A 3D Tissue Map of the Human Lymphatic System

人体淋巴系统 3D 组织图

基本信息

批准号：
10265654
负责人：
MARK A. ATKINSON
金额：
$ 43.37万
依托单位：
UNIVERSITY OF FLORIDA
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：
2020-09-08 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10265654
关键词：
3-Dimensional Address Advisory Committees Aging Algorithms Anatomy Antigen-Presenting Cells Architecture Atlases Autoimmune B-Lymphocytes Back Biological Assay Blood Blood Circulation Body Fluids Body fat Cardiovascular system Cell Separation Cells Chromium Chyle Clinical Data Collaborations Communities Confocal Microscopy Consultations Coupled Cytometry Data Data Analyses Data Files Dendritic Cells Diabetes Mellitus Discontinuous Capillary Disease Dissection Drainage procedure Erythrocytes Excision Exclusion Criteria Florida Flow Cytometry Fluorescence Microscopy Fluorescence-Activated Cell Sorting Fluorescent in Situ Hybridization Formalin Fostering Generations Goals Housing Human Human BioMolecular Atlas Program Human body IgG2 Image Immune Immune response Immune system Immunity Immunoglobulin M Immunohistochemistry Immunologic Surveillance In Situ Hybridization Infection Invaded Knowledge Laboratories Leukocytes Life Light Link Lobular Longevity Lymph Lymphatic System Lymphatic function Lymphocyte Magnetic Resonance Imaging Malignant Neoplasms Malpighian corpuscles Maps Messenger RNA Microscopy Morphology Normalcy Online Systems Optics Organ Organ Donor Organ Procurements Organism Pancreas Paraffin Embedding Pathologist Patients Periarteriolar Lymphoid Sheath Peripheral Physiological Positioning Attribute Pregnancy Procedures Production Proliferating Protocols documentation Publishing Quality Control Recording of previous events Reproducibility Research Research Personnel Resolution Reticular Cell Rodent Role Series Site Specimen Spleen Splenic Red Pulp Stains Stretching Structure Structure of germinal center of lymph node Structure of thymic cortex System T-Cell Development T-Lymphocyte Temperature Thymus Gland Tissues Transplantation United States National Institutes of Health Universities Vascular blood supply Veins Visualization Work absorption arteriole autoinflammatory base blood filter blood filtration cDNA Library cancer cell data sharing daughter cell experience fatty acid transport file format imaging modality improved insight lymph nodes lymphatic vessel lymphoid organ mRNA Expression macrophage magnetic field member millimeter multidisciplinary nanometer new technology novel operation optical imaging particle programs protein expression reconstruction scale up secondary lymphoid organ single molecule single-cell RNA sequencing spectroscopic imaging synergism tissue processing trafficking transcriptome sequencing web site

3-Dimensional Address Advisory Committees Aging Algorithms Anatomy Antigen-Presenting Cells Architecture Atlases Autoimmune B-Lymphocytes Back Biological Assay Blood Blood Circulation Body Fluids Body fat Cardiovascular system Cell Separation Cells Chromium Chyle Clinical Data Collaborations Communities Confocal Microscopy Consultations Coupled Cytometry Data Data Analyses Data Files Dendritic Cells Diabetes Mellitus Discontinuous Capillary Disease Dissection Drainage procedure Erythrocytes Excision Exclusion Criteria Florida Flow Cytometry Fluorescence Microscopy Fluorescence-Activated Cell Sorting Fluorescent in Situ Hybridization Formalin Fostering Generations Goals Housing Human Human BioMolecular Atlas Program Human body IgG2 Image Immune Immune response Immune system Immunity Immunoglobulin M Immunohistochemistry Immunologic Surveillance In Situ Hybridization Infection Invaded Knowledge Laboratories Leukocytes Life Light Link Lobular Longevity Lymph Lymphatic System Lymphatic function Lymphocyte Magnetic Resonance Imaging Malignant Neoplasms Malpighian corpuscles Maps Messenger RNA Microscopy Morphology Normalcy Online Systems Optics Organ Organ Donor Organ Procurements Organism Pancreas Paraffin Embedding Pathologist Patients Periarteriolar Lymphoid Sheath Peripheral Physiological Positioning Attribute Pregnancy Procedures Production Proliferating Protocols documentation Publishing Quality Control Recording of previous events Reproducibility Research Research Personnel Resolution Reticular Cell Rodent Role Series Site Specimen Spleen Splenic Red Pulp Stains Stretching Structure Structure of germinal center of lymph node Structure of thymic cortex System T-Cell Development T-Lymphocyte Temperature Thymus Gland Tissues Transplantation United States National Institutes of Health Universities Vascular blood supply Veins Visualization Work absorption arteriole autoinflammatory base blood filter blood filtration cDNA Library cancer cell data sharing daughter cell experience fatty acid transport file format imaging modality improved insight lymph nodes lymphatic vessel lymphoid organ mRNA Expression macrophage magnetic field member millimeter multidisciplinary nanometer new technology novel operation optical imaging particle programs protein expression reconstruction scale up secondary lymphoid organ single molecule single-cell RNA sequencing spectroscopic imaging synergism tissue processing trafficking transcriptome sequencing web site

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项目摘要

The lymphatic system serves five primary roles: 1) removal of excess body fluids; 2) absorption and transport of fatty acids/chyle to the circulatory system; 3) blood filtration; 4) mounting the primary defense against infections and cancer through immune cell production and activation; and 5) generation and activation of regulatory immune cells that protect against autoimmune/autoinflammatory disease. Given these key physiological functions, we deem it important to develop a three-dimensional (3D) tissue map for three major lymphoid organs of the human immune system: spleen, thymus and lymph nodes (Organ Specific Projects 1- 3, respectively). As one of our primary strengths, we have over 10 years of experience in the procurement of transplant-quality organs for research, including those of the lymphatic system. As part of this 24/7/365 effort, we have well-established relationships with the U.S. Organ Procurement Organizations (OPO), having referrals from 56/58 over the last decade. Our standard operating procedures (SOPs), both published and web-based, include assessment of normality, quality control assays, and systematic anatomical dissection/storage. We also participate in National QA/QC programs to evaluate tissue processing/banking procedures. Our approach to developing assay pipelines towards the common goal of a 3D tissue map will initially involve acquiring a macro image of the intact tissue and addressing tissue morphology, using our strengths in magnetic resonance imaging (MRI). The proposed optical microscopy pipeline will address microanatomical features using formalin fixed paraffin embedded (FFPE) and optimal cutting temperature (OCT) compound embedded sections as well as tissue optical clearing and expansion. These specimens will be studied from nm to mm resolution using stochastic optical reconstruction microscopy (STORM), confocal, multiphoton and light sheet fluorescence microscopies (LSFM), with all pipelines sharing a common file format for simplified 3D reconstruction. Based on the unique role for lymphatic organs in production and trafficking of immune cells, fluorescence activated cell sorting (FACS) of cells from blood and each lymphatic organ will provide a comparison of the patient- specific immune cell repertoire and serve in subsequent single cell RNA-seq analyses. To co-register biomolecules to their cognate cells, we will employ imaging mass cytometry (IMC), multiplex single molecule fluorescence in situ hybridization (smFISH) and multiplexed error-robust FISH (MERFISH) to map cellular protein and mRNA expression, ultimately on each 3D tissue atlas. Our experience in organ procurement, the availability of the National High Magnetic Field Laboratory (NHML) Advanced Magnetic Resonance Imaging and Spectroscopy (AMRIS) Facility at UF, multiphoton/confocal microscopy, LSFM, state of the art FACS, 10X GENOMICS Chromium controller, and high throughput cDNA library sequencing for RNA-Seq at the University of Florida, as well as world leading IMC and 3D data analysis facility at the University of Zurich, ideally positions us to provide the highest quality 3D Human BioMolecular Atlas (HuBMAP) of the lymphatic organs.

淋巴系统扮演五个主要角色：1）去除多余的体液； 2）吸收和运输循环系统的脂肪酸/chyle； 3）血液过滤； 4）安装主要防御通过免疫细胞的产生和激活感染和癌症； 5）产生和激活预防自身免疫/自身炎症疾病的调节免疫细胞。给定这些钥匙生理功能，我们认为为三个主要的三维组织图开发三维（3D）的组织很重要人免疫系统的淋巴器官：脾脏，胸腺和淋巴结（器官特定项目1- 分别为3）。作为我们的主要优势之一，我们拥有超过10年的采购经验用于研究的移植质量器官，包括淋巴系统的器官。作为这项24/7/365努力的一部分，我们与美国器官采购组织（OPO）建立了良好的关系，从过去十年中的56/58起。我们的标准操作程序（SOP），既发布又基于Web，包括评估正态性，质量控制测定和系统的解剖解剖/存储。我们还参加国家质量保证/QC计划，以评估组织加工/银行程序。我们的方法开发针对3D组织图的共同目标的测定管道最初将涉及获取完整组织的宏观图像和解决组织形态的宏观图像，使用我们在磁共振中的强度成像（MRI）。所提出的光学显微镜管道将使用福尔马林解决微解剖学特征固定的石蜡嵌入（FFPE）和最佳切割温度（OCT）化合物嵌入切片作为组织光学清除和膨胀。这些标本将使用NM到MM分辨率研究随机光学重建显微镜（风暴），共焦，多光子和光板荧光显微镜（LSFM），所有管道都共享用于简化3D重建的通用文件格式。基于关于淋巴器官在免疫细胞的生产和运输中的独特作用，荧光激活了来自血液和每个淋巴器官的细胞的细胞分选（FACS）将提供患者的比较特定的免疫细胞库，并在随后的单细胞RNA-seq分析中使用。共同注册生物分子对其同源细胞，我们将采用成像质量细胞仪（IMC），多重单分子荧光原位杂交（Smfish）和多路复用误射鱼（Merfish）以映射细胞蛋白质和mRNA表达，最终在每个3D组织地图集上。我们在器官采购方面的经验，国家高磁场实验室（NHML）的可用性高级磁共振成像在UF，多光子/共聚焦显微镜，LSFM，最新的FACS，10X 基因组学铬控制器和大学RNA-Seq的高吞吐量cDNA库测序佛罗里达大学以及苏黎世大学的世界领先的IMC和3D数据分析设施定位我们提供淋巴器官的最高质量3D人类生物分子图（Hubmap）。