Mechanism of regulation of CNS inflammation by microglia

描述(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，在维持正常组织中发挥功能。小胶质细胞可在疾病过程中被激活和/或调节失调，从而影响MS的疾病进展或解决方案，由于缺乏标志物和分子小胶质细胞信号，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，它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的可能性提供了深入的了解。基于这种新的分子标记，我们开发了独特的工具来研究EAE中的小胶质细胞，包括：1)鉴定独特的小胶质细胞标记，将用于研究复发与进行性EAE；2)产生新的小胶质细胞和单核细胞特异性单抗作为研究小胶质细胞的工具；3)制备纳米串微胶质细胞芯片；4)开发一种新的体外培养成年小胶质细胞的技术；5)建立FCRLS转基因小鼠，以研究小胶质细胞在体内的作用和功能；6)依赖CRE的特异性小胶质细胞基因表达的调控和EAE中小胶质细胞的条件性消融；7)在常驻小胶质细胞中寻找可作为药物治疗靶点的新靶点。根据我们的初步数据，我们假设驻留的小胶质细胞在中枢神经系统炎症的不同阶段有不同的表型，包括促进恢复的表型和持续慢性炎症的表型。这一假说将在以下目标进行验证：目标1：确定EAE中分子小胶质细胞信号的调节机制。目的2：探讨小胶质细胞对EAE不同阶段T细胞反应的调节作用。目的3：通过靶向和调节小胶质细胞来调节EAE。在修订后的应用中，我们加入了新的实验，根据我们获得的新的初步数据探索EAE中小胶质细胞功能障碍的机制，使应用不那么具有描述性。我们使用替代方法来特异性地耗尽驻留的小胶质细胞，并已确定载脂蛋白E/核因子κB途径是在神经炎症过程中改变动态平衡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重点无关的实验。

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.