Exploring metabolic governance of immune cell form and function

项目总结 在这个提案中，我们想要了解对组织驻留至关重要的代谢因素。 巨噬细胞的发育和功能。代谢组学技术的新进展 帮助破译了细胞新陈代谢如何帮助塑造免疫细胞的形态和功能，但 到目前为止，这些进展基本上没有对新陈代谢进行有意义的探索。 组织内的免疫细胞本身。对于像TRMS这样的细胞，它只驻留在 这些组织，在我们能够确定它们是如何 利用细胞代谢来支持其特定于组织的身份和功能。考虑到重要的是 TRMS在维持组织动态平衡和塑造致病环境方面的作用，它是 我们开始探索这些细胞的生物化学。我们提出了一种探索的方法 TRM在体内的代谢。通过使用体内代谢示踪剂和快速分离TRMS 从组织中，我们详细描述了如何使用这种方法来确定必要的代谢程序 对组织内TRM的发育和功能的影响。我们展示了原理证明测试是如何 这一框架揭示了多胺-亚硫氨酸轴作为一种新的代谢节点活跃在 单核细胞和巨噬细胞的分化。我们实施了体外和体内验证研究 在我们的框架中，包括骨髓嵌合体、异型共生和新的小鼠模型 创造，以表明这条途径对于TRMS的发展和维护是必不可少的 横跨多个器官。我们还提出了一种非常新颖的方法来研究trm。 人类的新陈代谢。使用常温灌注机，我们将为人体器官灌流 利用代谢示踪剂在原位了解人类TRM生物学。至关重要的是，我们将 使用这种方法对小鼠TRMS中被识别为重要的通路进行实验测试 在人的环境中。最后，该提案的一个主要焦点是探索体内的TRM新陈代谢 肿瘤。在动态平衡期间，使用上述针对TRMS的类似方法，我们将评估 用明确定义的小鼠肿瘤模型研究TAMs的代谢活性。我们将用我们的创新 人体系统，允许我们试验性地操纵人体器官来探测 人肝细胞癌病变和位于其中的TRMS的代谢。 通过这些跨物种的正交方法，我们希望建立一幅详细的图景 肿瘤巨噬细胞代谢活性，可用于识别新的途径，可以 有针对性地调节肿瘤内的这些细胞，以达到治疗的目的。

PROJECT SUMMARY In this proposal we want to understand the metabolic factors that are essential to tissue-resident macrophage (TRM) development and function. New advances in metabolomics technology have helped to decode how cellular metabolism helps to shape immune cell form and function, but these developments have so far generally stopped short of meaningfully probing the metabolism of immune cells within the tissues themselves. For cells like TRMs, that exclusively reside within the tissues, a full understanding the biology of these cells is missing until we can identify how they utilize cellular metabolism to support their tissue-specific identity and function. Given the important roles TRMs play in maintaining tissue homeostasis and shaping pathogenic environments, it is essential we begin to probe the biochemistry of these cells. We present an approach for exploring TRM metabolism in vivo. Through the use of metabolic tracers in vivo and rapid isolation of TRMs from the tissue, we detail how this approach will be used to identify metabolic programs essential to TRM development and function within the tissues. We show how proof of principle testing of this framework reveals the polyamine-hypusine axis as a novel metabolic node active in differentiating monocytes and TRMs. We implement in vitro and in vivo validation studies laid out in our framework, which include bone marrow chimera, parabiosis and novel mouse model creation, to show that this pathway is essential for the development and maintenance of TRMs across a multitude of organs. We also propose a highly novel approach to studying TRM metabolism in humans. Using normathemic perfusion machines, we will perfuse human organs with metabolic tracers to gain an understanding of human TRM biology in situ. Crucially, we will employ this approach to experimentally test the pathways identified as important in mouse TRMs in a human setting. Finally, a major focus of this proposal is to explore TRM metabolism within tumors. Using similar approaches outlined above for TRMs during homeostasis, we will evaluate the metabolic activity of TAMs using well defined murine tumor models. We will use our innovative human system that allows us to experimentally manipulate human organs to probe the metabolism of human hepatocellular carcinoma lesions and of the TRMs that reside inside them. Through these orthogonal approaches across species, we expect to build up a detailed picture of tumor macrophage metabolic activity that can be used to identify novel pathways that can be targeted to modulate these cells within tumors for therapeutic benefit.