Stanford Cooperative Research Center for Novel, Alternative Model Systems for Enteric Diseases

斯坦福肠道疾病新型替代模型系统合作研究中心

基本信息

批准号：
8855404
负责人：
MANUEL R AMIEVA
金额：
$ 100.96万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2020-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8855404
关键词：
3-Dimensional Acute Address Adherence Adherent Culture Animal Model Animals Architecture Bacterial Infections Bacterial Model Biological Models Biomedical Engineering Cell Communication Cell Count Cells Cellular Structures Characteristics Chronic Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Colon Communicable Diseases Cytometry Dendritic Cells Development Digestion Disease Disease model Effector Cell Engineering Ensure Enteral Epithelial Epithelial Cells Exhibits Experimental Models Extracellular Matrix Functional disorder Gastrointestinal tract structure Generations Genes Genetic Genetic study Goals Helicobacter Infections Helicobacter pylori Host resistance Human Human Cell Line Human Characteristics Human Development Image Analysis Immune Immune response Immunity Immunoassay Immunology In Vitro Infection Integration Host Factors Interferons Intestines Investigation Lead M cell Mesenchymal Methods Microfluidic Microchips Microfluidics Modeling Morbidity - disease rate Mus Normal tissue morphology Oncogenic Organ Organoids Pathogenesis Play Population Predisposition Process Production Proteomics Reproducibility Research Role Rotavirus Rotavirus Infections Route Salmonella typhi Salmonella typhimurium Small Intestines Source Stimulus Stomach Stromal Cells System Technology Tissue Engineering Tissues Transformed Cell Line Universities Viral Virus Virus Diseases Work absorption base direct application enteric pathogen flexibility gastrointestinal gastrointestinal bacteria gastrointestinal epithelium genetic manipulation human disease improved in vivo Model innovation macrophage mortality multidisciplinary novel novel strategies pathogen programs public health relevance response statistics tissue tropism

项目摘要

DESCRIPTION OF THE OVERALL U19 APPLICATION (provided by applicant): The human gastrointestinal tract exhibits diverse and essential roles in processes as widespread as digestion, absorption, secretion and immunity. However, these same crucial functions represent a significant vulnerability by which bacterial and viral gastrointestinal pathogens induce substantial morbidity and mortality in humans worldwide. While animal models are invaluable to the study of basic mechanisms of infectious disease, the interactions between some of the most prominent gastrointestinal pathogens with host tissues are species specific and host restricted, particularly for infections that have co-evolved with humans. Thus, the study of many pathogens has heretofore required animal-adapted strains that do not accurately recapitulate the human disease pathophysiology and host range restriction in mice, or alternatively oncogenically transformed human cell lines that do not model normal tissue characteristics. Recent advances in culture of primary tissues as 3-dimensional "organoids" that reproduce multilineage differentiation and organ architecture represent a promising technology to modeling the interaction of human enteric pathogens with human gastrointestinal epithelium in vitro. The overall goal of this application is to create alternative model systems for a variety of human enteric diseases and pathogen-associated specific immune responses using optimized 3D human gastrointestinal organoid cultures via the co-culture of (1) primary gastric and intestinal organoids with (2) bacterial/viral pathogens and/or (3) immune effector cells. We have thus assembled a multidisciplinary, collaborative, and synergistic team at Stanford University including Calvin Kuo (gastrointestinal organoid culture), Manuel Amieva (Helicobacter pylori), Harry Greenberg (Rotavirus), Sarah Heilshorn (microenvironment bioengineering), Sean Bendall (CyTOF), Elizabeth Mellins (immunology), and Denise Monack (Salmonella Typhi). Together, these PIs form the Stanford Novel, Alternative Models for Enteric Diseases Cooperative Research Center (Stanford NAMSED CRC) Program. The Stanford NAMSED CRC is composed of Cores A-C (Administration, Organoid Production, and Advanced Co-Culture Engineering and Single Cell Statistics of Gut Immunology "ACCESS-GI"), and Projects 1-2, which study bacterial and viral pathogen interactions with human GI organoids, respectively. Towards these goals, we employ human organoid technologies incorporating epithelial-only or epithelial/mesenchymal components to create tissue-specific models of bacterial (Helicobacter pylori, Salmonella Typhi, Salmonella Typhimurium) or viral (rotavirus) infections, with or without immune effector cells or stimuli. We deploy innovative supporting technologies including CyTOF mass cytometry, bioengineered extracellular matrices, microfluidics and CRISPR-based gene editing. Overall, the Stanford NAMSED represents a synergistic group devoted to the integrated modeling of enteric pathogens as an epithelial-immune organoid unit.

整体U19应用程序的描述（由适用提供）：人类胃肠道在像消化，滥用，分泌和免疫等宽度的过程中表现出潜水员和基本作用。然而，这些相同的关键功能代表了一个重要的脆弱性，细菌和病毒胃肠道病原体在全球范围内引起了大量的发病率和死亡率。虽然动物模型对于研究传染病的基本机制是无价的，但一些最突出的胃肠道病原体与宿主组织之间的相互作用是特异性的，受宿主受到限制，尤其是对于与人类共同发展的感染。这是对许多病原体的研究，迄今为止需要动物适应的菌株，这些菌株不能准确地概括小鼠的人体病理生理学和宿主范围限制，或者替代地肿瘤转化的人类细胞系，这些人细胞系未模拟正常组织特征。原代组织培养的最新进展为三维“器官”，从而再现了多节分区分和器官结构，这是一种有前途的技术，可以建模人类肠道病原体与人类胃肠道上皮的相互作用。该应用的总体目标是通过（1）原发性胃和（2）细菌/病毒性病原体和/或（3）免疫效应的免疫效应细胞，使用优化的3D人类胃肠道和肠机培养物，为各种人类肠道疾病创建替代模型系统和与病原体相关的特异性免疫反应。 We have thus assembled a multidisciplinary, collaborative, and synergistic team at Stanford University including Calvin Kuo (gastrointestinal organoid culture), Manuel Amieva (Helicobacter pylori), Harry Greenberg (Rotavirus), Sarah Heilshorn (microenvironment bioengineering), Sean Bendall (CyTOF), Elizabeth Mellins (immunology),和Denise Monack（沙门氏菌）。这些PI共同构成了斯坦福小说的肠道疾病合作研究中心（Stanford NAMSED CRC）计划的替代模型。 Stanford NAMSED CRC由核心A-C（施用，器官生产和晚期共培养工程和肠道免疫学“访问GI”）和单个细胞统计以及分别研究细菌和病毒病原体与人类GI类型的单细胞统计。为了实现这些目标，我们采用人体器官技术融合了仅上皮或上皮/间质成分来创建细菌（幽门螺杆菌，沙门氏菌，沙门氏菌Typhimurium）或病毒性（Rotavirus）（rotavirus）感染的细菌的组织特异性模型（幽门螺杆菌，沙门氏菌）或不含免疫细胞感染。我们部署了创新的支持技术，包括细胞质量细胞仪，生物工程细胞外材料，微流体和基于CRISPR的基因编辑。总体而言，斯坦福大学的名称代表了一个协同组，该群体致力于肠道病原体作为上皮免疫器官单元的综合建模。