Blood and Brain Based Biomarkers of Injury to Assess the Cerebrovascular Impact of Emerging Alternatives to Classic Cigarette Products

摘要： 在过去的十年中，几种替代电子烟产品已经上市，迅速赢得了成年人的消费者 尤其是青少年。电子尼古丁输送系统或电子烟（e-Cigs）已广泛普及 在吸烟者和以前不吸烟者中，因为人们相信它们比传统香烟安全得多。其中 JUUL已经成为包括中学生在内的年轻人追捧的产品。而且，还好 确定吸烟（TS）与血管内皮功能障碍有关，其原因和剂量 依赖方式主要与活性氧（ROS）、尼古丁和氧化应激的TS含量有关 （OS）驱动的炎症。当前的科学观点认为操作系统介导的途径在 这些疾病的发病机制，尤其是中风。临床前研究还表明，尼古丁（主要电子物质） 电子烟中使用的液体成分）也会导致OS、脑缺血加剧和继发性脑损伤。 同样，长期吸电子烟可能是脑血管损伤的前驱症状，并会促进脑血管损伤。 有利于中风发作和缺血后脑损伤恶化的条件。与相关的健康影响 慢性电子烟在很大程度上是未知的，尤其是在脑血管系统方面。广泛使用这些 替代产品强烈要求进行调查研究以确定其真正的健康影响以及更严格的要求 有关电子烟解决方案（电子液体）内容的监管指南。我们的新初步数据还表明 这些产品可能存在特定性别的敏感性。因此，根据计划范围和研究 FDA 烟草中心发布的呼吁采取行动调查电子烟对健康的影响和毒性的目标 产品 基于我们实验室已发表的大量初步数据，我们提出以下建议： 1) 评估电子烟和 Juuling 与 TS 暴露对脑血管的影响，并制定一组潜在的影响 生物标志物来确定这些产品的危害。这些研究的前提是并列比较研究 电子烟和 JUUL 与 TS 对 BBB 的危害或毒性。这包括评估不同电子烟油的影响 配方中的尼古丁含量以及用于产生蒸汽的线圈热量的影响。 2) 在体内评估和验证长期接触电子烟和 JUUL 与 TS 相比对中风风险的影响， 继发性脑损伤和缺血后神经功能障碍。这些研究的前提建立在 最新的临床发现和我们最近发布的数据表明，与 TS 类似，长期接触电子烟会促进 脑血管炎症、中风，并使缺血后继发性脑损伤恶化。也解决了这种可能性 对于特定性别的敏感性/风险和结果，我们将使用雄性和雌性小鼠的混合性别群体。 总的来说，我们将比较吸烟和电子烟（包括含有不同物质的电子烟液）的影响 尼古丁浓度和用于蒸发电子烟液的温度设置）对脑血管损伤的影响。我们的研究将 特别关注它们对中风风险和结果的影响，这是通过基于大脑和血液的生物标志物确定的。

ABSTRACT: In the past decade several alternative vaping products have hit the market, rapidly gaining consumers among adults and, especially, adolescents. Electronic nicotine delivery systems or e-cigarettes (e-Cigs) have become wide spread among both smokers and former non-smokers due to the belief that they are much safer than traditional cigarettes. Among them JUULs have become the sought-after product among youth including middle schoolers. Moreover, it is well established that tobacco smoking (TS) has been associated with vascular endothelial dysfunction in a causative and dose dependent manner primarily related to the TS content of reactive oxygen species (ROS), nicotine, and oxidative stress (OS) -driven inflammation. Current scientific opinion considers OS-mediated pathways to play a major role in the pathogenesis of these disorders, especially stroke. Preclinical studies have also shown that nicotine (the principal e- liquid's ingredient used in e-cigarettes) can also cause OS, exacerbation of cerebral ischemia and secondary brain injury. Likewise, chronic e-Cig vaping could be prodromal to cerebrovascular impairment and promote cerebrovascular conditions favoring the onset of stroke and worsening post-ischemic brain injury. The health impact associated with chronic e-Cig vaping is largely unknown especially concerning the cerebrovascular system. Wide spread use of these alternative products strongly calls for investigative studies to determine their real health impact as well as more stringent regulatory guidelines concerning the content of the vaping solutions (e-liquids). Our new preliminary data also suggests the possibility for gender specific sensitivities to these products. Thus, in response to the program scope and research objectives of this call for action to investigate e-Cig health effects and toxicity issued by the FDA Center for Tobacco Products Based and based on the substantial published and preliminary data from our Labs we propose the following: 1) Assess the cerebrovascular impact of e-Cig vaping and Juuling vs. TS exposure and develop a panel of potential biomarkers to determine harm of these products. The premise of these studies is to comparatively investigate side by side the harm or toxicity of e-Cigs and JUUL vs. TS on the BBB. This includes assessing the impact of different e-liquid formulations in respect to the nicotine content as well as impact of the coil heat used to generate the vapor. 2) Evaluate and validate in vivo the impact of chronic exposure to e-Cigs and JUULs vs. TS on the risk of stroke, secondary brain damage, and post-ischemic neurological impairments. The premise for these studies builds upon the latest clinical findings and our recent published data suggesting that similarly to TS, chronic e-Cigs exposure promotes cerebrovascular inflammation, stroke, and worsen post-ischemic secondary brain injuries. To also address the possibility for gender specific sensitivities/risks and outcomes we will use a mixed gender population of male and female mice. Overall, we will compare the impact of tobacco smoking and e-Cig vaping (including e-liquids containing different nicotine concentrations and temperature settings used to vaporize the e-liquid) on brain vascular damage. Our study will focus specifically on their impact on stroke risk and outcome, ascertained from brain and blood based biomarkers.