Molecular profiling of exosomes from multiple sclerosis B cells

项目摘要/摘要 B细胞是多发性硬化症(MS)发病机制的核心，包括与 产生免疫球蛋白(Ig)。灰质损伤，特别是硬膜下灰质损伤是一种病理现象。 多发性硬化症进展阶段的基础，许多人经历了认知和身体状况的恶化 病人。我们的假设是，外周免疫系统的B细胞进入脊髓液并 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)和缓慢演变的血管周围病变。我们的研究团队位于 韦恩州立大学和宾夕法尼亚大学正在合作研究这一假说。 MS患者外周血B细胞培养上清对OL和 神经细胞的体外培养；来自正常对照组(NC)的神经元表现出很少或没有毒性(利萨克等人)。2012、2017)。杀戮就是 与补体无关，且与血清中的免疫球蛋白、免疫球蛋白M或以下各项的任何单一或组合无关 大量的可溶性细胞因子等蛋白质。OL和神经元的死亡涉及细胞凋亡和IS 由MS B细胞Sup的&gt；300 kDa部分中的一个或多个因素引起(利萨克等人2017年)。我们最近 发现OL的毒性存在于富含外切体(Ex-en)的细胞外小泡(EVS)中 来自MS患者的B细胞(Benjamins等人)。2019年)。我们将利用蛋白质组学来鉴定候选因子/S， RNAseq和脂质组学。阻断因子的产生或作用/S可降低OL和神经元 损害皮质多发性硬化症病变，从而防止疾病进展和增强重新髓鞘形成的能力 和轴突保护。预期的新成果包括(1)全面的分子图谱和 MS B细胞释放的Ex-en EV的特征，这是以前从未做过的，以及(2) 外源性EVS中或其上可介导B细胞损伤的因子的鉴定 多发性硬化症的皮质损害我们提出的综合多重组学方法将产生丰富的数据集 将深入了解B细胞EV的基本生物学以及B细胞EV表型的差异 与正常对照组相比，MS患者的功能和功能有明显的改善。这个项目意义重大，因为它具有很强的 确定新的目标和策略，以减少MS患者中枢神经系统的损伤并促进修复 这种B细胞毒性可能与非复发的进行性病理生理学特别相关--这是一个尚未满足的主要问题 治疗需求。我们的项目是创新的，因为Ex-en电动汽车的功能、组件和角色 由多发性硬化症B细胞释放的，以前没有被研究过。

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.