Metabolic regulation of macrophage-dependent wound healing in vivo

项目摘要 巨噬细胞是炎症性疾病进展的基础，包括动脉粥样硬化、自身免疫性 疾病、慢性创伤和癌症。活化巨噬细胞的代谢重组改变其功能， 成为一种有吸引力的治疗策略，但缺乏体内证据支持其治疗潜力。 本文提出的研究将解决巨噬细胞效应功能的代谢调节， 通过代谢辅酶NAD（P）H和FAD的荧光寿命成像的生理相关设置， 使用斑马鱼作为炎症和伤口愈合的体内模型。重要的是，这种非侵入性成像 模态测量细胞内代谢状态同时将巨噬细胞维持在它们的天然微环境中， 不同于传统的方法。本研究将监测不同浓度下巨噬细胞代谢谱的变化， 随着时间的推移，伤口模型，并测试代谢调节剂，Irg 1，Stat 3和 线粒体ROS，在巨噬细胞依赖性伤口愈合。还将进行代谢组学分析 使用受伤的组织，以获得进一步的机制洞察巨噬细胞的代谢重编程， vivo.总的来说，这项研究将开发基于成像的工具，以真实的时间探测时间和空间 在活体动物中免疫细胞功能的代谢调节，可以为新疗法的开发提供信息， 减轻巨噬细胞介导的炎症。 在此职业发展奖申请的建议研究将在小学进行 安娜Huttenlocher博士的指导和梅丽莎Skala博士在K99阶段的共同指导， 奖威斯康星大学麦迪逊分校的Huttenlocher实验室是这些研究的理想环境， 在炎症和伤口愈合中的白细胞生物学以及斑马鱼活体成像策略方面的专业知识 胚胎Skala博士是开发代谢辅酶的自发荧光寿命成像及其 应用.我将从Huttenlocher博士和Skala博士的指导中受益匪浅，并将在 两个导师的专业知识，以阐明巨噬细胞的代谢重编程机制， 复杂的体内环境。我致力于作为一个学术独立调查员的职业生涯， 研究白细胞生物学的机构;特别是研究白细胞效应物的代谢调节 在炎症性疾病中起作用。除了已经很好的资源，我是由我的 导师，威斯康星大学麦迪逊分校提供了丰富的其他科学和职业发展机会，以支持我的 学术成长此外，我积极参与莫格里奇研究所的代谢社区， 研究，位于威斯康星大学麦迪逊分校校园，以进一步加强我在免疫代谢的训练。采取 一起，我的导师和威斯康星大学麦迪逊分校提供了一个理想的环境，以充分支持我的科学追求， 确保我实现长期职业目标。

PROJECT SUMMARY Macrophages underlie the progression of inflammatory diseases, including atherosclerosis, autoimmune diseases, chronic wounds and cancer. Metabolic rewiring of activated macrophages to alter their function has become an attractive therapeutic strategy, however in vivo evidence is lacking to support its therapeutic potential. The research proposed here will address the metabolic regulation of macrophage effector function in physiologically relevant settings by fluorescence lifetime imaging of metabolic coenzymes NAD(P)H and FAD, using zebrafish as an in vivo model of inflammation and wound healing. Importantly, this non-invasive imaging modality measures intracellular metabolic state while maintaining macrophages in their native microenvironment, unlike traditional approaches. This study will monitor changes in the metabolic profile of macrophages at various wound models over time, and test the functional requirement of metabolic regulators, Irg1, Stat3 and mitochondrial ROS, in macrophage-dependent wound healing. Metabolomics analysis will also be performed using wounded tissue to gain further mechanistic insight into the metabolic reprogramming of macrophages in vivo. Collectively, this study will develop imaging-based tools to probe in real time the temporal and spatial metabolic regulation of immune cell functions in live animals that can inform development of new therapies to mitigate macrophage-mediated inflammation. The proposed study in this career development award application will be conducted under the primary mentorship of Dr. Anna Huttenlocher and the co-mentorship of Dr. Melissa Skala during the K99 phase of the award. The Huttenlocher lab at UW-Madison is an ideal environment for these studies due to the leading expertise in leukocyte biology in inflammation and wound healing, as well as live imaging strategies in zebrafish embryos. Dr. Skala is a leader in developing autofluorescence lifetime imaging of metabolic coenzymes and its applications. I will benefit immensely from the mentorship of Drs. Huttenlocher and Skala, and will bridge the expertise of both mentors toward elucidating the mechanisms of metabolic reprogramming of macrophages in complex in vivo environments. I am committed to a career as an independent investigator at an academic institution studying leukocyte biology; in particular, studying the metabolic regulation of leukocyte effector functions in context of inflammatory disorders. In addition to the already excellent resources I am afforded by my mentors, UW-Madison provides a wealth of other scientific and career development opportunities to support my academic growth. Moreover, I am actively involved with the metabolic community at the Morgridge Institute for Research, located on UW-Madison campus, to further enhance my training in immunometabolism. Taken together, my mentors and UW-Madison provide an ideal environment to fully support my scientific pursuits and ensure that I achieve my long term career goals.