Developing an optimized cell based platform for assays of the gastrointestinal enteroendocrine system

开发用于胃肠道内分泌系统检测的优化细胞平台

• 批准号: 10080388
• 负责人: Christopher Eldridge Sims
• 金额: $ 71.3万
• 依托单位: ALTIS BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10080388
• 关键词: Age; Analytical Chemistry; Automobile Driving; Biological Assay; Biological Models; Biotechnology; Blood Circulation; Cardiovascular Diseases; Cell Line; Cell model; Cells; Cellular Assay; Characteristics; Communities; Complex; Data; Degenerative polyarthritis; Desire for food; Diabetes Mellitus; Diagnosis; Diet; Disease; Eating; Enteral; Enterochromaffin Cells; Enteroendocrine Cell; Feeding Patterns; Feeding behaviors; Food; Foundations; Funding; Goals; Health; Healthcare Systems; Hormone secretion; Hormones; Human; Hyperphagia; In Vitro; Industry; Ingestion; Intestinal Hormones; Intestines; Journals; L Cells; Laboratories; Legal patent; Letters; Libraries; Manuscripts; Measurement; Metabolic; Metabolic Diseases; Metabolism; Methods; Mission; Modeling; Molecular; Neuraxis; Neurotransmitters; Nutrient; Obesity; Organ; Peptide YY; Performance; Peripheral; Pharmacologic Substance; Phase; Physiological; Physiological Processes; Physiology; Play; Process; Production; Productivity; Recreation; Regulation; Reproducibility; Research; Role; Satiation; Scientist; Serotonin; Services; Ships; Signaling Molecule; Small Business Innovation Research Grant; Solid; Stroke; System; Techniques; Testing; Therapeutic; Tissues; Variant; Well in self; Work; base; blood glucose regulation; cell behavior; cell type; cellular engineering; comorbidity; cost; design; gastrointestinal; gastrointestinal system; genetically modified cells; glucagon-like peptide; high standard; high throughput screening; hormone regulation; human model; improved; in vitro Model; in vivo; innovation; interest; intestinal epithelium; monoamine; monolayer; neoplastic cell; neural network; novel; programs; relating to nervous system; response; screening; sex; stem cell differentiation; stem cells

Project Summary The human intestine is a remarkable organ which stores and secretes a variety of hormones from enteroendocrine (EEC) cells. These hormones play critical roles in regulating human feeding behavior and satiety, and their dysregulation leads to overeating and a host of other metabolic disorders. For these reasons, there is a need in the therapeutics marketplace for in vitro intestinal EEC cell platform that precisely recapitulates the physiology of in vivo intestines. To meet this need, Altis Biosystems Inc., an early stage biotechnology company, has collaborated with scientists at academic laboratories to develop a novel, primary-stem-cells- based, in vitro intestinal model (termed RepliGut). We have finished the SBIR Phase I program by optimizing the RepliGut platform to enrich enterochromaffin (EC) cells, a subtype of EEC cells, and increase barrier integrity. We have developed a simple but efficient method that significantly increases the formation of EEC cells compared with the starting culture conditions. We have investigated signaling molecules for forced differentiation towards EEC lineage allocation, quantified assays for serotonin, and investigated passage and donor variation. We validated the platform with a small-scale compound screen for serotonin secretion from EC cells. All proposed milestones in the Phase I SBIR were accomplished, thus providing a solid foundation for this Phase II SBIR application. The focus of this Phase II proposal is to continue the optimization of EEC formation to meet the market needs for high-throughput screening assays. Besides EC cells, we will extend our research to the other important subtype, enteroendocrine L-cells, which secrete GLP-1 and PYY in response to the ingestion of food. Additional characterizations will be focused on the uniformity of cell behaviors within a 96-well format. Low well-to-well and plate-to-plate variation will be confirmed before use as a cellular assay platform. Potential regional-, sex- and age-based variations will be further investigated by testing stem cells derived from 5 donors and all 6 sections of intestine. The platform will be validated by screening a large library of metabolic compounds. Performance characteristics of the platform will then be evaluated in comparisons of our in-house assays vs. kits shipped to collaborating laboratories.

项目总结