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.

项目摘要/摘要 B细胞是多发性硬化症（MS）发病机理的核心，包括与B细胞功能无关的B细胞功能 免疫球蛋白（IG）的产生。灰质损伤（GM），尤其是亚型GM，是一种病理 MS渐进阶段的基板，许多人经历了认知和身体上的忧虑 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， 脑膜和释放非避孕药（Ig）因素，损害寡头（OL）和神经元/轴突的因素 潜在的脑皮质灰质（GM）和缓慢发展的周围病变。我们的研究团队 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细胞培养物获得的上清液（SUP）都是细胞毒性的，既可以 体外神经元；正常对照（NC）的毒性很少或没有毒性（Lisak等，2012，2017）。杀人是 独立于完成，并且与IgG，IgM或任何单一或组合的SUP水平不相关 大量固体细胞因子和其他蛋白质。 OL和神经元的死亡涉及凋亡，IS 由MS B细胞SUP的> 300 kDa级分中的一个或多个因素引起（Lisak等，2017）。我们最近 发现在富含外泌体的（EX-EN）细胞外蔬菜（EV）中发现了OL毒性。 来自MS患者的B细胞（Benjamins等，2019）。我们将使用蛋白质组学确定候选因子/s， RNASEQ和脂肪组学。阻止因子的产生或作用可能会降低OL和神经元 皮质MS病变的损害，从而预防进行性疾病并增强remer髓的能力 和轴突保护。 MS B细胞释放的EX-EN EV的表征，此前尚未完成，（2） 识别可以介导B细胞诱导的损害的前EV中或前EV中的因素的识别 MS中的皮质病变。我们提出的综合多种方法方法将生成丰富的数据集 将洞悉B细胞电动汽车的基本生物学以及B细胞EV表型的差异 与NC相比，MS患者的功能和功能。该项目很重要，因为它的强大潜力 确定新的目标和策略，以减少MS患者中CNS的损害并促进CNS的修复。 该B细胞毒性可能与非伴随性的进行性病理生理学特别相关 - 一种主要的未满足 治疗需求。我们的项目具有创新性，因为前EV的功能，组成部分和角色 先前尚未研究MS B细胞发布。