Identification of molecular mechanisms of carbon nanoparticle-induced senescence and aging ex vivo and in vivo: Role of reactive oxygen species

碳纳米粒子诱导体内和体外衰老的分子机制的鉴定：活性氧的作用

• 批准号: 270971646
• 负责人: Professorin Dr. Judith Haendeler
• 金额: --
• 依托单位: Institut für Klinische Chemie und Laboratoriumsdiagnostik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/270971646?language=en
• 关键词: Identification; molecular; mechanisms; carbon; nanoparticle

The common principle of all combustion-derived carbon nanoparticles (CDNP) is the carbonaceous core with a primary diameter below 100 nm. Multiple studies have demonstrated that the effects of pure carbon nanoparticles can explain several adverse health outcomes to the lung and the cardiovascular system induced by inhaled CDNP. Epidemiologic data indicate that chronic exposure to CDNP leads to features of premature lung aging and age-related cardiovascular diseases. During the process of aging a functional decline of organs occurs resulting to multiple organ failure and death. One hallmark of aging is the occurrence of cellular senescence. Several studies demonstrated that senescent cells are present in vivo and are less functional than non-senescent cells. The Haendeler and Unfried groups together demonstrated for the first time that pure carbon nanoparticles induce features of cellular senescence in endothelial cells and lung epithelial cells. This exposure also increased reactive oxygen species (ROS), believed to be involved in aging and age-associated diseases. Therefore, the aims of this research proposal are to identify the so far unknown underlying molecular mechanisms leading to carbon nanoparticle-induced senescence in primary human lung epithelial cells and vessel endothelial cells ex vivo and to determine whether these mechanisms are also relevant in vivo. First, we will investigate whether ROS are causally involved in senescence induction by concurrent treatment with different antioxidants and subsequent measurement of senescence parameters. Furthermore, we will determine whether carbon nanoparticles change the expression and/or activity of oxidative and/or antioxidative systems. If so, the respective proteins will be permanently re-expressed or downregulated by a lentiviral approach. As mitochondria play an important role in aging processes and senescence, the functionality of these organelles will be measured after carbon nanoparticle treatment. One relevant protein for mitochondrial functions, aging and senescence is mitochondrial Telomerase Reverse Transcriptase (TERT), which is reduced by carbon nanoparticles. Thus, we will determine whether re-expression of mitochondrially localized TERT rescues lung epithelial cells and vessel endothelial cells from carbon nanoparticle-induced senescence. Another important feature of declined lung and vessel functions is impaired intercellular communication. Thus, we will determine the effect of carbon nanoparticles on intercellular communication with a specific focus on Connexin 43. To get further insights in so far unknown pathways, we will perform transcriptome and proteome analyses. Finally, we will investigate whether carbon nanoparticles in non-inflammatory doses also induce senescence in vivo. To test specific pathways we will use two mouse models, one overexpressing NADPH oxidase 4 specifically in the endothelium and one containing TERT exclusively in the mitochondria of all tissues.

所有燃烧法制备的碳纳米颗粒(CDNP)的共同原理是一次直径小于100 nm的碳核。多项研究表明，纯碳纳米颗粒的作用可以解释吸入CDNP对肺和心血管系统造成的几种不良健康后果。流行病学数据表明，慢性接触CDNP会导致肺早衰和年龄相关性心血管疾病的特征。在衰老过程中，器官功能衰退，导致多器官衰竭和死亡。衰老的一个标志是细胞衰老的发生。几项研究表明，衰老细胞存在于体内，其功能不如非衰老细胞。海德勒和昂弗里德的研究小组首次共同展示了纯碳纳米颗粒在内皮细胞和肺上皮细胞中诱导细胞衰老的特征。这种接触还会增加活性氧(ROS)，据信与衰老和与年龄相关的疾病有关。因此，本研究计划的目的是确定碳纳米颗粒在体外诱导原代人肺上皮细胞和血管内皮细胞衰老的潜在分子机制，并确定这些机制是否也与体内相关。首先，我们将通过同时处理不同的抗氧化剂和随后测量衰老参数来研究ROS是否与衰老诱导有关。此外，我们将确定碳纳米颗粒是否会改变氧化和/或抗氧化系统的表达和/或活性。如果是这样的话，相应的蛋白质将通过慢病毒方法永久地重新表达或下调。由于线粒体在衰老过程和衰老过程中起着重要作用，这些细胞器的功能将在碳纳米颗粒处理后进行测量。与线粒体功能、衰老和衰老相关的蛋白质之一是线粒体端粒酶逆转录酶(TERT)，它被碳纳米颗粒还原。因此，我们将确定线粒体定位的TERT的重新表达是否能将肺上皮细胞和血管内皮细胞从碳纳米颗粒诱导的衰老中拯救出来。肺和血管功能下降的另一个重要特征是细胞间通讯受损。因此，我们将确定碳纳米颗粒对细胞间通讯的影响，并特别关注连接蛋白43。为了进一步了解目前未知的途径，我们将进行转录组和蛋白质组分析。最后，我们将研究非炎症剂量的碳纳米颗粒是否也会在体内诱导衰老。为了测试特定的通路，我们将使用两个小鼠模型，一个在内皮细胞中过表达NADPH氧化酶4，另一个在所有组织的线粒体中专门表达TERT。