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.

项目摘要：维持人类肠道健康的生理学是药物的重点。研究模型缺乏药物/营养滥用，异生元，毒理学和临床前效率受到阻碍准确，可重现且易于使用细胞培养模型来评估此类主题。由于这些原因，强烈需要更好地体外模型来概括人类肠道的疾病状态，并更好的药物发现和验证平台。为了满足这一需求Altis Biosystems Inc.，早期舞台生物技术公司将与北卡罗来纳大学的科学家合作教堂山以开发可生成并应用于其肠道的新颖，快速筛选的细胞基于干细胞的技术称为Repligut™，它是人类上皮的单层器官供体的小肠或结肠，并包含所有增殖剂和分化的细胞类型在体内发现。这一阶段I研究将1）开发并优化新的方法以生成鲁棒通过转基因或CRISPR-GENE编辑基于荧光蛋白的细胞系，2）创建和验证两个可筛分细胞系报告与肠道健康和疾病相关的关键特性 - 上皮屏障功能和注射，以及3）执行概念验证的小型复合屏幕展示实用性并为扩大发展提供理由。这些可筛选的单元将是在创建可筛选细胞和遗传疾病的II期期间以事件目标开发 Repligut™上的型号，可以适应高通量和高内心筛选基础架构。这学术界，Altis和NIH之间的合作是进一步发展的理想机会 Repligut™用于将这种高度生理文化和筛查平台转换为一个不发达的市场。