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

基本信息

批准号：
10427388
负责人：
MARK DONOWITZ
金额：
$ 203.23万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10427388
关键词：
3-Dimensional Address Anaerobic Bacteria Attention Attenuated Vaccines Biological Response Modifiers Cell Communication Cell model Cell physiology Cells Cellular biology Childhood Clinical Coculture Techniques Collaborations Communicable Diseases Culture Media Dendritic Cells Development Diarrhea Diarrheagenic E. coli Disease Electronic Mail Enteral Enterotoxins Environment Epithelial Escherichia coli Exposure to Functional disorder Funding Gastroenterology Glycoproteins Goals Goblet Cells Human Human Resources Immune Immune Evasion Immune system Immunology International Intestines Laboratories Leukocytes M cell Maryland Measurement Methodology Methods Microbiology Modeling Molecular Mucins Mucous Membrane Organism Pathogenesis Peptide Hydrolases Phagocytes Physiology Production Program Research Project Grants Publications Reagent Recording of previous events Research Research Personnel Research Project Grants Role Serine Protease Shigella Shigella Infections Shigella Vaccines System T-Lymphocyte Telephone Universities Virginia Work antimicrobial peptide base cell motility cytokine diarrheal disease eVisit enteric pathogen enteroaggregative Escherichia coli enterotoxigenic Escherichia coli experience gut inflammation human model improved insight intraepithelial macrophage meetings model development monolayer neutrophil novel pathogen pathogenic bacteria pre-clinical programs response scaffold shear stress tool transcytosis

项目摘要

Pathogenesis of E. coli and Shigella Infections in Human Enteroid Models Overall Description / Summary This first renewal of a funded program Project Grant (PPG) is to continue progress in understanding the pathophysiology and identification of ways to interfere with diarrhea caused by Shigella and two pathotypes of diarrheagenic E. coli (enteroaggregative and enterotoxigenic E. coli). The substantial progress made has resulted from the close collaboration and combined expertise of investigators from the University of Maryland, Johns Hopkins University and the University of Virginia who are using shared models of normal human enteroid and colonoid monolayers exposed to enteric bacterial pathogens as well as human enteroid monolayers co- cultured with innate phagocytic cells present in the intestine. The group of investigators has internationally renowned experience in enteric diseases, as well as in microbiology, gastroenterology, molecular physiology, pediatric infectious diseases, cell biology, molecular pathogenesis, and mucosal immunology. This consortium has interacted and in a superbly collaborative manner sharing technical and scientific information through formal monthly face-to-face or Zoom-based meetings, frequent telephone or electronic communication and visits to laboratories, with the latter being the standard form of interaction of the U Maryland and JHU investigators. The Enteroid and Immunology Cores have developed new tools and methods that were rapidly transferred to the Project investigators, resulting in a significant number of collaborative publications from the PPG. As expected, progress was stepwise with transfer of methodology and reagents for propagating the enteroids, production of monolayers, development of the bacterial-enteroid models, and development of co-cultures with immune cells leading to new insights in the pathogen-enteroid interactions. The overall goal of the PPG remains to increase understanding of the pathophysiology and potential treatments of these three important pathogens. The proposed studies will continue to use normal human enteroids or colonoids, grown on monolayers, and extend this through co-culture to make the model more closely resemble human intestine. This will be accomplished by expanding the co-culture models to a) establish monolayers in a 3D scaffold matrix to facilitate epithelial-immune cell interaction and immune cell movement; b) incorporate macrophages, neutrophils and dendritic cells as well as to develop a new co-culture system that includes intraepithelial γδ +T lymphocytes (IEL); c) apply an enriched M cell model; d) expose the co-culture model to flow-based shear stress as occurs normally in the intestine; e) determine the contribution of an anaerobic environment. As during the past funding period, examination of pathophysiologic aspects common to the diseases studied will serve to integrate the projects. In addition to pathogen specific pathophysiologic aspects, these include the role of mucins, bacterial proteases called SPATES, enterotoxins, secreted cytokines and involvement of co-cultured innate immune cells.

人类肠样模型中大肠杆菌和志贺氏菌感染的发病机制总体描述/摘要项目补助金 (PPG) 资助计划的首次更新是为了继续了解志贺氏菌及其两种致病型引起的腹泻的病理生理学和干预方法的鉴定致腹泻性大肠杆菌（肠聚集性和产肠毒素性大肠杆菌）已取得实质性进展。来自马里兰大学研究人员的密切合作和综合专业知识的成果，约翰·霍普金斯大学和弗吉尼亚大学正在使用正常人类肠样的共享模型和暴露于肠道细菌病原体的结肠样单层细胞以及人肠样单层细胞共- 研究人员小组在国际上与肠道中存在的先天吞噬细胞一起培养。在肠道疾病以及微生物学、胃肠病学、分子生理学、儿科传染病、细胞生物学、分子发病机制和粘膜免疫学。通过正式的方式进行互动并以极好的协作方式共享技术和科学信息每月举行面对面或基于 Zoom 的会议、频繁的电话或电子通讯以及拜访实验室，后者是马里兰大学和约翰霍普金斯大学研究人员互动的标准形式。肠类和免疫学核心开发了新的工具和方法，并迅速转移到正如预期的那样，项目研究人员促成了 PPG 的大量合作出版物。随着传播肠类的方法和试剂的转让、生产单层细胞、细菌肠样模型的开发以及与免疫细胞共培养物的开发 PPG 的总体目标仍然是增加病原体与肠类相互作用的新见解。了解这三种重要病原体的病理生理学和潜在治疗方法。拟议的研究将继续使用在单层上生长的正常人类肠类或类结肠，并扩展这将通过共培养使模型更加类似于人类肠道来实现。将共培养模型扩展到 a) 在 3D 支架基质中建立单层以促进上皮免疫细胞相互作用和免疫细胞运动；b) 还包括巨噬细胞、中性粒细胞和树突状细胞开发包含上皮内 γδ + T 淋巴细胞 (IEL) 的新共培养系统； M 细胞模型；d) 将共培养模型暴露于肠道中通常发生的基于流动的剪切应力；与过去的资助期间一样，确定厌氧环境的贡献。除了所研究的疾病的共同病理生理学方面之外，还将有助于整合这些项目。病原体特定的病理生理学方面，包括粘蛋白、称为细菌蛋白酶的作用 SPATES、肠毒素、分泌细胞因子以及共培养先天免疫细胞的参与。