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.

概括 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 全球数百万人。它是年轻人神经功能障碍的主要原因。的贡献 针对这种疾病的免疫系统已经很完善。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 这种疾病仍不清楚。人类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，神经系统衍生的髓磷脂抗原在神经系统发育中发挥着重要作用。 疾病。然而，这些抗原信号的格式以及它们如何转运到外周淋巴组织 组织仍然知之甚少。最近的证据表明，成熟细胞分泌的外泌体 少突胶质细胞 (OL) 富含多种髓磷脂抗原。外泌体是分泌的囊泡 人们越来越认识到它们在细胞间通讯中发挥着重要作用。重要的是含有髓磷脂 据报道，多发性硬化症患者的外周血中存在外泌体。 OL 衍生的外泌体的具体作用 尚未对人类 MS 或啮齿动物 EAE 进行研究。我们假设髓鞘形成产生的外泌体 细胞，尤其是 OL，可以作为中枢和外周髓磷脂抗原呈递的早期来源， 最终促进中枢神经系统实质中自身反应性淋巴细胞的致病性积累。具体来说， 我们建议通过首先分析髓鞘细胞来源的原位动力学来研究这一假设 在目标 1 中使用细胞类型特异性外泌体在小鼠 EAE 中的中枢神经系统和外周免疫组织中检测外泌体 报告鼠系，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)，其中包括使用铜宗短期治疗破坏中央髓磷脂，然后使用 诱导外周免疫反应。然后我们将检查外泌体到达各种目标的能力 与 Cup-EAE 相比，外周组织及其受控制的中枢神经系统内相关细胞类型的摄取 状况。我们还计划扩展我们有希望的初步数据，以进一步描述保护措施 EAE 由外泌体释放抑制剂引起。在目标 2 中，我们将重点了解 由对照和 MS 相关因子（TNFa 和谷氨酸）处理的 OL 在体外产生的外泌体。各种各样的 将评估这些囊泡的特性，包括大小、货物、抗原呈递细胞 (APC) 的摄取、 以及这些外泌体在体外支持 APC 向髓磷脂特异性 T 细胞呈递髓磷脂抗原的能力。 总的来说，我们相信我们利用创新的外泌体报告小鼠的建议将提供重要且 信息丰富的数据开始了解髓鞘细胞产生的外泌体在多发性硬化症中的作用 特别强调 OL 衍生的含髓磷脂的外泌体。