MetabolGut: a rapid assay platform to evaluate the impact drugs on lipid-handlingpathways and chylomicron-associated drug distribution using stem cell-drivenhuman absorptive enterocytes.

MetabolGut：一个快速检测平台，使用干细胞驱动的人体吸收性肠上皮细胞来评估药物对脂质处理途径和乳糜微粒相关药物分布的影响。

• 批准号: 10766493
• 负责人: Bill Thelin
• 金额: $ 34.33万
• 依托单位: ALTIS BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10766493
• 关键词: 3-Dimensional; Address; Amino Acids; Apical; Applications Grants; Architecture; Biological Assay; Biological Availability; Biosensor; Biotechnology; Blood Circulation; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cells; Chimeric Proteins; Chylomicrons; Collaborations; Colon; Detection; Dietary Fats; Disease; Donor person; Drug Kinetics; Drug Modelings; Drug Screening; Drug Transport; Elements; Enterocytes; Epidemic; Epithelium; Etiology; Excretory function; Exposure to; Fatty Acids; Fatty acid glycerol esters; Flow Cytometry; Foundations; Gene Transfer Techniques; Goals; Health; Hepatic; Human; Hydrogels; Hyperlipidemia; Hypertension; Insulin Resistance; Intestines; Kinetics; Lipids; Lipoproteins; Liver Circulation; Lymphatic System; Metabolic; Metabolic Diseases; Metabolism; Methods; Modality; Modeling; Nonesterified Fatty Acids; North Carolina; Nutrient; Obesity; Occupations; Oral; Organ Donor; Organ Transplantation; Organoids; Pathway interactions; Pharmaceutical Preparations; Phase; Physiological; Physiology; Play; Pre-Clinical Model; Process; Proliferating; Property; Rapid screening; Reader; Reproducibility; Role; Scientist; Small Intestines; System; Technology; Thick; Thin Layer Chromatography; Tissues; Toxicology; Unhealthy Diet; Universities; Validation; Xenobiotics; absorption; cancer cell; cell type; commercialization; detection assay; detection limit; dietary; drug development; drug discovery; drug distribution; drug testing; high throughput screening; improved; in vitro Model; in vivo; innovation; lipophilicity; lymphatic circulation; metabolic engineering; metabolomics; monolayer; novel; novel strategies; nutrient absorption; preclinical efficacy; prototype; rapid detection; rapid test; sedentary; stem cells; sugar

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. Lipid-handling is central to human health conditions and pharmacokinetics. Lipid-related metabolic disorders (i.e. obesity, insulin resistance, hyperlipidemia, and hypertension) are a global epidemic and predicted to increase as sedentary jobs and unhealthy diets increase. Treatments for metabolic disorders are sparse with limited efficacy highlighting the need for more broadly effective drugs. Lipid-handling mechanisms by Absorptive Enterocytes (AEs) can strongly influence oral drug Absorption, Distribution, Metabolism and Excretion (ADME), and drug bioavailability can be negatively and positively regulated by AE lipid-handing as many lipid soluble drugs are associated with chylomicrons (CMs). CM-associated-drug export by AEs is a first-line metric of bioavailability for lipophilic drugs with no accurate preclinical model. New tactics that harness lipid- handling mechanisms have strong potential to improve drug engineering for metabolic diseases, bioavailability and efficacy. 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 Absorptive Enterocytes (AEs) on a high-throughput 96-Traswell format. ‘MetabolGut’ is a monolayer of differentiated AEs derived from the foundational technology, RepliGutTM, which is a stem cell-driven 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. The goal of this Phase I proposal is to develop four innovative elements: 1) rapid fluorescent readouts for lipid absorption and export, 2) simultaneous rapid detection of barrier integrity and fatty acid export, 3) rapid and highly sensitive identification of fatty acid metabolic species using non-radioisotope methods, 4) rapid fluorescent quantification of chylomicron (CM) export. While elements 1-3 have shown substantial utility for academic studies, they will be explored for commercial viability. Element 4 will be developed de novo as it represents a new assay for high-throughput detection of CM export and CM-associated drug quantification.

项目摘要 保持人体肠道的健康生理是Pharma的一大重点。模型来研究 药物/营养素吸收、外源性物质、毒理学和临床前功效由于缺乏 准确，可重复，易于使用的细胞培养模型来评估这些主题。基于这些理由， 强烈需要更好的体外模型，其再现人肠道的疾病状态，并且 更好的药物发现和验证平台。脂质处理是人类健康的核心 条件和药代动力学。脂质相关的代谢紊乱（即肥胖、胰岛素抵抗， 高脂血症和高血压）是一种全球流行病，预计随着久坐不动的工作和 不健康的饮食增加。代谢紊乱的治疗方法很少，突出显示疗效有限 需要更广泛有效的药物。吸收性肠细胞（AE）的脂质处理机制 可强烈影响口服药物的吸收、分布、代谢和排泄（ADME）， 生物利用度可以通过AE脂质处理负性和正性调节，因为许多脂溶性 药物与乳糜微粒（CM）有关。按AE列出的CM相关药物输出是一线指标 没有准确的临床前模型的亲脂性药物的生物利用度。利用脂质的新策略- 处理机制具有很强的潜力来改进用于代谢疾病的药物工程， 生物利用度和功效。为了满足这一需求，Altis Biosystems Inc.，一个早期的生物技术 该公司将与位于查佩尔山的北卡罗来纳州大学的科学家合作， 高通量96-Traswell格式上的吸收性肠细胞（AE）。“MetabolGut”是一种单层的 分化AE源自基础技术，即干细胞驱动的干细胞移植技术。 来源于器官捐献者小肠或结肠的单层人类上皮细胞，包含所有 体内发现的增殖和分化细胞类型。第一阶段提案的目标是 开发四个创新元素：1）快速荧光读数，用于脂质吸收和输出，2） 同时快速检测屏障完整性和脂肪酸输出，3）快速且高度灵敏 使用非放射性同位素方法鉴定脂肪酸代谢种类，4）快速荧光 乳糜微粒（CM）输出的定量。虽然要素1-3已经显示出对以下方面的实质性效用： 学术研究，将探讨它们的商业可行性。要素4将重新开发， 它代表了一种用于CM输出和CM相关药物的高通量检测的新方法 量化