Molecular profiling of exosomes from multiple sclerosis B cells

多发性硬化症 B 细胞外泌体的分子分析

• 批准号: 10040973
• 负责人: AMIT BAR-OR
• 金额: $ 44.7万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10040973
• 关键词: Affect; Apoptosis; Axon; B-Lymphocytes; Biology; Cell physiology; Cerebrospinal Fluid; Cerebrum; Cessation of life; Cognitive; Complement; Correlation Studies; Data; Data Set; Development; Disease; Disease Progression; Facilities and Administrative Costs; Functional disorder; Future; Goals; Health; Immune system; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; In Vitro; Income; Individual; Injury; Lead; Lesion; Lipids; Mediating; Medical; Meninges; MicroRNAs; Molecular; Molecular Profiling; Multiple Sclerosis; Multiple Sclerosis Lesions; Neurons; Oligodendroglia; Outcome; Pathogenesis; Pathologic; Pathway Analysis; Pathway interactions; Patients; Pennsylvania; Peripheral; Phase; Phenotype; Production; Progressive Disease; Proteins; Proteomics; RNA; Research; Role; Sampling; Systems Biology; Testing; Therapeutic; Toxic effect; Universities; cost; cytokine; cytotoxic; disability; exosome; experience; extracellular vesicles; gray matter; high reward; high risk; innovation; insight; multiple omics; multiple sclerosis patient; new therapeutic target; prevent; protein B; release factor; remyelination; repaired; transcriptome sequencing; vesicular release

Project Summary/Abstract B cells are central to the pathogenesis of multiple sclerosis (MS), including B cell functions unrelated to production of immunoglobulins (Ig). Injury to gray matter (GM), particularly subpial GM, is a pathologic substrate of the progressive stages of MS, with cognitive and physical worsening experienced by many patients. Our hypothesis is that B cells from the peripheral immune system enter the spinal fluid and meninges, and release non-immunoglobulin (Ig) factors that injure oligodendroglia (OL) and neurons/axons in underlying cerebral cortical gray matter (GM) and slowly evolving perivascular lesions. Our research teams at Wayne State University and the University of Pennsylvania are collaborating to investigate this hypothesis. Supernatants (Sup) obtained from cultures of circulating B cells from MS patients are cytotoxic to both OL and neurons in vitro; those from normal controls (NC) show little or no toxicity (Lisak et al. 2012, 2017). Killing is independent of complement, and does not correlate with Sup levels of IgG, IgM or any single or combination of a large number of soluble cytokines and other proteins. Death of OL and neurons involves apoptosis and is caused by one or more factors in the >300 kDa fraction of MS B cell Sup (Lisak et al. 2017). We recently discovered that the OL toxicity is found in exosome-enriched (Ex-En) extracellular vesicles (EVs) released by the B cells from patients with MS (Benjamins et al. 2019). We will identify candidate factor/s using proteomics, RNASeq and lipidomics. Blocking the production or action of the factor/s could decrease OL and neuronal damage in cortical MS lesions and thus prevent progressive disease and enhance capacity for remyelination and axonal protection. Anticipated new outcomes include (1) comprehensive molecular profiling and characterization of Ex-En EVs released by MS B cells, which has not been done before, and (2) identification of factors in or on the Ex-En EVs that could mediate the damage induced by B cells in cortical lesions in MS. The integrated multiple -omics approaches we propose will generate rich datasets that will provide insight into the fundamental biology of B cell EVs as well as the differences in B cell EV phenotype and function in MS patients compared to NC. This project is significant because of its strong potential for identifying new targets and strategies to decrease damage and promote repair of the CNS in patients with MS. This B cell toxicity may be particularly relevant to non-relapsing progressive pathophysiology – a major unmet therapeutic need. Our project is innovative because the function, components and roles of Ex-En EVs released by MS B cells have not been previously investigated.

项目总结/摘要 B细胞是多发性硬化（MS）发病机制的核心，包括与MS无关的B细胞功能。 产生免疫球蛋白（IG）。灰质（GM），特别是软膜下GM的损伤是一种病理性损伤， MS进展阶段的基质，许多人经历了认知和身体恶化 患者我们的假设是来自外周免疫系统的B细胞进入脊髓液， 脑膜，并释放非免疫球蛋白（IG）因子，损伤少突胶质细胞（OL）和神经元/轴突， 潜在的大脑皮质灰质（GM）和缓慢发展的血管周围病变。我们的研究团队在 韦恩州立大学和宾夕法尼亚大学正在合作调查这一假设。 从MS患者的循环B细胞培养物中获得的上清液（Sup）对OL和IL-10均具有细胞毒性。 体外神经元;来自正常对照（NC）的那些显示很少或没有毒性（Lisak等人，2012，2017）。杀人是 不依赖于补体，并且与IgG、IgM或任何单一或组合的Sup水平无关。 大量的可溶性细胞因子和其他蛋白质。OL和神经元的死亡涉及细胞凋亡， 由MS B细胞Sup>300 kDa部分中的一种或多种因子引起（Lisak et al. 2017）。我们最近 发现OL毒性存在于由细胞释放的富含外泌体（Ex-En）的细胞外囊泡（EV）中。 来自MS患者的B细胞（Benjamins等人，2019）。我们将使用蛋白质组学确定候选因子， RNASeq和脂质组学。阻断该因子的产生或作用可降低OL和神经元的功能， 皮质MS损伤，从而防止疾病进展并增强髓鞘再生能力 和轴突保护预期的新成果包括（1）全面的分子谱分析， 表征由MS B细胞释放的Ex-En EV，这在以前没有进行过，和（2） 鉴定Ex-En EV中或其上可介导由B细胞诱导的损伤的因子， 我们提出的综合多组学方法将产生丰富的数据集， 将提供对B细胞EV的基础生物学以及B细胞EV表型差异的深入了解 与NC相比，MS患者的功能。该项目意义重大，因为它具有强大的潜力， 确定新的靶点和策略，以减少MS患者的CNS损伤并促进其修复。 这种B细胞毒性可能与非复发性进行性病理生理学特别相关，这是一个主要的未满足的 治疗需要我们的项目是创新的，因为Ex-En EV的功能，组件和角色 以前没有研究过MS B细胞释放的。