Delineating the RNA cargo of exosomes from brain microenvironment

项目摘要 循环胞外小泡(EV)是细胞外颗粒(EP)的一个子集，存在于 所有组织和体液的细胞外空间。EPS由包括神经元和星形胶质细胞在内的所有细胞释放 并被认为在突触上具有调节功能。循环中的EPs含有分子足迹(脂类， 蛋白质、代谢物、RNA和DNA)。到目前为止，许多组织特异性标志物已经 在从唾液、尿液、脑脊液和血液等各种生物液体中提取的EVS中发现了这种病毒。 因此，电动汽车和其他EP越来越被认为是细胞间通信的关键参与者， 主要通过囊泡运输，促使它们被评估为预后和治疗反应的生物标志物。 我计划在这里解决的一个关键问题是，描述大脑所有EP的分子图景 微环境和血液。神经内分泌组是我在这里介绍的一个概念，它体现了 连接人类大脑和血液的循环分子。人们对神经分泌组知之甚少 仍然是知识方面的一个主要差距。这些分泌颗粒包括但不限于微泡、外体、 外显子、高密度脂蛋白和低密度脂蛋白、ArgAerte蛋白、核糖核蛋白和无细胞 蛋白质/DNA/RNA。 尽管它们的希望越来越大，但分离电动汽车和其他EP进行分析仍然是一项劳动密集型和时间密集型任务- 考虑到它们的纳米尺寸和低浮力密度，它们面临着巨大的挑战。这部分是由于缺乏 这种感兴趣的纳米颗粒的可重复性的分离和表征方法。我的目标是调查 所有脑微环境的EPs和匹配的血液。为了实现这一目标，我建议使用 我开发的从人体生物液中分离和鉴定电动汽车和其他EPs的技术。在… 这个项目的完成，我希望提供：1)神经分泌组的完整分子图谱 AD患者的大脑；2)血液中神经分泌体的完整分子图谱；3)重叠 来自脑和血液的所有EPs的转录组/蛋白质组的签名。

Project Summary Circulating extracellular vesicles (EVs) are a subset of extracellular particles (EPs) canopy present in the extracellular space of all tissues and bodily fluids. EPs are released by all cells including neurons and astrocytes and are implicated to have a regulatory function at synapses. Circulating EPs contain molecular footprints (lipids, proteins, metabolites, RNA, and DNA) from their cell of origin. To date, numerous tissue-specific markers have been discovered in the EVs derived from a variety of biofluids including saliva, urine, cerebrospinal fluid, and blood. Consequently, EVs and other EPs are increasingly recognized as key players in the cell-to-cell communication, mainly through vesicle trafficking, prompting their evaluation as prognostic and treatment response biomarkers. A key question that I plan to address here, is characterization of the molecular landscape of all EPs of the brain microenvironment and blood. The neurosecretome is a concept I introduce here to embody the compendium of circulating molecules that link the brain and blood in humans. The neurosecretome is poorly understood and remains a major gap in knowledge. These secretory particles include, but not limited to, microvesicles, exosomes, exomeres, high and low density-lipoproteins, Argonaute proteins, ribonucleic proteins, and cell free proteins/DNA/RNA. Despite their growing promise, isolation of EVs and other EPs for analysis remains a labor-intensive and time- consuming challenge given their nanoscale dimensions and low buoyant density. This in part is due to lack of reproducible isolation and characterization methods of such nanoparticles of interest. My objective is to investigate all EPs of the brain microenvironment and matching blood. To accomplish this goal, I propose to use the technology that I have developed to isolate and characterize the EVs and other EPs from human biofluids. At the completion of this project, I expect to provide; 1) A complete molecular profile of the neuro-secretome of the brain from AD patients; 2) A complete molecular profile of the neurosecretome in the blood; 3) An overlapping signature from the transcriptome/proteome of all EPs from brain and blood.