Immunomodulatory role of Pic in EAEC Pathogenesis

Pic 在 EAEC 发病机制中的免疫调节作用

• 批准号: 10745565
• 负责人: Fernando Ruiz
• 金额: $ 6.47万
• 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 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10745565
• 关键词: Age; Award; Binding; Biological Assay; CD14 gene; Cell Line; Cell physiology; Cell secretion; Cell surface; Cells; Chemotaxis; Chronic; Citrobacter; Coculture Techniques; Collaborations; Colon; Communicable Diseases; Complex; Data; Developed Countries; Developing Countries; Development; Diarrhea; Disease model; Epithelial Cells; Epithelium; Escherichia coli; Escherichia coli Infections; Event; Exocytosis; Family; Flow Cytometry; Gastroenterologist; Gene Silencing; Generations; Genes; Genetic; Glycoproteins; Goblet Cells; Growth; Human; Immunologist; Immunology; Impaired cognition; Impairment; Infection; Inflammation; Inflammatory; Intervention; Intestinal Mucosa; Intestines; Lectin; Leukocytes; MAP3K7 gene; Macrophage; Maryland; Mediating; Microscopy; Modeling; Mucin 1 protein; Mucin-2 Staining Method; Mucins; Mucous body substance; Outcome; Pathogenesis; Penetration; Play; Process; Productivity; Property; Protease Inhibitor; Research Personnel; Role; Salmonella; Serine Protease; Serine Proteinase Inhibitors; Shigella; Shigella Infections; Shigella flexneri; Signal Pathway; Signal Transduction; Specialist; System; Therapeutic Intervention; Thinness; Traveler' s diarrhea; Universities; Virginia; Virulence Factors; WFDC2 gene; Work; diarrheal disease; enteric pathogen; enteroaggregative Escherichia coli; enterotoxigenic Escherichia coli; gut colonization; gut inflammation; immunoregulation; insight; knock-down; migration; monolayer; mouse model; multidisciplinary; neutrophil; pathogen; pathogenic Escherichia coli; pharmacologic; receptor; synergism; transcriptomics

Abstract Enteroaggregative E. coli (EAEC) is an important pathogen of traveler's diarrhea, diarrhea in industrialized countries and growth faltering in developing countries. In this application, we propose to study the role of highly prevalent serine proteases from the SPATE family in EAEC pathogenesis, and their relationship with chronic inflammatory enteropathy using a newly established human leukocyte-colonoid model. This application comprises a consortium among investigators at Johns Hopkins University (JHU), who have pioneered the development of the enteroid/colonoid model, and researchers at the University of Maryland (UMB) and the University of Virginia (UVA), who are leaders in the study of the pathogenesis of diarrheal disease caused by diarrhegenic E. coli and Shigella. We have successfully used the colonoid model to characterize relevant features of EAEC pathogenesis, highlighting the important role of the lectin-like serine protease Pic in intestinal colonization and inflammation. In this project will leverage the existence of the newly developed leukocyte- colonoid monolayer model to characterize the modulatory activity of Pic and other serine proteases in goblet cells, leukocytes and epithelial cells in EAEC pathogenesis. This project will comprise three Specific Aims. In Aim 1, we will characterize the effect of Pic in goblet cell function by obtaining mechanistic insights into Pic- mediated MUC2 exocytosis and by establishing the role of the goblet-cell secreted serine protease inhibitor WFDC2 in EAEC pathogenesis by using unbiased transcriptomics and pharmacological approaches. In Aim 2, we will characterize the role of SPATEs in EAEC-induced epithelial inflammation by obtaining mechanistic insights into MUC1-mediated epithelial inflammation in EAEC infection, employing gene silencing and pharmacological approaches. In Aim 3, we will characterize the immunomodulatory property of Pic lectin and mucinolytic activity on leukocyte functions in EAEC pathogenesis employing neutrophils- and macrophages- colonoids co-cultures along with state-of-the-art time-lapse microscopy, flow cytometry and Luminex-based assays. This work will continue to advance our understanding of EAEC pathogenesis, generating fundamental insights that will illuminate aspects of pathogenesis relevant to other enteric pathogens, and could open up new avenues for therapeutic intervention. We will work closely and synergistically with investigators in other projects and the Core components.

抽象的 EnteroAggregative E. Coli（EAEC）是旅行者腹泻的重要病原体，工业化的腹泻 发展中国家的国家和增长步履蹒跚。在此应用中，我们建议研究高度的作用 EAEC发病机理中Spate家族的普遍丝氨酸蛋白酶及其与慢性的关系 使用新建立的人类白细胞棒棒模型的炎症肠病。此应用程序 约翰·霍普金斯大学（JHU）的调查员之间的财团是一个率先的 开发肠道/结肠型模型，马里兰大学（UMB）的研究人员和 弗吉尼亚大学（UVA）是研究腹泻病发病机理的领导者 腹泻大肠杆菌和志贺氏菌。我们已经成功使用了结肠模型来表征相关的 EAEC发病机理的特征，突出了肠蛋白样丝氨酸蛋白酶PIC的重要作用 定植和炎症。在这个项目中，将利用新开发的白细胞的存在 结肠单层模型以表征PIC和其他丝氨酸蛋白酶的调节活性 EAEC发病机理中的细胞，白细胞和上皮细胞。该项目将构成三个具体目标。在 AIM 1，我们将通过获得机械洞察到PIC-来表征PIC在Goblet细胞功能中的效果。 介导的MUC2胞吐作用，并通过建立杯状细胞分泌丝氨酸蛋白酶抑制剂的作用 通过使用公正的转录组学和药理方法，在EAEC发病机理中的WFDC2。在AIM 2中， 我们将通过获得机械性来表征SPATE在EAEC诱导的上皮炎症中的作用 对EAEC感染中MUC1介导的上皮炎症的见解，采用基因沉默和 药理方法。在AIM 3中，我们将表征PIC凝集素的免疫调节特性和 使用中性粒细胞和巨噬细胞的EAEC发病机理中白细胞功能的粘蛋白水解活性 - 结石型共培养以及最新的延时显微镜，流式细胞术和基于Luminex 测定。这项工作将继续提高我们对EAEC发病机理的理解，产生基本 可以阐明与其他肠道病原体有关的发病机理方面的见解，并且可以开放新的 治疗干预的途径。我们将与其他项目中的调查人员紧密和协同合作 和核心组件。