Rat and Canine Microphysiological Systems of the Kidney Proximal Tubule for Chemical Toxicity Screening

项目摘要 人类通过食物、家用产品、药品和其他化学品不断暴露于各种化学品中。 环境肾脏特别容易受到化学损伤;药物引起的肾毒性是主要的 严重急性肾功能衰竭的所有病例中有25%是由这种健康问题引起的。肾毒性也是一种 药物开发中的一个重要问题;它是过程后期流失的主要原因，占19% 失败的III期临床试验最先进的动物模型和体外预测工具的局限性 人类肾毒性是众所周知的，这引发了先进的体外3D技术的发展 人体组织和器官的模型，所谓的器官芯片系统（OOC）或微生理系统 （MPS）。虽然基于人类的MPS进展迅速，但MPS数据之间仍存在翻译差距 和体内数据。因此，将基于动物的MPS的观察结果与体内动物数据进行比较， 我们将人类MPS发现转化为临床医学的能力。 这项快速拨款申请提出了从两种动物中开发肾近端小管（KPT）MPS， 经常用于肾毒性筛选的种属：大鼠和犬。一旦研制成功， 经过验证和商业化，KPT-MPS将成为化学毒性筛选的重要新工具 和药物开发，允许交叉引用基于动物的MPS数据与体内动物数据，人类- 基于MPS数据和临床结局。它还有可能取代一些动物测试，这一点很重要 减少在临床前试验中使用活体动物。 该项目的第一阶段旨在从大鼠和犬中建立稳健的KPT-MPS模型。该项目将利用 市售Nortis MPS平台和预先建立的用于创建基于人类的KPT的方案- MPS。I期/AIM 1将证明来自大鼠和狗的近端小管细胞形成活性和结构 完整的近端小管，并且当暴露于 在体内对KPT有毒性（I期/AIM 2）。将在两个项目中的至少一个项目中确定可行性要求 在项目进入第二阶段之前，第二阶段将重点优化大鼠和/或犬KPT- MPS原型和开发用于测试潜在肾毒性化合物的试验（II期/AIM 1）。为 通过这些努力，我们将利用一种新的由注塑热塑性材料制成的微流体芯片， 是为基于人类的KPT-MPS开发的。在II期/AIM 2期间，一组五种化合物， 将在KPT-MPS中检测大鼠、犬和人的种属特异性肾毒性，目的是确定 所有三个种属的KPT-MPS数据与体内数据之间的相关性。第二阶段/目标3是证明， 大鼠/狗KPT-MPS可以作为即插即用的预接种产品运送给未来的客户。

Project Summary Humans are constantly exposed to a variety of chemicals via food, household products, medicines, and the environment. The kidney is particularly susceptible to chemical damage; drug-induced nephrotoxicity is a major health concern that contributes to 25% of all cases of severe acute kidney failure. Nephrotoxicity is also a significant problem in drug development; it is a major cause of attrition late in the process, accounting for 19% of failed Phase III clinical trials. The limitations of state-of-the-art animal models and in vitro tools for predicting human nephrotoxicity are well acknowledged, which has sparked the development of advanced 3D in vitro models of human tissues and organs, so called organ-on-chip systems (OOC) or microphysiological systems (MPS). While progress with human-based MPS has been rapid, a translational gap remains between MPS data and in vivo data. Therefore, comparing observations from animal-based MPS to in vivo animal data will inform our ability to translate human MPS findings to clinical medicine. This fast-track grant application proposes the development of kidney proximal tubule (KPT) MPS from two animal species that are frequently used in kidney toxicity screening: rat and dog. Once successfully developed, validated, and commercialized, the KPT-MPS will serve as an important new tool in chemical toxicity screening and drug development, allowing cross-referencing animal-based MPS data with in vivo animal data, human- based MPS data and clinical outcomes. It also has the potential to replace some animal testing, significantly reducing the use of live animals in preclinical testing. Phase I of the project is designed to establish robust KPT-MPS models from rat and dog. The project will leverage the commercially available Nortis MPS platform and pre-established protocols for creating human based KPT- MPS. Phase I/AIM 1 will demonstrate that proximal tubule cells from rat and dog form viable and structurally complete proximal tubules and that these tubules exhibit a stress response when exposed to compounds that are toxic to KPTs in vivo (Phase I/AIM 2). Feasibility requirements will be established in at least one of the two species before the project progresses into Phase II. Phase II will focus on optimizing the rat and or dog KPT- MPS prototypes and on developing assays for testing potentially nephrotoxic compounds (Phase II/AIM 1). For these efforts we will leverage a new microfluidic chip made of an injection-molded thermoplastic material that was developed for a human-based KPT-MPS. During Phase II/AIM 2, a panel of five compounds with published species-specific nephrotoxicity for rat, dog, and human will be tested in the KPT-MPS with the goal to establish correlation between KPT-MPS data and in vivo data in all three species. Phase II/AIM 3 is to demonstrate that rat/dog KPT-MPSs can be shipped to future customers as a plug & play pre-seeded product.