Genetic and metabolic regulation of macrophage activation at steady state

项目摘要 炎症对于组织修复和清除有害病原体和分子至关重要，但如果激活 先天性免疫系统不适当地触发自身炎性病症。宏观调控发挥关键作用 在炎症和对人类健康有巨大的影响，积极和消极的，但许多问题 关于这些细胞仍然没有答案。它们是异质的，并且采用一系列激活状态，两者都是 作为不同的组织驻留巨噬细胞和在循环中作为单核细胞。这些激活状态的驱动因素 仍然难以捉摸代谢起着重要作用，巨噬细胞在促凋亡过程中发生代谢变化， 炎症激活，包括从氧化磷酸化转换为糖酵解。但无论 代谢或其他细胞内过程单独可以决定激活状态仍然不清楚， 了解许多与不适当炎症相关的人类疾病。目前拟议的 一项研究计划解决了这个问题，并通过分析这些问题大大扩展了我们以前的工作。 在不同的器官系统（包括肝脏和肠道）的过程。 我们正在利用斑马鱼的独特优势，提供精致的遗传操作，高通量 筛选和体内成像，以剖析细胞内过程之间的复杂关系， 巨噬细胞活化在NOD样受体nlrc 3l中发现失活突变， 不适当的巨噬细胞激活和小胶质细胞的损失使我们在斑马鱼中建立了一种新的遗传筛选， 产生了几种新的巨噬细胞突变，我们将进行研究。这些新的突变导致不适当的 巨噬细胞活化、破坏小胶质细胞发育或两者。我们正在调查nlrc 3l和 相关基因以及我们从筛选中发现的新基因通过改变巨噬细胞的活性来调节巨噬细胞的活化。 代谢和其他细胞内过程。我们在多个层面采取综合方法， 差异转录组学、蛋白质组学和代谢组学，高功率体内单细胞和全身 成像，基于CRISPR的基因工程，蛋白质-蛋白质相互作用和跨物种 实验来解决这些问题。此外，我们以前的结果提出了关键的新问题， 关于激活状态是如何被调节以确保正常的组织巨噬细胞发育，以及是否 存在控制跨多个器官的巨噬细胞活化的常见机制。我们的研究计划将 使用一组不同的组织巨噬细胞（小胶质细胞、肠巨噬细胞）， 巨噬细胞和枯否细胞）。这些研究的目的是提供必要的见解，巨噬细胞 分化，可塑性和疾病相关背景下的潜力，并解决看似不同的 功能（发育和激活调节）编码在控制 巨噬细胞总之，破译调节免疫激活的分子网络对于 解决与不适当炎症相关的健康挑战，包括糖尿病、肥胖、癌症， 和自身免疫性疾病。我们的研究将带来新的知识和技术， 用于疾病预防和治疗的巨噬细胞的特性。

Project Summary Inflammation is essential for tissue repair and removal of harmful pathogens and molecules , but if activated inappropriately, the innate immune system triggers autoinflammatory conditions. Macrophages play a key role in inflammation and have tremendous impact on human health, both positive and negative, yet many questions about these cells remain unanswered. They are heterogeneous and adopt a spectrum of activation states, both as distinct tissue-resident macrophages and in circulation as monocytes. The drivers of these activation states remain elusive. Metabolism plays a role, with macrophages undergoing metabolic changes during pro- inflammatory activation, including switching from oxidative phosphorylation to glycolysis. However, whether metabolism or other intracellular processes alone can dictate activation states remains unclear and is critical to understanding a host of human diseases associated with inappropriate inflammation. The current proposed research program addresses this question and significantly expands our previous work by analyzing these processes in different organ systems (including the liver and the intestine). We are leveraging the unique advantages zebrafish provide for exquisite genetic manipulations, high throughput screening, and in vivo imaging to dissect the complex relationship between intracellular processes and macrophage activation. The discovery of an inactivating mutation in a NOD-like receptor, nlrc3l, that causes inappropriate macrophage activation and loss of microglia led us to create a new genetic screen in zebrafish that yielded several novel macrophage mutations that we will investigate. These new mutations cause inappropriate macrophage activation, disrupt microglia development or both. We are investigating whether nlrc3l and associated genes as well as new genes we identify from the screen regulate macrophage activation by altering metabolic and other intracellular processes. We are taking an integrated approach at multiple levels—using differential transcriptomics, proteomics, and metabolomics, high power in vivo single cellular and whole-body imaging, CRISPR based genetic engineering, protein-protein interactions, and cross-species experimentations—to address these questions. Furthermore, our previous results raise critical new questions regarding how activation states are regulated to ensure normal tissue macrophage development, and whether common mechanisms exist to control macrophage activation across multiple organs. Our research program will expand to address these key areas in vivo using a diverse set of tissue macrophages (microglia, intestinal macrophages, and Kupffer cells). These studies aim to provide necessary insights into macrophage differentiation, plasticity, and potential in a disease-relevant context, and address how seemingly disparate functions (development and activation regulation) are encoded in the molecular program governing macrophages. In summary, deciphering the molecular networks regulating immune activation is critical for addressing health challenges associated with inappropriate inflammation, including diabetes, obesity, cancer, and autoimmune diseases. Our research will lead to new knowledge and technologies needed to harness the properties of macrophages for disease prevention and treatment.