Nicotine, Airway Smooth Muscle and Asthma

尼古丁、气道平滑肌和哮喘

• 批准号: 10441276
• 负责人: Christina Maria Pabelick
• 金额: $ 51万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-05 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10441276
• 关键词: Acute; Adult; Affect; Agonist; Airway Resistance; Allergens; Asthma; Bathing; Biochemical; Biology; Cell Proliferation; Cell membrane; Cells; Cholinergic Receptors; Chronic; Cytokine Signaling; Data; Devices; Electronic cigarette; Electrophysiology (science); Elements; Epithelial; Exposure to; Extrinsic asthma; Genes; Goals; Grant; Human; In Vitro; Inflammation; Inflammatory; Interleukin-13; Interleukin-6; Knockout Mice; Lung; MAP Kinase Gene; Mediating; Metabolism; Mitochondria; Molecular Chaperones; Mus; Muscarinic Acetylcholine Receptor; Muscle; Neurons; Nicotine; Nicotinic Receptors; OPA1 gene; Organ; PI3 gene; Pathway interactions; Regulation; Respiration; Role; Sampling; Signal Transduction; Small Interfering RNA; Smoking; Smoking History; Smooth Muscle; Smooth Muscle Myocytes; Structure; System; TNF gene; Testing; Toxic effect; Traction; airway hyperresponsiveness; alpha-bungarotoxin receptor; antagonist; asthma model; asthmatic; asthmatic airway; asthmatic airway smooth muscle; clinically relevant; clinically significant; cytokine; endoplasmic reticulum stress; inhibitor; methacholine; mouse allergen; mouse model; nicotine exposure; nicotine inhalation; nicotine use; novel; patch clamp; protein metabolism; respiratory smooth muscle; response; sex; therapeutic target

ABSTRACT While e-cigarettes are advertised as safer alternatives to smoking for asthmatics, there is increasing evidence of their pulmonary toxicity, necessitating better understanding of airway nicotine biology: a clinically-relevant unmet need, and major goal of this grant. In asthma, inflammation enhances airway smooth muscle (ASM) Ca2+ ([Ca2+]cyt) and contractility (airway hyperreactivity; AHR), along with remodeling involving ASM proliferation. Given that inhaled nicotine can directly influence ASM, nicotine effects on asthmatic ASM are critical, but little is known about nicotinic receptor (nAChR) expression, its role in human ASM, or in asthma. Preliminary data in human ASM show functional α7 nAChRs that A) increase with inflammation and asthma, B) enhance ASM [Ca2+]cyt/contractility; C) enhance ER stress and mitochondrial fission, and respiration, and D) activate pro-proliferative pathways. Such effects are observed in a mixed-allergen mouse model of asthma exposed to acute or chronic inhaled nicotine, but blunted by α7 inhibition and in α7 KO mice. Thus, our hypothesis is that nicotine acts via ASM α7 to promote AHR and remodeling in asthma. Our Aims are: Aim 1: : In human ASM, determine mechanisms of α7 expression and regulation in inflammation and asthma. Aim 2: In human ASM of non-asthmatics vs. asthmatics, to determine mechanisms by which α7 contributes to nicotine enhancement of [Ca2+]cyt and contractility in the context of AHR. Aim 3: In human ASM of non-asthmatics vs. asthmatics, to determine mechanisms by which α7 contributes to nicotine enhancement of cell proliferation. Aim 4: In a mouse model of allergic asthma, to determine the role of α7 in effects of inhaled nicotine on AHR and remodeling. Studies use adult human ASM of non-asthmatics vs. mild-moderate asthmatics exposed to cytokines relevant to ASM and to asthma (TNFα, IL-6, IL-13), with/without nicotine. Aim 1 explores expression and localization of α7 nAChR, regulatory chaperones, and mechanisms by which α7 is increased in inflammation/asthma (e.g. MAPKs, PI3/Akt, NFκB, Stats). α7 functionality as a channel is tested using electrophysiology. Aim 2 explores acute vs. chronic nicotine effects on [Ca2+]cyt regulation (influx, SR Ca2+ release) and contractility (traction force, organ bath). Aim 3 explores nicotine effects on ASM proliferation, and the roles of ER stress, mitochondrial fission (Drp1, Fis1) vs. fusion (Mfn1/2, Opa1) and respiration (3a), and of cytokine-associated proliferative pathways (3b). In these Aims, role of α7 is determined using broad vs. subunit- specific agonists, antagonists or siRNAs. In vitro results are integrated in Aim 4 using the adult mouse model of asthma with/without acute vs. chronic inhaled nicotine. Alleviating effects of α7 inhibitor (MG624) and effects of α7 KO in smooth muscle are tested. Studies assess airway reactivity, remodeling, and ASM biochemical changes. Clinical significance lies in identifying ASM α7 as a novel target for alleviating AHR and remodeling of asthma as well as with nicotine.

抽象的 虽然电子烟是广告作为哮喘患者吸烟的安全替代品，但有越来越多的证据 它们的肺毒性，必须更好地了解气道尼古丁生物学：临床上的毒性 未满足的需求和这笔赠款的主要目标。在哮喘中，注射增强气道平滑肌（ASM） Ca2+（[Ca2+] Cyt）和收缩力（气道高反应性； AHR），以及涉及ASM的重塑 增殖。鉴于继承的尼古丁可以直接影响ASM，因此尼古丁对哮喘的影响是 关于烟碱受体（NACHR）的表达，其在人类ASM或哮喘中的作用鲜为人知，但知之甚少。 人ASM中的初步数据显示功能性α7NACHR，a）随着注射和哮喘的增加而增加 b）增强ASM [Ca2+] Cyt/收缩力； c）增强ER应力和线粒体裂变，呼吸，以及 d）激活促生产途径。在哮喘的混合过敏原小鼠模型中观察到这种影响 暴露于急性或慢性遗传的尼古丁，但受到α7抑制和α7KO小鼠的钝化。那，我们的 假设是尼古丁通过ASMα7作用以促进AHR和重塑哮喘。我们的目标是： AIM 1 ：：在人ASM中，确定炎症和哮喘中α7表达和调节的机制。 目的2：在非亚错的人类ASM中与哮喘患者，以确定α7有助于的机制 在AHR的情况下，[Ca2+] Cyt和收缩力的尼古丁增强。目的3：在非asthmatics的人类ASM中 vs.哮喘患者，以确定α7有助于尼古丁增强的机制。 AIM 4：在过敏性哮喘的小鼠模型中，确定α7在遗传尼古丁对AHR的影响中的作用 和重塑。研究使用非亚错的成人人类ASM与暴露于轻度的哮喘患者 与ASM和哮喘相关的细胞因子（TNFα，IL-6，IL-13），具有/没有尼古丁。 AIM 1探索表达 以及α7NACHR，调节伴侣的定位以及α7在中增加的机制 炎症/哮喘（例如MAPK，PI3/AKT，NFκB，Stats）。使用α7功能作为通道进行测试 电生理学。 AIM 2探索急性与慢性尼古丁对[Ca2+] Cyt调节的影响（涌入，SR CA2+ 释放）和收缩力（牵引力，器官浴）。 AIM 3探索尼古丁对ASM增殖的影响，并 ER应力，线粒体裂变（DRP1，FIS1）与融合（MFN1/2，OPA1）和呼吸（3A）的作用（3A） 细胞因子相关的增殖途径（3B）。在这些目标中，使用广泛的和亚基 - 特定的激动剂，拮抗剂或siRNA。体外结果使用成人小鼠模型集成在AIM 4中 哮喘，有/没有急性与慢性遗传的尼古丁。 α7抑制​​剂（MG624）的减轻作用和 测试平滑肌中的α7KO。研究评估气道反应性，重塑和ASM生化 更改。临床意义在于将ASMα7识别为减轻AHR和重塑的新目标 哮喘和尼古丁。

项目成果

Aging increases senescence, calcium signaling, and extracellular matrix deposition in human airway smooth muscle.

• DOI: 10.1371/journal.pone.0254710
• 发表时间: 2021
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Wicher SA;Roos BB;Teske JJ;Fang YH;Pabelick C;Prakash YS
• 通讯作者: Prakash YS

Glial-derived neurotrophic factor in human airway smooth muscle.

• DOI: 10.1002/jcp.30489
• 发表时间: 2021-12
• 期刊: Journal of cellular physiology
• 影响因子: 5.6
• 作者: Bhallamudi S;Roos BB;Teske JJ;Wicher SA;McConico A;M Pabelick C;Sathish V;Prakash YS
• 通讯作者: Prakash YS

Mechanosensitive channels in lung disease.

• DOI: 10.3389/fphys.2023.1302631
• 发表时间: 2023
• 期刊: Frontiers in physiology
• 影响因子: 4

Nicotinic α7 acetylcholine receptor (α7nAChR) in human airway smooth muscle.

• DOI: 10.1016/j.abb.2021.108897
• 发表时间: 2021-07-30
• 期刊: Archives of biochemistry and biophysics
• 影响因子: 3.9
• 作者: Borkar NA;Roos B;Prakash YS;Sathish V;Pabelick CM
• 通讯作者: Pabelick CM

Network and co-expression analysis of airway smooth muscle cell transcriptome delineates potential gene signatures in asthma.

• DOI: 10.1038/s41598-021-93845-x
• 发表时间: 2021-07-13
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Banerjee P;Balraj P;Ambhore NS;Wicher SA;Britt RD Jr;Pabelick CM;Prakash YS;Sathish V
• 通讯作者: Sathish V