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

项目摘要 维持人体肠道的健康生理是医药的一大重点。要研究的模型 药物/营养吸收、外源物质、毒理学和临床前疗效因缺乏 准确、可重复性和易于使用的细胞培养模型来评估这样的主题。出于这些原因， 迫切需要更好的体外模型来概括人类肠道的疾病状态，以及 为药物发现和验证提供更好的平台。脂质的处理是人类健康的核心 条件和药代动力学。与脂质相关的代谢紊乱(如肥胖、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 不健康的饮食增加了。代谢紊乱的治疗方法很少，疗效有限。 需要更广泛有效的药物。肠吸收细胞(AEs)的脂质调节机制 可强烈影响口服药物的吸收、分布、代谢和排泄(ADME)，以及药物 AE脂类处理可以负向和正向调节生物利用度 药物与乳糜粒(CMS)有关。AEs的CM相关药物出口是一线指标 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- 处理机制在改善新陈代谢疾病的药物工程方面具有很强的潜力， 生物利用度和有效性。为了满足这一需求，早期生物技术公司Altis BiosSystems Inc. 公司将与北卡罗来纳大学教堂山分校的科学家合作开发 在高通量96-Traswell格式上培养可吸收的肠细胞(AEs)。新陈代谢GUT是单层的 分化的AE源自基础技术RepliGutTM，这是一种干细胞驱动的 人体上皮的单层，来源于器官捐赠者的小肠或结肠，包含所有 体内发现的增殖和分化细胞类型。此第一阶段提案的目标是 开发四个创新元素：1)用于脂肪吸收和输出的快速荧光读数，2) 同时快速检测屏障完整性和脂肪酸输出，3)快速和高灵敏度 用非放射性同位素方法鉴定脂肪酸代谢物种，4)快速荧光 乳胶粒(CM)出口的量化。虽然元素1-3已经显示出对 在学术研究方面，将探索它们的商业可行性。元素4将从头开发为 这为中药出口和中药相关药物的高通量检测提供了一种新的方法 量化。

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.