The UCLA - Boston University Lung Cancer Biomarker Development Laboratory

说明(由申请人提供)：迫切需要制定评估新的和新兴的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。我们的建议基于“伤害场”范式，在该范式中，1)吸入烟草烟雾等毒素会改变整个呼吸道衬里上皮细胞的基因表达，2)测量容易接触到的上呼吸道上皮细胞中的这些变化可以深入了解这种暴露的生理效应。我们以前已经使用这一概念来确定对烟草烟雾暴露的生理反应，并开发出一种支气管呼吸道基因表达特征，这是肺癌的早期检测生物标志物。这项建议的总体目标是：1)证明呼吸道上皮细胞图谱用于识别对其他烟草产品的生理反应；2)建立体外暴露系统，准确模拟生理暴露，并快速评估与烟草相关的产品在体外的潜在致癌性。我们将这一建议集中在电子烟(ECigs)上，这是一种新兴的FDA监管的烟草相关产品，原因是：1)它们越来越受欢迎，2)它们与烟草香烟(TCIGS)的化学成分不同，以及3)缺乏关于它们潜在健康影响的数据。我们提出了三项同步研究，重点是确定eCigs对呼吸道上皮的生理和细胞反应。前两项研究涉及eCigs使用者的支气管和鼻腔上皮刷子中的基因表达谱，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看法，并将评估是否可以使用其中一种或两种呼吸道组织来容易地评估eCigs暴露的生理影响。第三项研究将在两个体外模型系统中评估eCigs暴露的影响。一种模型通过在器官型培养中培养原代呼吸道上皮细胞来概括呼吸道的生理学；另一种模型使用转基因的呼吸道上皮细胞，这些细胞被致敏后经历致癌转化。这些模型将使我们能够使用基因表达来评估对体外暴露的反应的保真度，并快速测试eCigs暴露在促进癌症发生方面的效果。使用eCigs成功完成这些目标将为将这些方法用于各种其他烟草产品奠定基础，并将改变评估新烟草产品的范式，使快速评估其潜在的健康风险和致癌性成为可能。 与公共健康相关：迫切需要迅速评估新烟草产品的生理影响，以确定那些构成严重健康风险的产品。我们正在开发一种系统，根据这些产品的使用者对它们如何影响呼吸道细胞生物学的详细分子画像来评估这些产品，并将其与实验室中生长的细胞对这些产品的暴露做出反应进行比较，以确定基于实验室的快速产品评估的可行性。作为我们的测试案例，我们将研究电子烟：一种吸入性尼古丁释放装置，尽管缺乏关于其安全性和长期健康影响的数据，但仍有超过3%的美国成年人使用过这种装置。

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.