Human Intestinal Organoids: Pre-Clinical Models of Non-Inflammatory Diarrhea

人类肠道类器官：非炎症性腹泻的临床前模型

基本信息

批准号：
8414961
负责人：
MARK DONOWITZ
金额：
$ 49.3万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-24 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8414961
关键词：
APPBP2 gene Adult Adverse effects Affect Aging Animal Model Anions Area Back Biological Assay Biological Models Biopsy Brush Border Carrier Proteins Cause of Death Cell Culture Techniques Cessation of life Child Childhood Cholera Cholera Toxin Clinical Trials Cystic Fibrosis Transmembrane Conductance Regulator Dehydration Developing Countries Development Diarrhea Digestive Physiology Disease Drug Delivery Systems Electrolytes Epigenetic Process Epithelial Epithelial Cells Escherichia coli EHEC Failure Fluorescence Functional disorder Genetic Polymorphism Goals Growth Homeostasis Human Human Development In Vitro Individual Infant Inflammatory International Intestines Ion Transport Ions Location Medicine Methods Modeling Modification Morbidity - disease rate NHE1 NHE2 Na(+)-K(+)-Exchanging ATPase Operative Surgical Procedures Oral Organoids Patients Pharmaceutical Preparations Pharmacotherapy Physiological Physiology Pluripotent Stem Cells Population Pre-Clinical Model Preclinical Drug Evaluation Proteins Regulation Rehydration Solutions Reporting Reproducibility Rotavirus Sodium Chloride Staging Stem cells Techniques Testing Time Tissues Translational Research Transport Process Villus Water absorption aged base cell bank cell type drug development drug metabolism human disease intestinal crypt intestinal epithelium mortality novel secretion process species difference

项目摘要

DESCRIPTION (provided by applicant): Understanding of human intestinal physiology, pathophysiology of diarrheal diseases and development of anti-diarrheal drug therapy has been held back by lack of an available and easy to use model of human intestine. Recently two methods (from LGR5 positive intestinal stem cells and from isolated intact intestinal crypts) have been developed to allow development of human small intestinal organoids that form normal appearing villus/crypt axes which contain all normally occurring epithelial cells (Na absorptive, Cl secretory, goblet, enteroendocrine and Paneth). We propose to compare the two types of human small intestinal organoids and use them to advance understanding of normal intestinal digestive physiology and the pathophysiology of several important human diarrheal diseases. We will 1) compare the two types of human small intestinal organoids for a) how they develop polarity and differentiation over time and define the distribution and changes with organoid differentiation of transport proteins involved in Na absorption and Cl secretion, processes which become abnormal in diarrhea. b) Test the function in the organoids of two transport proteins that are important for the pathophysiology of diarrhea, NHE3 and CFTR under basal and regulated conditions which mimic normal digestive physiology and become abnormal in diarrhea. c) Use the organoids and understanding of Na absorption and Cl secretion to increase understanding of three important human diarrheal diseases caused by cholera toxin, rotavirus and enterohemorrhagic E. coli. Other important potential uses of the organoids are for a) high throughput drug screening based on the fluorescence assays for NHE3 and CFTR and b) personalized medicine, since the organoids can be developed from individual patients. PUBLIC HEALTH RELEVANCE: Understanding of human intestinal physiology, pathophysiology of diarrheal diseases and development of anti-diarrheal drug therapy has been held back by lack of an available and easy to use model of human intestine. There are now two methods allowing growth of intestinal organoids which consist of all normally present intestinal epithelial cells organized into a crypt/villus axis. We will compare these two methods as to how they polarize, differentiate, and the location of transport proteins the abnormal function of which contribute to the pathophysiology of diarrhea (NHE3 and CFTR); evaluate the function of these transport proteins under normal conditions; and use the organoid models to study pathophysiology, including function of these transport proteins, of three important diarrheal diseases, cholera, rotavirus and enterohemorrhagic E. coli.

描述（由申请人提供）：由于缺乏可用且易于使用的人类肠道模型，对人类肠道生理学、腹泻疾病病理生理学的理解以及抗腹泻药物疗法的开发受到阻碍。最近开发了两种方法（来自 LGR5 阳性肠干细胞和来自分离的完整肠隐窝），以允许开发人类小肠类器官，形成正常的绒毛/隐窝轴，其中包含所有正常存在的上皮细胞（Na 吸收、Cl 分泌、高脚杯、肠内分泌和潘氏）。我们建议比较两种类型的人类小肠类器官，并利用它们来加深对正常肠道消化生理学和几种重要人类腹泻疾病病理生理学的了解。我们将 1) 比较两种类型的人类小肠类器官，a) 它们如何随时间发展极性和分化，并定义参与 Na 吸收和 Cl 分泌（腹泻时变得异常的过程）的转运蛋白的分布和类器官分化变化。 b) 在模拟正常消化生理学并在腹泻中变得异常的基础和调节条件下测试对腹泻病理生理学重要的两种转运蛋白 NHE3 和 CFTR 的类器官功能。 c) 利用类器官以及对 Na 吸收和 Cl 分泌的了解，加深对由霍乱毒素、轮状病毒和肠出血性大肠杆菌引起的三种重要人类腹泻疾病的了解。类器官的其他重要潜在用途包括：a) 基于 NHE3 和 CFTR 荧光分析的高通量药物筛选；b) 个性化医疗，因为类器官可以从个体患者身上开发出来。公共卫生相关性：由于缺乏可用且易于使用的人体肠道模型，对人类肠道生理学、腹泻疾病病理生理学的理解以及抗腹泻药物治疗的开发受到阻碍。现在有两种方法允许肠道类器官的生长，这些类器官由组织成隐窝/绒毛轴的所有正常存在的肠上皮细胞组成。我们将比较这两种方法如何极化、分化以及功能异常的转运蛋白的定位。有助于腹泻的病理生理学（NHE3 和 CFTR）；评估这些转运蛋白在正常条件下的功能；并利用类器官模型研究霍乱、轮状病毒和肠出血性大肠杆菌这三种重要腹泻疾病的病理生理学，包括这些转运蛋白的功能。