Development of pharmacokinetic assays utilizing an organ-on-chip model of the human kidney proximal tubule

利用人肾近端小管的器官芯片模型开发药代动力学测定

• 批准号: 10210318
• 负责人: Thomas Neumann
• 金额: $ 102.48万
• 依托单位: NORTIS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10210318
• 关键词: Animal Model; Animal Testing; Apical; Area; Biological Assay; Blood; Carrier Proteins; Cell Compartmentation; Cell Line; Cell membrane; Cell physiology; Cells; Cimetidine; Clinical; Clinical Trials; Cultured Cells; Development; Disease model; Drug Kinetics; Drug Screening; Drug toxicity; Environmental Health; Epithelial; Evaluation; Exposure to; Filtration; Generations; Hematologic Agents; Human; Impairment; Industry; Innovation Corps; Kidney; Laboratories; Laboratory Research; Laboratory culture; Liquid substance; Measures; Mediating; Metformin; Modeling; Organ; Organic Anion Transport Protein 1; POU2F2 gene; Performance; Perfusion; Permeability; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Play; Pre-Clinical Model; Preparation; Proximal Kidney Tubules; Qualifying; Regenerative Medicine; Reproducibility; Research Contracts; Running; Safety; Sampling; Shipping; Ships; Side; Site; Small Business Innovation Research Grant; Source; Surface; System; Technology; Testing; Time; Tissue Engineering; Toxicity Tests; Tubular formation; United States National Institutes of Health; Urine; adefovir; cell immortalization; chromatin immunoprecipitation; cost efficient; design; drug candidate; drug development; in vitro Model; in vivo; in vivo Model; inhibitor/antagonist; kidney cell; microphysiology system; nephrotoxicity; organ on a chip; pre-clinical; pre-clinical assessment; precision medicine; programs; prospective; screening; shear stress; side effect; success

Project Summary The proposed application focuses on the development of a plug-and-play living human kidney-on-chip for assaying secretion of drug candidates by the kidney proximal tubule. Once commercialized, the chip will enable drug developers to better screen pre-clinical drug candidates for nephrotoxic side effects. This stands to reduce the high attrition rate of drugs in clinical trials, given that about 20% fail because of nephrotoxicity, and would make drug development faster, more cost efficient, and safer. Conventional preclinical models are of limited use for assessing tubular secretion. In animal models, transporter proteins in the kidney epithelium differ significantly from those present in human kidney; 2D in-vitro models, even when using human cells, fail to provide appropriate culture conditions that enable appropriate cell function. The proposed kidney chip will leverage the commercially available Nortis organ-on-chip platform that is currently in use for a number of organ applications--including a first-generation kidney model that contains a human kidney proximal tubule, tissue-engineered from primary cells. In order to make the kidney chip useful for wide-spread use in pre-clinical assessment of renal secretion, substantial improvements are necessary. A marketable product requires a stable source of cells with sufficient performance, precision delivery of compounds to the basolateral side of the proximal tubule in the chip, and precise fluid sampling from the chip. Further, chips arriving pre-loaded with living proximal tubules will represent a significant value increase to the customer. Phase I of the project will aim to demonstrate that specific transporter proteins OAT1/3 and OCT2, which are crucial to renal secretion, function correctly (Phase I/AIM1). To prove this, proximal tubules in the chips will be exposed to specific substrates (adefovir for OAT1/3, cimetidine for OCT2), in conjunction with specific inhibitors of these transporters (para-aminohippurate for adefovir and metformin for cimetidine). The cells used for growing the tubules will be derived from an immortalized human kidney proximal tubule cell line that has been shown to express OAT1/3 and OCT2 in the correct cell compartments when cultured in the Nortis chip. Phase I/AIM2 will be to implement chip features for precise sampling of microliter-scale fluid volumes upstream and downstream of the tubule, which is required for assessing the secretory flux parameter termed the permeability-surface area product (Pa). During Phase II the chips will be equipped with features for tight control of compound delivery to the basolateral side of the proximal tubule (Phase II/AIM1). The following aim (Phase II/AIM2) will then focus on qualifying assays of kidney proximal tubule secretory clearance for OAT1/3 and OCT2 with sufficient statistical power to establish robustness and reproducibility. Phase II/AIM3 is designed to establish that living kidney chips can be shipped to customers with uncompromised assay performance, which will be validated by an independent laboratory. Once fully developed and commercialized the proposed kidney chip might be suitable for applications beyond the pharmaceutical sector, such as environmental health & safety, precision medicine, and regenerative medicine.

项目摘要 拟议的申请专注于开发插件的生命人类肾脏片 通过肾脏近端小管对候选药物的分泌。一旦商业化，芯片将启用 药物开发人员可以更好地筛选临床前候选药物，以实现肾毒性副作用。这将减少 临床试验中药物的高损耗率很高，鉴于由于肾毒性而失败了20％，并且会失败，并且会失败 使药物开发更快，更具成本效益和更安全。常规的临床前模型的使用有限 用于评估管状分泌。在动物模型中，肾上皮中的转运蛋白有很大差异 从人类肾脏中的那些;即使使用人类细胞，2D含量模型也无法提供适当的 培养条件，使能够适当的细胞功能。拟议的肾脏芯片将在商业上利用 可用的Nortis器官片平台，目前正在用于许多器官应用程序 - 包括 第一代肾脏模型，其中含有人类肾脏近端小管，由原发性设计 细胞。为了使肾脏芯片可用于广泛使用肾脏分泌的临床预测， 有必要改进。可销售的产品需要足够的稳定的细胞来源 性能，将化合物的精确递送到芯片近端小管的基底外侧侧， 精确的流体采样从芯片中进行。此外，预先装载有活近端小管的芯片将代表 对客户的重大价值增加。该项目的第一阶段将旨在证明特定的 转运蛋白OAT1/3和OCT2对肾脏分泌至关重要，功能正确（I/AIM1阶段）。 为了证明这一点，芯片中的近端小管将暴露于特定的底物（OAT1/3的adefovir，cimetidine，cimetidine 对于OCT2），与这些转运蛋白的特异性抑制剂（用于adefovir和Adefovir和 二甲苯胺的二甲双胍）。用于生长小管的细胞将源自永生的人 肾脏近端小管细胞系已显示在正确的细胞中表达OAT1/3和OCT2 在诺蒂斯芯片中培养时隔室。 I/AIM2阶段将是实现精确的芯片功能 小管的上游和下游的微尺度流体体积的采样，这是必需的 评估称为渗透率 - 表面面积产品（PA）的分泌通量参数。在第二阶段期间 芯片将配备具有对复合递送到近端基底外侧的紧密控制的功能 小管（II阶段/AIM1）。然后，以下目标（II/AIM2）将专注于肾脏近端的合格测定法 OAT1/3和OCT2的小管分泌清除具有足够的统计能力，可以建立鲁棒性和 可重复性。 II阶段/AIM3旨在确定可以通过 不妥协的测定性能将由独立实验室验证。一旦完全开发 并将拟议的肾脏芯片商业化可能适用于药品以外的应用 领域，例如环境健康与安全，精密医学和再生医学。