Modeling immunity with a hybrid lymph node tissue-chip

使用混合淋巴结组织芯片模拟免疫

• 批准号: 10059169
• 负责人: Rebecca R Pompano
• 金额: $ 45.13万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-20 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10059169
• 关键词: Acute; Affect; Alzheimer' s Disease; Animals; Anti-Inflammatory Agents; Antigens; Autoimmunity; Biodistribution; Biological Models; Biological Process; Biomedical Research; Brain; Buffers; Caliber; Cell Communication; Cell Culture Techniques; Cells; Chemistry; Chronic; Clinical; Coupled; Development; Devices; Disease; Drug Targeting; Event; Experimental Models; Gases; Goals; Hour; Human; Hybrids; Immune response; Immune system; Immunity; Immunology; Immunotherapy; In Vitro; Infection; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Label; Lateral; Liquid substance; Location; Lymph Node Tissue; Malignant Neoplasms; Manuals; Measures; Membrane; Metabolic; Methods; Microfluidics; Modeling; Movement; Multiple Sclerosis; Mus; Physiologic pulse; Population; Production; Publishing; Research; Research Personnel; Resolution; Response to stimulus physiology; Rest; Rheumatoid Arthritis; Role; Signal Transduction; Signaling Protein; Slice; Stimulus; Stream; Suspensions; System; Systems Analysis; TNF gene; TNFRSF1A gene; Technology; Testing; Time; Tissue Microarray; Tissue Sample; Tissue Slice Technology; Tissues; Tonsil; Vaccines; Work; adaptive immunity; behavioral response; cell motility; chronic inflammatory disease; cytokine; design; experimental study; extracellular; fighting; human tissue; improved; in vivo; in vivo Model; inflammatory marker; innovation; interest; lymph nodes; macromolecule; middle age; nanoparticle; novel; novel therapeutics; preservation; prevent; response; therapy design; tool; tumor; tumor necrosis factor-alpha inhibitor; young adult

PROJECT SUMMARY/ABSTRACT Local interactions in the immune system determine whether an immune response is protective or destructive, fighting infection or initiating autoimmunity. Adaptive immunity begins in the lymph node (LN), a highly organized and dynamic tissue. Currently, it is difficult to parse the role of inflammatory mediators or rationally design therapies for chronic inflammatory disease such as artherosclerosis, rheumatoid arthritis and multiple sclerosis, which together affect 5 – 7% of the Western population. We hypothesize that analyzing local responses ex vivo in intact tissue will provide information not easily obtained from current methods (in vitro/in vivo). Such experiments require new tools to analyze dynamics in the immune system, which we develop by combining expertise in bioanalytical chemistry, microfluidics, and immunology. In this project, we will develop a novel ex vivo model of immunity, using a hybrid of microfluidic culture and LN slices. In Aim 1, we will establish long-term culture coupled with analysis methods for live murine and human LN slices. Slice culture offers the advantage of preservation of the extracellular microenvironment and any matrix-bound signals. We will optimize long-term culture (7-21 days) for murine LN slices and human tonsil slices to maintain high viability, low cellular activation markers at rest, and ability to respond to inflammatory and antigen-specific stimuli. In Aim 2, we will develop a novel microfluidic system for on-demand local stimulation of LN slices. We will improve the spatial resolution of our previously developed device, to target clusters 2 – 10 cells in diameter (20 – 100 μm lateral resolution) using short- and long-term stimulation. We will also enable on-demand selection of delivery zone by using a mobile port, making the whole tissue accessible with minimal handling. In Aim 3, we will validate the hybrid microfluidic-tissue slice system for analysis of inflammatory responses and anti-inflammatory therapies. We will compare the inflammatory response to a pro-inflammatory cytokine, TNF-α, in slices versus cell cultures and in vivo systems. Finally, we will test the extent to which the model provides new information to guide immunotherapy, by using the hybrid tissue-chip system with mouse and human tissue to compare the effects of competing TNF-α inhibitors (anti-TNF-α monoclonals or soluble TNF-α receptor). Combining local microfluidic stimulation with tissue slice technology produces the first experimental platform for analysis of spatially organized signaling and cell-cell interactions in live LN tissue. This innovative platform will advance both basic and translational biomedical research: locally delivered cytokines will serve as a much-needed model of acute or chronic inflammation, and locally delivered immunotherapies will guide the design of targeted drug-loaded nanoparticles. This technology is broadly applicable for a host of inflammatory diseases, including rheumatoid arthritis, Chron’s disease, multiple sclerosis, Alzheimer’s disease, and cancer.

项目摘要/摘要 免疫系统中的局部相互作用确定免疫响应是否受到保护或 破坏性，打击感染或引发自身免疫。自适应免疫组织化学始于淋巴结（LN），A 高度组织和动态组织。目前，很难解析炎症介体的作用或 合理设计用于慢性炎性疾病的疗法，例如白盘状性，类风湿关节炎和 多发性硬化症，共同影响了5 - 7％的西方人口。我们假设分析了本地 完整组织中的离体反应将提供不容易从当前方法获得的信息（体外/IN 体内）。这样的实验需要新工具来分析免疫系统中的动态，我们通过 结合生物分析化学，微流体和免疫学方面的专业知识。 在这个项目中，我们将使用微流体的杂种开发一种新型的免疫力模型 文化和LN切片。在AIM 1中，我们将建立长期文化以及现场分析方法 鼠和人类LN切片。切片文化提供了保存细胞外的优势 微环境和任何矩阵结合信号。我们将为鼠LN优化长期培养（7-21天） 切片和人扁桃片切片，以维持高的生存能力，静止的低细胞激活标记以及能够 在AIM 2中，我们将开发一个新型的微流体系统 按需局部模拟LN切片。我们将改善先前开发的空间分辨率 设备，使用短期和长期的直径2 - 10个单元格（20 - 100μm横向分辨率） 刺激。我们还将使用移动端口启用按需选择交付区域的选择 全组织可通过最小的处理来访问。在AIM 3中，我们将验证杂交微流体组织切片 用于分析炎症反应和抗炎疗法的系统。我们将比较 切片与细胞培养和体内促炎性细胞因子TNF-α的炎症反应 系统。最后，我们将测试该模型提供的新信息以指导免疫疗法， 通过使用小鼠和人组织的混合组织芯片系统比较竞争的影响 TNF-α抑制剂（抗TNF-α单克隆或固体TNF-α受体）。 将局部微流体刺激与组织切片技术相结合，产生了第一个实验 用于分析活LN组织中空间组织的信号传导和细胞 - 细胞相互作用的平台。这种创新 平台将推进基本和翻译的生物医学研究：当地传递的细胞因子将作为 急性或慢性感染的急需模型，以及局部传递的免疫疗法将指导 靶向药物纳米颗粒的设计。该技术广泛适用于大量炎症 包括类风湿关节炎，Chron病，多发性硬化症，阿尔茨海默氏病和癌症在内的疾病。