Analyzing Oligodendrocytes-Derived Exosomes in Mouse Models of Multiple Sclerosis

分析多发性硬化症小鼠模型中少突胶质细胞来源的外泌体

• 批准号: 10622532
• 负责人: ALEXEI DEGTEREV
• 金额: $ 24.75万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10622532
• 关键词: Affect; Antigen Presentation; Antigen-Presenting Cells; Antigens; Autoimmune; Autoimmune Responses; Autoimmunity; Biogenesis; CD4 Positive T Lymphocytes; Cell Communication; Cells; Central Nervous System; Central Nervous System Diseases; Chronic; Circulation; Clinical; Cuprizone; Data; Demyelinating Diseases; Demyelinations; Dendritic Cells; Development; Disease; Disease remission; Dose; Endosomes; Enteral; Epitopes; Etiology; Experimental Autoimmune Encephalomyelitis; Foundations; Functional disorder; Future; Generations; Glutamates; Human; Immune; Immune response; Immune system; Immunization; In Situ; In Vitro; Injury; Investigation; Knowledge; Label; Lead; Lymphocyte; Lymphoid Tissue; Macrophage; Mediating; Microglia; Modeling; Modification; Multiple Sclerosis; Mus; Myelin; Myelin Proteins; Nerve Degeneration; Nervous System; Neuroglia; Neurologic; Neurons; Oligodendroglia; Outcome; Pathogenesis; Pathogenicity; Pathologic; Peptides; Peripheral; Persons; Phosphatidylserines; Play; Property; Recurrence; Reporter; Reporting; Rodent; Role; Schwann Cells; Serum; Signal Transduction; Solid; Source; Spleen; Surface; T-Cell Activation; T-Lymphocyte; Tissues; Vesicle; autoreactivity; cell type; central nervous system injury; disability; draining lymph node; exosome; experimental study; extracellular vesicles; glutamatergic signaling; in vivo; inhibitor; innovation; interest; mouse model; multiple sclerosis patient; neuroinflammation; novel therapeutics; oligodendrocyte-myelin glycoprotein; peripheral lymphoid organ; protective effect; protein aggregation; response; uptake; young adult

Summary Multiple Sclerosis is a progressive multifocal demyelinating disease which affects approximately 2.5 million people worldwide. It is the leading cause of neurologic disability in young adults. The contribution of the immune system to this disease is well established. Multiple treatments targeting B and T lymphocytes have been developed against the most prevalent Recurring Remitting form of the disease (RRMS). However, etiology of the disease remains unclear. Studies from human MS and rodent Experimental Autoimmune Encephalomyelitis (EAE) models suggest that nervous system-derived myelin antigens are important in the development of the disease. However, the format of these antigenic signals and how they are transported to the peripheral lymphoid tissues remain poorly understood. Recent evidence suggest that exosomes secreted by mature oligodendrocytes (OLs) are enriched in a variety of myelin antigens. Exosomes are the secreted vesicles that are increasingly appreciated to play important roles in cell-cell communications. Importantly, myelin-containing exosomes have been reported peripherally in MS patients. The specific roles of OL-derived exosomes either in human MS or rodent EAE has not been investigated. We hypothesize that exosomes produced by myelinating cells, especially OLs, may serve as early sources for central and peripheral myelin antigen presentation, ultimately facilitating pathogenic accumulation of autoreactive lymphocytes in the CNS parenchyma. Specifically, we propose to investigate this hypothesis by first analyzing in situ dynamics of the myelinating cell-derived exosomes in CNS and peripheral immune tissues in mouse EAE in Aim 1 using a cell-type specific exosome reporter mouse line, recently developed by the MPI lab. For these experiments we will use a modified EAE model (Cup-EAE) which includes both disruption of central myelin using short treatment with cuprizone followed by the induction of the peripheral immune response. We will then examine both the ability of exosomes to reach various peripheral tissues as well as their uptake by relevant cell types within the CNS under control versus Cup-EAE conditions. We also plan to expand on our promising preliminary data to further characterize the protection from EAE by the exosome release inhibitor. In Aim 2, we will focus on understanding the functional properties of exosomes produced in vitro by control and MS-associated factors (TNFa and glutamate)-treated OLs. Various properties of these vesicles will be assessed including size, cargo, uptake by antigen presenting cells (APCs), and ability of these exosomes to support myelin antigen presentation by APCs to myelin-specific T cells in vitro. Overall, we believe that our proposal utilizing an innovative exosome reporter mouse will provide important and informative data to begin the understanding of the roles of exosomes produced by myelinating cells in MS with specific emphasis on OL-derived myelin-containing exosomes.

摘要 多发性硬化症是一种进行性多灶性脱髓鞘疾病，影响约2.5 全世界有一百万人。它是导致年轻人神经功能障碍的主要原因。《纽约时报》的贡献 对这种疾病的免疫系统很好地建立了。针对B和T淋巴细胞的多种治疗方法 针对最流行的复发缓解型疾病(RRMS)开发的。然而，病因学 这种疾病目前仍不清楚。人多发性硬化症和啮齿动物实验性自身免疫性脑脊髓炎的研究 (EAE)模型表明，神经系统来源的髓鞘抗原在慢性粒细胞白血病的发生发展中起重要作用。 疾病。然而，这些抗原信号的格式以及它们是如何运输到外周淋巴系统的 人们对组织仍然知之甚少。最近的证据表明，成熟细胞分泌的外切体 少突胶质细胞富含各种髓鞘抗原。Exosome是分泌的囊泡， 在细胞间的通讯中扮演着重要的角色。重要的是，含有髓鞘的 在多发性硬化症患者中，已有外周外周的报道。OL来源的外切体在体内的特殊作用 尚未对人类多发性硬化症或啮齿动物EAE进行调查。我们假设外体是通过髓鞘作用产生的 细胞，尤其是OL，可能是中枢和外周髓鞘抗原呈递的早期来源， 最终促进自体反应性淋巴细胞在中枢神经系统实质中的病理性积聚。具体来说， 我们建议首先通过分析髓鞘细胞来源的原位动力学来研究这一假说。 AIM-1小鼠EAE中枢神经系统和外周免疫组织中的Exosome 记者鼠标线，最近由MPI实验室开发。对于这些实验，我们将使用修改后的EAE模型 (CUP-EAE)，包括使用铜酮短期治疗破坏中央髓鞘，然后 诱导外周免疫反应。然后我们将检查外切体到达各种不同的 对照与CUP-EAE下中枢神经系统内相关细胞类型对周围组织的摄取 条件。我们还计划扩大我们有希望的初步数据，以进一步描述保护免受 外切体释放抑制剂治疗EAE。在目标2中，我们将重点了解 对照和多发性硬化症相关因子(TNFa和谷氨酸)处理的OL体外产生的外切体。五花八门 这些囊泡的性质将被评估，包括大小，体积，抗原提呈细胞(APC)的摄取， 以及这些外切体在体外支持APC向髓鞘特异性T细胞递呈髓鞘抗原的能力。 总体而言，我们相信，我们的建议利用创新的外切体记者鼠标将提供重要的和 开始了解髓鞘细胞产生的外体在多发性硬化症中的作用的有用数据 特别强调OL来源的含有髓磷脂的外切体。