Project 2-Thymus

项目2-胸腺

• 批准号: 10211115
• 负责人: Todd Michael Brusko
• 金额: $ 46.75万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-14 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10211115
• 关键词: 3-Dimensional; Age; Anatomy; Asthma; Autoimmunity; Avidity; Benchmarking; Binding; Biologic Development; Biological Response Modifier Therapy; CD8-Positive T-Lymphocytes; Canis familiaris; Cell Therapy; Cells; Chromium; Clonal Deletion; Collection; Computer software; Coupled; Cryopreserved Tissue; Cytometry; Dendritic Cells; Deposition; Disease; Emigrations; Event; Expression Profiling; FOXP3 gene; Fluorescence Microscopy; Fluorescence-Activated Cell Sorting; Fluorescent in Situ Hybridization; Formalin; Gene Expression; Genetic Transcription; Histocompatibility; Human; Hypersensitivity; Image; Immune; Immune Tolerance; Immune response; Immunity; Immunofluorescence Immunologic; Immunohistochemistry; Immunologic Memory; Immunologics; Immunophenotyping; Investigation; Label; Life; Light; Lymphatic System; Magnetic Resonance Imaging; Maps; Measurement; Mediating; Medical; Microanatomy; Microscopy; Modernization; Modification; Molecular; Mus; Optics; Organ; Organ Donor; Pancreas; Paraffin; Paraffin Embedding; Pathology; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phase; Physiological; Polymers; Population; Procedures; Proteins; Protocols documentation; RNA; Race; Reagent; Recovery; Regulatory Element; Regulatory T-Lymphocyte; Reporting; Research Personnel; Resolution; Rodent Model; Role; Sampling; Secondary to; Source; Specimen; Stains; Standardization; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Technology; Temperature; Thinking; Three-Dimensional Image; Thymus Gland; Tissues; Training; Vaccines; Validation; base; cell type; central tolerance; confocal imaging; data de-identification; density; experience; hematopoietic cell transplantation; human disease; human imaging; imaging system; magnetic field; microscopic imaging; multiphoton imaging; preservation; reconstruction; scale up; secondary lymphoid organ; sex; single cell analysis; single-cell RNA sequencing; spectroscopic imaging; thymocyte; 3-Dimensional; Age; Anatomy; Asthma; Autoimmunity; Avidity; Benchmarking; Binding; Biologic Development; Biological Response Modifier Therapy; CD8-Positive T-Lymphocytes; Canis familiaris; Cell Therapy; Cells; Chromium; Clonal Deletion; Collection; Computer software; Coupled; Cryopreserved Tissue; Cytometry; Dendritic Cells; Deposition; Disease; Emigrations; Event; Expression Profiling; FOXP3 gene; Fluorescence Microscopy; Fluorescence-Activated Cell Sorting; Fluorescent in Situ Hybridization; Formalin; Gene Expression; Genetic Transcription; Histocompatibility; Human; Hypersensitivity; Image; Immune; Immune Tolerance; Immune response; Immunity; Immunofluorescence Immunologic; Immunohistochemistry; Immunologic Memory; Immunologics; Immunophenotyping; Investigation; Label; Life; Light; Lymphatic System; Magnetic Resonance Imaging; Maps; Measurement; Mediating; Medical; Microanatomy; Microscopy; Modernization; Modification; Molecular; Mus; Optics; Organ; Organ Donor; Pancreas; Paraffin; Paraffin Embedding; Pathology; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phase; Physiological; Polymers; Population; Procedures; Proteins; Protocols documentation; RNA; Race; Reagent; Recovery; Regulatory Element; Regulatory T-Lymphocyte; Reporting; Research Personnel; Resolution; Rodent Model; Role; Sampling; Secondary to; Source; Specimen; Stains; Standardization; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Technology; Temperature; Thinking; Three-Dimensional Image; Thymus Gland; Tissues; Training; Vaccines; Validation; base; cell type; central tolerance; confocal imaging; data de-identification; density; experience; hematopoietic cell transplantation; human disease; human imaging; imaging system; magnetic field; microscopic imaging; multiphoton imaging; preservation; reconstruction; scale up; secondary lymphoid organ; sex; single cell analysis; single-cell RNA sequencing; spectroscopic imaging; thymocyte

The thymus has long been considered an enigmatic and mysterious organ; having suffered to some extent from a widespread notion, especially amongst those with a background in medical training, that it has only of eminent importance in early life. Indeed, this line of thinking originates from its critical role in establishing central immune tolerance in early life subsequently followed by a waning period of involution. However, an emerging body of evidence has suggested this “dogma” is, in effect, a “dog” in that many of the notions related to thymic function in humans are, in fact, limited in terms of their confidence as to physiological activities. Much of our understanding of the lymphatic system and its role in immunity has been based on studies of rodent models that appear to differ in considerable ways and cannot be extrapolated directly to humans. On top of this, there are clear voids in terms of the relationship between anatomy and function as well as the influences of age, sex, and race on thymic function. Hence, with this disconnect, and given the importance of the immune response to so many aspects of human disease, we believe it begs the notion of studying the anatomy of the human thymus, from normal organ donors (i.e., unaffected by thymic or immunological pathology), in an attempt to eventually impact our understanding of human disease. To be clear, in terms of targeted organs, given the role of the thymus in establishing central tolerance, there is perhaps no organ more relevant that should be subject to such anatomical investigation. Specifically, we believe a biomolecular mapping of the human thymus is essential to identify where and how human T cell selection occurs, T cell receptor (TCR) reactivities/binding avidities that pass selection or are subject to clonal deletion, and the specific regulatory elements governing T lymphocyte development prior to emigration to secondary lymphoid organs. To achieve this, we posit the following specific aims: AIM I: Establish systematic, anatomical best practices/benchmarks for collection, preservation, validation and standardization of QC protocols (SOPs) for acquiring and handling organ donor grade human thymus; AIM II: Reconstruct the three-dimensional (3D) human thymus; AIM III: Immunophenotype human thymus cells by fluorescence activated cell sorting (FACS). As the thymus is uniquely involved in T lymphocyte development, thymocyte and dendritic cell (DC) populations from dispersed thymus and peripheral blood mononuclear cells from the same patient will be defined by FACS and the immune repertoires compared. Isolated cell subsets will serve as the source for RNA expression profiling (AIM IVB) to guide the selection of markers used in AIMs II and IV; and AIM IV; ultimately, to determine the molecular composition and gene expression within the normal 3D human thymus.

长期以来，胸腺一直被认为是神秘和神秘的器官。在一定程度上遭受了 从广泛的概念中，尤其是在接受医学培训背景的人中，只有 在早期生活中的重要性。确实，这种思维方式源于其在建立中的关键作用 早期的中央免疫耐受性随后发生了衰落时期。但是，一个 新兴的证据表明，这种“教条”实际上是一种“狗”，因为许多笔记都与 实际上，人类对胸腺功能的信心限制了对体育活动的信心。很多 我们对淋巴系统及其在免疫力中的作用的理解是基于对啮齿动物的研究 似乎以相当大的方式不同并且不能直接推断到人类的模型。在 这是关于解剖学与功能之间的关系以及影响的明显空隙 年龄，性别和胸腺功能的种族。因此，与这种断开连接，并鉴于免疫的重要性 对人类疾病的许多方面的反应，我们认为这是一种研究的概念 来自正常器官供体的人胸（即不受胸腺或免疫病理学影响） 试图最终影响我们对人类疾病的理解。明确地说，就有针对性的器官而言 鉴于胸腺在建立中心容忍中的作用，可能没有更相关的器官 应受到这种解剖学投资的约束。具体而言，我们相信 人百里香对于确定人类T细胞选择的何处以及如何发生T细胞受体（TCR）至关重要 通过选择或受到克隆缺失和特定调节的反应性/结合狂热 在移民到继发性淋巴器官之前，有关T淋巴细胞发育的元素。实现 这是我们海报以下特定目的：目标I：建立系统的，解剖的最佳实践/基准 用于收集，保存，验证和标准化QC协议（SOP），用于处理和处理 器官捐赠者级人类胸腺； AIM II：重建三维（3D）人百里香； AIM III： 通过荧光激活细胞分选（FACS），免疫表型人胸腺细胞。由于胸腺是 独特地参与T淋巴细胞发育，胸腺细胞和树突状细胞（DC）种群。 来自同一患者的胸腺和外周血单核细胞将由FACS和 比较免疫曲目。孤立的细胞子集将作为RNA表达分析的来源（AIM IVB）指导AIMS II和IV中使用的标记的选择；和目标IV；最终，确定 正常3D人胸腺中的分子组成和基因表达。