理解提供了重要的基础 先天免疫系统如何受到控制的遗传和代谢基础，并将影响 我实验室的方向远远超出了 MIRA 资助的 5 年。