Perfused organ panel as an animal surrogate for chemical toxicity testing

该项目响应NIEHS RFA-ES-20-005“从实验中开发的器官型培养模型” 动物化学毒性筛选，”这是符合NTP的跨机构中心的需要， 替代毒理学方法评价（NICEATM），用于开发和评价新的、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）开发并商业化了一种器官- 芯片式灌注器官面板，具有专有的液体呼吸技术， 间质灌注、上级氧气输送和稳定pH值，48个统计学上独立的器官培养物。 在该项目中，Lena Biosciences将使用灌注器官面板开发生理相关的体外 筛选系统，其使用来源于通常用于毒理学测试的动物物种的细胞。下一篇：LB 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，LB 产生适合于体外和体内动物毒理学研究之间比较的测定数据，以及 Tox 21 HTS数据。最终，该项目将提供彻底表征和验证的替代体外 具有高特异性和灵敏度的测试系统，以减少或取代在毒性测试中使用动物。 在第一阶段，LB将开发具有生物学和外源性代谢能力的啮齿动物肝脏和大脑模型 具有体内类似细胞呼吸代谢以实现最佳线粒体反应性， 对毒物的敏感性肝脏模型将提供高活性的药物代谢酶用于原位 产生反应性代谢物，并模拟母体药物缺失和代谢物 形成更好的模型和预测毒理学结果。大脑模型将模仿大脑的先天 对药物过量有强烈毒理学反应的免疫力，急性乙酰胆碱酯酶抑制 神经毒性（I期）和导致迟发性神经病的神经病靶向酯酶抑制（II期） 暴露于有机磷（OP）化学品及其有毒代谢物后。 灌注器官组和3组体外对照将用对乙酰氨基酚（APAP）和马拉硫磷处理， 一种含有神经毒性代谢物Malaoxon的OP杀虫剂，与特定组织的测试相关 模型和物种匹配的体内数据已经存在。这将有助于基准测试和显示 灌注器官小组作为美国环境保护局目前使用的体内模型的替代品的效用 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 肝脏和大脑先进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（Morgan博士，Emory，药理学和化学系 生物学），APAP毒性（Jaeschke博士，堪萨斯大学医学中心），脑和肝毒理学（Dr. Caudle，Emory，环境卫生部，Jaeschke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).