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

通过基因编辑和转基因产生可药物筛选的原代人肠上皮

基本信息

批准号：
10009998
负责人：
Christopher Eldridge Sims
金额：
$ 24.59万
依托单位：
ALTIS BIOSYSTEMS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2021-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10009998
关键词：
Academia Acute Affect Age Apical Bacterial Toxins Biological Assay Biotechnology CRISPR/Cas technology Cell Culture Techniques Cell Differentiation process Cell Line Cells Childhood Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Colon Cystic Fibrosis Defect Dehydration Development Diet Disease Disease model Drug Screening Drug toxicity Elderly Environmental Risk Factor Epithelial Epithelium Etiology Gastrointestinal Diseases Gastrointestinal tract structure Gender Gene Transfer Techniques Genes Genetic Diseases Goals Health Healthcare Systems Human Hydration status Hydrogels Impairment Inflammation Inflammatory Inflammatory Bowel Diseases Infrastructure Intestines Ion Transport Measures Methods Mission Modeling Mucous body substance Natural regeneration North Carolina Organ Donor Organoids Outcome Pathologic Patients Pharmaceutical Preparations Pharmacologic Substance Phase Physiological Physiology Pre-Clinical Model Property Proteins Race Radiation exposure Reporting Reproducibility Research Scientist Small Intestines Source Technology Tissues Toxicology Translations United States National Institutes of Health Universities Validation Virus Xenobiotics base cell type cost design diarrheal disease driving force drug candidate drug discovery epithelial stem cell gastrointestinal gastrointestinal epithelium genetic makeup high throughput screening human disease in vitro Model in vivo intestinal epithelium microbiota monolayer new therapeutic target novel novel therapeutics nutrient absorption phase 1 study preclinical efficacy prevent productivity loss response screening stem cells

项目摘要

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.

项目概要：维持人体肠道的健康生理是制药模型研究的一个重点。药物/营养吸收、异生物质、毒理学和临床前疗效因缺乏准确、可重复且易于使用的细胞培养模型来评估此类主题。强烈需要更好的体外模型来概括人类肠道的疾病状态，并且为了满足这一需求，Altis Biosystems Inc. 成立了一个早期的更好的药物发现和验证平台。 stage 生物技术公司将与北卡罗来纳大学的科学家合作教堂山开发新型、可快速筛选的细胞，这些细胞可以在肠道上生成并应用于肠道基于干细胞的技术，称为 RepliGut™，它是源自人类上皮细胞的单层器官捐献者的小肠或结肠，包含所有增殖和分化的细胞类型该第一阶段研究将 1) 开发和优化新方法以产生稳健的结果。通过转基因或 CRISPR 基因编辑基于荧光蛋白的细胞系，2) 创建并验证两个可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) 进行概念验证小规模复合筛选证明这些可筛选细胞的实用性并为扩大开发提供理由。在第二阶段开发的最终目标是创造可筛选细胞和遗传疾病 RepliGut™ 上的模型可以适应高通量和高内涵的筛选基础设施。学术界、Altis 和 NIH 之间的合作为进一步发展提供了理想的机会 RepliGut™ 将这种高度生理性的培养和筛选平台转化为市场不发达。