The UCLA - Boston University Lung Cancer Biomarker Development Laboratory

描述（由申请人提供）：迫切需要制定评估新型和新兴烟草相关产品的生物影响的方法，以便确定那些最有可能对人类健康造成严重风险的产品。我们的建议是基于“损伤场”范式，其中1)吸入的毒素如烟草烟雾改变了整个呼吸道上皮细胞的基因表达；2)在容易接近的上呼吸道上皮细胞中测量这些变化，可以深入了解这种暴露的生理效应。我们之前已经使用这一概念来识别烟草烟雾暴露的生理反应，并开发了支气管气道基因表达特征，这是肺癌的早期检测生物标志物。本提案的总体目标是1)证明气道上皮谱分析在识别对其他烟草制品的生理反应方面的效用，2)建立体外暴露系统，可以准确模拟生理暴露并快速评估烟草相关产品的体外潜在致癌性。我们将本提案的重点放在电子烟（ECIGS）上，这是一种新兴的fda监管的烟草相关产品，因为：1)它们越来越受欢迎，2)它们与烟草卷烟（TCIGS）的化学不同，以及3)缺乏关于其潜在健康影响的数据。我们提出三项同时进行的研究，重点是确定气道上皮对ECIGS的生理和细胞反应。前两项研究涉及ECIGS使用者支气管和鼻上皮刷牙的基因表达谱分析，目前和以前的TCIGS吸烟者作为对照组。这些研究将对ECIGS的生理影响提供一个全面的观点，并将评估是否可以使用任一或两种气道组织来容易地评估ECIGS暴露的生理影响。第三项研究将评估ECIGS暴露在两个体外模型系统中的影响。一个模型通过在器官型培养中培养原代气道上皮细胞来概括气道的生理；而另一种则使用基因修饰的气道上皮细胞，使其敏化以进行致癌转化。这些模型将使我们能够使用基因表达来评估体外暴露反应的保真度，并快速测试ECIGS暴露在促进致癌作用方面的影响。利用ECIGS成功实现这些目标将为在各种其他烟草制品上使用这些方法奠定基础，并将改变评价新烟草制品的范例，从而能够快速评估其潜在的健康风险和致癌性。

DESCRIPTION (provided by applicant): There is a significant need to develop approaches for evaluating the biological impact of new and emerging tobacco-related products in order to identify those that are most likely to cause serious risks to human health. Our proposal is based on the "field of injury" paradigm in which 1) inhaled toxins such as tobacco smoke alter gene-expression in epithelial cells lining the entire respiratory tract and 2) measuring these alteratios in readily accessible upper airway epithelial cells provides insight into the physiological effect f that exposure. We have previously used this concept to identify physiologic responses to tobacco smoke exposure and develop a bronchial airway gene-expression signature that is an early detection biomarker for lung cancer. The overall goals of this proposal are to 1) demonstrate the utility of airway epithelium profiling to identify physiologic responses to other tobacco products and 2) establish in vitro exposure systems that can accurately model physiological exposures and rapidly assess the potential carcinogenicity of tobacco-related products in vitro. We have focused this proposal on E-cigarettes (ECIGS), an emerging FDA-regulated tobacco-related product, due to: 1) their growing popularity, 2) their chemical dissimilarity to tobacco cigarettes (TCIGS), and 3) the absence of data regarding their potential health effects. We propose three simultaneous studies focused around identifying physiologic and cellular responses of airway epithelium to ECIGS. The first two studies involve gene-expression profiling in bronchial and nasal epithelial brushings from ECIGS users with current and former TCIGS smokers serving as comparative groups. These studies will provide a comprehensive view of the physiologic impact of ECIGS, and will assess whether either or both airway tissues can be used to readily assess the physiological effects of ECIGS exposure. The third study will evaluate the effect of ECIGS exposure in two in vitro model systems. One model recapitulates the physiology of the airway by growing primary airway epithelial cells in organotypic culture; while the other uses genetically modified airway epithelial cells that are sensitized to undergo carcinogenic transformation. These models will allow us to use gene expression to evaluate the fidelity of the response to in vitro exposure, and rapidly test the effects of ECIGS exposure in promoting carcinogenesis. Successful completion of these aims using ECIGS will set the stage for using these approaches with a variety of other tobacco products and will alter the paradigm for evaluation of new tobacco products, enabling rapid assessment of their potential health risks and carcinogenicity. PUBLIC HEALTH RELEVANCE: There is an urgent need to rapidly assess the physiologic impact of new tobacco products to identify those that pose serious health risks. We are developing a system to evaluate such products based on detailed molecular portraits of how they impact the biology of airway cells from users of these products and comparing this to how cells grown in the laboratory respond to exposures to these products to determine the feasibility of rapid lab-based product assessment. As our test case, we will study E- cigarettes: an inhaled nicotine delivery device that has been used by over 3% of the adult US population despite a paucity of data about their safety and long term health impact.