Eye Organ Specific Project

眼器官特定项目

• 批准号: 10254372
• 负责人: Kevin L Schey
• 金额: $ 40.26万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10254372
• 关键词: 3-Dimensional; Address; Affect; African American; Age; Age Factors; Age related macular degeneration; American; Analytical Chemistry; Anterior; Antibodies; Aqueous Humor; Architecture; Atlases; Biological Assay; Brain; Cataract; Cell Count; Cell Separation; Cells; Cessation of life; Ciliary Body; Clinical; Collection; Cornea; Cytometry; Data; Data Analyses; Data Science; Defect; Deposition; Detection; Development; Diagnostic Imaging; Disease; Dissociation; Emerging Technologies; Ensure; Epidemiology; Extracellular Matrix; Eye; Eye diseases; Fluorescence; Foundations; Gene Expression Profiling; Genetic; Glaucoma; Glean; Goals; Heterogeneity; Histocytochemistry; Histologic; Hour; Human; Human BioMolecular Atlas Program; Human Resources; Image; Immunofluorescence Immunologic; Investigation; Ions; Kidney; Knowledge; Life; Light; Link; Lipids; Liquid substance; Location; Maps; Mass Spectrum Analysis; Metabolic; Methods; Microscopy; Molecular; Molecular Analysis; Molecular Profiling; Morphology; Multimodal Imaging; Normalcy; Ophthalmology; Optical Coherence Tomography; Optics; Organ; Pancreas; Patients; Photons; Preparation; Property; Proteins; Quality Control; Quality of life; Race; Recording of previous events; Research; Research Personnel; Resolution; Retina; Retinal Photoreceptors; Sampling; Sensory; Signal Transduction; Specificity; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Stains; Standardization; Structure; Structure of retinal pigment epithelium; Sunlight; Surface; System; Technology; Time; Tissue Banks; Tissues; Trabecular meshwork structure; Vision; Visual; Visual impairment; Vitreous humor; Work; absorption; base; biobank; cell type; clinical diagnostics; data quality; demographics; design; experimental study; high intraocular pressure; high risk; human imaging; imaging modality; indexing; insight; instrumentation; lens; macula; molecular imaging; multimodality; novel therapeutics; professor; programs; reconstruction; sex; spatial relationship; three dimensional structure

PROJECT SUMMARY – Eye Organ Specific Project. Comprehensive 3-dimensional biomolecular analysis within tissues requires the coordination of data from many analytical technologies. We will develop a multimodal strategy to obtain spatially specific ‘omics’ information directly from human whole eye tissue. This work will leverage the unique instrumentation capabilities and expertise of the Mass Spectrometry Research Center (MSRC) in which Professors Spraggins, Caprioli, Schey, Gutierrez and Van de Plas are key personnel. Building from previous work developing atlases of human kidney tissue, we will integrate ultra-high speed, high spatial resolution (<10μm) MALDI timsTOF IMS with high mass resolution (>40,000 Resolving Power, <2 ppm mass accuracy) for molecular imaging of human eyes with an unprecedented combination of spatial fidelity and molecular specificity. The high-throughput capabilities of MALDI timsTOF IMS will be critical for generating high- spatial resolution 3-D molecular images from statistically relevant numbers of normal pancreas samples. MALDI IMS will provide the specificity and mass accuracy necessary to link ion images to orthogonal LC-based fragmentation experiments for molecular identification. Fragmentation will be performed using spatially specific surface sampling approaches to produce LC-MS/MS data from tissues in a manner that is compatible with IMS and capable of being integrated into HuBMAP tissue atlases. To maximize information gleaned from imaging experiments and lay the groundwork for data-driven image fusion and 3-D reconstruction outlined in the Data Analysis Core Research Strategy, we will also develop methods for collecting various microscopy-based image modalities (e.g. autofluorescence and stained microscopy) and pancreas specific, highly multiplexed immunofluorescence using CODEX. Whole eye atlases will be constructed by combining our multimodal IMS/Microscopy pipelines with ex vivo 3-D optical coherence tomography (OCT) imaging. Through the use of our multimodal pipeline, we propose to map spatial relationships of eye cell types from all regions of the eye within healthy tissue across a multitude of demographics. Aim 1: To create an eye- specific biospecimen collection and management plan. Aim 2: To establish a pre-analytical pipeline for standardizing sample preparation, determining tissue normalcy, and providing quality control metrics. Aim 3: To develop a multimodal characterization pipeline for eye-specific 3-D molecular imaging. Aim 4: To scale and standardize eye-specific 3-D molecular imaging. To accomplish these aims, we have assembled a highly interactive and established team of investigators consisting of complementary expertise in ophthalmology, analytical chemistry, and data science.

项目摘要 - 眼器特定项目。综合3维生物分子分析 在组织中需要许多分析技术的数据协调。我们将开发多模式 直接从人类全眼组织中获取经常特定的“ OMICS”信息的策略。这项工作将 利用质谱研究中心的独特仪器功能和专业知识 （MSRC）其中Spraggins，Caprioli，Schey，Gutierrez和Van de Plas教授是关键人员。建筑 从以前开发人类肾脏组织的地图的工作中，我们将整合超高速度，高空间 分辨率（<10μm）MALDI TIMSTOF IMS具有高质量分辨率（> 40,000分辨率，<2 ppm质量 精度），用于人眼的分子成像，其空间保真度和 分子特异性。 MALDI TIMSTOF IMS的高通量功能对于产生高 来自统计相关数量的正常胰腺样品的空间分辨率3-D分子图像。马尔迪 IMS将提供将离子图像与基于正交LC连接的特异性和质量精度 分子鉴定的碎片实验。碎片将使用空间特定的 表面采样方法以与IMS兼容的方式从TIMS产生LC-MS/MS数据 并能够被整合到Hubmap组织地图集中。从成像中最大化信息 实验并为数据驱动的图像融合和数据中概述的3-D重建奠定了基础 分析核心研究策略，我们还将开发用于收集各种基于显微镜图像的方法 模态（例如自动荧光和染色显微镜）和胰腺特异性，高度多路复用 使用法典免疫荧光。将通过组合我们的多模式来构建全眼图 IMS/显微镜管道，带有体内3-D光学相干断层扫描（OCT）成像。 通过使用我们的多模式管道，我们建议映射眼部电池类型的空间关系 来自健康组织内的所有区域，跨越了许多人口统计学。目标1：创建眼睛 - 特定的生物循环收集和管理计划。目标2：建立用于分析前管道 目标3： 为眼睛特异性3-D分子成像开发多模式表征管道。目标4：扩展和 标准化眼睛特异性的3-D分子成像。为了实现这些目标，我们组装了一个高度的 互动和成熟的研究人员团队，包括眼科专业知识， 分析化学和数据科学。