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：
10745560
负责人：
MARK DONOWITZ
金额：
$ 12.94万
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/10745560
关键词：
3-Dimensional Acceleration Address 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 Epithelium 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 Macrophage Maryland Measurement Methodology Methods Microbiology Modeling Molecular Mucins Mucous Membrane Organism Pathogenesis Peptide Hydrolases Phagocytes Physiology Porosity 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 cell motility cytokine diarrheal disease eVisit enteric pathogen enteroaggregative Escherichia coli enterotoxigenic Escherichia coli experience gut inflammation human model improved insight intraepithelial meetings 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.

大肠杆菌和志贺氏菌感染的发病机理总体描述 /摘要资助计划项目赠款（PPG）的第一个续约是继续在理解病理生理学和识别志贺氏菌引起的腹泻和两种病原体的方法的鉴定腹泻性大肠杆菌（肠ofgreg型和肠毒素大肠杆菌）。取得的实质进展已有由马里兰州大学的调查人员的密切合作和联合专业知识造成约翰·霍普金斯大学和弗吉尼亚大学使用共享的正常人肠内模型暴露于肠细菌病原体以及人类肠片单层共同的结肠单层。用肠道中的先天吞噬细胞培养。一组调查员在国际上拥有肠道疾病以及微生物学，胃肠病学，分子生理学的著名经验，小儿传染病，细胞生物学，分子发病机理和粘膜免疫学。这个财团通过公式进行了互动，并以极好的协作方式共享技术和科学信息每月面对面或基于缩放的会议，经常电话或电子通信以及访问实验室，后者是U马里兰州和JHU调查人员相互作用的标准形式。这肠和免疫学核心开发了新的工具和方法，这些工具和方法已迅速转移到项目调查人员，导致PPG的大量合作出版物。不出所料，随着方法和试剂的传播，进步是逐步的单层，开发细菌 - 侵入模型，并开发具有免疫细胞的共培养导致对病原体 - 输入相互作用的新见解。 PPG的总体目标仍在增加了解这三种重要病原体的病理生理学和潜在治疗。拟议的研究将继续使用正常的人类肠toi或结肠素，在单层上生长，并扩展通过共培养使该模型更加类似于人类肠道。这将通过将共培养模型扩展到a）在3D支架矩阵中建立单层细胞相互作用和免疫细胞运动； b）还结合巨噬细胞，中性粒细胞和树突状细胞为了开发一个新的共培养系统，其中包括上皮内γδ +T淋巴细胞（IEL）； c）应用富集 M细胞模型； d）将共培养模型暴露于肠中通常发生的基于流动的剪切应力； e）确定厌氧环境的贡献。与过去的资金期一样研究疾病常见的病理生理方面将有助于整合项目。此外病原体特定的病理生理方面，其中包括粘蛋白的作用，细菌蛋白酶称为 Spates，肠毒素，分泌的细胞因子和共同培养的先天免疫球的参与。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Protocol for measuring transcytosis and recycling of IgG in intestinal epithelial Caco-2 cells and primary human intestinal organoids.

DOI：
10.1016/j.xpro.2023.102335
发表时间：
2023-05-25
期刊：
STAR PROTOCOLS
影响因子：
0
作者：
Maeda, Keiko;Gwilt, Katlynn Bugda;Schmieder, Stefanie S.;Zachos, Nicholas C.;Lencer, Wayne I.
通讯作者：
Lencer, Wayne I.

Co-culturing Human Intestinal Enteroid Monolayers with Innate Immune Cells.

人肠肠样单层细胞与先天免疫细胞共培养。

DOI：
10.1007/978-1-0716-3076-1_16
发表时间：
2023
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Staab,JanetF;Lemme-Dumit,JoseM;Latanich,Rachel;Pasetti,MarcelaF;Zachos,NicholasC
通讯作者：
Zachos,NicholasC

The role of the minor colonization factor CS14 in adherence to intestinal cell models by geographically diverse ETEC isolates.

DOI：
10.1128/msphere.00302-23
发表时间：
2023-10-24
期刊：
MSPHERE
影响因子：
4.8
作者：
Smith, Emily M.;Papadimas, Antonia;Gabor, Caitlin;Cooney, Ceanna;Wu, Tao;Rasko, David;Barry, Eileen M.
通讯作者：
Barry, Eileen M.

Human Breast Milk Enhances Intestinal Mucosal Barrier Function and Innate Immunity in a Healthy Pediatric Human Enteroid Model.

DOI：
10.3389/fcell.2021.685171
发表时间：
2021
期刊：
Frontiers in cell and developmental biology
影响因子：
5.5
作者：
Noel G;In JG;Lemme-Dumit JM;DeVine LR;Cole RN;Guerrerio AL;Campbell JD;Kovbasnjuk O;Pasetti MF
通讯作者：
Pasetti MF

Highly-conserved regulatory activity of the ANR family in the virulence of diarrheagenic bacteria through interaction with master and global regulators.