Mechanism of regulation of CNS inflammation by microglia

描述（由申请人提供）：小胶质细胞是CNS中的常驻髓系细胞，具有维持正常组织的功能。小胶质细胞可以在疾病期间被激活和/或失调，从而影响MS中的疾病进展或消退。由于缺乏标记物和分子小胶质细胞特征，理解小胶质细胞的生物学是一个挑战。最近，我们确定了一个独特的分子小胶质细胞的签名，提供了深入了解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的可能性。基于这种新的分子标记，我们已经开发了独特的工具来研究EAE中的小胶质细胞，包括1）鉴定将用于研究复发缓解型与进行性EAE的独特分子小胶质细胞标记; 2）产生新的小胶质细胞和单核细胞特异性mAb作为研究小胶质细胞的工具; 3）产生Nanostring小胶质细胞芯片; 4）建立了一种新的成人小胶质细胞体外培养技术，5）建立了FCRLS转基因小鼠，研究了小胶质细胞在体内的作用和功能，6）Cre依赖的小胶质细胞基因表达调控和EAE中小胶质细胞的条件性消融; 7）在常驻小胶质细胞中鉴定可用作治疗药物靶标的新靶标。基于我们的初步数据，我们假设，居民小胶质细胞在CNS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永久化的表型。该假设将在以下目标中进行测试：目标1：确定EAE中分子小胶质细胞签名的调节机制。目的2：探讨小胶质细胞在EAE发病不同阶段对T细胞免疫应答的调节作用。目的3：通过靶向和调节小胶质细胞来调节EAE。在修订后的申请中，我们纳入了新的实验，探索EAE中小胶质细胞功能障碍的机制，基于我们获得的新的初步数据，使申请描述性更少。我们使用替代方法来特异性地消耗驻留的小胶质细胞，并且已经确定APOE/NFκB通路是在神经炎症期间改变稳态小胶质细胞特征的特定机制。我们已经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.