Impact of pathogenic MS patient-derived IgG on oligodendrocytes: bystanders or participants in remyelination failure?

项目摘要/摘要 多发性硬化症(MS)是一种中枢神经系统(CNS)的自身免疫性脱髓鞘疾病 最常见的特点是神经学上可测量和持续下降的间歇期 先起作用后缓解。可用干预措施通过干预针对炎性损伤 炎性免疫功能，但通常不刺激髓鞘修复或恢复神经功能。 促进多发性硬化症髓鞘再生的有针对性的方法将对其管理产生重大影响： 脱髓鞘干扰了通常由少突胶质细胞(OG)提供的对轴突的代谢支持。结果, 剥离的轴突经常退化，从而加重MS患者的残疾。尽管最近对这一问题的研究 神经胶质细胞在中枢神经系统病理中的不同作用已确定在损伤和损伤过程中表达的独特转录本 恢复后，OG对抗体介导的脱髓鞘/重新髓鞘形成的反应仍不清楚。这项建议 结合了一种创新和跨学科的方法来剖析髓鞘失效期间的OG反应 修理。我们已经开发了独特的体外和体外使用疾病特异性髓鞘结合的检测方法。 首次从多发性硬化症患者脑脊液中克隆的重组抗体(RAB)概括了一些 多发性硬化症中的再髓鞘障碍与其他脱髓鞘/再髓鞘模型不同，我们以患者为基础 再髓鞘模型阻断了OGS从早期髓鞘到活跃状态的再髓鞘形成 髓鞘阶段，而不是在OPC分化期间。本申请中描述的实验旨在 通过以下方式解开OGs导致RAB诱导的再髓鞘失败的内在和外在因素 使用活细胞成像、免疫组织化学、转基因小鼠、 创新的髓鞘形成分析和新一代RNA测序模式。要最大限度地提高翻译效率 在这个模型中产生的结果的相关性，重新髓鞘形成阻断的生物标记物将被识别和 在多发性硬化症的病变中问诊。因此，通过利用连续的实验和翻译方法，这 提案将确定B细胞体液自身免疫和OGs之间的特定相互作用如何有助于 人类疾病中的再髓鞘衰竭。首先，我们将测试MS RAB如何通过以下方式直接影响少突胶质细胞 观察它们对成骨细胞分化、形成新髓鞘的能力和信号转导途径的影响 拉布斯写的。然后我们将确定在分化过程中RAB对少突胶质细胞基因表达的影响 和髓鞘形成的实验，并评估人类重新髓鞘形成阻断的候选生物标记物 斑块女士。这些研究不仅将扩大对多发性硬化症神经免疫学的科学理解，它们还将 还阐明了可用于治疗多发性硬化症和其他疾病的导致再髓鞘形成的因素 中枢神经系统脱髓鞘疾病。

Project Summary/Abstract Multiple sclerosis (MS) is an autoimmune demyelinating disease of the central nervous system (CNS) that is most commonly characterized by discreet periods of measurable and sustained reduction in neurologic function followed by remission. Available interventions target inflammatory injury by interfering with inflammatory immune functions but typically do not stimulate myelin repair or restore neurologic function. Targeted approaches that promote remyelination in MS would have a substantial impact on its management: demyelination interrupts metabolic support to axons normally provided by oligodendrocytes (OGs). As a result, denuded axons often degenerate, thereby worsening disability in MS patients. Although recent studies into the diverse role of glia in CNS pathologies have identified unique transcriptomes expressed during injury and recovery, OG responses to antibody-mediated demyelination/remyelination remain unclear. This proposal incorporates an innovative and cross-disciplinary approach to dissect OG responses during failed myelin repair. We have developed unique ex vivo and in vitro assays using disease-specific myelin binding recombinant antibodies (rAbs) first cloned from MS patient cerebrospinal fluid that recapitulate some of the barriers to remyelination in MS. In contrast to other demyelination/remyelination models, our patient-based remyelination model blocks remyelination during the transition of OGs from the early myelinating to actively myelinating stage, not during OPC differentiation. Experiments described in this application are designed to disentangle factors intrinsic & extrinsic to OGs that contribute to rAb-induced remyelination failure by thoroughly delineating OG-rAb responses using live cell imaging, immunohistochemistry, transgenic mice, innovative myelination assays, and next generation RNA sequencing modalities. To maximize the translational relevance of results generated in this model, biomarkers of remyelination blockade will be identified and interrogated in MS lesions. Thus, by utilizing sequential experimental and translational approaches, this proposal will identify how specific interactions between B cell humoral autoimmunity and OGs contribute to remyelination failure in human disease. Initially, we will test how MS rAbs affect oligodendrocytes directly by observing their effect on OG differentiation, ability to elaborate new myelin, and signaling pathways transduced by rAbs. We will then identify how oligodendroglial gene expression is influenced by rAbs during differentiation and myelinogenesis experimentally and evaluate candidate biomarkers for remyelination blockade in human MS plaques. These studies will not only expand scientific understanding of MS neuroimmunology, they will also illuminate factors contributing to remyelination that may be harnessed for the treatment of MS and other CNS demyelinating diseases.