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）炎症的分子机制对我们理解包括多发性硬化症、阿尔茨海默病和帕金森病在内的广泛疾病至关重要。这些疾病是造成严重发病率和死亡率的原因，迫切需要新的预防和治疗方法。这一建议基于我们最近的发现，神经胶质成熟因子（GMF），一种大脑特有的高度保守蛋白，作为一种重要的免疫调节剂，通过促进促炎细胞因子的产生和分泌，导致小胶质细胞（大脑中的抗原呈递细胞）的激活。人类基因组序列联盟已在第14号染色体上鉴定出GMF基因。在本课题中，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），并假设GMF参与了EAE的发病机制。本研究将包括对现有转基因基因敲除小鼠、野生型小鼠和转基因转基因小鼠（处于开发的最后阶段）的研究，在星形细胞特异性GFAP启动子的影响下，将转基因基因重新引入转基因基因敲除小鼠。这将首次为研究无GMF、GMF正常量和GMF高表达情况下EAE的发病机制提供一个独特的机会。目的1：研究GMF对参与中枢神经系统炎症和免疫过程的两个主要信号转导级联，即p38 MAP激酶和JAK/STAT通路的影响。我们将利用转基因基因敲除小鼠培养的脑细胞、星形胶质细胞、小胶质细胞和少突胶质细胞与野生型小鼠进行对比。目的II：研究转基因蛋白在EAE病理生理中的作用，并确定缺乏转基因蛋白可减轻疾病的严重程度。(A) GMF敲除和野生型小鼠用适当抗原激发诱导EAE。在疾病的各个阶段大脑和脊髓的发展和组织病理学将进行比较。(B)评价脑、脊髓和分离的单核细胞中EAE进展过程中细胞因子和自由基生成介质的表达谱。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，如阿尔茨海默病和帕金森病。

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.