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Role of Mucus in EHEC colonization of human colonoids and intestinal disease devleopment

粘液在肠出血性大肠杆菌定植于人类结肠和肠道疾病发展中的作用

基本信息

批准号：
9351785
负责人：
OLGA KOVBASNJUK
金额：
$ 14.3万
依托单位：
JOHNS HOPKINS UNIVERSITY
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：
2016-09-23 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9351785
关键词：
Adherent Culture Adopted Affect Animal Model Antibiotics Apical Bacteria Binding Biological Assay Biopsy Cell Communication Cell Line Cessation of life Cleaved cell Clinical Data Colon Complex Data Development Diarrhea Disease Distal Energy-Generating Resources Enteral Enzymes Epithelial Epithelial Attachment Epithelial Cells Epithelium Escherichia coli EHEC Escherichia coli Infections Face Flagella Goals Grant Hemolytic-Uremic Syndrome Hemorrhagic colitis Hospitalization Human In Vitro Infection Intestinal Diseases Intestines Life Mediating Medical Modeling Molecular Mucin-2 Staining Method Mucins Mucous body substance Oryctolagus cuniculus Pathogenesis Pathology Patients Phase Play Production Risk Role Septic Toxemia Shiga Toxin Site Staging Stem cells Surface Systemic disease Technology Testing Thick Time Toxin Virulence Virulence Factors Work adult stem cell base colon cancer cell line effective therapy enteric pathogen foodborne human stem cells human subject improved individual patient insight intestinal epithelium microbial monolayer mutant new therapeutic target novel novel therapeutic intervention research study stem cell biology sugar therapeutic target uptake

项目摘要

Summary The proposed work will advance understanding of enteric disease caused by food-borne, Shiga-toxin- producing enterohemorrhagic E. coli (EHEC) constitute the major US cause of life-threatening hemorrhagic colitis and hemolytic uremic syndrome (HUS). While shiga toxin 2a provides the main virulence factor for intestinal and extra-intestinal disease manifestations, clinical data strongly suggest that Stx-toxemia occurs early in illness and is short-lived. We thus focus on early EHEC-host interactions, especially the complex interactions between EHEC and intestinal mucus. Intestinal mucus represents the first line of defense against enteric pathogens and is the first site for the interaction of EHEC with the human colonic epithelium. Our exciting preliminary data suggest that mucus play an important role in EHEC colonization of the human colon. These data were obtained using a novel model of EHEC infection, termed human colonoids, which are primary colonic epithelial cultures derived from adult stem cells isolated from biopsies of healthy donors. Upon differentiation, human colonoids produce a thick layer of mucus similar to that which is normally present in the human colon. EHEC infection results in the destruction of the mucus layer allowing EHEC to gain access to the colonocyte surface. These data suggest that human colonoids recapitulate EHEC infection and provide unique insights into pathogenesis that differ from other studies performed in human colon cancer cell lines, allowing improved appreciation of the roles of virulence factors and assessment of novel therapeutic targets. We hypothesize that EHEC interaction with colonic mucus is a necessary step in initial colonization of colonic epithelium leading to Stx production and full manifestation of intestinal disease. To test this hypothesis, we propose to use two complementary models of EHEC infection: human colonoids and the rabbits, which is the best animal model that can recapitulate EHEC-induced human intestinal pathologies, including hemorrhagic colitis, to achieve the following Aims: 1. Determine whether mucus degradation by EHEC is necessary to colonize the human epithelium. 2. Determine whether direct interaction between EHEC flagella and mucin(s) provides an initial anchor for EHEC-colonocyte attachment. 3. Determine whether the interaction of EHEC with mucus affects the production and release of the major EHEC virulence factor, Stx2a. The results gained from the proposed experiments will further elucidate the molecular mechanisms for EHEC interactions with human colonic epithelium, particularly the mucus layer, and establish the role of mucus in EHEC pathogenesis.

总结