In Vitro Systems: Predict Drug-induced Pulmonary Injury

描述(由申请人提供)：国家癌症研究所需要开发、标准化和验证能够快速且廉价地确定或预测潜在癌症治疗药物的器官特异性毒性的分析方法。该项目的目标是开发一种体外肺细胞分析方法，以确定或预测抗癌药物对人体造成的潜在剂量限制性肺损伤。R21阶段的研究将集中于一个或多个能够满足这一目标的原代多细胞系统的开发、评估和优化，特别是精密切割的肺切片和肺内皮细胞与上皮细胞的共培养。待开发的检测方法最终将能够区分抗癌药物引起的肺部损伤的主要类型、毒性效力的差异以及物种间的敏感性差异。在这项工作中，将以大鼠为模型，以临床化学和先进的蛋白质组学和激光-MS(Jet-REMPI)技术为基础，研究已知的和新的指标来评估药物在体外造成的损伤的程度和性质，以与体内反应相对应。用于开发和验证检测的参考化合物将是在动物和人类中具有特定的、明确的、具有代表性的毒性的抗癌药物。R33阶段将专注于开发该分析的全部能力，作为NCI筛选靶器官毒性并帮助预测临床试验的安全剂量水平和方案所需的体外毒性测试组件的重要和关键部分。

DESCRIPTION (provided by applicant): The National Cancer Institute has a need for development, standardization, and validation of assays that can quickly and cheaply determine or predict organ-specific toxicities of potential cancer therapeutic agents. The objective of the proposed project is to develop an in vitro lung cell assay to determine or predict potential dose-limiting pulmonary injury by anticancer agents in humans. Research in the R21 phase will focus on the development, evaluation, and optimization of one or more primary multicell systems that can meet that objective, in particular precision-cut lung slices and co-cultures of lung endothelial and epithelial cells. The assay to be developed will ultimately be able to discriminate between the major types of injury induced in lung by anticancer agents, differences in toxic potency, and interspecies differences in sensitivity. In this work, known and novel indicators will be investigated for assessing the degree and nature of the injury produced by the agents in vitro for correspondence with in vivo response, using the rat as a model and clinical chemistry and advanced proteomic and laser-MS (Jet-REMPI) techniques. Reference compounds used to develop and validate the assay will be anticancer agents with specific, well- defined, representative toxicities in animals and humans. The R33 phase will focus on developing the full capabilities of the assay as an important and critical part of the in vitro toxicity test battery needed by the NCI to screen for target organ toxicities and to aid in predicting safe dose levels and regimens for clinical trials.