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

• 批准号: 8668192
• 负责人: MARK DONOWITZ
• 金额: $ 9.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-24 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8668192
• 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 Targeting; 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

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.

描述(由申请人提供)：由于缺乏可用的和易于使用的人体肠道模型，对人类肠道生理学、腹泻疾病的病理生理学和止泻药物治疗的发展的了解一直受到阻碍。最近，两种方法(来自LGR5阳性的肠干细胞和来自分离的完整肠隐窝)被开发出来，以使人类小肠类器官形成正常外观的绒毛/隐窝轴，其中包含所有正常出现的上皮细胞(钠吸收细胞、氯离子分泌细胞、杯状细胞、肠内分泌细胞和Paneth细胞)。我们建议对这两种类型的人类小肠器官进行比较，并利用它们来促进对正常肠道消化生理和几种重要人类腹泻疾病的病理生理的了解。我们将1)比较这两种类型的人类小肠类器官a)它们如何随着时间的推移发展极性和分化，并确定参与钠吸收和氯分泌的转运蛋白的分布和随器官分化的变化，这些过程在腹泻中变得异常。B)在模拟正常消化生理和在腹泻中变得异常的基础和调节条件下，测试对腹泻的病理生理重要的两种转运蛋白NHE3和CFTR在有机体中的功能。C)利用有机物质和对钠吸收和氯分泌的了解，增加对霍乱毒素、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。有机化合物的其他重要潜在用途是a)基于NHE3和CFTR荧光分析的高通量药物筛选，以及b)个性化药物，因为有机化合物可以从个体患者身上开发出来。