SBIR-STTR Award

The human intestine is a remarkable organ which stores 90% of the body’s important neurotransmitter, serotonin, in enteroendocrine cells (EECs). Serotonin and other EECs-secreted hormones play critical roles in regulating human feeding behavior and satiety, and their dysregulation leads to overeating and a host of other diseases. For these reasons, there is a need in the therapeutics marketplace for in vitro intestinal EECs platform that precisely recapitulates the physiology of in vivo intestines. 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 a novel, primary-stem-cells-based, in vitro intestinal model (termed RepliGut) that contains sufficient EECs for studying serotonin secretion. The platform will be designed with the eventual goal during Phase II of creating systems for assaying a variety of intestinal hormones in a high-content screening. The goal is to validate the RepliGut product and bring this technology to therapeutics market. The collaboration represents an ideal opportunity for the translation of an academic technology to the marketplace through the NIH sponsored SBIR program. In this Phase I SBIR, this collaboration will optimize the RepliGut 96- well platform for EECs cell lineage allocation, investigate cell variation, quantify serotonin secretion, and validate the platform with a small-scale compound screen for serotonin.

Public Health Relevance Statement:
The novel enteroendocrine cell-rich RepliGut platform will have broad applications in screening and validating therapeutics to manipulate feeding behavior and satiety which will revolutionize the treatment of many diseases in humans.

Project Terms:
Age; base; biobank; Biological Assay; Biological Models; Biotechnology; Cardiovascular Diseases; Cell Line; Cell Lineage; Cell model; Cell physiology; Cell secretion; cell type; Cells; Cellular Assay; Collaborations; Communities; Comorbidity; Complex; cost; Degenerative polyarthritis; demographics; design; Desire for food; Diabetes Mellitus; Diagnosis; Disease; Enteral; Enterochromaffin Cells; Enteroendocrine Cell; Epithelial; expectation; Feeding behaviors; Feeding Patterns; gastrointestinal system; Gastrointestinal tract structure; Gender; Goals; Health; Healthcare Systems; high throughput screening; hormone regulation; Hormone secretion; Hormones; Human; human model; Hyperphagia; improved; In Vitro; in vitro Model; in vivo; Industry; interest; intestinal epithelium; Intestinal Hormones; Intestines; Legal patent; Letters; matrigel; Measurement; Measures; Mission; Modeling; monolayer; Natural regeneration; neoplastic cell; Nervous system structure; neural network; Neuraxis; Neurotransmitters; North Carolina; novel; Nutrient; Obesity; Obesity associated disease; off-patent; Organ; Organoids; Peripheral; Pharmacologic Substance; Phase; Physiological; Physiology; Play; prevent; Process; Productivity; programs; Recreation; Regulation; relating to nervous system; Research; response; Role; Satiation; Scientist; screening; Serotonin; Small Business Innovation Research Grant; Source; Stem cells; Stroke; Surface; System; Techniques; Technology; Therapeutic; Tissues; Translations; Tumor Cell Line; United States National Institutes of Health; Universities; Variant

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.

Public Health Relevance Statement:
Project Narrative The novel enteroendocrine cell-rich RepliGut platform will have broad applications in screening and validating therapeutics to manipulate feeding behavior and satiety which will revolutionize the treatment of many metabolic disorders in humans.

Project Terms:
Age; Analytical Chemistry; Automobile Driving; base; Biological Assay; Biological Models; Biotechnology; Blood Circulation; blood glucose regulation; Cardiovascular Diseases; cell behavior; Cell Line; Cell model; cell type; Cells; Cellular Assay; cellular engineering; Characteristics; Communities; comorbidity; Complex; cost; Data; Degenerative polyarthritis; design; Desire for food; Diabetes Mellitus; Diagnosis; Diet; Disease; Eating; Enteral; Enterochromaffin Cells; Enteroendocrine Cell; Feeding behaviors; Feeding Patterns; Food; Foundations; Funding; gastrointestinal; gastrointestinal system; genetically modified cells; glucagon-like peptide; Goals; Health; Healthcare Systems; high standard; high throughput screening; hormone regulation; Hormone secretion; Hormones; Human; human model; Hyperphagia; improved; In Vitro; in vitro Model; in vivo; Industry; Ingestion; innovation; interest; intestinal epithelium; Intestinal Hormones; Intestines; Journals; L Cells; Laboratories; Legal patent; Letters; Libraries; Manuscripts; Measurement; Metabolic; Metabolic Diseases; Metabolism; Methods; Mission; Modeling; Molecular; monoamine; monolayer; neoplastic cell; neural network; Neuraxis; Neurotransmitters; novel; Nutrient; Obesity; Organ; Peptide YY; Performance; Peripheral; Pharmacologic Substance; Phase; Physiological; Physiological Processes; Physiology; Play; Process; Production; Productivity; programs; Recreation; Regulation; relating to nervous system; Reproducibility; Research; response; Role; Satiation; Scientist; screening; Serotonin; Services; sex; Ships; Signaling Molecule; Small Business Innovation Research Grant; Solid; stem cell differentiation; stem cells; Stroke; System; Techniques; Testing; Therapeutic; Tissues; Variant; Well in self; Work