Integration of mononuclear phagocytes into the human gastrointestinal GOFlowChip for investigation of luminal antigen sampling

将单核吞噬细胞整合到人胃肠道 GOFlowChip 中用于腔内抗原采样研究

• 批准号: 10263208
• 负责人: Diane Bimczok
• 金额: $ 59.93万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10263208
• 关键词: 3-Dimensional; Address; Antigen-Presenting Cells; Antigens; Apical; Apoptosis; Bacteria; Bacterial Antigens; Bacterial Translocation; Biological; Biological Assay; Biological Models; Biological Process; Biology; Biomedical Engineering; Biotechnology; Cell Communication; Cells; Coculture Techniques; Collaborations; Complex; Dendrites; Dendritic Cells; Development; Disease; Drops; Drug Delivery Systems; E-Cadherin; Engineering; Epithelial; Epithelial Cells; Escherichia coli; Extracellular Matrix; Fc Receptor; Gastrointestinal tract structure; Health; Helicobacter pylori; Homeostasis; Human; Human Experimentation; Human Microbiome; Image; Immune; Immune Tolerance; Immune system; Immunity; Immunology; Intestines; Investigation; Liquid substance; Microbe; Microbiology; Microfluidics; Modeling; Mononuclear; Movement; Mucosal Immunity; Mucous Membrane; Neonatal; Organoids; Oxygen; Phagocytes; Population; Positioning Attribute; Reproducibility; Research; Research Personnel; Sampling; Sentinel; Side; Sorting - Cell Movement; Stomach; Surface; System; Testing; Therapeutic; Thick; Tight Junctions; Time; Walking; Work; adaptive immunity; base; beneficial microorganism; cell type; clinical application; design; field study; fluid flow; gastric organoids; gastrointestinal; gastrointestinal epithelium; gut microbes; gut microbiota; host-microbe interactions; human tissue; immune activation; in vitro Model; inhibitor/antagonist; innovation; intestinal homeostasis; matrigel; mechanotransduction; member; microbial; microbial colonization; microbiome; microbiome research; microbiota; microorganism; microorganism antigen; migration; monocyte; mucosal vaccine; neutralizing antibody; novel; novel vaccines; organ on a chip; pathogenic microbe; prototype; receptor; recruit; residence; response; shear stress; temporal measurement; therapeutic development; tissue culture; tool; transcriptome sequencing; transcytosis; uptake; vaccine delivery; vaccine development

PROJECT SUMMARY This project will develop novel in vitro models of the human gastrointestinal (GI) tract for understanding natural cellular responses to microbes and the induction of immune tolerance and activation. The development of gastrointestinal organoids, 3-D permanent cultures of complex primary epithelial cell populations embedded in an extracellular matrix, has revolutionized research in gastrointestinal development, microbiology and immunology in the past 5 years. For our project, we have assembled a trans-disciplinary team of investigators with expertise in bioengineering (Wilking, Chang), immunology (Bimczok, Jutila), and human microbiome research (Walk) to significantly advance 3-D gut organoid-microbiome co-culture systems. Our team has recently established a millifluidic gut-on-a chip-platform, the GoFlowChip, that recapitulates luminal and basal flow in human intestinal organoids. In parallel investigations, we have established co-cultures of primary human monocyte-derived DCs and human gastric spheroids that we have successfully infected with H. pylori. Here, we seek to leverage the unique capabilities of our two models and combine them into a single analytical platform to study antigen sampling from the gastrointestinal lumen for the induction of adaptive mucosal immunity or tolerance. Specifically, we seek to define and quantify the contributions of candidate mechanisms including transepithelial dendrite formation and Fc-receptor-dependent transcytosis that enable mononuclear phagocytes (MNPs) to acquire luminal antigens. We hypothesize that distinct mechanisms of MNP antigen acquisition are reproduced and can be quantitatively analyzed using the GOFlowChip platform and that colonizing bacteria and fluid dynamics regulate epithelial antigen transport. To test our hypotheses, we will (1) Develop and validate a chip-based organoid-DC co-culture system with luminal and basolateral flow capacity. (2) Quantify the net effect of biologic complexity on GI organoid biology and MNP interaction. (3) Elucidate the mechanisms involved in bacterial antigen sampling from the gastrointestinal lumen by human MNPs. This approach will enable us to optimize our integrated GoFlowChip co-culture system as a powerful new tool for the field for studies on vaccine or drug delivery and on the impact of intestinal microbiota on antigen sampling. The proposed research is conceptually innovative, because it integrates all three necessary cell types (epithelial, microbial, and immune) involved in mucosal host-microbe interactions. The GoFlowChip platform is technologically innovative, because it replicates a complex, oxygen-utilizing epithelium and a microbially colonized lumen, is the first to incorporate fluidics into 3-D organoid cultures, and reproduces the intimate interactions that naturally occur between the gastrointestinal epithelium and sentinel MNPs. The proposed research is significant, because it will provide an incredibly powerful new tool to address fundamental, mechanistic questions in human mucosal biology, microbiology, and immunology in the context of human health and disease.

项目摘要 该项目将开发人类胃肠道（GI）的新型体外模型，以理解 对微生物的天然细胞反应以及免疫耐受性和激活的诱导。发展 胃肠道器官，复杂原代上皮细胞种群的3-D永久培养物嵌入 在细胞外基质中，彻底改变了胃肠道发展，微生物学和 过去5年的免疫学。对于我们的项目，我们组建了一个跨学科的调查员团队 具有生物工程专业知识（Wilking，Chang），免疫学（Bimczok，Jutila）和人类微生物组 研究（步行）可显着促进3-D肠杆菌 - 微生物组共培养系统。我们的团队有 最近建立了一个毫米肠道的芯片平台，Goflowchip，概括了腔和基础 人类肠癌中的流动。在平行研究中，我们建立了主要人类的共同文化 我们成功地感染了幽门螺杆菌的单核细胞衍生的DC和人类胃球体。在这里，我们 寻求利用我们的两种型号的独特功能，并将它们结合到一个分析平台中 研究胃肠道腔中的抗原采样，以诱导适应性粘膜免疫或 宽容。具体而言，我们试图定义和量化候选机制的贡献 旋转的树突形成和FC受体依赖性转胞病，使单核吞噬细胞能够 （MNP）获取腔抗原。我们假设MNP抗原采集的不同机制是 复制并可以使用Goflowchip平台进行定量分析，并定居细菌和 流体动力学调节上皮抗原转运。为了检验我们的假设，我们将（1）开发和验证 基于芯片的类器官-DC共培养系统，具有腔内和基底外侧流量。 （2）量化净效果 GI器官生物学和MNP相互作用的生物复杂性。 （3）阐明涉及的机制 人类MNP从胃肠道腔中采样的细菌抗原采样。这种方法将使我们能够 优化我们集成的Goflowchip共同培养系统，作为该领域的强大新工具，用于研究疫苗 或药物输送以及肠道菌群对抗原采样的影响。拟议的研究是 从概念上创新，因为它整合了所有三种必要的细胞类型（上皮，微生物和免疫） 参与粘膜宿主 - 微叶相互作用。 Goflowchip平台在技术上具有创新性，因为 它复制复杂的，氧气利用的上皮和微生物​​定殖的管腔，是第一个结合的 流体化成3-D器官培养物，并再现自然发生的亲密相互作用 胃肠道上皮和哨兵MNP。拟议的研究很重要，因为它将提供 令人难以置信的强大新工具来解决人类粘膜生物学中的基本机械问题， 在人类健康和疾病的背景下，微生物学和免疫学。