Neutrophil Microscopy and Quantitative Imaging Core B

中性粒细胞显微镜和定量成像 Core B

• 批准号: 10651782
• 负责人: Bill William Kiosses
• 金额: $ 29.89万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-16 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10651782
• 关键词: 3-Dimensional; Actins; Adhesions; Apolipoprotein E; Apoptosis; Apoptotic; Arterial Fatty Streak; Autoimmunity; Azurophilic Granule; Bacteria; Biological Assay; Biology; Cardiovascular Diseases; Cell Death; Cell membrane; Cells; Cellular biology; Cessation of life; Collaborations; Coronary Arteriosclerosis; Cytoplasmic Granules; Cytoskeleton; Dedications; Development; Dimensions; Docking; Electron Microscopy; Ensure; Event; Exocytosis; Fibrosis; Fluorescence Microscopy; Gelatinases; Generations; Goals; High Fat Diet; Human; Hyperlipidemia; Image; Inflammasome; Inflammation; Inflammatory; Light; Longevity; MMP9 gene; Mediating; Microscopy; Mitochondria; Modeling; Molecular; Mouse Strains; Multivesicular Body; Mus; Neutrophil Activation; Organelles; Pathway interactions; Patients; Performance; Phagocytosis; Play; Process; Production; Productivity; Program Research Project Grants; Proteins; Quality Control; Quantitative Microscopy; Reactive Oxygen Species; Research; Resolution; Resources; Role; Sampling; Stimulus; Technology; Time; Vasculitis; Vesicle; Visualization; Work; analytical tool; autoinflammatory diseases; cost; data sharing; design; extracellular; fungus; in vivo; innovation; intravital microscopy; light microscopy; live cell microscopy; microscopic imaging; migration; mouse model; nanometer; neutrophil; novel strategies; progenitor; quantitative imaging; single molecule; superresolution microscopy; trafficking; ultra high resolution; western diet

SUMMARY Neutrophils are the first line of defense against bacteria and fungi and induce inflammation. The study of the molecular mechanisms mediating neutrophil activation is therefore of fundamental importance. Visualizing these mechanisms requires unique, state-of-the-art technologies and approaches. To achieve this, Core B is designed to coordinate, design, develop and execute new technical approaches applied to the understanding of neutrophil biology in inflammation. Core B will work with all three Projects and will provide relevant imaging and microscopy assays in mice and human samples. The core utilizes available cutting-edge technology to implement new experimental approaches and to help elucidate the intricate cell biology processes proposed in research plans of Projects 1 (Dr. Hedrick), Project 2 (Dr. Catz) and Project 3 (Dr. Hoffman). To this end, we will have the dedicated effort of a Technology Core director (Dr. Kiosses) who will allocate specific time and efforts to support the goals of the PPG. Core B optimizes the workflows of neutrophil imaging, image quantification and data sharing for all projects. It provides stringent quality controls and ensures uniformity of performance of multiple key technologies applied to the study of neutrophils and precursors in the setting of coronary artery disease. Core B will implement unique live-cell approaches to study neutrophil vesicular trafficking, cytoskeleton remodeling and cell death pathways. Core B is technically and conceptually innovative because it utilizes, neutrophil-dedicated, novel approaches that include super-resolution microscopy and quantitative analyses of neutrophil dynamics to elucidate neutrophil functions and to elucidate their role in inflammation. In relationship with Project 1, Core B will identify neutrophils and their precursors in atherosclerotic plaques during NETosis and pyroptosis in conditions of inflammasome activation at various stages of plaque development in high-fat diet, both ex-vivo and in vivo using the ApoE-/- Western diet model of CAD mice. In association with Project 2, Core B will utilize single-molecule super-resolution microscopy and Total Internal Reflection Fluorescence Microscopy to study molecular mechanisms regulating vesicular trafficking and actin remodeling in neutrophil precursors. Core B will also utilize Correlated-Light Electron Microscopy (CLEM) to characterize the newly identified precursor-specific granules (PSGs). In collaboration with Project 3, Core B will examine three-dimensional localization and spatial changes in the mitochondrial network as the cells undergo apoptotic or necroptotic cell death in both neutrophils and their precursors from mouse models and human neutrophils from patients with CAD. In conclusion, Core B is uniquely designed to use the valuable technology available in La Jolla to the fullest extent possible to maximize neutrophil-dedicate research and productivity associated with the PPG.

概括 中性粒细胞是针对细菌和真菌的第一道防线，并诱发炎症。研究 因此，介导嗜中性粒细胞活化的分子机制至关重要。可视化这些 机制需要独特的最先进的技术和方法。为了实现这一目标，核心B的设计 协调，设计，开发和执行用于理解中性粒细胞的新技术方法 炎症中的生物学。核心B将与所有三个项目一起工作，并提供相关的成像和显微镜检查 小鼠和人类样品的测定。核心利用可用的尖端技术来实施新的 实验方法并帮助阐明研究计划中提出的复杂的细胞生物学过程 项目1（Hedrick博士），项目2（Catz博士）和项目3（Hoffman博士）。为此，我们将拥有 技术核心总监（Kiosses博士）的奉献精力，他们将分配特定的时间和支持 PPG的目标。 Core B优化中性粒细胞成像，图像定量和数据的工作流程 共享所有项目。它提供了严格的质量控制，并确保了多重性能的均匀性 关键技术应用于冠状动脉疾病的中性粒细胞和前体的研究。 核心B将实施独特的活细胞方法来研究中性粒细胞囊肿，细胞骨架 重塑和细胞死亡途径。核心在技术上和概念上都是创新的，因为它利用了， 中性粒细胞的新型方法包括超分辨率显微镜和定量分析 中性粒细胞动力学以阐明中性粒细胞功能并阐明其在炎症中的作用。在关系中 使用项目1，核心B将在Netosis期间识别动脉粥样硬化斑块中的中性粒细胞及其前体 在高脂饮食中斑块发育的各个阶段炎性体激活条件下的凋亡， 前体和体内都使用了CAD小鼠的apoE - / - 西方饮食模型。与项目2联合核心 B将利用单分子超分辨率显微镜和总内反射荧光显微镜 研究调节中性粒细胞前体中囊泡运输和肌动蛋白重塑的分子机制。 Core B还将利用相关的光学显微镜（CLEM）来表征新鉴定的 前体特异性颗粒（PSGS）。与项目3合作，Core B将检查三维 线粒体网络中的定位和空间变化，因为细胞经历凋亡或坏死细胞 嗜中性粒细胞的死亡及其前体来自小鼠模型和人类嗜中性粒细胞的死亡 卡德。总之，核心B的设计独特地使用La Jolla中可用的有价值的技术 最大化与PPG相关的中性粒细胞研究和生产力的可能程度。