INTRODUCTION Like all nucleated cells, glioblastoma (GBM) cells shed small membrane-encapsulated particles called extracellular vesicles (EVs). EVs can transfer oncogenic components and promote tumor growth by transferring short non-coding RNAs, altering target cell gene expression. Furthermore, GBM-derived EVs can be detected in blood and have potential to serve as liquid biopsies. METHODS EVs were harvested from culture supernatants from human GBM cell lines, purified via sequential centrifugation, and quantified by nanoparticle tracking. RNA was isolated and short non-coding RNA was sequenced. Data was analyzed via the OASIS-2.0 platform using HG38. MirTarBase and MirDB interrogated validated/predicted miRNA-gene interactions respectively. RESULTS Many short non-coding RNA's were identified within GBM EV's. In keeping with earlier reports utilizing GBM EV micro-RNA (miRNA) arrays, these included abundant micro-RNA's including miR-21. However, RNA sequencing revealed a total of 712 non-coding RNA sequences most of which have not been associated with GBM EV's previously. These included many RNA species (piRNA, snoRNA, snRNA, rRNA and yRNAs) in addition to miRNA's. miR-21-5p, let-7b-5p, miR-3182, miR-4448, let-7i-5p constituted highest overall expression. Top genes targeted by non-coding RNA's were highly conserved and specific for cell cycle, PI3K/Akt signaling, p53 and Glioma curated KEGG pathways. CONCLUSIONS Next generation short non-coding RNA sequencing on GBM EV's validates findings from earlier studies using miRNA arrays but also demonstrates expression of many additional non-coding RNA sequences and classes previously unassociated with GBM. This may yield important insights into pathophysiology, point to new therapeutic targets, and help develop new biomarkers for disease burden and treatment response.

中文翻译：

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胶质母细胞瘤细胞外囊泡的短非编码RNA测序。

简介与所有有核细胞一样，胶质母细胞瘤（GBM）细胞会散发出称为细胞外囊泡（EVs）的膜包裹小颗粒。电动汽车可通过转移短的非编码RNA改变靶细胞基因表达，从而转移致癌成分并促进肿瘤生长。此外，可以在血液中检测到GBM衍生的EV，并且有潜力用作液体活检。方法从人GBM细胞系的培养上清液中收获EV，通过连续离心纯化，并通过纳米粒子跟踪进行定量。分离RNA，并对短的非编码RNA进行测序。使用HG38通过OASIS-2.0平台分析数据。MirTarBase和MirDB分别询问已验证/预测的miRNA-基因相互作用。结果GBM EV中鉴定出许多短的非编码RNA。与先前利用GBM EV微型RNA（miRNA）阵列的报道保持一致，其中包括丰富的微型RNA，包括miR-21。但是，RNA测序揭示了总共712个非编码RNA序列，其中大多数以前与GBM EV无关。除miRNA外，这些还包括许多RNA种类（piRNA，snoRNA，snRNA，rRNA和yRNA）。miR-21-5p，let-7b-5p，miR-3182，miR-4448，let-7i-5p构成最高的整体表达。非编码RNA靶向的顶级基因是高度保守的，对细胞周期，PI3K / Akt信号转导，p53和胶质瘤调控的KEGG通路具有特异性。结论GBM EV'上的下一代短非编码RNA短测序 s验证了使用miRNA阵列进行的早期研究的结果，但也证明了以前与GBM不相关的许多其他非编码RNA序列和类别的表达。这可能会产生重要的病理生理学见解，指向新的治疗靶标，并有助于开发新的生物标记物以减轻疾病负担和治疗反应。