Eye Organ Specific Project

眼器官特定项目

基本信息

批准号：
10704490
负责人：
Kevin L Schey
金额：
$ 82.26万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
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/10704490
关键词：
3-Dimensional Address Affect African American population 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 Dimensions Disease Dissociation Emerging Technologies 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 Tissue atlas Tissues Trabecular meshwork structure Vision Visual Visual impairment Vitreous humor Work absorption biobank cell type clinical diagnostics data quality demographics design experimental study fortification high intraocular pressure high risk human imaging imaging facilities imaging modality indexing insight instrumentation lens macula mass spectrometric imaging molecular imaging multimodality novel therapeutics professor programs reconstruction sex spatial relationship three dimensional structure tissue mapping transmission process

项目摘要

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 – 眼器官特定项目。综合 3 维生物分子分析。组织内需要协调来自许多分析技术的数据，我们将开发一种多模式。这项工作将直接从人类整个组织眼睛中获取空间特定的“组学”信息。利用质谱研究中心独特的仪器能力和专业知识 (MSRC)，其中 Spraggins、Caprioli、Schey、Gutierrez 和 Van de Plas 教授是大楼的关键人员。根据之前开发的人体肾脏组织工作图谱，我们将整合超高速、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），用于人眼分子成像，前所未有地结合了空间保真度和 MALDI timsTOF IMS 的高通量能力对于产生高通量至关重要。来自技术相关数量的正常胰腺样本的空间分辨率 3D 分子图像。 IMS 将提供将离子图像链接到基于正交 LC 所需的特异性和质量准确度将使用空间特异性进行碎片化实验。表面采样方法，以与 IMS 兼容的方式从组织中生成 LC-MS/MS 数据并能够集成到 HuBMAP 组织图谱中，以最大化从成像中收集的信息。实验并为数据驱动的图像融合和 3D 重建奠定基础分析核心研究策略，我们还将开发收集各种基于显微镜的图像的方法模式（例如高度自发荧光和染色显微镜）和胰腺特异性、多重检测使用 CODEX 的免疫荧光将通过结合我们的多模式构建全眼图谱。具有离体 3D 光学相干断层扫描 (OCT) 成像的 IMS/显微镜管道。通过使用我们的多模式管道，我们建议绘制眼细胞类型的空间关系来自众多人群健康组织内眼睛的所有区域目标 1：创造一只眼睛具体的生物样本采集和管理计划目标 2：建立预分析管道。标准化样品制备、确定组织正常情况并提供质量控制指标。开发用于眼睛特异性 3D 分子成像的多模态表征流程。目标 4：规模化和规模化。标准化眼睛特异性 3D 分子成像为了实现这些目标，我们组装了一个高度互动且已建立的研究团队由眼科领域互补的专业知识组成，分析化学和数据科学。