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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要人体肠道是储存和分泌多种激素的重要器官来自肠内分泌（EEC）细胞。这些激素在调节人体内分泌方面发挥着重要作用喂养行为和饱腹感，它们的失调会导致暴饮暴食和许多其他问题代谢紊乱。由于这些原因，治疗市场需要体外肠道 EEC 细胞平台，精确再现体内肠道的生理学。为了满足这一需求，早期生物技术公司 Altis Biosystems Inc. 与学术实验室的科学家合作开发一种新型的原代干细胞基于体外肠道模型（称为 RepliGut）。我们已经完成了SBIR第一阶段计划通过优化 RepliGut 平台来富集肠嗜铬 (EC) 细胞（EEC 的一种亚型）细胞，并增加屏障完整性。我们开发了一种简单但有效的方法与起始培养条件相比，EEC 细胞的形成显着增加。我们研究了强制分化为 EEC 谱系的信号分子分配，血清素定量分析，并研究传代和供体变异。我们通过小规模复合筛选验证了 EC 分泌血清素的平台细胞。第一阶段 SBIR 中所有拟议的里程碑均已完成，从而提供了坚实的基础第二阶段 SBIR 应用的基础。第二阶段提案的重点是继续优化EEC形成以满足高通量筛选的市场需求化验。除了EC细胞之外，我们还将把我们的研究扩展到其他重要的亚型，肠内分泌 L 细胞，响应食物的摄入而分泌 GLP-1 和 PYY。其他表征将集中于 96 孔内细胞行为的均匀性格式。在用作细胞之前，将确认孔与孔之间以及板与板之间的低变异性分析平台。潜在的地区、性别和年龄差异将进一步调查测试来自 5 名捐赠者和所有 6 个肠道部分的干细胞。该平台将是通过筛选大型代谢化合物库进行验证。性能特点然后将通过我们的内部检测与运送到的试剂盒的比较来评估该平台合作实验室。