A 3D biomimetic liver sinusoid construct for predicting physiology and toxicity

描述（由申请人提供）：用于预测生理和毒性的3D仿生肝窦结构大约90%的候选药物进入i期临床试验失败，药物失败的主要原因之一是意外毒性。肝脏在人体中起着中心作用，具有体内平衡和生物转化、药物代谢等重要功能。肝脏也是药物毒性最常见的靶点。现有的体外模型和体内动物模型对人类肝毒性的预测能力有限。该项目的目标是构建一个模拟人类肝脏功能和反应的微流控肝脏模块，其读数旨在显示正常肝功能和毒性反应。该人体肝脏模型有望成为模拟人体暴露的基本消除器官，为药物诱导的肝脏毒性提供改进的预测，也可作为药物发现的疾病模型。我们的方法是用人类肝细胞、库普弗细胞、星状细胞和内皮细胞开发一个3D微流控系统，来模拟肝腺泡——肝脏最小的功能单位。该模型的一个独特之处在于介质的氧化，以及氧梯度的建立，这被认为是正常人类肝脏正弦空间中肝细胞重要的代谢、基因表达和功能异质性的原因。富氧区的肝细胞在氧化代谢、脂肪酸氧化、糖异生、胆汁酸提取、氨解毒到尿素和谷胱甘肽偶联方面效率高，而缺氧区的肝细胞在糖酵解、脂肪生成和细胞色素P-450生物转化方面效率高。该模型的另一个独特之处在于结合了“哨兵”生物传感器细胞，这是一小部分带有工程生物传感器的细胞，可以指示细胞功能的变化。当与其他荧光探针、标准生化和质谱读数相结合时，该模型将提供实时高含量分析（HCA）剖面，以监测器官功能和反应。该模型的读数和性能的选择和验证将基于一组具有可用临床数据的参考药物进行评估。为了便于比较，将建立一个包含临床数据以及其他体外和体内研究数据的药物数据库。该项目的最终目标是开发一种人体肝脏功能的微流控模型，该模型将与一系列其他人体器官模块相结合，创建一个微生理学平台，可以再现人体临床试验结果，并为药物开发提供更好的暴露、安全性和有效性预测。作为药物代谢的结果，肝脏在人类药物相互作用中起着核心作用，无论是在肝脏内还是在其他器官中。肝脏模块的性能是微生理学平台性能的核心。我们相信这里提出的设计将在该平台上最佳地概括人类肝脏功能。

DESCRIPTION (provided by applicant): A 3D biomimetic liver sinusoid construct for predicting physiology and toxicity Approximately 90% of drug candidates entering Phase 1 clinical trials fail, and one of the main reasons for drug failure is unexpected toxicity. The liver plays a centra role in the human body, contributing to homeostasis and important functions such as biotransformation and metabolism of drugs. The liver is also the most common target for drug-induced toxicity. Existing in vitro models and in vivo animal models have limited predictive power for human liver toxicity. The goal of this project is to construct a microfluidic liver modul which mimics the functions and responses of the human liver, with readouts designed to indicate both normal liver function and toxic responses. This human liver model is expected to be the essential elimination organ for modeling human exposure, provide improved predictions of drug induced liver toxicity, and also serve as a disease model for drug discovery. Our approach will be to develop a 3D microfluidic system with human hepatocyte, kupffer, stellate and endothelial cells, to mimic the liver acinus - the smallest functional unit of the liver. A uniue feature of the model will be the oxygenation of the media, and the establishment of an oxygen gradient, which is believed to account for important metabolic, gene expression and functional heterogeneity of the hepatocytes in the sinusoidal space of normal human liver. Hepatocytes in the oxygen rich zone are efficient in oxidative metabolism, fatty acid oxidation, gluconeogenesis, bile acid extraction, ammonia detoxification to urea and glutathione-conjugation while hepatocytes in the oxygen depleted zone are efficient in glycolysis, liponeogenesis and Cytochrome P-450 biotransformation. Another unique feature of the model will be the incorporation of 'sentinel' biosensor cells, a small fraction of cells with engineered biosensors that indicate changes in cellular functions. When combined with other fluorescent probes, standard biochemical and mass spectroscopy readouts, the model will provide a real-time High Content Analysis (HCA) profile to monitor organ function and response. The selection and validation of readouts and performance of the model will be evaluated based on a panel of reference drugs with available clinical data. To facilitate that comparison, a database of drugs with clinical data, and data from other in vitro and in vivo studies will be constructed. The ultimate goal of this project is to develop a microfluidic model of human liver function that will integrate with a series of other human organ modules, to create a microphysiology platform that reproduces human clinical trial results and provides improved predictivity of exposure, safety and efficacy for drug development. The liver plays a central role in human drug interactions, both within the liver and in other organs, as a result of drug metabolism. The performance of the liver module is central to the performance of the microphysiology platform. We believe the design proposed here will optimally recapitulate human liver function on that platform.