Regulation of IL4 signal transduciton by poly (ADP-ribose) polymerase-1.

聚 (ADP-核糖) 聚合酶 1 对 IL4 信号转导的调节。

• 批准号: 7750870
• 负责人: Rahul Datta
• 金额: $ 2.86万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7750870
• 关键词: Abate; Adenovirus Vector; Air Pollutants; Allergens; Animal Model; Animals; Asthma; Cell Nucleus; Cells; Chronic Disease; Cytokine Receptors; Data; Development; Disease; Enzymes; Eosinophilia; Fluorescent Antibody Technique; Gene Expression; Genes; Genetic Transcription; Human; IL4 gene; In Vitro; Incidence; Inflammation; Inflammation Mediators; Inflammatory Response; Interleukin 4 Receptor; Interleukin-4; Interleukin-5; Interruption; Intranasal Administration; JAK1 gene; JAK3 gene; Knockout Mice; Laboratories; Link; Lung diseases; Mediating; Molecular; Nuclear; Nuclear Export; Pathogenesis; Patients; Phenotype; Phosphorylation; Pneumonia; Pollen; Poly(ADP-ribose) Polymerases; Process; Production; Proteins; Public Health; Publishing; Refractory; Regulation; Role; STAT6 gene; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Suspension substance; Suspensions; System; TP53 gene; Testing; United States; Western Blotting; Work; airway inflammation; base; improved; inhibitor/antagonist; insight; interest; mRNA Expression; mouse model; novel; oxidant stress; protein expression; therapeutic target; trafficking; transcription factor; urban area

DESCRIPTION (provided by applicant): The rising incidence of asthma in the United States is an increasingly onerous public health concern. The disease is disproportionately problematic in urban areas. As a disease that has been linked to environmental triggers such as pollen and air pollutants, it is crucial that we study its pathogenesis. While there are treatments available for asthma today, they do not deal with refractory cases and the lack of treatments often prove fatal to the patients suffering from these refractory cases. The enzyme poly(ADP- ribose)polymerase-1 (PARP-1) has been established as a mediator of inflammation and a player in asthmic eosinophilia. We study PARP-1 inhibition using PARP-1 knockout mice or a novel PARP-1 inhibitor, TiQ. Increased PAR levels in the disease state shows that PARP-1 is activated in both the mouse model of asthma as well as in human asthma. Furthermore, PARP-1 inhibition curtails the inflammatory response in the mouse model of asthma as eosinophilia is dramatically reduced. Preliminary data show PARP-1 is involved in the signal transduction pathway of the IL-4 cytokine receptor. Intranasal administration of IL-4 fails to restore eosinophilia to PARP-1 knockout mice, but administration of IL-5 does restore airway eosinophilia. Thus, PARP-1 inhibition results in the interruption of the signal transduction pathway between IL-4 and IL-5. My first specific aim is to examine the effects of PARP-1 inhibition on the activation and expression of the various signal transduction components of IL-4 signal transduction pathway including JAK1, JAK3, STAT6, and GATA3. Western blotting will be used to examine levels of protein expression as well as phosphorylation. This will show us whether there is an interruption in the IL-4 signal propagation. We will also look at mRNA expression of the various genes of interest. We hypothesize that PARP-1 has a specific effect on STAT6 expression. My second specific aim is to examine the effects of PARP-1 inhibition on the translocation of STAT6 into the nucleus. Translocation is a critical step in the transduction of the IL-4 signal and PARP-1 may have a role in the process. We will examine STAT6 translocation using immunofluorescence techniques. The findings from each of the specific aims will be confirmed using an adenoviral vector to restore PARP-1 to the animals or cell suspension and looking for phenotype reversal. Environmental conditions have been shown to correlate chronic disease incidence as well as triggering actual asthmatic episodes. Given the data demonstrating that PARP-1 inhibition abates of pulmonary inflammation in the mouse model of asthma, it is important to study the role of PARP-1 in inflammation in order to develop strategies in dealing with the disease.

描述(由申请人提供)：美国哮喘发病率的上升是一个日益沉重的公共卫生问题。这种疾病在城市地区是不成比例的问题。作为一种与环境因素(如花粉和空气污染物)有关的疾病，研究其发病机制至关重要。虽然目前有治疗哮喘的方法，但它们不处理难治性病例，缺乏治疗往往证明这些难治性病例的患者是致命的。多聚腺苷二磷酸核糖聚合酶-1(PARP-1)是一种炎症介质，在哮喘嗜酸性粒细胞增多症中起重要作用。我们使用PARP-1基因敲除小鼠或一种新的PARP-1抑制剂TIQ来研究PARP-1的抑制作用。疾病状态下PAR水平的增加表明，PARP-1在哮喘小鼠模型和人类哮喘模型中都被激活。此外，抑制PARP-1可抑制哮喘小鼠模型的炎症反应，因为嗜酸性粒细胞增多显著减少。初步研究表明，PARP-1参与了IL-4细胞因子受体的信号转导途径。鼻腔给药IL-4不能恢复PARP-1基因敲除小鼠的嗜酸性粒细胞增多，但给药IL-5确实能恢复呼吸道嗜酸性粒细胞增多症。因此，抑制PARP-1导致IL-4和IL-5之间的信号转导通路中断。我的第一个特定目的是研究抑制PARP-1对IL-4信号转导途径中各种信号转导成分的激活和表达的影响，包括JAK1、JAK3、STAT6和GATA3。Western blotting将用于检测蛋白质表达水平和磷酸化水平。这将向我们显示IL-4信号传播是否中断。我们还将观察各种感兴趣基因的mRNA表达。我们假设PARP-1对STAT6的表达有特异性的影响。我的第二个特定目标是研究PARP-1抑制对STAT6转位到细胞核中的影响。易位是IL-4信号转导的关键步骤，PARP-1可能在这一过程中发挥作用。我们将使用免疫荧光技术检测STAT6易位。使用腺病毒载体将PARP-1恢复到动物或细胞悬液中，并寻找表型逆转，将证实来自每个特定目标的发现。环境条件已被证明与慢性病发病率以及引发实际哮喘发作有关。鉴于有数据表明，在哮喘小鼠模型中，PARP-1抑制可以减轻肺部炎症，因此研究PARP-1在炎症中的作用，以制定应对疾病的策略是很重要的。