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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

基本信息

批准号：
9893635
负责人：
Diane Bimczok
金额：
$ 56.54万
依托单位：
MONTANA STATE UNIVERSITY - BOZEMAN
依托单位国家：
美国
项目类别：
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/9893635
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 Epithelium 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 gastrointestinal gastrointestinal epithelium gut microbes gut microbiota host-microbe interactions human model 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

项目摘要

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.

项目概要该项目将开发新型人体胃肠道体外模型，以了解细胞对微生物的自然反应以及免疫耐受和激活的诱导。发展历程胃肠道类器官，嵌入复杂原代上皮细胞群的 3D 永久培养物在细胞外基质中，彻底改变了胃肠道发育、微生物学和近5年免疫学。对于我们的项目，我们组建了一个跨学科的研究团队拥有生物工程（Wilking、Chang）、免疫学（Bimczok、Jutila）和人类微生物组方面的专业知识研究（Walk）显着推进 3D 肠道类器官-微生物组共培养系统。我们的团队有最近建立了一个微流体肠道芯片平台 GoFlowChip，它概括了管腔和基础在人体肠道类器官中流动。在平行研究中，我们建立了原始人类的共培养物我们已成功感染幽门螺杆菌的单核细胞来源的 DC 和人胃球体。在这里，我们寻求利用我们两个模型的独特功能并将它们组合到一个分析平台中研究从胃肠道腔采集抗原以诱导适应性粘膜免疫或宽容。具体来说，我们寻求定义和量化候选机制的贡献，包括跨上皮树突形成和 Fc 受体依赖性转胞吞作用，使单核吞噬细胞成为可能（MNP）来获取管腔抗原。我们假设 MNP 抗原获取的不同机制是可以使用 GOFlowChip 平台进行复制和定量分析，并且定植的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) 开发并验证基于芯片的类器官-DC 共培养系统，具有管腔和基底侧流能力。 (2) 量化净效应胃肠道类器官生物学和 MNP 相互作用的生物复杂性。 (3) 阐明相关机制通过人类 MNP 从胃肠道腔中取样细菌抗原。这种方法将使我们能够优化我们的集成 GoFlowChip 共培养系统，将其作为疫苗研究领域的强大新工具或药物输送以及肠道微生物群对抗原采样的影响。拟议的研究是概念上的创新，因为它整合了所有三种必要的细胞类型（上皮细胞、微生物细胞和免疫细胞）参与粘膜宿主-微生物相互作用。 GoFlowChip平台具有技术创新性，因为它复制了复杂的耗氧上皮和微生物定植的管腔，是第一个将将流体学引入 3-D 类器官培养物中，并重现自然发生在类器官之间的密切相互作用胃肠道上皮和前哨 MNP。拟议的研究意义重大，因为它将提供非常强大的新工具，可以解决人类粘膜生物学中的基本机械问题，人类健康和疾病背景下的微生物学和免疫学。