Perfused organ panel as an animal surrogate for chemical toxicity testing

该项目响应了NIEHS RFA-ES-20-005“器官类型培养模型，该模型由实验发展而来 动物用于化学毒性筛查，这与NTP的跨部门中心的需求一致 用于开发和评估新的、修订的、 以及确定对人类健康和环境的潜在危害的替代方法。 通过我们正在进行的NCATS SBIR第二阶段，Lena Biosciences(LB)开发了一种器官并将其商业化- 像芯片一样的灌流器官面板，具有专有的液体呼吸技术，提供统一的 对48个统计独立的器官培养，间质灌流、卓越的氧气输送和稳定的pH。 在这个项目中，莉娜生物科学公司将使用灌流器官面板在体外开发与生理相关的 使用来自动物物种的细胞的筛选系统，通常用于毒理学测试。接下来，德州分校 将复制在动物组织中观察到的生物相互作用和毒理学反应。最后，德意志银行将 产生适合于比较体外和体内动物毒理学研究的化验数据； Tox21高温超导数据。最终，该项目将在体外提供完全表征和验证的替代方案 具有高特异性和敏感性的测试系统，以减少或取代在毒性测试中使用动物。 在第一阶段，兰德堡将开发具有生物和异种生物代谢能力的啮齿动物肝脏和大脑模型 在体内具有类似细胞的呼吸代谢，以实现最佳的线粒体响应和 对毒物的敏感性。肝脏模型将为原位提供高活性的药物代谢酶 生成反应性代谢物，并模拟母体药物缺失和代谢物的并行过程 编队以更好地模拟和预测毒理学结果。大脑模型(S)将模仿大脑固有的 对药物过量有强烈毒理学反应的免疫，乙酰胆碱酯酶抑制与急性 神经毒性(第一阶段)，以及导致迟发性神经病变的以酯酶抑制为靶点的神经病变(第二阶段) 在暴露于有机磷(OP)化学品及其有毒代谢物之后。 灌流器官面板和3组体外对照将用扑热息痛(APAP)和马拉硫磷治疗， 一种含有神经毒性代谢物马拉氧磷的OP杀虫剂，与特定组织的测试有关 模型和物种匹配的活体数据已经存在。这将有助于进行基准测试和展示 灌流器官面板作为美国环境保护署目前使用的活体模型的替代工具 保护局。为了成功地进行这些研究并确保项目的成功，我们召集了一个 肝脏和大脑高级细胞培养模型的专家团队(LB的Pi和CSO，Shoemaker博士)，药物 代谢和代谢物形成(摩根博士，埃默里，药理和化学系 生物学)、APAP毒性(Jeschke博士，堪萨斯大学医学中心)以及脑和肝脏毒理学(Dr。 Caudle、Emory、环境卫生部、Jeschke博士和Morgan博士)。

This project responds to the NIEHS RFA-ES-20-005 “Organotypic culture models developed from experimental animals for chemical toxicity screening,” that is aligned with the needs of the NTP’s Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) for development and evaluation of new, revised, and alternative methods to identify potential hazards to human health and the environment. Through our ongoing NCATS SBIR Phase II, Lena Biosciences (LB) developed and commercialized an organ- on-a-chip-like, Perfused Organ Panel, with a proprietary liquid breathing technology that provides uniform interstitial perfusion, superior delivery of oxygen and stable pH to 48 statistically independent organ cultures. In this project, Lena Biosciences will use the Perfused Organ Panel to develop physiologically-relevant in vitro screening systems using cells derived from animal species typically utilized for toxicological testing. Next, LB will replicate biological interactions and toxicological responses observed in animal tissues. Lastly, LB will produce assay data that is suitable for comparisons between in vitro and in vivo animal toxicology studies, and Tox21 HTS data. Ultimately, this project will provide thoroughly characterized and validated, alternative in vitro test systems with high specificity and sensitivity to reduce or replace the use of animals in toxicity testing. In Phase I, LB will develop biologically and xenobiotic-metabolically competent rodent liver and brain models having in vivo like cellular respiratory metabolism to achieve optimal mitochondrial responsiveness and susceptibility to toxicants. The liver model will provide high activity of drug metabolizing enzymes for in situ generation of reactive metabolites, and mimic parallel processes of parent drug deletion and metabolite formation to better model and predict toxicological outcomes. The brain model(s) will mimic the brain’s innate immunity with robust toxicological responses to drug overdose, Acetylcholinesterase inhibition with acute neurotoxicity (Phase I), and neuropathy target esterase inhibition that cause delayed neuropathy (Phase II) following the exposure to organophosphorus (OP) chemicals and their toxic metabolites. Perfused Organ Panel and 3 sets of in vitro controls will be treated with Acetaminophen (APAP) and Malathion, an OP insecticide with a neurotoxic metabolite, Malaoxon, that are relevant to the testing of specific tissue models and for which species-matched in vivo data already exists. This will facilitate benchmarking and show the Perfused Organ Panel’s utility as an alternative to in vivo models currently used by the U.S. Environmental Protection Agency. To successfully carry out these studies and ensure the project’s success, we assembled a team of experts in advanced cell culture models of liver and brain (LB’s PI and CSO, Dr. Shoemaker), drug metabolism and metabolite formation (Dr. Morgan, Emory, Department of Pharmacology and Chemical Biology), APAP toxicity (Dr. Jaeschke, University of Kansas Medical Center), and brain and liver toxicology (Dr. Caudle, Emory, Department of Environmental Health, Dr. Jaeschke, and Dr. Morgan).