GMF in CNS inflammation

摘要 多发性硬化症（MS）是一种中枢神经系统的致残性炎性脱髓鞘疾病 估计有35万美国人和全球超过一百万人受到影响。由于发病机制 MS的发病率尚不清楚，目前尚无明确的治疗方法。我们目前对 MS的促发因素是基于实验性自身免疫性脑脊髓炎（EAE）， 女士MS发病机制的几种理论存在并涉及浸润性T细胞、促炎性细胞因子和免疫抑制剂。 细胞因子、趋化因子、活化的小胶质细胞和星形胶质细胞。近年来我们对胶质细胞的研究 成熟因子（GMF）是我们实验室分离、测序和克隆的一种蛋白质， GMF的主要免疫调节功能。最近，我们已经建立了转基因食品依赖生产的 小胶质细胞中的炎性细胞因子/趋化因子和随后的少突胶质细胞（髓鞘）的破坏 产生细胞）和神经元。基于转基因食品的能力，激活小胶质细胞和诱导几个良好的- 建立促炎介质，我们假设GMF参与MS/EAE的发病机制。 我们还使用GMF缺陷小鼠提供了强有力的实验证据， 内源性转基因食品延迟发病，并大大降低EAE的严重程度，在主动和适应性 转移模型我们的研究结果首次为靶向转基因食品的治疗提供了新的理论依据。 这项建议的目的是研究转基因食品抑制EAE的影响， 阐明转基因食品作为一个候选人的治疗干预MS。这项建议将调查一种新的 有效抑制EAE中GMF功能的治疗方法。基于我们最近在 在体外抑制转基因食品的表达，我们将使用RNA干扰（RNAi）来抑制转基因食品的表达， 中和EAE小鼠中内源性GMF蛋白的抗GMF抗体。我们的假设是 抑制GMF功能将被证明是减缓甚至逆转致病性 EAE中的过程。我们将追求两个具体目标。在目标1A中，我们将研究RNAi抑制 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小鼠递送中和性抗GMF抗体将预防或逆转 EAE。在目标2中，我们将在CNS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.