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

摘要 由于尼古丁的高度成瘾性，10-12%的美国女性在怀孕期间吸烟。 母亲在怀孕期间吸烟改变了正常的肺发育，导致后代终生减少 肺功能和呼吸系统疾病的风险增加。我们实验室和其他人的工作 研究表明，几乎所有母亲在怀孕期间吸烟对肺部发育的影响都是由以下因素介导的： 尼古丁穿过胎盘与发育中的肺中的尼古丁受体相互作用。这有力地表明 怀孕期间使用电子烟将对肺部发育产生重大不利影响， 后代肺部疾病;更糟糕的是，尼古丁成瘾会驱使怀孕的电子烟使用者继续使用 孕期此外，除了引起后代肺功能的终身变化外， 尼古丁暴露似乎也会改变后代的肺功能，最有可能是通过 表观遗传机制这使得描述子宫内电子烟的影响至关重要 对后代肺发育和功能的影响。因此，本申请的总体目标是使用 小鼠模型，以表征围产期电子烟暴露对后代肺功能的影响， 疾病和宫内电子烟呼吸危害代际传播的可能性 exposure.这个应用程序代表了两个杰出团体之间的合作，平克顿 实验室是世界领先的呼吸暴露和Spindel实验室，世界领先的 子宫内尼古丁暴露对肺部的影响。我们的具体目标如下： 目标1.描述母体宫内电子烟暴露对第一代的直接影响 (F1)后代肺功能、呼吸系统疾病和表观遗传变化。会来做这项工作 将怀孕的BALB/c小鼠暴露于过滤空气、不含尼古丁的电子烟和含尼古丁的电子烟中， 妊娠第1天至出生后第7天，并在8周龄时分析对肺的影响。此外， 将使用对屋尘螨抗原的致敏作用分析子宫内暴露对哮喘易感性的影响。 目标2.描述祖母-母亲宫内电子烟暴露的代际效应 对第二代（F2）后代肺功能、呼吸系统疾病和表观遗传变化的影响。 这将与aim 1一样进行，但将确定对F2代的代际影响。 目标3。描述外祖母和母亲的累加多代效应， 宫内电子烟暴露对后代肺功能、呼吸系统疾病和表观遗传的影响 变化吸烟者的孩子比非吸烟者的孩子更容易吸烟，这是一个不幸的事实。 吸烟者;因此，我们将测试多代宫内电子烟暴露的潜在附加危害。 总之，这些目标将提供一个全面的分析，宫内电子烟暴露的影响， 后代肺功能和潜在的多代增强的影响。

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.