INHIBITING JNK, A NEW ANTI-INFLAMMATORY STRATEGY

描述（改编自研究者摘要）：本研究的主要目标 建议是表征促炎性细胞因子TNF和IL-1激活 血管内皮细胞（EC）中的信号转导，以识别独特的 炎症中TNF介导的通路。炎症是一个必不可少的过程 宿主防御和组织修复。然而，防御可能会引起致病变化 导致血管疾病如动脉粥样硬化。TNF触发生存 通过激活NF-κ B信号传导，通过激活炎症（和凋亡 关于JFK这一提议的主要假设是， NF-kappaB激活的破坏将提供一种有效的方法， 抗炎治疗。我们已经阐明，不同的促炎因子 细胞因子激活独特的以及常见的细胞内信号传导途径， 通过同源脱敏（HMD）和异源脱敏（HMD）证实 脱敏（HTD）。用TNF预处理的EC变得对TNF难治 但IL-1可再刺激激活JNK并表达 E-选择素，反之亦然（即JNK活化和E-选择素表达显示 同源脱敏- HMD）。相反，动脉粥样硬化保护层流 抑制TNF和IL-1诱导的JNK和粘附分子（异源的 脱敏-HTD）。我们认为HMD调节了一种特异性的 炎症，而HTD调节所有细胞因子诱导的炎症。我们将 使用HMD和HTD作为模型来剖析TNF激活的JNK激活， NF-κ B和IL-1信号通路。具体的假设是 调查结果是：1）。HMD是通过TNF受体相关因子2（TRAF 2）， 在TNF信号传导中作为JNK分叉点的衔接蛋白， NF-κ B。2）。HTD是通过MAP激酶ASK 1，一个会聚点 TNF和IL-1信号通路在EC中的作用。令人兴奋的初步数据表明， TRAF 2在EC中的过表达使TNF诱导的JNK和E-选择素脱敏， TRAF 2的特异性结构域和天然切割产物在肿瘤的发生发展中发挥作用， JNK激活。提出了三个具体目标。1）。HMD通过TRAF 2 专注于表征TRAF 2独特的残基，结合蛋白和切割 JNK激活的产物。2)HTD通过ASK 1专注于表征流动 对TNF和IL-1诱导的ASK活化的影响。3)描述角色 TRAF 2和ASK 1在TNF诱导的小鼠模型中的炎症中的基因递送 TRAF 2或ASK 1进入角膜内皮。该提案应提供候选人 作为TNF介导的炎症反应的特异性或独特靶点的蛋白质 促进新的治疗方法的发展，以控制 炎症在各种疾病的设置。

DESCRIPTION (Adapted from Investigator's Abstract): The primary goal of this proposal is to characterize proinflammatory cytokines TNF and IL-1 activated signal transduction in vascular endothelial cells (EC) to identify unique TNF-mediated pathways in inflammation. Inflammation is a process essential for host defense and tissue repair. However, defense may cause pathogenic changes leading to vascular diseases such as atherosclerosis. TNF triggers survival signal via activation of NF-kappaB, inflammation (and apoptosis) via activation of JFK. The major hypothesis of this proposal is that inhibition of JNK without disruption of NF-kappaB activation would provide a valid approach for anti-inflammatory therapy. We have elucidated that different proinflammatory cytokines activate unique as well as common intracellular signaling pathways as demonstrated by homologous desensitization (HMD) and heterologous desensitization (HTD). EC pre-treated with TNF become refractory to TNF restimulation but can be restimualted by IL-1 to active JNK and express E-selectin and vice versa (i.e. JNK activation and E-selectin expression show homologous desensitization - HMD). In contrast, atheroprotective laminar flow inhibits both TNF and IL-1-induced JNK and adhesion molecules (heterologous desensitization-HTD). We believe that HMD modulates a cytokine-specific inflammation, whereas HTD modulates all-cytokine-induced inflammation. We will use HMD and HTD as models to dissect TNF-activated JNK activation from NF-kappaB and from IL-1 signaling pathway. The specific hypothesis to be investigated is that: 1). HMD is via TNF receptor-associated factor 2 (TRAF2), an adaptor protein in TNF signaling as a bifurcation point for JNK and NF-kappaB. 2). HTD is via the MAP kinase kinase kinase ASK1, a convergent point of TNF and IL-1 signaling in EC. Exciting preliminary data indicate that overexpression of TRAF2 in EC desensitizes TNF-induced JNK and E-selectin, and that the specific domains and the natural cleavage products of TRAF2 play roles in JNK activation. Three specific aims are proposed. 1). HMD is via TRAF2 focusing on characterizing TRAF2 unique residues, binding proteins and cleavage products in JNK activation. 2) HTD is via ASK1 focusing on characterizing flow effect on TNF and IL-1-induced ASK lactivation. 3) Characterizing roles of TRAF2 and ASK1 in TNF-induced inflammation in murine models by gene delivery of TRAF2 or ASK1 into cornea endothelium. This proposal should provide candidate proteins that are specific or unique targets for TNF-mediated inflammatory events and facilitate development of new therapeutic approaches to control inflammation in various disease settings.