Cytokines in Glial Cells and EAE Brain

描述（由申请人提供）：多发性硬化症（MS）是一种炎症性脱髓鞘疾病，导致少突胶质细胞、髓鞘和轴突的损失以及神经退行性病变的形成。目前针对免疫调节机制开发的药物疗效有限，因为尽管治疗，中枢神经系统疾病仍在继续进展。包括我们在内的各个实验室的研究已经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中中枢神经系统疾病病理的原因的证据，但很少，如果有的话，导致这些病理的机制。本研究旨在了解线粒体和过氧化物酶体功能障碍的机制，并确定针对这些病理的药物对EAE中神经保护和神经修复的疗效。最近，我们报道了RhoA-Rock-PPAR介导的他汀类药物活性以及AMP活化蛋白激酶（AMPK）的激活保护EAE少突胶质细胞免受炎症损伤，洛伐他汀与特异性激活剂AMPK AICAR联合使用可大大提高EAE疾病的疗效，提示这些药物对EAE的中枢神经系统疾病有潜在的疗效。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，最近一项针对140名进展型多发性硬化症患者的ii期双盲安慰剂对照研究报告了辛伐他汀治疗两年内脑萎缩减少50%的疗效，也支持了他汀类药物治疗多发性硬化症中枢神经系统疾病的疗效。我们假设针对EAE/MS疾病的RhoA-Rock-PPAR信号机制（洛伐他汀）和AMPK/PGC-1α-mTOR信号机制与AMPK激活剂（AICAR）的药物可以提供更大的神经保护和加速神经修复，从而提高EAE/MS的疗效。因此，本研究将利用培养的神经元和少突胶质细胞，以及EAE小鼠模型，研究EAE疾病条件下线粒体和过氧化物酶体异常的机制。目的1：探讨EAE条件下培养的神经元和少突胶质细胞线粒体和过氧化物酶体生物发生和自噬清除的机制。目的2：评价洛伐他汀介导的RhoA/ ppar机制和AMPK/ pgc -1α-介导的AICAR机制对EAE小鼠的神经保护和神经修复的治疗潜力。该研究的新颖性是一种针对EAE和ms的中枢神经系统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，这种他汀类药物单独或与AMPK激活剂联合使用的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.