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; 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）：已开发并用于各种基础研究和行业药物输送应用程序，并使用了商业上人类传播的体外模型（Epiairway）。但是，该模型尚未在药物开发和风险评估中的毒理学应用中正式验证。本赠款提案的目的是在验证替代方法验证（ICCVAM）和欧洲替代方法验证中心（ECVAM）指南后，验证这些目的的Epiairway体外人类气道模型。将进行实验，以确定用20种已知体内导管有毒电位的已知化学物质处理后，Epiairway模型的体外反应。将从基于人体和动物吸入数据的职业安全与健康管理（OSHA）建立的具有允许暴露限制的人（PEL）中选择测试化学品。 Epiairway模型中的体外终点将是通过MTT和Toxilight细胞毒性测定法测量的细胞毒性，以及炎症性细胞因子/趋化因子（介绍性细胞因子/趋化因子（Ilthleukin-8，介菌素6和颗粒细胞巨噬细胞刺激因子）的产生。整个实验集将重复两次。然后，将对数据进行统计分析，以确定经验室内的可重复性和实验室间可转移性。体外细胞毒性和细胞因子/趋化因子生产数据将与OSHA为测试化学物质建立的体内（PEL）相关。最后，将对数据进行统计评估，以产生用于将体外细胞毒性结果转化为体内pel的预测的预测模型。这些阶段的目标构成了建立进行充分正式验证的可行性的流行过程。提高到第二阶段的里程碑是可接受的可重复性，协议的间隙转移性以及可接受的预测模型的发展。在第二阶段，将按照ICCVAM指南进行形式验证研究。测试化合物和配方的数量和类型将在4个独立实验室的盲试验中进行扩展和测试。然后，将提交研究结果以进行独立评估和监管接受。