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NF κ B和蛋白激酶的级联反应， 应激激活蛋白激酶（SAPK，也称为 JNK）。 虽然一些激活SAPK的上游激酶被表征， 中间事件，其在应激剂之间传输应激信号 受体和蛋白激酶级联还不清楚。 活性氧 物种（ROS）被认为是参与，因为高水平的 含硫醇的化学物质可阻断NF κ B和SAPK的活化。 的 调查人员已经完成了一个两个混合互动屏幕，以确定 这些蛋白质可以与SAPK激活剂MEKK 1相互作用， 实验室鉴定为SAPK蛋白激酶的近似激活剂 级联。 令人惊讶的是，NAD（P）H-醌还原酶NQO 1 （DT-心肌黄酶）被鉴定为MEKK 1结合蛋白。 NQO 1是 被化合物（通常是醌类）抑制， 底物抑制剂 他们的初步数据还显示， 强烈抑制SAPK的活化和NF κ B的活化， 并将其称为应激抑制化合物（SICs）。 这些试剂 在抑制SAPK和NF κ B方面， 是含硫醇的试剂。 本申请的最初目的是 通过合理的药物设计，开发更低IC 50的改良SICs (aim 1）。 其次，在酵母中使用基因选择，其中SIC可以防止 在压力下生存，酵母和人类基因克服了这一点 将分离SIC和胁迫致死组合（目的2）。 这些 是SIC行动的潜在目标。 炎症是 环境引起的疾病的发病机制，包括 广泛称为过敏性肺炎，由广泛的 包括谷物粉尘和脂多糖（LPS）在内的各种制剂 污染这些灰尘 对这些环境的炎症反应 刺激伴随着细胞因子的分泌，细胞因子反过来可以强烈地 激活SAPK和NF κ B。 调查人员的初步数据显示， 抑制LPS介导人这两种应激途径的活化 肺泡巨噬细胞是炎症和损伤介质的主要来源 在肺过敏性肺炎的发展过程中。 因此 研究人员假设，SICs可能代表了一种新的手段， 中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 他们将采用两种体外模型， 肺泡巨噬细胞（目的3）和LPS和谷物粉尘诱导的小鼠模型 肺损伤（目的4），以测试SICs中断生物化学反应的能力。 以及这些物质的生物学后果。

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.