Brain-wide quantitative mapping of microglia activation

小胶质细胞激活的全脑定量图谱

• 批准号: 9908302
• 负责人: Damian G. Wheeler
• 金额: $ 45万
• 依托单位: TRANSLUCENCE BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-23 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9908302
• 关键词: AIDS dementia; Affect; Aftercare; Algorithms; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein; Amyloid deposition; Anti-inflammatory; Antibodies; Atlases; Biological Assay; Biological Markers; Brain; Brain Diseases; Brain region; Cell Density; Cells; Deposition; Detection; Development; Dimensions; Disease model; Dose; Encephalitis; Evaluation; Exhibits; Genes; Genetic Transcription; Goals; Gold; Histologic; Histology; Human; Image; Imagery; Imaging Techniques; Immune; Individual; Infection; Inflammation; Inflammatory; Injury; Label; Light; Lipopolysaccharides; Machine Learning; Maps; Measures; Mediating; Methods; Microglia; Modeling; Molecular; Monitor; Morphology; Multiple Sclerosis; Mus; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neurodevelopmental Disorder; Parkinson Disease; Pathologic; Pathology; Pattern; Phase; Phenotype; Physical shape; Play; Positioning Attribute; Process; Proliferating; Property; Protocols documentation; Publishing; Resolution; Rest; Role; Schizophrenia; Senile Plaques; Site; Speed; Stains; Techniques; Technology; Testing; Therapeutic; Three-Dimensional Imaging; Time; Tissues; Transgenic Mice; Traumatic Brain Injury; abeta deposition; autism spectrum disorder; base; brain cell; chemobrain; cytokine; density; design; experimental study; imaging Segmentation; imaging system; improved; in vitro Assay; in vivo; inflammatory marker; interest; mouse model; nervous system disorder; neuroinflammation; novel; protein biomarkers; response; selective expression; therapeutic candidate; therapeutic evaluation; tool

SUMMARY Recent technological developments such as tissue clearing and light sheet imaging have allowed for the three- dimensional visualization of the entire brain at cellular resolution. Translucence Biosystems has advanced this technology further by developing a) a proprietary Mesoscale Imaging System that allows visualization of an entire mouse brain much faster than prior techniques, b) machine learning-based algorithms that identify individual cells across the entire intact mouse brain and c) routines that rapidly determine cell densities in >1,200 brain regions defined by the annotated Allen Brain Atlas. In the present proposal we will show the power of our technique by visualizing microglia density across the entire mouse brain as a biological marker for neuroinflammation. Microglia are the resident immune cells in the brain. While they play important roles in healthy brain function, they also mediate neuroinflammatory processes that have a significant impact in multiple neurodegenerative diseases such as Alzheimer's, Parkinson's, and multiple sclerosis as well as play possible roles in several neurodevelopmental and neurological disorders (e.g., schizophrenia, autism spectrum disorder, chemo brain). A tool for three-dimensional imaging of neuroinflammation patterns across the whole brain will help advance our understanding of neuroinflammatory processes in neurological diseases and aid in the evaluation of therapeutic approaches. To visualize microglia, we will evaluate multiple strategies, including the CX3CR1-GFP mouse line, which expresses GFP in microglia, Iba1 immunolabeling and other antibody markers of activated microglia. The utility of our approach for monitoring neuroinflammation will be demonstrated in CX3CR1-GFP mice by treating them with the inflammatory agent lipopolysaccharide (LPS). Mice will be injected with three different LPS doses, and then microglia will be visualized and automatically counted across >1,200 brain regions. We will confirm the neuroinflammatory response to LPS by measuring protein markers of inflammation. The applicability of our technique to neurodegenerative disease will be demonstrated by studying a mouse line used as a model of Alzheimer's disease that presents pronounced patterns of neuroinflammation. These experiments are designed to prove that our approach can provide reliable and detailed information describing neuroinflammation and that our results are better in terms of speed, resolution, and richness of information than any other technique available. Once the goals for Phase I are met, we will be positioned to develop our microglia activation assay into a new gold standard for precise and complete histological detection of neuroinflammation. During phase II, we plan to 1) develop machine learning tools to establish morphological criteria that differentiate resting from activated microglia, 2) characterize microglial signatures in various mouse models of diseases with known or suspected neuroinflammation components, and 3) validate the microglial targeting of anti-inflammatory therapeutic candidates.

概括 最近的技术发展，例如组织清理和轻板成像，已允许三 整个大脑在细胞分辨率下的尺寸可视化。半透明生物系统已经提高了这一点 通过开发a）专有的中尺度成像系统进一步技术，该系统允许可视化整个 鼠标大脑比以前的技术快得多，b）识别个体的基于机器学习的算法 整个完整的小鼠脑和c）迅速确定> 1,200大脑中细胞密度的例程 由注释的艾伦脑图定义的区域。 在本提案中，我们将通过可视化整个小胶质细胞密度来显示我们技术的力量 整个小鼠大脑作为神经炎症的生物学标记。小胶质细胞是驻留的免疫细胞 脑。尽管它们在健康的大脑功能中起重要作用，但它们也介导了神经炎症过程 对多种神经退行性疾病产生重大影响，例如阿尔茨海默氏症，帕金森氏症和 多发性硬化症以及在几种神经发育和神经系统疾病中起可能的作用（例如 精神分裂症，自闭症谱系障碍，化学脑大脑）。用于三维成像的工具 整个大脑的神经炎症模式将有助于提高我们对神经炎症的理解 神经系统疾病的过程并有助于评估治疗方法。 为了可视化小胶质细胞，我们将评估多种策略，包括CX3CR1-GFP鼠标系列 在小胶质细胞，IBA1免疫标记和其他活化小胶质细胞的其他抗体标记中表达GFP。实用程序 我们的监测神经炎症的方法将在CX3CR1-GFP小鼠中证明 与炎症剂脂多糖（LPS）。小鼠将以三种不同的LP剂量注入 然后，将对小胶质细胞进行可视化并自动计算在> 1200个大脑区域。我们将确认 通过测量炎症的蛋白质标志物对LPS的神经炎症反应。 我们的技术对神经退行性疾病的适用性将通过研究小鼠来证明 线条用作阿尔茨海默氏病模型，表现出明显的神经炎症模式。这些 实验旨在证明我们的方法可以提供可靠的详细信息描述 神经炎症，我们的结果在速度，解决方案和信息丰富方面比 任何其他可用的技术。 一旦实现了第一阶段的目标，我们将被定位于将我们的小胶质细胞激活分析发展为 用于神经炎症的精确和完整组织学检测的新金标准。在第二阶段，我们 计划到1）开发机器学习工具以建立区分休息与休息的形态标准 活化的小胶质细胞，2）在各种具有已知或已知或 可疑的神经炎症成分，3）验证抗炎的小胶质细胞靶向 治疗候选人。