Effects of e-cigarette exposure during pregnancy on offspring lung function and disease: Characterization of pulmonary, intergenerational, and epigenetic effects

摘要 10%-12%的美国女性在怀孕期间吸烟，这反映了尼古丁高度上瘾的本质。 母亲怀孕期间吸烟会改变正常的肺部发育，导致后代终生减少 肺功能和呼吸系统疾病风险增加。我们实验室和其他实验室的工作已经 研究表明，怀孕期间母亲吸烟对肺部发育的几乎所有影响都是通过 尼古丁穿过胎盘与发育中的肺中的尼古丁受体相互作用。这强烈地表明 怀孕期间使用电子烟会对肺部发育和 儿童肺部疾病；更糟糕的是，尼古丁成瘾会促使怀孕的电子烟使用者继续使用 在怀孕期间。此外，还会导致子代肺功能的终身变化，在子宫内 尼古丁暴露似乎也会改变后代的肺功能，很可能是通过 表观遗传机制。因此，确定宫内电子烟的影响至关重要。 暴露对子代肺发育和功能的影响。因此，此应用程序的总体目标是使用 围产期电子烟暴露对子代肺功能和肺功能影响的小鼠模型 疾病和宫内电子烟对呼吸危害的代际传递的可能性 曝光。这个应用程序代表了两个杰出的团体平克顿之间的合作 Spindel实验室是呼吸暴露领域的世界领先者，Spindel实验室是世界领先的 宫内尼古丁暴露对肺部的影响。我们的具体目标如下： 目的1.研究母体宫内电子烟暴露对第一代的直接影响 (F1)子代肺功能、呼吸系统疾病和表观遗传学改变。这将通过以下方式完成 将怀孕的BALB/c小鼠暴露在过滤空气、不含尼古丁的电子烟和含有来自 分析妊娠第1天至出生后第7天对8周龄小鼠肺组织的影响。此外， 宫内暴露对哮喘易感性的分析将使用对尘螨抗原的致敏作用。 目的2.研究孕母宫内电子烟暴露的代际效应 对第二代(F2)后代的肺功能、呼吸系统疾病和表观遗传学改变。 这将在aim1中完成，但将确定对F2代的代际影响。 目的3.表征大母体和母体的加性多世代效应 宫内电子烟暴露对子代肺功能、呼吸系统疾病和表观遗传学的影响 改变。吸烟者的孩子比不吸烟的孩子更有可能吸烟，这是一个不幸的事实 因此，我们将测试多代宫内电子烟暴露的附加危害的可能性。 这些目标将综合分析宫内接触电子烟对健康的影响 子代肺功能和多代效应增强的可能性。

Abstract Reflecting the highly addictive nature of nicotine, 10-12% of American women smoke during pregnancy. Maternal smoking during pregnancy alters normal lung development to produce lifelong decreases in offspring pulmonary function and increased risk of respiratory diseases. Work from our laboratory and others has shown that almost all the effects of maternal smoking during pregnancy on lung development are mediated by nicotine crossing the placenta to interact with nicotinic receptors in developing lung. This strongly suggests that use of e-cigarettes during pregnancy will have significant detrimental effects on lung development and offspring lung disease; and worse, that nicotine addiction will drive pregnant e-cigarette users to continue use during pregnancy. Further, as well as causing lifelong changes in offspring pulmonary function, in-utero nicotine exposure also appears to alter pulmonary function in subsequent generations, most likely through epigenetic mechanisms. This makes it critically important to characterize the effects of in-utero e-cigarette exposure on offspring lung development and function. Thus, the overall objective of this application is to use a mouse model to characterize the effects of perinatal e-cigarette exposure on offspring pulmonary function and disease and the potential for intergenerational transmittal of the respiratory harms of in-utero e-cigarette exposure. This application represents a collaboration between two outstanding groups, the Pinkerton laboratory which is a world leader in respiratory exposures and the Spindel laboratory, a world leader in the pulmonary effects of in utero nicotine exposure. Our specific aims are as follows: Aim 1. To characterize the direct effect of maternal in-utero e-cigarette exposure on first generation (F1) offspring pulmonary function, respiratory disease and epigenetic changes. This will be done by exposing pregnant BALB/c mice to filtered air, e-cigarettes without nicotine and e-cigarettes with nicotine from gestation day 1 to postnatal day 7 and effects on lung analyzed at 8 weeks of age. In addition, the effects of in-utero exposures on asthma susceptibility will be analyzed using sensitization to house dust mite antigen. Aim 2. To characterize the intergenerational effect of grand-maternal in-utero e-cigarette exposure on second generation (F2) offspring pulmonary function, respiratory disease and epigenetic changes. This will be done as for aim1, but intergenerational effects on the F2 generation will be determined. Aim 3. To characterize the additive, multigenerational effect of both grand-maternal and maternal in-utero e-cigarette exposure on offspring pulmonary function, respiratory disease and epigenetic changes. It is an unfortunate fact that children of smokers are more likely to smoke than children of non- smokers; thus, we will test for the potential of additive harms of multigenerational in-utero e-cigarette exposure. Together these aims will provide a comprehensive analysis of the effects of in-utero e-cigarette exposure on offspring pulmonary function and the potential for multigenerational potentiation of the effects.