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: 10245283
• 负责人: Christopher Eldridge Sims
• 金额: $ 69.93万
• 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 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10245283
• 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; 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; dietary; 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.

项目摘要 人类肠道是一个杰出的器官，可存储和分泌多种激素 来自肠内分泌（EEC）细胞。这些激素在调节人类方面起关键作用 喂养行为和饱腹感及其失调导致暴饮暴食和许多其他 代谢障碍。由于这些原因，在治疗市场中需要 精确概括体内肠的生理学的体外肠EEC细胞平台。 为了满足这一需求，早期生物技术公司Altis Biosystems Inc.已有 与学术实验室的科学家合作，开发了一种新颖的，主要的茎细胞 - 基于体外肠模型（称为复制）。我们已经完成了SBIR I阶段计划 通过优化富集肠球菌蛋白（EC）细胞的复制平台，EEC的亚型 细胞，并增加屏障完整性。我们已经开发了一种简单但有效的方法 与起始培养条件相比，显着增加了EEC细胞的形成。 我们已经研究了向EEC谱系强迫分化的信号分子 分配，5-羟色胺的量化测定以及研究通过和供体变化。我们 用EC的小尺度复合屏幕验证了平台 细胞。在I阶段SBIR中所有提议的里程碑均已完成，从而提供了固体 此II阶段SBIR应用的基础。该第二阶段提案的重点是继续 EEC组的优化以满足高通量筛查的市场需求 测定。除EC细胞外，我们还将我们的研究扩展到其他重要的亚型， 肠内分泌L细胞，分泌GLP-1和PYY，以响应食物的摄入。 其他特征将集中于96孔内细胞行为的均匀性 格式。在使用之前，将确认低井到孔和板到板的变化 测定平台。潜在的区域，性别和年龄的变化将由 测试来自5个供体和所有6个肠道的干细胞。平台将是 通过筛选大型代谢化合物库来验证。性能特征 然后，将在我们内部测定法与运送到的套件的比较中评估该平台 合作实验室。