Generating drug-screenable primary human intestinal epithelium by gene-editing and transgenesis

项目摘要： 维持人体肠道的健康生理是Pharma的一大重点。模型来研究 药物/营养素吸收、外源性物质、毒理学和临床前功效由于缺乏 准确，可重复，易于使用的细胞培养模型来评估这些主题。由于这些原因， 强烈需要更好的体外模型，其再现人肠道的疾病状态，并且 更好的药物发现和验证平台。为了满足这一需求，Altis Biosystems Inc.，早日 阶段生物技术公司，将与科学家在北卡罗来纳州大学在 查佩尔山开发新的，可快速筛选的细胞，可以产生，并施加到他们的肠道 基于干细胞的技术称为干肠™，这是一个单层的人上皮细胞来源于 器官供体的小肠或结肠，并且包含所有增殖和分化的细胞类型 在vivo中发现这项I期研究将1）开发和优化新的方法， 2）通过转基因或CRISPR基因编辑创建并验证两种基于荧光蛋白的细胞系， 筛选细胞系以报告与肠道健康和疾病相关的关键特性-上皮屏障 功能和炎症，以及3）进行概念验证的小规模化合物筛选， 展示实用性并为扩展开发提供理论依据。这些可筛选的细胞将 第二阶段的最终目标是创造可筛选的细胞和遗传疾病 适用于高通量和高内容筛选基础设施的<$Gut ™模型。这 学术界，Altis和NIH之间的合作是进一步发展的理想机会 <$Gut ™将这种高度生理性的培养和筛选平台转化为 欠发达市场

Project Summary: Maintaining healthy physiology of the human gut is a large focus of Pharma. Models to study drug/nutrient absorption, xenobiotics, toxicology, and preclinical efficacies are hampered by the lack of accurate, reproducible, and easy to use cell culture models to evaluate such topics. For these reasons, there is a strong need for better in vitro models that recapitulate disease states of the human gut, and better platforms for drug discovery and validation. To meet this need Altis Biosystems Inc., an early stage biotechnology company, will collaborate with scientists at the University of North Carolina at Chapel Hill to develop novel, rapidly screenable cells that can be generated on, and applied to, their gut stem cell-based technology called RepliGut™, which is a monolayer of human epithelium derived the small intestine or colon of organ donors, and contains all of the proliferative and differentiated cell types found in vivo. This Phase I study will 1) develop and optimize novel methods to for generating robust fluorescent protein based cell lines by transgenesis or CRISPR-gene editing, 2) create and validate two screenable cell lines to report key properties associated with gut health and disease - epithelial barrier function and inflammation, and 3) perform a proof-of-concept small scale compound screen to demonstrate utility and provide rationale for expanded development. These screenable cells will be developed with the eventual goal during Phase II of creating screenable cells and genetic disease model on RepliGut™ that can fit into high-throughput and high-content screening infrastructure. This collaboration between academia, Altis, and the NIH represents an ideal opportunity to further develop RepliGut™ for translation of this highly physiologic culture and screening platform into an underdeveloped marketplace.