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：
9022400
负责人：
MANUEL R AMIEVA
金额：
$ 100.79万
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/9022400
关键词：
3-Dimensional Acute Address Adherence Adherent Culture Animal Model Animals Architecture Bacterial Infections Bacterial Model Biological Models Biomedical Engineering CRISPR/Cas technology 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 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 Virus Replication 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.

U19整体应用的描述（由申请人提供）：人类胃肠道在消化、吸收、分泌和免疫等广泛的过程中表现出多样化且重要的作用，然而，这些相同的关键功能代表了细菌和病毒胃肠道的显着脆弱性。虽然动物模型对于研究传染病的基本机制非常有价值，但一些最重要的胃肠道病原体与病原体之间的相互作用。宿主组织是物种特异性和宿主的，特别是对于与人类共同进化的感染来说，迄今为止，许多病原体的研究需要动物适应的菌株，这些菌株不能准确地重现人类疾病的病理生理学和小鼠的宿主范围限制。或者致癌转化的人类细胞系不模拟正常组织特征，作为再现多谱系分化和器官结构的3维“类器官”的培养的最新进展代表了模拟人类相互作用的有前途的技术。该应用的总体目标是利用优化的 3D 人类胃肠道类器官培养物，通过 (1) 的共培养，为各种人类肠道疾病和病原体相关的特异性免疫反应创建替代模型系统。）初级胃和肠类器官与（2）细菌/病毒病原体和/或（3）免疫效应细胞因此，我们在斯坦福大学组建了一个包括卡尔文在内的多学科、协作和协同团队。 Kuo（胃肠类器官培养）、Manuel Amieva（幽门螺杆菌）、Harry Greenberg（轮状病毒）、Sarah Heilshorn（微环境生物工程）、Sean Bendall（CyTOF）、Elizabeth Mellins（免疫学）和 Denise Monack（伤寒沙门氏菌）。 PI 组成了斯坦福新型肠道疾病替代模型合作研究中心（斯坦福 NAMSED）斯坦福大学 NAMSED CRC 项目由核心 A-C（管理、类器官生产、肠道免疫学高级共培养工程和单细胞统计“ACCESS-GI”）以及研究细菌和病毒的项目 1-2 组成。为了实现这些目标，我们采用仅包含上皮成分或上皮/间质成分的人类类器官技术来创建病原体的组织特异性模型。细菌（幽门螺杆菌、伤寒沙门氏菌、鼠伤寒沙门氏菌）或病毒（轮状病毒）感染，无论有或没有免疫效应细胞或刺激，我们都部署创新的支持技术，包括 CyTOF 质谱流式细胞术、生物工程细胞外基质、微流体和基于 CRISPR 的基因编辑。斯坦福大学 NAMSED 代表了一个协同小组，致力于将肠道病原体作为上皮免疫类器官单元进行综合建模。