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

描述（由申请人提供）：在美国，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尤为严重。作为一种与花粉和空气污染物等环境因素有关的疾病，研究其发病机制至关重要。虽然目前有治疗哮喘的方法，但它们并不针对难治性病例，而缺乏治疗往往对患有这些难治性病例的患者是致命的。酶聚（ADP-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水平升高表明PARP-1在哮喘小鼠模型和人哮喘中均被激活。此外，PARP-1抑制抑制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通路。鼻内给药IL-4不能恢复PARP-1敲除小鼠的嗜酸性细胞，但给药IL-5可以恢复气道嗜酸性细胞。因此，PARP-1抑制导致IL-4和IL-5之间的信号转导通路中断。我的第一个具体目标是研究PARP-1抑制对IL-4信号转导途径中各种信号转导成分（包括JAK1、JAK3、STAT6和GATA3）的激活和表达的影响。Western blotting将用于检测蛋白表达水平和磷酸化水平。这将向我们显示IL-4信号传播中是否存在中断。我们还将研究各种感兴趣基因的mRNA表达。我们假设PARP-1对STAT6的表达有特异性影响。我的第二个具体目标是研究PARP-1抑制对STAT6转位进入细胞核的影响。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查STAT6易位。通过腺病毒载体将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引发实际的哮喘发作有关。鉴于有数据表明PARP-1抑制可减轻哮喘小鼠模型中的肺部炎症，因此研究PARP-1在炎症中的作用对于制定治疗该病的策略具有重要意义。

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.