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

基本信息

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
项目类别：
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

项目摘要

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影响结果包括细胞衰老和与全身相关的肺部毒理学影响我们最近报道了 ETS 扰乱了生物钟。 REV-ERBα 是一种重要的功能，诱导氧化应激并刺激异常炎症反应。分子时钟的组成部分，调节核心时钟基因、促炎和我们的初步数据表明 ETS 介导的细胞衰老是依赖于肺部时钟基因核受体 REV-ERB 的不规则活动。数据进一步显示分子钟缺陷 REV-ERBα 中 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 激动剂可降低 ETS 诱导的促炎和促衰老介质的水平。然而，分子时钟功能障碍在慢性 ETS 介导的 DNA 损伤/修复、细胞 Sirtuin 1 (SIRT1)（一种蛋白质/组蛋白脱乙酰酶）的衰老和毒理学作用仍不清楚。 ETS 减少，导致细胞衰老、氧化应激和异常炎症反应。初步数据显示 SIRT1 调节 REV-ERBα 丰度，但尚不清楚 SIRT1 是否调节。通过 REV-ERBα 依赖性机制调节肺细胞衰老和炎症反应。我们目睹 ETS 破坏分子钟功能，特别是 REV-ERBα 丰度，导致 DNA 损伤通过 SIRT1 依赖性肺毒性机制引发细胞衰老。建议通过确定 ETS 诱导的 REV-ERB破坏对细胞的影响来检验这一假设基于以下三个方面的小鼠模型中的衰老、DNA 损伤/修复和毒理学反应目的：目标 1：确定 ETS 介导的 REV- 破坏的分子机制 ERB分子时钟功能导致细胞衰老和衰老相关炎症表型 (SASP)；目标 2：确定 REV-ERB 参与非同源末端的 DNA 修复 ETS 暴露后细胞衰老期间加入 (NHEJ)；以及目标 3：确定 SIRT1 在细胞衰老过程中的作用；调节 REV-ERB 和 ETS 后 DNA 损伤诱导的肺细胞衰老。该提案的结果将揭示分子钟在调节 DNA 损伤诱导中的作用在 ETS 暴露的异生素反应过程中，通过 SIRT1 依赖性机制抑制肺细胞衰老。反过来，我们的研究结果将对药物疗法的开发具有巨大的转化潜力基于靶向分子时钟功能来改善肺细胞衰老和 DNA 损伤长期接触 ETS。