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: 10526691
• 负责人: ALEXEI DEGTEREV
• 金额: $ 20.63万
• 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/10526691
• 关键词: Affect; Antigen Presentation; Antigen-Presenting Cells; Antigens; Attenuated; Autoimmune; Autoimmune Responses; Autoimmunity; B-Lymphocytes; Biogenesis; Blood Circulation; CD4 Positive T Lymphocytes; Cell Communication; Cells; Central Nervous System Diseases; Chronic; Clinical; Cuprizone; Data; Demyelinating Diseases; Demyelinations; Dendritic Cells; Development; Disease; Dose; 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; Mediating; Microglia; Modeling; Modification; Multiple Sclerosis; Mus; Myelin; Myelin Proteins; Nerve Degeneration; Nervous system structure; Neuraxis; Neuroglia; Neurologic; Neurons; Oligodendroglia; Outcome; Pathogenesis; Pathogenicity; Pathologic; Peptides; Peripheral; Persons; Phosphatidylserines; Play; Property; Reporter; Reporting; Rodent; Role; Schwann Cells; Serum; Signal Transduction; Solid; Source; Spleen; Surface; T-Cell Activation; T-Lymphocyte; Tissues; Vesicle; Yang; autoreactivity; cell type; central nervous system injury; disability; draining lymph node; exosome; experimental study; extracellular vesicles; glutamatergic signaling; in vivo; inhibitor; innovation; interest; macrophage; mouse model; multiple sclerosis patient; neuroinflammation; novel therapeutics; oligodendrocyte-myelin glycoprotein; peripheral lymphoid organ; protective effect; protein aggregation; response; serine containing aminolipid; 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 多发性硬化是一种进行性多灶性脱髓鞘疾病，影响约2.5 全球百万人。它是年轻人神经疾病的主要原因。贡献 该疾病的免疫系统已经确定。靶向B和T淋巴细胞的多种治疗方法已经 针对最普遍的疾病（RRMS）反复恢复形式而开发。但是，病因 该疾病尚不清楚。人类MS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开发中很重要 疾病。但是，这些抗原信号的格式以及它们如何转运到周围淋巴样 组织仍然了解不足。最近的证据表明，成熟分泌的外泌体 少突胶质细胞（OLS）富含各种髓磷脂抗原。外泌体是分泌的囊泡 越来越多地赞赏在细胞电池通信中扮演重要角色。重要的是，含有髓磷脂 MS患者的外围据报道外泌体。 OL衍生外泌体的特定作用在 尚未研究人类MS或啮齿动物EAE。我们假设髓鞘产生的外泌体 细胞，尤其是OLS，可以作为中央和周围髓磷脂抗原表现的早期来源， 最终促进了CNS实质中自动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衍生的原位动力学来研究这一假设 CNS中的外泌体和AIM 1中小鼠EAE的外周免疫组织使用细胞型特异性外泌体 记者鼠标线，最近由MPI实验室开发。对于这些实验，我们将使用修改的EAE模型 （杯-EEE），包括使用cuprizone的短疗法，然后是髓鞘中央纤维的破坏 周围免疫反应的诱导。然后，我们将检查外泌体达到各种能力的能力 外围组织以及它们在控制下的中枢神经系统中相关细胞类型与杯-EAE的吸收 状况。我们还计划扩展我们有希望的初步数据，以进一步表征 EAE通过外泌体释放抑制剂。在AIM 2中，我们将专注于了解 由对照和MS相关因子（TNFA和谷氨酸）处理的OLS在体外产生的外泌体。各种各样的 将评估这些囊泡的特性，包括尺寸，货物，抗原呈递细胞的吸收（APC）， 这些外泌体在体外对髓磷脂特异性T细胞支持髓磷脂抗原表现的能力。 总体而言，我们认为，使用创新的外部记者鼠标使用创新的建议将提供重要的和 提供信息的数据，以开始了解MS中髓鞘细胞产生的外泌体的作用 对含OL的含髓磷脂外泌体的特定强调。