Cytokines in Glial Cells and EAE Brain

描述（由申请人提供）：多发性硬化症（MS）是一种炎性脱髓鞘疾病，导致少突胶质细胞、髓鞘和轴突损失以及神经退行性病变形成。目前开发的靶向免疫调节机制的药物疗效有限，因为尽管进行了治疗，CNS疾病仍在继续进展。来自包括我们在内的各个实验室的研究提供了线粒体和过氧化物酶体功能异常作为EAE（实验性自身免疫性脑脊髓炎）和MS中CNS疾病病理学原因的证据，但很少（如果有的话）关于导致这些病理学的机制。本提案旨在了解线粒体和过氧化物酶体功能障碍的机制，并确定针对EAE中神经保护和神经修复的这些病理的药物的疗效。最近，我们已经描述了RhoA-Rock-PPAR介导的他汀类药物的活性以及AMP活化的蛋白激酶（AMPK）的活化保护少突胶质细胞免受EAE中的炎性损伤，并且洛伐他汀和AICAR（一种特异性活化剂AMPK）的组合极大地改善了对EAE疾病的功效，从而表明这些药物对EAE的CNS疾病的潜在功效。AMPK是一种细胞能量传感器，在过氧化物酶体和线粒体的生物发生、动力学和自噬清除中发挥重要作用。此外，最近对140名患有进展型MS（一种没有批准药物的疾病）的患者进行的II期双盲安慰剂对照研究报告辛伐他汀治疗的功效为在两年内脑萎缩减少50%，这也支持他汀类药物对MS的CNS疾病的功效。我们假设，针对RhoA-Rock-PPAR信号机制的药物（洛伐他汀）和AMPK/PGC-1α-mTOR信号转导机制与AMPK激活剂（AICAR）在EAE/MS疾病提供更大的神经保护和加速的神经修复，因此改善了EAE/MS的疗效。因此，这些研究将利用培养的神经元和少突胶质细胞来研究线粒体和过氧化物酶体异常的机制在EAE疾病条件下，和EAE小鼠模型。目标1：探讨EAE条件下神经元和少突胶质细胞线粒体和过氧化物酶体的生物合成及自噬清除机制。目标二：评价使用洛伐他汀的RhoA/PPAR介导机制和使用AICAR的AMPK/PGC-1α介导机制对EAE小鼠的神经保护和神经修复的治疗潜力。该研究的新奇是针对EAE和MS的CNS疾病机制的新治疗方法的前提。拟议的研究将使用最先进的方法来解决拟议的目标。他汀类药物是治疗高胆固醇血症最常用的药物;因此，他汀类药物单独或与AMPK激活剂联合使用的这种新型口服疗法的转化潜力相当高。

DESCRIPTION (provided by applicant): Multiple sclerosis (MS) is an inflammatory demyelinating disease leading to loss of oligodendrocytes, myelin and axons and neurodegenerative lesion formation. Present day drugs developed to target immunomodulatory mechanisms provide limited efficacy since CNS disease progression continues despite treatment. Studies from various laboratories including ours have provided evidence of functional abnormality of mitochondria and peroxisomes as a cause of CNS disease pathologies in EAE (experimental autoimmune encephalomyelitis) and MS but very little, if any, about the mechanisms leading to these pathologies. The present proposal is designed to understand the mechanisms of mitochondrial and peroxisomal dysfunction and to determine the efficacy of drugs targeting these pathologies for neuroprotection and neurorepair in EAE. Recently, we have described that RhoA-Rock-PPAR mediated activity of statins as well as activation of AMP activated protein kinase (AMPK) protects oligodendrocytes against inflammatory insult in EAE and that a combination of lovastatin and AICAR, a specific activator AMPK, greatly improved the efficacy against EAE disease thus indicating the potential efficacy of these drugs against the CNS disease of EAE. AMPK is a cellular energy sensor playing an essential role in biogenesis, dynamics, and autophagic clearance of peroxisomes and mitochondria. Moreover, a recent phase-II double blind placebo controlled study of 140 patients with the progressive form of MS, a disease with no approved drug, reporting efficacy of simvastatin treatment as 50% reduction in brain atrophy over two years also supports the efficacy of statins against CNS disease of MS. Based on these findings, we hypothesize that drugs targeting RhoA-Rock-PPAR signaling mechanisms (lovastatin) and AMPK/PGC-1α-mTOR signaling mechanisms with AMPK activator (AICAR) in EAE/MS disease provides greater neuroprotection and accelerated neurorepair and hence improved efficacy in EAE/MS. Therefore, the proposed studies will investigate the mechanisms underlying mitochondrial and peroxisomal abnormalities using both cultured neurons and oligodendrocytes under EAE disease conditions, and mouse models of EAE. Aim 1: To investigate the mechanisms of mitochondrial and peroxisomal biogenesis and autophagic clearance in cultured neurons and oligodendrocytes under EAE conditions. Aim 2: To evaluate the therapeutic potential of RhoA/PPAR-mediated mechanisms using lovastatin and AMPK/PGC-1α-mediated mechanisms using AICAR for neuroprotection and neurorepair in EAE mice. The novelty of the study is the premise of a new therapeutic approach targeting CNS disease mechanisms of EAE and MS. The proposed studies will use state-of-the-art methodologies to address the proposed goals. Statins are the most commonly used drugs for hypercholesterolemia; therefore, translational potential of such a novel oral therapy of statin alone or in combination with activator of AMPK is reasonably high.