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.

 整体U19应用程序的描述（由适用提供）：该提案将使用人类胃肠道器官作为模型系统。我们将将最新方法与胃肠道疾病研究的协作和协同研究方法相结合。我们将开发体外胃肠道模型，以复制人类肠道的生物学结构。这些组织将概括人类的生理学和疾病病理学，并结合对疾病和人类宿主反应至关重要的成分。我们的重点将跨越从基础研究（发病机理）到产品开发的管道。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辅助研究的模型系统。我们将通过以下特定目标推进该领域：目标1。标题：使用人类胃癌的幽门螺杆菌宿主感染的机制。研究项目1将使用新开发的胃癌系统研究幽门螺杆菌。提出的研究有可能提高我们对幽门螺杆菌改变胃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的作用的理解。目标2。标题：人肠癌，研究毒细菌病原体。研究项目II将使用人肠癌作为一种新型模型系统来研究两种产生大肠杆菌（STEC）的毒素肠道病原体，艰难梭菌和志贺毒素（STX）（STX）。拟议的研究有可能提高我们对艰难梭菌和STEC的致病过程的理解，并创建模型系统，以评估潜在的治疗剂。 AIM 3。标题：人类胃肠道和结肠镜作为胃肠道感染的4D模型。 Research Project III will develop ex vivo models to determine age-dependent and temporary GI epithelial responses to: a) H. pylori infection in human gastric organoids (HGOs) and gasroids and b) Clostridium difficile (CD) and Shiga toxin-producing Escherichia coli (STEC) infection and their major toxins in human colonoids (HCOs) and colonoids.目标4。技术传播。我们将促进并鼓励与目前隶属此提案的调查人员的教育和培训活动，通过其他名称奖项资助的调查人员以及一般的科学界成员。 AIM 5。器官工程核心。人体器官核心将开发报告基因线，其中荧光蛋白在特定的谱系中表达。 ShRNA敲低和CRISPR靶向线将导致缺乏特定谱系的人类胃肠道类器官，而四环素调节的系统将用于扩展特定的谱系。