Accelerating remyelination using lanthionine ketimine derivatives

这个项目的主要目标是寻找能够加速中枢神经系统内髓鞘再生的新化合物。 多发性硬化症(MS)患者。虽然已经开发了许多治疗多发性硬化症的方法，但几乎所有的方法都是针对 免疫系统正在努力减少持续的损害；相比之下，以再生为目标的相对较少 产生髓鞘的少突胶质细胞(OLG)恢复功能的能力。在正在进行的研究中， 对于半合成氨基酸衍生物--羊硫氨酸凯蒂亚胺酯(LKE)的有益作用，我们发现 LKE不仅减少了MS小鼠模型的临床症状，而且还加速了随后的髓鞘再生 化学诱导的、非炎症性脱髓鞘的铜比林(CPZ)。新型LKE的体外筛选 衍生品表明，一些化合物比母体化合物更有效。这提出了LKE的主要假设 新的衍生品将增加髓鞘再生，恢复功能。这一问题将在 以下目标： 目的1：利用CPZ模型扩展我们对LKE益处的初步研究，包括优化LKE剂量和 持续时间；以及中度(CPZ为2周)和广泛(CPZ为5-9周)阶段LKE的比较 脱髓鞘的症状。评估将包括髓鞘、轴突和结节的电子显微镜和共聚焦显微镜。 OPC成熟度分析；神经元损伤；神经胶质细胞炎症。 目的2：描述LKE治疗引起的功能结果的改善，包括对 通过记录复合动作电位的神经传导；和行为学 使用旋转臂评估运动和平衡改善的分析；评估记忆和平衡的新对象识别 焦虑；以及评估记忆的y-迷宫。 目的3：完成对4种铅LKE-衍生物的筛选，以确定那些具有最高疗效(最低剂量， 更快速或更强劲的增长)，以诱导OPC成熟，最好的代谢稳定性和最高的膜 渗透性。最佳候选者将在CPZ模型中进行测试，并直接与LKE进行比较。 目的：通过体内和体外实验，探讨LKE及其衍生物的作用机制。这将是 包括电生理方法和钙显像法来检测LKE对CRMP2的影响，以及其 OPC中与电压门控钙通道(VGCC)的相互作用可用于条件基因敲除小鼠 CRMP2将允许测试OPC中的CRMP2是否调解LKE行动。

The major goal of this project is to identify novel compounds which will accelerate remyelination within the CNS of patients with Multiple Sclerosis (MS). While many therapies have been developed to treat MS, almost all target the immune system in efforts to reduce ongoing damage; in contrast relatively few target the regenerative capacity of myelin producing Oligodendrocytes (OLGs) to restore function. In ongoing studies to characterize the beneficial effects of Lanthionine Ketimine Ester (LKE), a semi-synthetic amino acid derivative, we found that LKE not only reduces clinical signs in a mouse model of MS, but also accelerates remyelination following chemically induced, non-inflammatory demyelination by cuprizone (CPZ). In vitro screening of new LKE derivatives suggests some are more potent than the parent compound. This raises the main hypothesis that LKE and new derivatives will increase remyelination and restore functional outcomes. This will be addressed in the following aims: Aim 1: Extend our initial studies of LKE benefit using the CPZ model, including optimization of LKE dosage and duration; and comparisons of LKE following modest (CPZ for 2 weeks) or extensive (CPZ for 5-9 weeks) stages of demyelination. Assessments will include electron and confocal microscopy of myelin, axons, and Nodes of Ranvier; analysis of OPC maturation; neuronal damage; and glial inflammation. Aim 2: Characterize LKE-treatment induced improvement on functional outcomes, including improvement in nerve conductance by recording compound action potentials across the corpus callosum; and behavioral analysis to assess motor and balance improvement with rotarod; novel object recognition to assess memory and anxiety; and y-maze to assess memory. Aim 3: Complete screening of 4 lead LKE-derivatives to identify those having the highest efficacy (lowest dosage, more rapid or robust increase) to induce OPC maturation, best metabolic stability, and highest membrane permeability. The best candidate will be tested in the CPZ model and directly compared to LKE. Aim 4: Use in vivo and in vitro experiments to explore mechanisms of action of LKE and derivatives. This will include electrophysiological methods and calcium imaging to examine effects of LKE on CRMP2, and its interactions with Voltage Gated Ca2+ Channels (VGCCs) in OPCs. Available conditional knockout mice for CRMP2 will allow testing if CRMP2 in OPCs mediates LKE actions.