Glia Maturation Factor in CNS Inflammation

描述（由申请人提供）：阐明调节中枢神经系统（CNS）炎症的分子机制对于我们理解广泛的疾病（包括多发性硬化症、阿尔茨海默病和帕金森病）至关重要。这些疾病造成了严重的发病率和死亡率，迫切需要新的预防和治疗方法。这一建议建立在我们最近的发现，神经胶质成熟因子（GMF），一个高度保守的蛋白质独特的大脑，作为一个重要的免疫调节剂，通过促进生产和分泌的促炎细胞因子，导致激活小胶质细胞，在大脑中的抗原呈递细胞。人类基因组序列联盟已经确定了14号染色体上的转基因食品基因。在这个建议中，我们将研究实验性自身免疫性脑脊髓炎（EAE），多发性硬化症的标准动物模型，假设GMF有助于EAE的发病机制。这项研究将包括对已经获得的转基因食品敲除小鼠、野生型小鼠和转基因食品转基因小鼠（处于开发的最后阶段）的研究，其中转基因食品转基因将在星形胶质细胞特异性GFAP启动子的影响下重新引入转基因食品敲除小鼠。这将首次提供一个独特的机会，研究EAE的发病机制，在对比设置无转基因食品，正常量的转基因食品，转基因食品的高表达。目标一：研究转基因食品对中枢神经系统炎症和免疫过程中两个主要的信号转导通路，即p38 MAP激酶和JAK/STAT通路的影响。我们将利用培养的脑细胞、星形胶质细胞、小胶质细胞和少突胶质细胞，这些细胞来源于GMF敲除小鼠与野生型小鼠。目的II：研究转基因食品在EAE的病理生理学中的作用，并确定转基因食品的缺乏减轻了疾病的严重程度。(A)将用适当的抗原攻击GMF敲除和野生型小鼠以诱导EAE。将比较疾病不同阶段的脑和脊髓的发育和组织病理学。(B)探讨EAE进展过程中细胞因子和自由基生成介质在脑、脊髓和单个核细胞中的表达谱。从这项研究中获得的知识也可以应用于小胶质细胞激活起重要作用的大脑退行性疾病，如阿尔茨海默氏症和帕金森氏症。

DESCRIPTION (provided by applicant): Elucidation of the molecular mechanisms that regulate inflammation in the central nervous system (CNS) is critical to our understanding of a broad spectrum of diseases, including multiple sclerosis, Alzheimer's, and Parkinson's. These diseases are responsible for significant morbidity and mortality, and new prevention and treatment approaches are urgently needed. This proposal builds on our recent finding that glia maturation factor (GMF), a highly conserved protein unique to the brain, serves as an important immune modulator by promoting the production and secretion of pro-inflammatory cytokines leading to the activation of microglia, the antigen presenting cells in the brain. The Human Genome Sequence Consortium has identified the gene of GMF in chromosome 14. In this proposal, we will study experimental autoimmune encephalomyelitis (EAE), the standard animal model for multiple sclerosis, with the hypothesis that GMF contributes to the pathogenesis of EAE. This research will include investigations on the already available GMF knockout mice, wild type mice, and GMF transgenic mice (in the final stages of development) in which GMF transgene will be reintroduced in GMF knockout mice under the influence of astrocyte-specific GFAP promoter. This will provide, for the first time, a unique opportunity to study the pathogenesis of EAE in the contrasting settings of no GMF, normal amount of GMF, and high expression of GMF. AIM I: To study the effects of GMF on the two major signal transduction cascades involved in CNS inflammation and immune processes, namely, the p38 MAP kinase and JAK/STAT pathways. We will utilize cultured brain cells, astrocytes, microglia and oligodendroglia, derived from GMF knockout versus wild type mice. AIM II: To study the role of GMF in the pathophysiology of EAE and to establish that the absence of GMF mitigates the severity of the disease. (A) GMF knockout and wild type mice will be challenged with the proper antigen to induce EAE. The development and the histopathology of the brain and spinal cord at various stages of the disease will be compared. (B) To evaluate the expression profiles of cytokines and the free radical generating mediators during the progression of EAE in the brain, spinal cord and in isolated mononuclear cells. Knowledge obtained from this study can also be applied to degenerative diseases of the brain where microglial activation plays an important role, such as Alzheimer's and Parkinson's.