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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.

项目总结