Molecular clock dysfunction in lung cellular senescence by environmental tobacco smoke

环境烟草烟雾导致肺细胞衰老的分子钟功能障碍

• 批准号: 10330545
• 负责人: IRFAN RAHMAN
• 金额: $ 34.65万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10330545
• 关键词: Adverse effects; Agonist; Attenuated; Behavior; Cell Aging; Cells; Chronic; Circadian Dysregulation; Circadian Rhythms; Clock protein; DNA Damage; DNA Repair; DNA Repair Gene; Data; Deacetylase; Development; Disease; Environmental Impact; Epithelial; Epithelial Cells; Excision; Exhibits; Functional disorder; G22P1 gene; Gene Expression; Genes; Genetic; Growth; Histone Deacetylase; Human; Impairment; Inflammation; Inflammatory; Inflammatory Response; Injury; Irritants; Knock-out; Knockout Mice; Luciferases; Lung; Lung Compliance; Mediating; Mediator of activation protein; Molecular; Molecular Target; Mouse Strains; Mus; Nonhomologous DNA End Joining; Nuclear Receptor Gene; Nuclear Receptors; Outcome; Oxidative Stress; Pathogenesis; Patients; Pharmacology; Phenotype; Physiology; Proteins; Reporter; Reporting; Role; SIRT1 gene; Signal Transduction; Stress; Testing; Toxic effect; Toxicology; Transgenic Mice; Wild Type Mouse; Xenobiotics; adverse outcome; airway epithelium; antagonist; anti aging; base; circadian; circadian pacemaker; environmental tobacco smoke; environmental tobacco smoke exposure; genetic corepressor; molecular clock; mouse model; mutant; premature; premature lung aging; recruit; response; rev Protein; senescence; translational impact; translational potential

SUMMARY Environmental tobacco smoke (ETS) or “secondhand smoke” exposure leads to stress-induced adverse outcomes including cellular senescence and toxicological effects on the lungs associated with systemic injury and inflammation in airway disorders. We have recently reported that ETS disrupts circadian clock function, induces oxidative stress and stimulates abnormal inflammatory responses. REV-ERBα is a critical component of the molecular clock, which regulates the expression of core clock genes, pro-inflammatory and pro-senescent mediators. Our preliminary data suggest that ETS-mediated cellular senescence is dependent on irregular activity of the clock gene nuclear receptor REV-ERB in the lungs. Our preliminary data further show an impaired DNA repair and inflammatory response in molecular clock deficient REV-ERBα KO mice, suggesting the involvement of the clock protein REV-ERB in regulating DNA damage/repair. Further, REV-ERBα agonists reduce ETS-induced levels of pro-inflammatory and pro-senescent mediators. However, the role of molecular clock dysfunction in chronic ETS-mediated DNA damage/repair, cellular senescence, and toxicological effects remains unknown. Sirtuin 1 (SIRT1), a protein/histone deacetylase, is reduced by ETS, resulting in cellular senescence, oxidative stress and abnormal inflammatory responses. Our preliminary data show that SIRT1 regulates REV-ERBα abundance. However, it is not known whether SIRT1 regulates lung cellular senescence and inflammatory responses through a REV-ERBα-dependent mechanism. We hypothesize that ETS disrupts molecular clock function, specifically REV-ERBα abundance, resulting in DNA damage-initiated cellular senescence through a SIRT1-dependent mechanism in pulmonary toxicity. We propose to test this hypothesis by determining the impact of ETS-induced REV-ERBdisruption on cellular senescence, DNA damage/repair, and toxicological responses in a mouse model based on the following three Aims: Aim 1: Determine the molecular mechanism through which ETS-mediated disruption of REV- ERBmolecular clock function leads to cellular senescence and senescence-associated inflammatory phenotype (SASP); Aim 2: Determine the involvement of REV-ERB in DNA repair by non-homologous end joining (NHEJ) during cellular senescence following ETS exposure; and Aim 3: Determine the role of SIRT1 in regulating REV-ERB and DNA damage-induced lung cellular senescence following ETS. The outcome of this proposal will unravel the role of the molecular clock in regulating DNA damage-induced lung cellular senescence via a SIRT1-dependent mechanism during the xenobiotic response to ETS exposure. In turn, our findings will have great translational potential for the development of pharmacological therapies based on targeting molecular clock function to ameliorate lung cellular senescence and DNA damage following chronic exposure to ETS.

概括 环境烟草烟雾（ETS）或“二手烟”暴露会导致压力引起的不利 结局包括细胞感应和毒理学对与全身性相关的肺的影响 气道疾病的伤害和感染。我们最近报告说ETS破坏了昼夜节律的时钟 功能，诱导氧化应激并刺激异常的炎症反应。 Rev-erbα是关键 分子时钟的成分，该分子时钟调节核心时钟基因的表达，促炎和 亲渗的介体。我们的初步数据表明ETS介导的细胞感应是 取决于肺中时钟基因核接收器Rev-erb的不规则活性。我们的初步 数据进一步显示了分子时钟缺陷REV-ERBα的DNA修复和炎症反应受损 KO小鼠，表明时钟蛋白Rev-erb参与调节DNA损伤/修复。 此外，Rev-ERBα激动剂降低了ETS诱导的促炎和促介质的水平。 但是，分子时钟功能障碍在慢性ETS介导的DNA损伤/修复，细胞中的作用 感应和毒理作用仍然未知。 SIRTUIN 1（SIRT1），一种蛋白质/组蛋白脱乙酰基酶，为 通过ETS降低，导致细胞感应，氧化应激和异常炎症反应。我们的 初步数据表明，SIRT1调节Rev-ERBα抽象。但是，尚不知道SIRT1是否 通过Rev-ERBα依赖性机制调节肺细胞感应和炎症反应。 我们假设ET破坏了分子时钟功能，特别是Rev-erbα抽象，导致 通过肺毒性中的SIRT1依赖机制，DNA损伤引起的细胞感染。我们 通过确定ETS诱导的Rev-erb的影响对细胞的影响来检验这一假设的建议 基于以下三种 目的：目标1：确定ETS介导的Rev-中断的分子机制 ERB分子时钟功能导致细胞感应和感应相关的炎症 表型（SASP）;目标2：确定Rev-erb的参与，通过非理论末端进行DNA修复 ETS暴露后的细胞感应期间（NHEJ）连接（NHEJ）；和目标3：确定SIRT1在 调节REV-ERB和DNA损伤诱导的ETS后肺细胞感应。 该提案的结果将揭示分子时钟在调节DNA损伤引起的 在异种生物对ETS暴露的反应期间，通过SIRT1依赖机制的肺部细胞感染。 反过来，我们的发现将具有巨大的翻译潜力来开发药物疗法 基于靶向分子时钟功能，以改善肺细胞感应和DNA损伤 长期暴露于ETS。