STRESS INHIBITORY COMPOUNDS IN ENVIRONMENTAL DISEASE

描述：环境应激和炎性细胞因子激活 转录因子NFKB和一系列蛋白激酶导致 应激激活蛋白激酶(SAPK，也称为 JNK)。虽然对一些激活SAPK的上游激酶进行了表征， 在应激剂之间传递应激信号的中间事件 受体和蛋白激酶级联尚不清楚。活性氧 物种(RO)被认为参与其中，因为高水平的 含有硫醇的化学物质可以阻断NFKB和SAPK的激活。这个 调查人员已经完成了一个双杂交互动屏幕，以确定 可以与SAPK激活剂MEKK1相互作用的蛋白质，我们的 实验室确认为SAPK蛋白激酶的近似激活剂 卡斯卡德。令人惊讶的是，一种名为NQO1的NAD(P)H-苯二酚还原酶 DT-黄递酶是一种MEKK1结合蛋白。NQO1是 被可能起竞争作用的化合物(通常是苯二酚)抑制 底物抑制剂。他们的初步数据还表明，这些特工 对SAPK和NFKB的激活有强烈的抑制作用， 并将其命名为应激抑制化合物(SICS)。这些特工是 在抑制SAPK和NFKB方面的效果比 都是含有硫醇的试剂。此应用程序的初始目标是 通过合理的药物设计开发更低IC50的改良SICS (目标1)。其次，在酵母中使用SICS阻止的基因选择 在压力、酵母和人类基因克服这种压力的情况下存活 碳化硅和应力的致命组合将被隔离(目标2)。这些 代表SIC行动的潜在目标。炎症是疾病的基础 环境诱发疾病的发病机制，包括 广义的过敏性肺炎，由广泛的 各种制剂，包括谷物粉尘和脂多糖(LPS) 污染了这些尘埃。对这些环境的炎症反应 刺激伴随着细胞因子的分泌，而细胞因子又可能强烈地 激活SAPK和NFKB。调查人员的初步数据显示，SICS 在人类中抑制内毒素介导的这两条应激途径的激活 肺泡巨噬细胞，炎症和损伤性介质的主要来源 在过敏性肺炎的发展过程中，在肺中。因此， 调查人员推测，SICS可能代表了一种新的 阻断导致限制性肺的病理生理过程 过敏性肺炎的疾病特征，无论 环境刺激。他们将使用这两种体外模型，使用人类 肺泡巨噬细胞(AIM 3)与内毒素和谷物粉尘诱导的小鼠模型 肺损伤(目标4)测试SICS中断生化的能力 以及这些毒剂的生物学后果。

DESCRIPTION: Environmental stress and inflammatory cytokines activate the transcription factor NFKB and a cascade of protein kinases that result in the activation of the Stress Activated Protein Kinase (SAPK, also known as JNK). While some upstream kinases that activate SAPK are characterized, intermediate events that transmit stress signals between the stress agent receptor and the protein kinase cascade are unclear. Reactive oxygen species (ROS) are thought to be involved because high levels of thiol-containing chemicals can block activation of NFKB and SAPK. The investigators have completed a two hybrid interaction screen to identify proteins that can interact with the SAPK activator MEKK1, which our laboratory identified as a proximate activator of the SAPK protein kinase cascade. Surprisingly, a NAD(P)H-quinone reductase termed NQO1 (DT-diaphorase) was identified as a MEKK1 binding protein. NQO1 is inhibited by compounds (generally quinones) that may serve as competitive substrate inhibitors. Their preliminary data also show that these agents are strongly inhibitory of the activation of SAPK and activation of NFKB, and have termed them Stress Inhibitory Compounds (SICs). These agents are approximately 1000 fold more effective towards inhibiting SAPK and NFKB than are thiol-containing agents. Initial aims in this application are to develop improved SICs with still lower IC50s through rational drug design (aim 1). Secondly, using a genetic selection in yeast in which SICs prevent survival in situations of stress, yeast and human genes that overcome this lethal combination of SIC and stress will be isolated (aim 2). These represent potential targets of SIC action. Inflammation underlies the pathogenesis of environmental-induced diseases including those broadly-termed hypersensitivity pneumonitis which is caused by a wide variety of agents including grain dusts and the lipopolysaccharide (LPS) contaminating these dusts. The inflammatory response to these environmental stimuli is accompanied by secretion of cytokines which in turn may strongly activate SAPK and NFKB. The investigators preliminary data show that SICs inhibit LPS-mediated activation of both of these stress pathways in human alveolar macrophages, a major source of inflammatory and injurious mediators in the lung during development of hypersensitivity pneumonitis. Thus, the investigators hypothesize that SICs may represent a novel means of interrupting the pathophysiologic processes leading to the restrictive lung disease characteristic of hypersensitivity pneumonitis, regardless of the environmental stimulus. They will employ both in vitro models using human alveolar macrophages (aim 3) and mouse models of LPS and grain dust-induced lung damage (aim 4) to test the ability of SICs to interrupt the biochemical and biological consequences of these agents.