Coord Core - Atkinson

坐标核心 - 阿特金森

• 批准号: 10440809
• 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
• 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/10440809
• 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

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.

淋巴系统有五个主要作用：1)清除多余的体液；2)吸收和运输