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：
10318511
负责人：
Diane Bimczok
金额：
$ 46.23万
依托单位：
MONTANA STATE UNIVERSITY - BOZEMAN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-23 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318511
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 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 innovation intestinal homeostasis matrigel mechanotransduction member microbial microbial colonization microbiome microbiome research microbiota microorganism microorganism antigen migration monocyte mucosal vaccine neonatal Fc receptor neutralizing antibody novel novel vaccines organ on a chip pathogenic microbe prototype recruit residence response shear stress temporal measurement therapeutic development tissue culture tool transcriptome sequencing transcytosis uptake vaccine delivery vaccine development

项目摘要

Summary The emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly infectious human respiratory tract pathogen that has the capacity to infect many different organ systems, including the gastrointestinal (GI) tract. The overall goal of this project is to elucidate the role of the GI mucosa in SARS- CoV-2 infection and transmission. Specifically, we seek to identify mucosal immune mechanisms involved in COVID-19 pathogenesis and compare epithelial responses to SARS-CoV-2 in the gut of humans and bats. Investigating bat cells is important, because bats are considered the original hosts for SARS-CoV-2, but do not develop overt disease upon infection. Our recently developed GOFlowChip platform, which integrates 3- dimensional GI organoids and immune cells on a millifluidic chip with luminal and basolateral flow capacity, is ideally suited to probe the mechanisms involved in SARS-CoV-2 infection of the gut mucosa. Here, we propose to utilize the GOFlowChip to elucidate mechanisms of viral infection, replication and spread involved in SARS-CoV-2 infection of the GI tract. Specifically, we seek to (1) Define how SARS-CoV-2 infection of the GI epithelium contributes to viral spread. (2) Determine to what extent GI mucosal immune mechanisms regulate SARS-CoV-2 infection and spreading in the GI tract. (3) Compare GI epithelial responses to SARS- CoV-2 between bats and humans. For Specific Aim 1, we will optimize the GOFlowChip for use under BSL-3 laboratory conditions, followed by SARS-CoV-2 infection experiments that will elucidate susceptibility and viral replication dynamics in different gut compartments. Specific Aim 2 will leverage our organoid-mononuclear phagocyte (MNP) co-culture system to elucidate whether MNP-dependent transport mechanisms impact viral invasion of the GI mucosa, and we will screen SARS-CoV-2 reactive patient sera for their ability to modulate gut infection. For Specific Aim 3, we will establish organoid lines from bat GI tissues in order to compare epithelial responses to SARS-CoV-2 in bats and humans using RNA sequencing in order to understand differences in viral pathogenicity between humans and bats. The proposed work is directly integrated with our existing project, because we will use out GOFlowChip to investigate how SARS-CoV-2 interacts with epithelial cells and immune system components to cross the GI barrier. Our research is technologically innovative, because we are using gut organoid-immune cell co-cultures in a fully contained tissue chip design to study a BSL-3 level pathogen. Our project is conceptually innovative, because it is the first to compare gut organoids from Jamaican fruit bats to human organoids for their ability to sustain SARS-CoV-2 infection and to mount cellular antiviral responses. The proposed research is significant because it will yield crucial information on the role of the GI mucosa in COVID-19 pathogenesis that may inform future strategies to identify viral carriers, prevent viral transmission, and design novel antiviral treatments and vaccines.

概括新出现的严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 具有高度传染性人类呼吸道病原体能够感染许多不同的器官系统，包括胃肠 (GI) 道。该项目的总体目标是阐明胃肠道粘膜在 SARS 中的作用 CoV-2 感染和传播。具体来说，我们试图确定参与的粘膜免疫机制 COVID-19 发病机制以及人类和蝙蝠肠道上皮对 SARS-CoV-2 的反应比较。研究蝙蝠细胞很重要，因为蝙蝠被认为是 SARS-CoV-2 的原始宿主，但并不感染后出现明显的疾病。我们最近开发的 GOFlowChip 平台，集成了 3- 具有管腔和基底外侧流动能力的微流体芯片上的三维胃肠道类器官和免疫细胞，是非常适合探索肠道粘膜 SARS-CoV-2 感染的机制。在这里，我们提议利用 GOFlowChip 阐明病毒感染、复制和传播的机制胃肠道 SARS-CoV-2 感染。具体来说，我们寻求 (1) 定义 SARS-CoV-2 感染如何胃肠道上皮有助于病毒传播。 (2)确定胃肠道粘膜免疫机制的程度调节 SARS-CoV-2 感染和在胃肠道中的传播。 (3) 比较胃肠道上皮对 SARS 的反应蝙蝠和人类之间的 CoV-2。对于具体目标 1，我们将优化 GOFlowChip 以在 BSL-3 下使用实验室条件，然后进行 SARS-CoV-2 感染实验，以阐明易感性和病毒不同肠道区室的复制动态。具体目标 2 将利用我们的类器官-单核吞噬细胞 (MNP) 共培养系统阐明 MNP 依赖性转运机制是否影响病毒入侵胃肠道粘膜，我们将筛选 SARS-CoV-2 反应性患者血清，以确定其调节的能力肠道感染。对于具体目标 3，我们将从蝙蝠胃肠道组织中建立类器官系，以便进行比较使用 RNA 测序来了解蝙蝠和人类上皮细胞对 SARS-CoV-2 的反应人类和蝙蝠之间病毒致病性的差异。拟议的工作直接与我们的整合现有项目，因为我们将使用 GOFlowChip 来研究 SARS-CoV-2 如何与上皮细胞相互作用细胞和免疫系统成分穿过胃肠道屏障。我们的研究具有技术创新性，因为我们正在完全包含的组织芯片设计中使用肠道类器官-免疫细胞共培养物来研究 BSL-3级病原体。我们的项目在概念上是创新的，因为它是第一个比较肠道类器官的项目从牙买加果蝠到人类类器官，它们能够维持 SARS-CoV-2 感染并增加细胞抗病毒反应。拟议的研究意义重大，因为它将产生有关胃肠道粘膜在 COVID-19 发病机制中的作用可能为未来识别病毒携带者的策略提供信息，防止病毒传播，并设计新型抗病毒治疗方法和疫苗。