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: 9381124
• 负责人: Celia E Shiau
• 金额: $ 35.57万
• 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/9381124
• 关键词: Address; Alpha Cell; 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; Human; Immune signaling; Immune system; Inflammation; Inflammatory; Innate Immune System; Knowledge; Lipids; Logic; Macrophage Activation; Malignant Neoplasms; Metabolic; Metabolic Pathway; Modeling; Molecular; Mus; 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; immune activation; in vivo; in vivo imaging; innovation; macrophage; metabolomics; 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.

项目摘要 了解细胞如何关闭并保持其静止状态，从根本上讲与了解细胞如何关闭并保持其静止状态一样重要。 被激活了这项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活化状态。因为失调和无端激活 免疫系统导致了许多与不适当的炎症有关的人类疾病， 调节免疫激活的分子网络通常对于解决这些健康问题至关重要。 挑战这将导致新的知识和技术需要利用的性质， 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， 易于处理的脊椎动物模型系统，斑马鱼，用于精细的遗传操作，高通量筛选， 和体内成像来剖析内在代谢信号和巨噬细胞之间的复杂关系， activation. 该提案包括一系列项目，这些项目共同定义了基本的负面调节剂及其 其功能是控制先天免疫系统，以维持巨噬细胞的正常平衡。 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 通路交叉形成巨噬细胞中的免疫激活，以及在巨噬细胞中发现无效突变。 斑马鱼细胞内NOD样受体（NLR）。这种新型NLR中的基因失活，nlrc 3l，导致 可能由于代谢失调引起的无端巨噬细胞活化。该提案旨在界定 nlrc 3l的分子相互作用网络来理解这个非常重要的机制， 在正常生物学条件下检查巨噬细胞。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nlrc 3l的可能的相互作用物和效应物的途径，并使用 基因突变和生化研究。该提案还将利用遗传筛查的力量， 发现类似于nlrc 3l的其他基因，其在稳定状态下防止巨噬细胞活化， 或全新的路径。我们设计了一种创新的检测方法来评估巨噬细胞 使用活细胞报告子对活化标记IRG 1进行活化。最后，该提案将审查 脂质和葡萄糖代谢途径对斑马鱼巨噬细胞活化的影响， 化学筛选这项工作将受益于与一个专家组的合作， 小鼠和人类模型。综合起来，这些项目为理解 先天免疫系统如何保持控制的遗传和代谢基础，并将影响 我的实验室的方向远远超出了5年的MIRA资金。