Perfused organ panel as an animal surrogate for chemical toxicity testing

灌注器官组作为化学毒性测试的动物替代品

基本信息

批准号：
10699787
负责人：
Jelena Vukasinovic
金额：
$ 88.12万
依托单位：
LENA BIOSCIENCES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10699787
关键词：
3-Dimensional Acetaminophen Acetylcholine Acetylcholinesterase Ache Acute Agrochemicals Analgesics Animal Experiments Animal Testing Animals Benchmarking Biochemical Biological Biological Assay Biological Sciences Biology Black race Blood Substitutes Boronic Acids Brain Breathing Carbamates Cell Culture Techniques Cell Respiration Cells Cessation of life Chemicals Competence Complex Data Development Dose Drug Interactions Drug Metabolic Detoxication Ensure Environment Environmental Health Enzyme Inhibition Enzymes Ethics Evaluation Exposure to Foundations Free Radicals Galactose Generations Glycolysis Goals Health Hemoglobin HepG2 Hepatocyte Human In Situ In Vitro Insecta Insecticides Kansas Lactate Dehydrogenase Liquid substance Liver Malathion Medical center Metabolic Metabolism Methods Mitochondria Modeling Mus N-acetyl-4-benzoquinoneimine National Center for Advancing Translational Sciences National Institute of Environmental Health Sciences Natural Immunity Neuromuscular Junction Neuropathy Normal Cell Organ Organ Culture Techniques Organophosphates Outcome Overdose Oxidative Phosphorylation Oxidative Stress Oxygen Paralysed Parents Perfusion Pesticides Pharmaceutical Preparations Pharmacology Phase Physiological Plasma Predisposition Process Production Protein Isoforms Publishing Pyruvate Qualifying Reporting Research Resistance Risk Rodent Rotenone Sensitivity and Specificity Small Business Innovation Research Grant System Technology Testing Tissue Model Toxic Environmental Substances Toxic effect Toxicity Tests Toxicology United States Environmental Protection Agency Universities Validation Xenobiotic Metabolism Xenobiotics acetaminophen overdose animal tissue carboxylesterase commercialization cost drug induced liver injury drug metabolism drug testing esterase hazard high throughput screening in vitro Model in vitro testing in vivo in vivo Model inhibitor interstitial invention malaoxon microphysiology system mitochondrial metabolism multiplex assay neuropathy target esterase neurotoxic neurotoxicity organ on a chip organophosphorus insecticide oxidation programs respiratory response screening success three-dimensional modeling toxicant

项目摘要

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).

该项目响应 NIEHS RFA-ES-20-005“从实验开发的有机培养模型” 动物进行化学毒性筛查”，这符合 NTP 跨机构中心的需求替代毒理学方法评估 (NICEATM)，用于开发和评估新的、修订的、以及识别对人类健康和环境潜在危害的替代方法。通过我们正在进行的 NCATS SBIR 二期，Lena Biosciences (LB) 开发了一种器官并将其商业化类似芯片的灌注器官面板，采用专有的液体呼吸技术，可提供均匀的 48 个统计独立的器官培养物具有间质灌注、优质的氧气输送和稳定的 pH 值。在这个项目中，Lena Biosciences 将使用灌注器官面板来开发生理相关的体外使用通常用于毒理学测试的动物物种细胞的筛选系统。最后，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。产生适合体外和体内动物毒理学研究之间比较的测定数据，以及最终，该项目将提供经过彻底表征和验证的体外替代数据。具有高特异性和敏感性的测试系统，可减少或替代毒性测试中动物的使用。在第一阶段，LB 将开发具有生物学和异生素代谢能力的啮齿动物肝脏和大脑模型具有体内类似的最佳细胞呼吸代谢，以实现线粒体反应性和对毒物的敏感性肝脏模型将提供高活性的原位药物代谢酶。产生反应性代谢物，并模拟母体药物缺失和代谢物的并行过程形成更好的模型和预测毒理学结果的大脑模型将模仿大脑的先天特征。对药物过量产生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（第一阶段）和神经病变靶向酯酶抑制导致迟发性神经病变（第二阶段）接触有机磷 (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) 和马拉硫磷进行处理，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以及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。灌注器官面板作为美国环境署目前使用的体内模型的替代品的实用性为了成功开展这些研究并确保项目的成功，我们组建了一个先进的肝脏和大脑细胞培养模型专家团队（LB 的 PI 和 CSO，Shoemaker 博士）、药物代谢和代谢物形成（摩根博士，埃默里大学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）以及脑和肝脏毒理学（Dr. Jaeschke，堪萨斯大学医学中心）。 Caudle，埃默里大学环境卫生系，Jaeschke 博士和 Morgan 博士）。