Human models of the particulate-induced inflammatory/antioxidant axis in aging

衰老过程中颗粒物诱导的炎症/抗氧化轴的人体模型

• 批准号: 9011527
• 负责人: HENRY Jay FORMAN
• 金额: $ 47.51万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-09 至 2019-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9011527
• 关键词: Age; Aging; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Biological; Biological Assay; Breathing; California; Cardiopulmonary; Cardiovascular system; Cells; Cerebellum; Characteristics; Chemicals; Depressed mood; Elderly; Enzyme Induction; Enzymes; Epithelial Cells; Equilibrium; Exposure to; Genetic Transcription; Glutamate-Cysteine Ligase; Glutathione; Goals; Health; Human; Human Cell Line; Individual; Inflammation; Inflammatory; Inflammatory Response; Injury; Interleukin-10; Knockout Mice; Knowledge; Learning; Link; Liver; Lung; Mediating; MicroRNAs; Microscopic; Microscopy; Modeling; Mus; NF-kappa B; Oxidants; Oxidative Stress; Oxidoreductase; Particulate; Pathology; Population; Production; Proteins; Proto-Oncogene Proteins c-myc; Quinones; Reduced Glutathione; Regulation; Research; Resolution; Risk; Shapes; Spectrum Analysis; TNF gene; Testing; Text; age effect; age related; antioxidant enzyme; attenuation; base; bronchial epithelium; c-myc Genes; chromatin immunoprecipitation; cytokine; design; glutathione diethyl ester; heme oxygenase-1; inhibitor/antagonist; macrophage; middle age; model design; monocyte; nanoparticle; nanoparticulate; novel; overexpression; particle; peripheral blood; response; transcription factor; young adult

DESCRIPTION (provided by applicant): Elderly individuals are particularly susceptible to cardiopulmonary pathology induced by airborne nanoparticulate matter (nPM). Antioxidant enzymes are induced by the transcription factor Nrf2 upon nPM inhalation in young but not older mice. The loss of antioxidant inducibility correlates with age-related elevation of c-Myc and Bach1, both of which inhibit Nrf2-regulation of antioxidant enzyme transcription. In aging, inflammatory cytokine production increases both basally and in response to nPM exposure. We hypothesize that the increase in Nrf2-inhibitory proteins in response to nPM during aging is responsible for suppressing antioxidant enzyme inducibility. We further hypothesize that the decrease in Nrf2-dependent antioxidant response in aging permits a greater nPM-induced, NF-kB-regulated increase in inflammatory cytokines. Primary human bronchial epithelium (NHBE) and M1 and M2 macrophages differentiated from peripheral blood monocytes (PBMC) from individuals in four age ranges will be used to model human nPM exposure. Ambient nPM is a mixture of different particle shapes, sizes, and compositions. Thus, we will manufacture defined, reproducible model nPM with physico-chemical characteristics derived from high-resolution microscopy and spectroscopy of ambient nPM from a region with one of the highest nPM health risks. Aim 1 is to demonstrate that the loss in Nrf2-regulated inducibility of antioxidant enzymes (glutamate cysteine ligase, NAD(P)H:quinone oxidoreductase 1, and heme oxygenase-1) in aging is caused by elevation of Nrf2- inhibitory proteins. Bach1 and/or c-Myc will be silenced or overexpressed in NHBE or M1 or M2 macrophages, and expression of Nrf2-regulated antioxidant enzymes in response to nPM determined. Aim 2 is to demonstrate that the decreased ability to induce Nrf2-regulated antioxidant enzymes in aging significantly contributes to elevated inflammatory cytokine production in response to nPM. We will determine the effects of silencing or overexpressing Nrf2, c-Myc and/or Bach1 on NF-kB activation of pro-inflammatory cytokines in response to nPM in NHBE and M1 cells from donors of different ages. The effects of aging and nPM exposure on TNF alpha-induced anti-inflammatory IL-10 expression in M2 cells will also be examined. Aim 3 is to demonstrate that reduced glutathione diethyl ester (GSH-E) or microRNAs (miRNA) that target Bach1 can reverse the attenuation of inducible antioxidant defense in aging. GSH-E, which is converted to glutathione in cells or Bach1-directed miRNAs will be used in NHBE and M1 and M2 macrophages of different ages. Basal and nPM-induced NF-kB activation and induction of cytokines will be determined. The use of human models and representative, reproducible nPM will provide a new mechanistic framework to resolve links among nPM-induced inflammation, antioxidant defense and aging. Acquiring this new knowledge is critical to achieving the ultimate goal of identifying novel means to reduce nPM-induced oxidative injury and inflammation in susceptible individuals, among who are the increasing US elderly population.

描述（由申请人提供）：老年人特别容易受到空中纳米核电物质（NPM）诱导的心肺病理的影响。抗氧化剂酶是由年轻小鼠但不是老鼠的NPM吸入后的转录因子NRF2诱导的。抗氧化剂诱导性的丧失与年龄相关的C-MYC和BACH1升高有关，这两者都抑制了抗氧化酶转录的NRF2调节。在衰老中，炎症性细胞因子产生基本上增加并响应NPM暴露。我们假设在衰老过程中响应NPM的NRF2抑制蛋白的增加是负责抑制抗氧化酶诱导性的。我们进一步假设，衰老中NRF2依赖性抗氧化剂反应的减少允许NPM诱导的NF-KB调节的炎症细胞因子增加。原发性人支气管上皮（NHBE）和M1和M2巨噬细胞与外周血单核细胞（PBMC）与四个年龄范围内的个体区分开来，用于对人NPM暴露进行建模。环境NPM是不同颗粒形状，大小和组成的混合物。因此，我们将使用来自NPM健康风险最高的区域之一的环境NPM的高分辨率显微镜和环境NPM的光谱来制造定义的，可重复的模型NPM。目的1是证明抗氧化剂酶（谷氨酸半胱氨酸连接酶，NAD（P）H：奎因酮氧化还原酶1和血红素氧酶-1）在衰老中的NRF2调节诱导性丧失是由NRF2-抑制蛋白的升高引起的。在NHBE，M1或M2巨噬细胞中，BACH1和/或C-MYC将沉默或过表达，并响应于确定的NPM响应NRF2调节的抗氧化剂酶的表达。 AIM 2是证明衰老中诱导NRF2调节的抗氧化酶的能力降低，显着促进了响应NPM的炎性细胞因子产生的升高。我们将确定沉默或过表达NRF2，C-MYC和/或BACH1对NHBE中NPM的促炎细胞因子的NF-KB激活NHBE的NF-KB激活的影响，来自不同年龄的NHBE和M1细胞。还将检查衰老和NPM暴露对M2细胞中TNFα诱导的抗炎IL-10表达的影响。 AIM 3是证明靶向Bach1可以逆转衰老中诱导抗氧化剂防御的衰减，证明减少了谷胱甘肽二乙基酯（GSH-E）或microRNA（miRNA）。在不同年龄的NHBE，M1和M2巨噬细胞中，将使用将其转化为细胞或BACH1指导的miRNA中的谷胱甘肽的GSH-E。将确定基底和NPM诱导的NF-KB激活以及细胞因子的诱导。 使用人类模型和代表性，可重复的NPM将提供一个新的机械框架，以解决NPM引起的炎症，抗氧化剂防御和衰老之间的联系。获取这一新知识对于确定新的方法是减少NPM诱导的氧化损伤和易感人群的炎症的最终目标至关重要，而美国老年人群的人口不断增加。