Molecular mechanisms for E-cigarette vapor-induced mucociliary dysfunction: role of nicotinic acetylcholine and TRP receptors

电子烟蒸汽引起的粘膜纤毛功能障碍的分子机制：烟碱乙酰胆碱和 TRP 受体的作用

• 批准号: 9756113
• 负责人: Samuel Chung
• 金额: $ 6.64万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9756113
• 关键词: Acetylcholine; Acute; Address; Adolescent; Adopted; Adoption; Adult; Aerosols; Affect; Agonist; Air; Apical; Belief; Biological; Calcium; Career Mobility; Cations; Cell Differentiation process; Chronic; Cinnamon - dietary; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Dehydration; Electronic cigarette; Environment; Epithelial Cells; Flavoring; Functional disorder; Future; Gene Expression Profile; Goals; Health; Health Policy; Human; Hydration status; Hydrogen Peroxide; Impairment; In Vitro; Inflammatory Response; Inhalation; Inhalation Exposure; Lead; Liquid substance; Lung; Lung diseases; Measures; Mediating; Menthol; Mentors; Methodology; Methods; Molecular; Mucociliary Clearance; Mucous body substance; Neuraxis; Nicotine; Nicotinic Receptors; Nose; Oxidants; Oxidases; Phase; Physiological; Production; Protein Isoforms; Public Health; Reporter; Reporting; Research; Research Personnel; Research Proposals; Role; Scientist; Signal Pathway; Signal Transduction; Smoker; Smoking History; Surface; System; TRPA1 Channel; TRPV1 gene; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Tobacco smoke; Toxic effect; Toxicology; Training; Translational Research; Variant; aerosolized; airway epithelium; cinnamic aldehyde; electronic cigarette use; electronic cigarette user; large-conductance calcium-activated potassium channels; mRNA Expression; member; never smoker; never smoking; nicotine exposure; nicotine inhalation; nicotine vapor; novel; post-doctoral training; receptor; respiratory; respiratory smooth muscle; smoking cessation; social stigma; tool; toxicant; vanillin; vaping; vapor; voltage

Use of e-cigarettes (E-cigs), known as “vaping”, is becoming widely adopted amongst adolescents who are attracted by their novelty, synthetic flavors, and belief of reduced toxicity. Previous smokers also use E-cigs as a smoking cessation tool that is an alternative nicotine delivery system with milder social stigma. However, “vaping” is an emerging public health problem, especially concerning young people with no smoking history, due to the lack of informative respiratory studies on E-cig vapor components. Past inhaled nicotine studies examined its effects mainly on airway smooth muscle and the central nervous system as opposed to the ciliated airway epithelia, the major barrier for inhaled E-cig vapor, and few studies investigated nicotine in isolation of tobacco smoke constituents. Additionally, toxicology studies of most E-cig flavorings were never completed for inhalation exposure. This research proposal, which doubles as a mentored postdoctoral training plan, aims to elucidate effects of E-cig vapor on the functional human ciliated airway epithelium. The airway epithelium expresses several isoforms of nicotinic acetylcholine receptors (nAChRs), transient receptor potential (TRP) cation channel member A1 (TRPA1), and other TRP channels, which are all calcium (Ca2+)-permeant. Nicotine is reported to stimulate both nAChRs and TRPA1, and some E-cig flavors are TRP agonists, such as cinnamaldehyde (TRPA1), vanillin (TRPV1), and menthol (TRPM8). The basic hypothesis of this proposal is that E-cig vapor components nicotine and cinnamaldehyde compromise mucociliary function by stimulating nAChRs and TRPA1 to cause sustained Ca2+ influx. Research goals for this study are to identify a molecular mechanism(s) for how E-cig vapor impairs mucociliary function and to develop novel high-throughput methodology for establishing continuous mucociliary transport in vitro. The latter will be used to probe adverse/therapeutic effects of aerosolized compounds. These goals will be tested in primary human bronchial epithelial cells differentiated at the air-liquid interface, using physiologically-relevant exposures, and measuring in vitro parameters of mucociliary function such as mucociliary transport, ciliary beating, and airway surface hydration. Training goals for this proposal are to develop expertise in studying pulmonary effects of inhaled toxicants and to become immersed in translational research by working in a diverse team composed of clinicians and scientists. Completion of this study will fulfill the unmet need for clarification and information regarding effects of inhaled nicotine and E-cig flavorings. Use of relevant exposures and primary cultures will provide a compelling biological rationale to adapt public health policy. It will also inform clinicians on the `pro & cons' of E-cigs as smoking cessation tools as well as the impact on young never-smoking subjects who take up “vaping”. Lastly, completion of this training plan will generate data and expertise necessary for career transition from postdoctoral trainee to independent investigator.

电子烟（E-cigs）的使用，被称为“vaping”，正在青少年中广泛采用。 被它们的新颖性、合成风味和毒性降低的信念所吸引。以前的吸烟者也使用电子烟作为 一种戒烟工具，是一种替代性尼古丁输送系统，具有较温和的社会耻辱感。然而， “吸电子烟”是一个新兴的公共卫生问题，特别是对于没有吸烟史的年轻人来说， 由于缺乏有关电子烟蒸气成分的翔实的呼吸研究。过去的吸入尼古丁研究 研究了其主要对气道平滑肌和中枢神经系统的影响，而不是对气道平滑肌和中枢神经系统的影响。 纤毛气道上皮细胞是吸入电子烟蒸汽的主要屏障，很少有研究调查尼古丁在 分离烟​​草烟雾成分。此外，大多数电子烟香料的毒理学研究从未进行过 完成吸入暴露。这项研究计划同时也是指导性博士后培训 计划，旨在阐明电子烟蒸汽对功能性人类纤毛气道上皮的影响。 气道上皮表达烟碱乙酰胆碱受体 (nAChR) 的几种亚型，瞬时 受体电位 (TRP) 阳离子通道成员 A1 (TRPA1) 和其他 TRP 通道，它们都是钙 (Ca2+)-渗透。据报道，尼古丁会刺激 nAChR 和 TRPA1，某些电子烟口味中含有 TRP 激动剂，例如肉桂醛 (TRPA1)、香草醛 (TRPV1) 和薄荷醇 (TRPM8)。基本假设 该提案的主要内容是电子烟蒸汽成分尼古丁和肉桂醛会损害粘膜纤毛 通过刺激 nAChR 和 TRPA1 引起持续的 Ca2+ 流入来发挥作用。本研究的研究目标 旨在确定电子烟蒸汽如何损害粘膜纤毛功能的分子机制，并开发新的 用于建立体外连续粘液纤毛运输的高通量方法。将使用后者 探讨雾化化合物的不良/治疗效果。这些目标将在初级人类身上进行测试 使用生理相关的暴露，在气液界面分化支气管上皮细胞，以及 测量粘膜纤毛功能的体外参数，例如粘膜纤毛运输、纤毛跳动和气道 表面水合作用。本提案的培训目标是培养研究肺部影响的专业知识 吸入有毒物质，并通过在由以下人员组成的多元化团队中工作，沉浸在转化研究中： 临床医生和科学家。 本研究的完成将满足对有关影响的澄清和信息的未满足的需求 吸入尼古丁和电子烟调味剂。使用相关的接触和原代文化将提供令人信服的 适应公共卫生政策的生物学原理。它还将告知临床医生电子烟的“优点和缺点” 戒烟工具以及对吸食“电子烟”的年轻不吸烟者的影响。最后， 完成该培训计划将产生职业转型所需的数据和专业知识 独立研究员的博士后实习生。