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

描述（由申请人提供）：美国哮喘发病率的上升是一个日益繁重的公共卫生问题。这种疾病在城市地区的问题尤其严重。作为一种与花粉和空气污染物等环境触发因素有关的疾病，我们研究其发病机制至关重要。虽然今天有治疗哮喘的方法，但它们不处理难治性病例，并且缺乏治疗通常证明对患有这些难治性病例的患者是致命的。聚腺苷二磷酸核糖聚合酶-1（PARP-1）已被确定为炎症介质和哮喘嗜酸性粒细胞增多症的参与者。我们使用PARP-1敲除小鼠或新型PARP-1抑制剂TiQ研究PARP-1抑制。在疾病状态下增加的PAR水平表明，PARP-1在哮喘小鼠模型以及人哮喘中都被激活。此外，PARP-1抑制减少了哮喘小鼠模型中的炎症反应，因为嗜酸性粒细胞显著减少。初步数据显示PARP-1参与IL-4细胞因子受体的信号转导途径。鼻内给予IL-4不能恢复PARP-1敲除小鼠的嗜酸性粒细胞增多，但给予IL-5确实恢复了气道嗜酸性粒细胞增多。因此，PARP-1抑制导致IL-4和IL-5之间的信号转导途径中断。我的第一个具体目标是检查PARP-1抑制对IL-4信号转导途径的各种信号转导组分（包括JAK 1、JAK 3、STAT 6和GATA 3）的激活和表达的影响。蛋白质印迹法将用于检查蛋白质表达水平以及磷酸化。这将显示IL-4信号传播是否中断。我们还将研究各种感兴趣基因的mRNA表达。我们假设PARP-1对STAT 6表达具有特异性影响。我的第二个具体目标是检查PARP-1抑制对STAT 6易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可能在该过程中发挥作用。我们将使用免疫荧光技术检查STAT 6易位。将使用腺病毒载体将PARP-1恢复到动物或细胞悬液中并寻找表型逆转来确认每个特定目的的发现。环境条件已被证明与慢性疾病的发病率以及触发实际哮喘发作相关。鉴于数据表明PARP-1抑制减轻了哮喘小鼠模型中的肺部炎症，研究PARP-1在炎症中的作用以开发应对该疾病的策略是重要的。

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.