Pathogenesis of E. coli and Shigella infections in human enteroid models

人肠模型中大肠杆菌和志贺氏菌感染的发病机制

• 批准号: 9982173
• 负责人: MARK DONOWITZ
• 金额: $ 158.2万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9982173
• 关键词: Adherence; Animal Model; Attention; Attenuated Live Virus Vaccine; Attenuated Vaccines; Brush Border; Cancer cell line; Cells; Cellular biology; Childhood; Coculture Techniques; Communicable Diseases; Culture Media; Data; Dendritic Cells; Development; Diarrhea; Diarrheagenic E. coli; Disease; Enteral; Enterotoxins; Epithelial Cells; Escherichia coli; Escherichia coli EHEC; Etiology; Functional disorder; Gastroenterology; Goals; Goblet Cells; Human; Human Resources; Immune response; Immunology; Infection; Innate Immune System; Institution; International; Intestinal Mucosa; Intestines; Investigation; Joints; Kinetics; Laboratories; Lead; Leadership; Maryland; Measures; Microbiology; Modeling; Molecular; Mucins; Mucous Membrane; Natural Immunity; Organism; Pathogenesis; Pathogenicity; Pathway interactions; Peptide Hydrolases; Physiology; Process; Program Research Project Grants; Proteins; Recording of previous events; Research; Research Personnel; Research Project Grants; Role; Shigella; Shigella Infections; Therapeutic Agents; Tight Junctions; Training; Transport Process; Universities; Virginia; Virulence Factors; Work; absorption; chemokine; cytokine; diarrheal disease; enteric pathogen; enteroaggregative Escherichia coli; enterotoxigenic Escherichia coli; enterotoxin STa; human disease; immunoregulation; insight; macrophage; meetings; monolayer; multidisciplinary; neutrophil; new therapeutic target; novel; novel therapeutics; pathogen; programs; vaccine candidate; vaccine development

Abstract The development of human enteroids as a model to study the human intestine offers tremendous opportunities to study the pathogenesis of infectious enteric disease. We propose to combine the substantial expertise from three institutions with long and impressive histories in the investigation of enteric diseases to exploit this powerful model to study the pathogenesis of four major etiologic agents of diarrheal disease: three pathotypes of diarrheagenic E.coli (enteroaggregative, enterohemorrhagic and enterotoxigenic E. coli) and Shigella. Investigators from the University of Maryland, Johns Hopkins University and the University of Virginia will collaborate in this Program Project Grant (PPG). The leadership team of this PPG comprises three internationally renowned experts in enteric diseases, with complementary training in microbiology, gastroenterology, molecular physiology, and pediatric infectious diseases. They will direct a multidisciplinary team of co-investigators with expertise in cell biology, molecular pathogenesis, and mucosal immunology. The overall goal is to increase understanding of the pathophysiology and potential treatments of these four important pathogens. The proposed studies will use normal human mini-intestines, called enteroids or colonoids, grown on monolayers to develop models that mimic human disease. Examination of pathophysiologic aspects common to the diseases studied will serve to integrate the projects. These aspects include the role of mucins, bacterial proteases called SPATES, enterotoxins and secreted cytokines and are in addition to pathophysiologic aspects specific to each infection. In addition, the contribution of cells involved in innate immunity will be examined by co-culture of the enteroids/colonoids with human macrophages, neutrophils and dendritic cells. These studies using human mini-intestines offer the possibility of revealing insights in disease pathophysiology that are specific to normal human intestine rather than the animal models and cancer cell line models used until now. Each of the four projects focuses on a specific pathogen: enteroaggregative E. coli, Shigella, enterohemorrhagic E. coli, entertoxigenic E. coli. Besides an Administrative Core, there is an Enteroid Core that provides human enteroids/colonoids and growth media, instructs all projects on how to produce enteroid/colonoid monolayers and an Immunology Core that measures cytokines and chemokines, and isolates human macrophages, neutrophils and dendritic cells. The Enteroid and Immunology Cores work together to develop co-culture systems of innate immune cells and enteroids/colonoids that will be used by the projects. The investigators will regularly interact by monthly joint laboratory meetings. The proposed project will yield many significant new insights into enteric disease caused by these important pathogens.

抽象的 人类肠to的发展作为研究人类肠道的模型，提供了巨大的机会 研究传染性肠道疾病的发病机理。我们建议将大量专业知识结合在一起 在调查肠道疾病的情况下，三个具有长期且令人印象深刻的历史的机构来利用这一点 研究腹泻病的四种主要病因学的发病机理的强大模型：三种病原体 腹泻性e.coli（肠道，肠肠癌和肠毒素大肠杆菌）和志贺氏菌。 马里兰大学，约翰·霍普金斯大学和弗吉尼亚大学的调查人员将 在此计划项目赠款（PPG）中合作。该PPG的领导团队包括三个 国际知名的肠道疾病专家，以及微生物学的互补培训， 胃肠病学，分子生理学和小儿传染病。他们将指导一个多学科 在细胞生物学，分子发病机理和粘膜免疫学方面具有专业知识的共同投资者团队。这 总体目标是提高对这四种病理生理学和潜在治疗的理解 重要的病原体。拟议的研究将使用普通的人类迷你构句，称为肠toi或 结肠镜，生长在单层上，以开发模仿人类疾病的模型。检查 所研究疾病常见的病理生理方面将有助于整合项目。这些方面 包括粘蛋白，称为SPATE，肠毒素和分泌细胞因子的细菌蛋白酶的作用，并在 除病理生理方面，每种感染。另外，涉及的细胞的贡献 天生的免疫将通过与人类巨噬细胞的共同文化进行检查， 中性粒细胞和树突状细胞。这些使用人类迷你构之类的研究提供了揭示的可能性 针对正常人类肠道而不是动物模型的疾病病理生理学的见解 迄今为止，使用的癌细胞系模型。四个项目中的每个项目都集中在特定的病原体上： 肠肠球菌大肠杆菌，志贺氏菌，肠肠菌大肠杆菌，肠毒素大肠杆菌。除了行政 核心，有一个肠核提供人类的肠动物/结肠素和增长媒体，指示所有人 有关如何生产肠片/肠结肠单层的项目和测量细胞因子的免疫学核心 和趋化因子，并分离人类巨噬细胞，中性粒细胞和树突状细胞。肠和 免疫学核心共同开发先天性免疫细胞和 项目将使用的肠to虫/结肠体。调查人员将定期按每月联合进行互动 实验室会议。拟议的项目将对引起的肠道疾病产生许多重要的新见解 通过这些重要的病原体。