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

致病性 MS 患者来源的 IgG 对少突胶质细胞的影响：髓鞘再生失败的旁观者还是参与者？

• 批准号: 10285891
• 负责人: Andrew S Lapato
• 金额: $ 6.6万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10285891
• 关键词: Impact; pathogenic; MS; patient; derived

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.

项目总结/摘要 多发性硬化（MS）是中枢神经系统（CNS）的自身免疫性脱髓鞘疾病， 最常见的特征是神经系统的可测量和持续的减少 功能缓解后。可用的干预措施通过干扰 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MS髓鞘再生的靶向方法将对其管理产生重大影响： 脱髓鞘中断了通常由少突胶质细胞（OG）提供的对轴突的代谢支持。因此，在本发明中， 裸露的轴突通常退化，从而使MS患者的残疾恶化。虽然最近的研究 神经胶质在CNS病理学中的不同作用已经鉴定了在损伤期间表达的独特转录组， 恢复，OG对抗体介导的脱髓鞘/髓鞘再生的反应仍然不清楚。这项建议 结合了创新和跨学科的方法来解剖失败的髓鞘OG反应 修复.我们已经开发了独特的离体和体外试验，使用疾病特异性髓鞘结合 重组抗体（rAbs）首先从MS患者脑脊液中克隆， 与其他脱髓鞘/髓鞘再生模型相比，我们基于患者的 髓鞘再生模型在OG从早期髓鞘形成到活跃髓鞘形成的过渡期间阻断髓鞘再生。 髓鞘形成阶段，而不是在OPC分化期间。本申请中描述的实验被设计成 通过以下方式解开OG的内在和外在因素，这些因素有助于rAb诱导的髓鞘再生失败： 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下一代RNA测序模式。为了最大化平移 在该模型中产生的结果的相关性，将鉴定髓鞘再生阻断的生物标志物， 在MS病变中询问。因此，通过利用连续的实验和翻译方法， 一项提案将确定B细胞体液自身免疫和OG之间的特异性相互作用如何有助于 髓鞘再生失败的人类疾病。最初，我们将通过以下方式测试MS rAbs如何直接影响少突胶质细胞 观察它们对OG分化、产生新髓鞘的能力和转导的信号通路的影响。 rAbs的。然后我们将确定少突胶质细胞基因表达在分化过程中如何受到rAbs的影响 和髓鞘形成的研究，并评估人类髓鞘再生阻断的候选生物标志物。 MS斑块。这些研究不仅将扩大对MS神经免疫学的科学理解， 还阐明了有助于髓鞘再生的因素，这些因素可用于治疗MS和其他疾病。 中枢神经系统脱髓鞘疾病。