Glia Maturation Factor in CNS Inflammation

描述(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了解广泛的疾病至关重要，包括多发性硬化症、阿尔茨海默氏症和帕金森氏症。这些疾病导致显著的发病率和死亡率，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基础上，即胶质成熟因子(GMF)是大脑特有的一种高度保守的蛋白质，通过促进促炎细胞因子的产生和分泌而发挥重要的免疫调节作用，从而激活小胶质细胞，小胶质细胞是大脑中的抗原递呈细胞。人类基因组序列联合会已经在14号染色体上发现了GMF基因。在这项计划中，我们将研究多发性硬化症的标准动物模型-实验性自身免疫性脑脊髓炎(EAE)，假设GMF参与了EAE的发病机制。这项研究将包括现有的GMF基因敲除小鼠、野生型小鼠和转基因小鼠(处于发育的最后阶段)，其中GMF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GMF基因敲除小鼠。这将第一次提供一个独特的机会，在没有转基因食品、正常转基因食品数量和高表达转基因食品的对比环境下研究EAE的发病机制。目的：研究GMF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p38MAPK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，来自GMF基因敲除小鼠和野生型小鼠。目的II：研究转基因食品在EAE病理生理学中的作用，并证实转基因食品的缺失减轻了疾病的严重程度。(A)基因敲除小鼠和野生型小鼠会接受适当的抗原攻击，以诱发EAE。将比较疾病不同阶段的大脑和脊髓的发育和组织病理学。(B)评估细胞因子和自由基产生介质在EAE发展过程中在脑、脊髓和分离的单个核细胞中的表达情况。从这项研究中获得的知识也可以应用于脑退行性疾病，在这些疾病中，小胶质细胞的激活发挥着重要作用，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.