GMF in CNS inflammation

摘要 多发性硬化症(MS)是一种致残的中枢神经系统炎性脱髓鞘疾病 据估计，这影响了35万美国人和全球100多万人。自发病机制以来 多发性硬化症的治疗尚不清楚，目前还没有明确的治疗方法。我们目前所了解的大部分关于 多发性硬化症的致病因素基于实验性自身免疫性脑脊髓炎(EAE)，这是一种 MS的发病机制存在几种学说，并与浸润性T细胞、促炎因子有关 细胞因子、趋化因子、激活的小胶质细胞和星形胶质细胞。我们近年来对神经胶质细胞的研究进展 成熟因子(GMF)是本实验室分离、测序和克隆的一种蛋白质，已证实具有一定的生物学活性。 转基因食品的主要免疫调节功能。最近，我们已经建立了依赖转基因食品的生产 小胶质细胞中的炎性细胞因子/趋化因子与随后的少突胶质细胞(髓鞘)的破坏 产生细胞)和神经元。基于GMF能够激活小胶质细胞并诱导多个良好的 建立了促炎介质，我们假设GMF参与了MS/EAE的发病。 我们还提供了使用转基因小鼠的强有力的实验证据，即缺乏 内源性转基因食品延缓了EAE的发病，并显著降低了EAE的严重程度，无论是活动性的还是适应性的 转移模型。我们的结果首次为以转基因食品为靶点进行治疗提供了一种新的理性 本建议的目的是研究抑制转基因食品对EAE的影响，并 阐明转基因食品是治疗多发性硬化症的候选方案这项提案将调查一项新的 有效抑制EAE中GMF功能的治疗途径。在我们最近取得成功的基础上 在体外抑制GMF的表达，我们将使用RNA干扰(RNAi)来抑制GMF的表达和 抗GMF抗体中和EAE小鼠内源性GMF蛋白。我们的假设是有效的 抑制转基因食品的功能将被证明是减缓甚至逆转致病的有效策略 EAE中的进程。我们将追求两个具体目标。在目标1A中，我们将研究RNAi抑制的能力 GMF在EAE小鼠中的表达，并确定RNAi是否可以预防EAE。在目标1B中，我们将测试 向EAE小鼠注射中和抗GMF抗体将预防或逆转EAE小鼠的病理特征 EAE。在目标2中，我们将在中枢神经系统炎症的背景下比较这两种GMF抑制策略。

Abstract Multiple sclerosis (MS) is a disabling inflammatory demyelinating disease of the central nervous system that affects an estimated 350,000 Americans and over a million individuals worldwide. Since the pathogenesis of MS is not clear, no definitive treatment is as of yet available. Much of our current knowledge about contributing factors of MS is based on experimental autoimmune encephalomyelitis (EAE), an animal model of MS. Several theories for the pathogenesis of MS exist and implicate infiltrating T cells, pro-inflammatory cytokines, chemokines, activated microglia and astrocytes. Our research efforts in recent years on glia maturation factor (GMF), a protein isolated, sequenced and cloned in our laboratory, have demonstrated a major immunomodulatory function of GMF. Recently, we have established the GMF-dependent production of inflammatory cytokines/chemokines in microglia and the subsequent destruction of oligodendroglia (myelin producing cells) and neurons. Based on GMF's ability to activate microglia and induce several well- established pro-inflammatory mediators, we hypothesize that GMF is involved in the pathogenesis of MS/EAE. We have also provided strong experimental evidence using GMF-deficient mice, that an absence of endogenous GMF delays the onset and drastically reduces the severity of EAE, in both active and adaptive transfer models. Our results provided for the first time a novel rational for targeting GMF for therapeutic intervention in MS. The objective of this proposal is to study the effect of GMF inhibition in EAE and to elucidate GMF as a candidate for therapeutic intervention in MS. This proposal will investigate a novel therapeutic approach to effectively suppress GMF-function in EAE. Building on our recent success in suppressing GMF expression in vitro, we will use RNA interference (RNAi) to suppress GMF expression and an anti-GMF antibody to neutralize endogenous GMF protein in EAE mice. Our hypothesis is that the effective suppression of GMF-function will prove to be an effective strategy to slow, and perhaps reverse pathogenic processes in EAE. We will pursue two Specific Aims. In Aim 1A, we will study the ability of RNAi to suppress GMF expression in EAE mice and determine whether RNAi can prevent EAE. In Aim 1B, we will test whether the delivery of a neutralizing anti-GMF antibody to EAE mice will prevent or reverse pathological hallmarks of EAE. In Aim 2, we will compare these two GMF-suppression strategies in the context of CNS inflammation.