Mechanism of regulation of CNS inflammation by microglia

描述（由申请人提供）：小胶质细胞是中枢神经系统中常驻的髓系细胞，具有维持正常组织的功能。小胶质细胞在疾病期间可以被激活和/或失调，从而影响ms的疾病进展或消退，由于缺乏标记物和分子小胶质细胞特征，了解小胶质细胞的生物学是一个挑战。最近，我们发现了一个独特的小胶质细胞分子特征，这为小胶质细胞生物学和靶向小胶质细胞治疗中枢神经系统疾病提供了新的见解。基于这种新的分子特征，我们开发了独特的工具来研究EAE中的小胶质细胞，包括：1)鉴定一种独特的分子小胶质细胞特征，用于研究复发缓解型与进展型EAE；2)新一代小胶质细胞和单核细胞特异性单克隆抗体作为研究小胶质细胞的工具；3)纳米链小胶质细胞芯片的制备；4)成人小胶质细胞体外培养新技术的建立；5)制备fcrls转基因小鼠，在体内研究小胶质细胞的作用和功能；6) EAE小胶质细胞特异性cre依赖性基因表达调控及小胶质细胞条件消融；7)在常驻小胶质细胞中发现可作为药物治疗靶点的新靶点。根据我们的初步数据，我们假设常驻小胶质细胞在中枢神经系统炎症的不同阶段具有不同的表型，包括促进恢复的表型和使慢性炎症持续存在的表型。我们将在以下几个方面对这一假设进行验证：目的1：确定小胶质细胞分子特征在脑电损伤中的调控机制。目的2：探讨小胶质细胞在EAE不同阶段对T细胞反应的调控。目的3：通过靶向和调节小胶质细胞调控EAE。在修订后的应用程序中，我们根据获得的新的初步数据纳入了新的实验，探讨了EAE中小胶质细胞功能障碍的机制，使应用程序更具描述性。我们使用替代方法特异性地消耗驻留的小胶质细胞，并确定APOE/NFκB途径是在神经炎症期间改变稳态小胶质细胞特征的特定机制。我们删除了与提案的主要焦点无关的实验。

DESCRIPTION (provided by applicant): Microglia are resident myeloid-lineage cells in the CNS and function in the maintenance of normal tissue. Microglia can become activated and/or dysregulated during disease, and thus affect disease progression or resolution in MS. Understanding the biology of microglia is a challenge due to absence of markers and molecular microglia signatures. Recently, we identified a unique molecular microglia signature which provides insights into microglial biology and the possibility of targeting microglia for the treatment of CNS disease. Based on this novel molecular signature we have developed unique tools to investigate microglia in EAE including 1) Identification of a unique molecular microglia signature that will be used to investigate relapsing- remitting vs. progressive EAE; 2) Generation of novel microglia and monocyte specific mAbs as tools to investigate microglia; 3) Generation of a Nanostring microglial chip; 4) Development of a new technique to culture adult microglia in vitro; 5) Generation of FCRLS-transgenic mice to study the role and function of microglia in vivo; 6) Specific Cre-dependent manipulation of gene expression in microglia and conditional ablation of microglia in EAE; 7) Identification of novel targets in resident microglia that can serve as dru targets for therapy. Based on our preliminary data we hypothesize that resident microglia have different phenotypes during different stages of CNS inflammation including phenotypes that facilitate recovery and phenotypes that perpetuate chronic inflammation. The hypothesis will be tested in the following aims: Aim 1: Identify the mechanism of regulation of molecular microglia signature in EAE. Aim 2: Investigate regulation of T cell responses by microglia during different stages in EAE. Aim 3: Regulation of EAE by targeting and modulation of microglia. In the revised application, we incorporate new experiments that probe the mechanisms of microglial dysfunction in EAE based on new preliminary data we have obtained making the application less descriptive. We use alternative approaches to specifically deplete resident microglia and have identified the APOE/NFκB pathway as a specific mechanism by which the homeostatic microglial signature is altered during neuroinflammation. We have removed experiments not related to the primary focus of the proposal.