GMF in CNS inflammation

描述(申请人提供)：多发性硬化症(MS)是一种致残性中枢神经系统炎症性脱髓鞘疾病，估计有35万美国人和全球100多万人受到影响。由于MS的发病机制尚不清楚，到目前为止还没有明确的治疗方法。目前对MS发病因素的认识大多建立在实验性自身免疫性脑脊髓炎(EAE)的基础上，EAE是MS的动物模型，MS的发病机制有多种学说，涉及浸润性T细胞、促炎细胞因子、趋化因子、活化的小胶质细胞和星形胶质细胞。近年来，我们对本实验室分离、测序和克隆的胶质细胞成熟因子(GMF)进行了研究，证明了GMF具有重要的免疫调节功能。最近，我们已经建立了依赖GMF的小胶质细胞产生炎性细胞因子/趋化因子以及随后的少突胶质细胞(髓鞘产生细胞)和神经元的破坏。基于GMF激活小胶质细胞和诱导多种公认的促炎介质的能力，我们推测GMF参与了MS/EAE的发病机制。我们还利用转基因食品缺陷小鼠提供了强有力的实验证据，证明在主动和适应性转移模型中，内源性转基因食品的缺失推迟了EAE的发生，并显著降低了EAE的严重程度。我们的结果首次为针对GMF治疗MS提供了一个新的理论基础。本研究的目的是研究GMF抑制在EAE中的作用，并阐明GMF作为治疗干预的候选药物。本建议将探索一种新的治疗方法，有效抑制EAE中的GMF功能。在我们最近在体外成功抑制GMF表达的基础上，我们将使用RNA干扰(RNAi)来抑制EAE小鼠的GMF表达，并使用抗GMF抗体来中和内源性GMF蛋白。我们的假设是，有效抑制GMF功能将被证明是减缓甚至逆转EAE致病过程的有效策略。我们将追求两个具体目标。在目标1A中，我们将研究RNAi抑制EAE小鼠GMF表达的能力，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抗体是否可以预防或逆转EAE的病理特征。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。公共卫生相关性：MS是一种致残的慢性炎症性脱髓鞘疾病，缺乏有效的治疗方法代表着治疗这种毁灭性疾病的能力的显著差距。这项建议的目的是阐明GMF，一种公认的促炎介质，作为MS/EAE治疗干预的新候选者。这些研究可能为开发一种新的无毒治疗MS提供科学依据。

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The lack of effective treatments for MS, a disabling chronic inflammatory demyelinating disease, represents a significant gap in the ability to treat this devastating disease. The aim of this proposal is to elucidate GMF, a well-established pro-inflammatory mediator, as a novel candidate for therapeutic intervention in MS/EAE. These studies may provide the scientific rationale for the development of a novel non-toxic therapy for MS.