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气中纳米颗粒物质（nPM）引起的心肺病变的影响。年轻小鼠吸入 nPM 后，转录因子 Nrf2 会诱导抗氧化酶，但老年小鼠则不会。抗氧化诱导能力的丧失与年龄相关的 c-Myc 和 Bach1 升高相关，这两者都抑制抗氧化酶转录的 Nrf2 调节。在衰老过程中，炎症细胞因子的产生在基础上和对 nPM 暴露的反应中都会增加。我们假设衰老过程中 Nrf2 抑制蛋白的增加对 nPM 的反应是抑制抗氧化酶诱导的原因。我们进一步假设，衰老过程中 Nrf2 依赖性抗氧化反应的减少使得 nPM 诱导的、NF-kB 调节的炎症细胞因子增加更大。人类原代支气管上皮 (NHBE) 以及由四个年龄段个体的外周血单核细胞 (PBMC) 分化而来的 M1 和 M2 巨噬细胞将用于模拟人类 nPM 暴露。环境 nPM 是不同颗粒形状、尺寸和成分的混合物。因此，我们将制造定义的、可重复的 nPM 模型，其物理化学特性源自高分辨率显微镜和来自 nPM 健康风险最高地区之一的环境 nPM 光谱。目标 1 是证明衰老过程中 Nrf2 调节的抗氧化酶（谷氨酸半胱氨酸连接酶、NAD(P)H：醌氧化还原酶 1 和血红素加氧酶 1）诱导能力的丧失是由 Nrf2 抑制蛋白的升高引起的。 Bach1 和/或 c-Myc 将在 NHBE 或 M1 或 M2 巨噬细胞中沉默或过表达，并确定响应 nPM 的 Nrf2 调节的抗氧化酶的表达。目标 2 是证明，在衰老过程中诱导 Nrf2 调节的抗氧化酶的能力下降，会显着导致响应 nPM 的炎症细胞因子的产生增加。我们将确定沉默或过表达 Nrf2、c-Myc 和/或 Bach1 对来自不同年龄供体的 NHBE 和 M1 细胞中 nPM 响应的促炎细胞因子 NF-kB 激活的影响。还将检查衰老和 nPM 暴露对 M2 细胞中 TNF α 诱导的抗炎 IL-10 表达的影响。目标 3 是证明靶向 Bach1 的还原型谷胱甘肽二乙酯 (GSH-E) 或 microRNA (miRNA) 可以逆转衰老过程中诱导性抗氧化防御的减弱。 GSH-E在细胞中转化为谷胱甘肽或Bach1定向的miRNA，将用于不同年龄的NHBE以及M1和M2巨噬细胞。将确定基础和 nPM 诱导的 NF-kB 激活以及细胞因子的诱导。 使用人体模型和具有代表性的、可重复的 nPM 将为解决 nPM 引起的炎症、抗氧化防御和衰老之间的联系提供一个新的机制框架。获得这一新知识对于实现最终目标至关重要，即找到新的方法来减少易感人群（其中包括不断增加的美国老年人口）由 nPM 引起的氧化损伤和炎症。