Validation of an In Vitro Human Airway Model

体外人体气道模型的验证

• 批准号: 6934284
• 负责人: PATRICK J HAYDEN
• 金额: $ 19.99万
• 依托单位: MATTEK CORPORATION
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934284
• 关键词: biological models; chemokine; colony stimulating factor; computational biology; cytokine; cytotoxicity; dosage; enzyme linked immunosorbent assay; immunocytochemistry; mathematical model; model design /development; occupational hazard; polymerase chain reaction; pulmonary respiration; respiratory airway pressure; respiratory function; statistics /biometry; tissue /cell culture; toxicant screening

DESCRIPTION (provided by applicant): A commercially available in vitro model of human conducting airways (EpiAirway) has been developed and utilized for a variety of basic research and industry drug delivery applications. However, the model has not been formally validated for toxicology applications in drug development and risk assessment. The goal of the present grant proposal is to validate the EpiAirway in vitro human airway model for these purposes following Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and European Center for the Validation of Alternative Methods (ECVAM) guidelines. Experiments will be performed to determine in vitro responses of the EpiAirway model after treatment with 20 chemicals of known in vivo conducting airway toxic potentials. Test chemicals will be chosen from those with permissible exposure limits (PELs) established by Occupational Safety and Health Administration (OSHA) based upon in vivo human and animal inhalation data. In vitro endpoints in the EpiAirway model will be cytotoxicity as measured by the MTT and ToxiLight cytotoxicity assays, and inflammatory cytokine/chemokine (interleukin-8, interleukin 6 and granulocyte macrophage stimulating factor) production as measured by enzyme linked immunosorbant assays. The entire set of experiments will be repeated two separate times. The data will then be statistically analyzed to determine intralaboratory reproducibility and interlaboratory transferability. The in vitro cytotoxicity and cytokine/chemokine production data will be correlated with in vivo (PELs) established for the test chemicals by OSHA. Finally, the data will be statistically evaluated to produce a prediction model for converting in vitro cytotoxicity results into a prediction of in vivo PELs. These Phase I goals constitute the prevalidation process for establishing the feasibility of proceeding to full formal validation. Milestones for advancement to Phase II are acceptable reproducibility, interlaboratory transferability of protocols and development of an acceptable prediction model. In Phase II, formal validation studies will be conducted following ICCVAM guidelines. The number and types of test compounds and formulations will be expanded and tested in blinded trials in 4 independent laboratories. The study results will then be submitted for independent assessment and regulatory acceptance.

描述（由申请人提供）：已开发出商业化的人体传导气道体外模型（EpiAirway）并用于各种基础研究和工业药物输送应用。然而，该模型尚未在药物开发和风险评估中的毒理学应用中得到正式验证。本拨款提案的目标是根据替代方法验证机构间协调委员会 (ICCVAM) 和欧洲替代方法验证中心 (ECVAM) 指南，验证 EpiAirway 体外人体气道模型的有效性。将进行实验以确定 EpiAirway 模型在用 20 种已知体内传导气道潜在毒性的化学物质处理后的体外反应。测试化学品将从职业安全与健康管理局 (OSHA) 根据人体和动物体内吸入数据制定的允许接触限值 (PEL) 中选择。 EpiAirway 模型的体外终点将是通过 MTT 和 ToxiLight 细胞毒性测定测量的细胞毒性，以及通过酶联免疫吸附测定测量的炎症细胞因子/趋化因子（白细胞介素 8、白细胞介素 6 和粒细胞巨噬细胞刺激因子）的产生。整组实验将单独重复两次。然后对数据进行统计分析，以确定实验室内的再现性和实验室间的可转移性。体外细胞毒性和细胞因子/趋化因子产生数据将与 OSHA 为测试化学品建立的体内 (PEL) 相关。最后，将对数据进行统计评估，以生成预测模型，将体外细胞毒性结果转换为体内 PEL 的预测。这些第一阶段目标构成了确定进行全面正式验证的可行性的预验证过程。进入第二阶段的里程碑是可接受的重现性、协议的实验室间可转移性以及可接受的预测模型的开发。在第二阶段，将按照 ICCVAM 指南进行正式验证研究。测试化合物和配方的数量和类型将扩大并在 4 个独立实验室进行盲法试验。研究结果将提交给独立评估和监管机构接受。