Intestinal Organoids as a model system for studying enteric disease

肠类器官作为研究肠道疾病的模型系统

• 批准号: 8856069
• 负责人: Alison A. Weiss
• 金额: $ 93.13万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8856069
• 关键词: Address; Advanced Development; Age; Animal Model; Award; Basic Science; Biological; Biological Models; Carcinogens; Clostridium difficile; Clustered Regularly Interspaced Short Palindromic Repeats; Communicable Diseases; Communities; Disease; Educational Activities; Engineering; Enteral; Epithelial; Epithelium; Escherichia coli; Escherichia coli EHEC; Escherichia coli Infections; Evaluation; Funding; Gastrointestinal Diseases; Goals; Grant; Helicobacter Infections; Helicobacter pylori; Human; Immune response; In Vitro; Infection; Intestines; Methodology; Modeling; Molecular; Morbidity - disease rate; Organoids; Pathogenesis; Pathology; Pathway interactions; Patient Rights; Physiology; Process; Proteins; Reagent; Recruitment Activity; Reporter; Research; Research Personnel; Research Project Grants; Risk Factors; Role; Stomach; Structure; System; Technology; Tetracyclines; Therapeutic; Time; Tissues; Toxin; Training Activity; age related; enteric pathogen; gastrointestinal; gastrointestinal infection; human disease; insight; interest; meetings; member; mortality; novel; novel therapeutics; pathogen; product development; public health relevance; research and development; response; small hairpin RNA; stem cell fate

 DESCRIPTION OF THE OVERALL U19 APPLICATION (provided by applicant): This proposal will use human gastrointestinal organoids as a model system. We will integrate state-of-the-art methodologies, with collaborative and synergistic research approaches to the study of gastrointestinal diseases. We will develop in vitro gastrointestinal models that replicate the biological structures of the human intestinal tract. These tissues will recapitulate human physiology and disease pathology, and incorporate components critical to disease and human host response. Our focus will span the pipeline from basic research (pathogenesis) to product development. For gastrointestinal pathogens that have no relevant animal models, another long-term goal is to develop model systems that can be used in IND-enabling studies. We will advance the field through the following Specific Aims: Aim 1. Title: Mechanisms of H. pylori-host infection using human gastric organoids. Research Project 1 will use the newly developed gastric organoid system to study Helicobacter pylori. The proposed studies have the potential to advance our understanding of the mechanisms by which H. pylori alters gastric stem cell fate, and advance our understanding of the role of H. pylori as a carcinogen. Aim 2. Title: Human Intestinal Organoids to Study Toxigenic Bacterial Pathogens. Research Project II will use human intestinal organoids as a novel model system to study two toxigenic enteric pathogens, Clostridium difficile and Shiga toxin (Stx) producing Escherichia coli (STEC). The proposed studies have the potential to advance our understanding of the pathogenic processes of C. difficile and STEC and create model systems for the evaluation of potential therapeutics. Aim 3. Title: Human gastroids and colonoids as 4D models of gastrointestinal infection. Research Project III will develop ex vivo models to determine age-dependent and temporal GI epithelial responses to: a) H. pylori infection in human gastric organoids (HGOs) and gastroids and b) Clostridium difficile (CD) and Shiga toxin-producing Escherichia coli (STEC) infection and their major toxins in human colonic organoids (HCOs) and colonoids. Aim 4. Technology dissemination. We will advance and encourage educational and training activities to investigators currently affiliated with this proposal, investigators funded through other NAMSED awards, and interested members of the scientific community in general. Aim 5. Organoid Engineering Core. The human organoid core will develop reporter lines in which fluorescent proteins are expressed in specific lineages. shRNA knockdown and CRISPR targeted lines will give rise to human gastro-intestinal organoids lacking specific lineages, and tetracycline-regulated systems will be used to expand specific lineages.

 U19 总体应用描述（由申请人提供）：该提案将使用人类胃肠道类器官作为模型系统。我们将把最先进的方法与协作和协同的研究方法相结合来研究胃肠道疾病。我们将开发复制人类肠道生物结构的体外胃肠道模型。这些组织将重现人类生理学和疾病病理学，并包含对疾病和人类宿主反应至关重要的成分。我们的重点将涵盖从基础研究（发病机制）到产品开发的整个流程。对于没有相关动物模型的胃肠道病原体，另一个长期目标是开发可用于IND研究的模型系统。我们将通过以下具体目标推进该领域的发展： 目标 1. 标题：利用人胃类器官感染幽门螺杆菌宿主的机制。研究项目1将利用新开发的胃类器官系统来研究幽门螺杆菌。拟议的研究有可能促进我们对幽门螺杆菌改变胃干细胞命运的机制的理解，并促进我们对幽门螺杆菌作为致癌物作用的理解。 目标 2。标题：研究产毒细菌病原体的人体肠道类器官。研究项目 II 将使用人类肠道类器官作为新型模型系统来研究两种产毒肠道病原体：艰难梭菌和产生志贺毒素 (Stx) 的大肠杆菌 (STEC)。拟议的研究有可能增进我们对艰难梭菌和 STEC 致病过程的理解，并创建用于评估潜在疗法的模型系统。 目标 3。标题：人类胃窦和结肠作为胃肠道感染的 4D 模型。研究项目 III 将开发离体模型，以确定年龄依赖性和时间性胃肠道上皮反应：a) 人胃类器官 (HGO) 和胃样中的幽门螺杆菌感染，b) 艰难梭菌 (CD) 和产志贺毒素大肠杆菌 (STEC) 感染及其在人结肠类器官 (HCO) 和类结肠中的主要毒素。 目标 4. 技术传播。我们将推动和鼓励对目前参与该提案的研究人员、通过其他 NAMSED 奖项资助的研究人员以及整个科学界感兴趣的成员开展教育和培训活动。 目标 5. 类器官工程核心。人类类器官核心将开发报告系，其中荧光蛋白以特定谱系表达。 shRNA敲低和CRISPR靶向细胞系将产生缺乏特定谱系的人类胃肠道类器官，而四环素调节系统将用于扩展特定谱系。