Intravital Imaging and Physiology Core

活体成像和生理学核心

• 批准号: 10706508
• 负责人: Yoshikazu Tsukasaki
• 金额: $ 32.31万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10706508
• 关键词: Algorithms; Animals; Biogenesis; Blood Cells; Blood Vessels; Cell Surface Receptors; Cell physiology; Cells; Endoplasmic Reticulum; Endothelial Cells; Endothelium; Image; Immune; In Situ; Individual; Inflammatory; Injury; Innate Immune System; Lung; Macrophage; Measures; Methods; Mitochondria; Molecular; Monitor; Morphologic artifacts; Motion; Mus; Myeloid Cells; Outcome; Pathway interactions; Permeability; Phagocytes; Phagocytosis; Phase; Phenotype; Phosphorylation; Physiological; Physiology; Pseudomonas aeruginosa; Regulation; Resolution; Ring Finger Domain; Role; Signal Transduction; Sphingosine-1-Phosphate Receptor; Vascular Endothelium; Visual; cadherin 5; imaging approach; imaging study; in vivo; in vivo imaging; injury and repair; innate immune function; interest; intravital imaging; lung imaging; lung injury; lung repair; migration; monocyte; mouse model; neutrophil; novel; novel strategies; programs; respiratory; response; spatiotemporal; success; trafficking; two-photon; ubiquitin-protein ligase; vascular injury

ABSTRACT The focus of this Program Project is to investigate the molecular and signaling mechanisms underlying the immune regulatory role of the pulmonary endothelial niche during inflammatory lung injury and resolution of injury. The Intravital Imaging and Physiology Core (Core D) is essential to precisely define how dynamic changes in the pulmonary endothelium during injury and repair programs the immune cells to either promote or resolve inflammatory lung injury. Each Project makes clear that spatio-temporal signaling regulated by endothelial cell-surface receptors and mitochondria is fundamentally important in regulating the endothelial immune niche and defining the outcome of lung injury and resolution. Thus, all Projects, using endothelial- specific genetically altered living mouse models, will address how signals emanating from basal and activated endothelial cells alter neutrophil/monocyte/macrophage fate in response to lung injury and contribute to resolution. Respiratory motion in the live animal represents a major obstacle for obtaining meaningful results from in vivo imaging studies of the normally respiring lung. We have developed a novel imaging approach to monitor pulmonary microvessels and immune cell trafficking during injury and repair. Thus, a major Core D function will be to provide advanced two-photon intravital lung imaging expertise to reliably quantify the activation of endothelium, to measure lung vascular leak, and to identify the role of activated pulmonary vascular niche in regulating transendothelial migration of myeloid cells in the living mouse lung. Each Project in conjunction with Core D will examine the functional effects of their pathways of interest during inflammatory lung injury and repair phases. Imaging analyses will include the assessment of transmigration of phagocytic cells and in situ phagocytic and efferocytotic activities of immune cells as programmed by endoplasmic reticulum-localized S1PR1 (sphingosine-1-phosphate receptor-1) in endothelial cells in Project 1; CHFR (an E3 ligase, checkpoint with Forkhead and ring finger domains)-induced degradation of VE-cadherin in endothelial cells in Project 2; and activation of endothelial cell mitophagy and mitochondrial biogenesis in Project 3. By providing visual in vivo examination of endothelial activity and its impact on myeloid cell functions and physiological assessments during inflammatory injury and repair, Core D will be essential for the Program’s success.

摘要 本项目的重点是研究细胞凋亡的分子和信号机制。 肺内皮细胞龛在炎症性肺损伤中的免疫调节作用及免疫调节机制的研究 损伤活体成像和生理学核心（核心D）对于精确定义动态 损伤和修复过程中肺内皮细胞的变化使免疫细胞要么促进 或解决炎症性肺损伤。每个项目都明确指出，时空信号调节由 内皮细胞表面受体和线粒体在调节内皮细胞增殖中具有根本重要性。 免疫生态位和定义肺损伤和解决的结果。因此，所有项目，使用内皮细胞- 特定的基因改变的活小鼠模型，将解决如何从基础和激活的信号发出， 内皮细胞响应肺损伤改变中性粒细胞/单核细胞/巨噬细胞的命运， 分辨率活体动物中的呼吸运动代表了获得有意义的呼吸运动的主要障碍。 结果来自正常呼吸肺的体内成像研究。我们开发了一种新的成像技术 监测损伤和修复期间肺微血管和免疫细胞运输的方法。因此 核心D的主要功能是提供先进的双光子活体肺成像专业知识， 定量内皮细胞的活化，测量肺血管渗漏，并确定活化的内皮细胞的作用。 肺血管生态位在调节活鼠肺髓样细胞跨内皮迁移中的作用。 每个项目与核心D一起将检查其感兴趣的途径的功能效果 在炎性肺损伤和修复阶段。影像学分析将包括以下评估 吞噬细胞的迁移和免疫细胞的原位吞噬和吞噬活性， 内皮细胞中由内质网定位的S1 PR 1（鞘氨醇-1-磷酸受体-1）编程 项目1中的细胞; CHFR（一种E3连接酶，具有叉头和环指结构域的检查点）诱导 在项目2中，内皮细胞中VE-钙粘蛋白的降解;以及内皮细胞线粒体自噬和 项目3中的线粒体生物合成。通过提供内皮活性的视觉体内检查及其 炎症损伤和修复对骨髓细胞功能和生理学评估的影响， D将是该计划成功的关键。