Metabolic regulation of macrophage-dependent wound healing in vivo

项目总结 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 疾病、慢性伤口和癌症。激活的巨噬细胞的代谢重联以改变其功能 成为一种有吸引力的治疗策略，然而体内证据缺乏支持其治疗潜力。 这项研究将解决巨噬细胞效应器功能的代谢调节。 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， 使用斑马鱼作为体内炎症和伤口愈合的模型。重要的是，这种非侵入性成像 通道测量细胞内的代谢状态，同时保持巨噬细胞在其自然微环境中， 与传统方法不同。这项研究将监测不同时期巨噬细胞代谢的变化。 随着时间的推移建立创伤模型，并测试代谢调节因子IRG1、STAT3和 线粒体ROS，在巨噬细胞依赖的伤口愈合中。还将进行代谢组学分析 利用损伤组织进一步了解巨噬细胞代谢重编程的机制 活着。总的来说，这项研究将开发基于成像的工具来实时探测时间和空间 活体动物免疫细胞功能的代谢调节，可为开发新的治疗方法提供信息 减轻巨噬细胞介导的炎症。 本次职业发展奖申请中的拟议研究将在小学 在K99阶段对Anna Huttenlocher博士和Melissa Skala博士的共同指导 获奖。威斯康星大学麦迪逊分校的Huttenlocher实验室是进行这些研究的理想环境，因为 在炎症和伤口愈合方面的白细胞生物学以及斑马鱼的活体成像策略方面的专业知识 胚胎。斯卡拉博士是新陈代谢辅酶的自体荧光寿命成像研究领域的领军人物。 申请。我将从Huttenlocher博士和Skala博士的指导中受益匪浅，并将在 两位导师在阐明巨噬细胞代谢重编程机制方面的专业知识 复杂的活体环境。我致力于成为一名学术机构的独立调查员 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 在炎症性疾病背景下的功能。除了已经很好的资源之外，我的 导师们，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 学业上的成长。此外，我还积极参与了莫里奇研究所的新陈代谢研究 位于威斯康星大学麦迪逊分校的研究，进一步加强了我在免疫代谢方面的培训。已被占用 我的导师和密歇根大学麦迪逊分校共同提供了一个理想的环境，完全支持我的科学追求和 确保我实现我的长期职业目标。

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.