Department of Neurology, Brigham and Women's Hospital and Harvard Medical School, HMS Initiative for RNA Medicine, Boston, MA, 02115, USA.
Vishuo Biomedical, #3-33 Teletech Park, 20 Science Park Road, Singapore, 117674, Singapore.
Nat Commun. 2017 Oct 26;8(1):1145. doi: 10.1038/s41467-017-01196-x.
Tumor-released RNA may mediate intercellular communication and serve as biomarkers. Here we develop a protocol enabling quantitative, minimally biased analysis of extracellular RNAs (exRNAs) associated with microvesicles, exosomes (collectively called EVs), and ribonucleoproteins (RNPs). The exRNA complexes isolated from patient-derived glioma stem-like cultures exhibit distinct compositions, with microvesicles most closely reflecting cellular transcriptome. exRNA is enriched in small ncRNAs, such as miRNAs in exosomes, and precisely processed tRNA and Y RNA fragments in EVs and exRNPs. EV-enclosed mRNAs are mostly fragmented, and UTRs enriched; nevertheless, some full-length mRNAs are present. Overall, there is less than one copy of non-rRNA per EV. Our results suggest that massive EV/exRNA uptake would be required to ensure functional impact of transferred RNA on brain recipient cells and predict the most impactful miRNAs in such conditions. This study also provides a catalog of diverse exRNAs useful for biomarker discovery and validates its feasibility on cerebrospinal fluid.
肿瘤释放的 RNA 可能介导细胞间通讯，并作为生物标志物。在这里，我们开发了一种定量、最小偏倚的方法，用于分析与微泡、外泌体（统称为 EV）和核糖核蛋白（RNP）相关的细胞外 RNA（exRNA）。从患者来源的神经胶质瘤干细胞样培养物中分离出的 exRNA 复合物具有不同的组成，其中微泡最能反映细胞转录组。exRNA 富含小分子 ncRNA，如外泌体中的 miRNA，以及 EV 和 exRNP 中精确加工的 tRNA 和 Y RNA 片段。EV 包裹的 mRNA 大多是碎片化的，UTR 富集；然而，也存在一些全长 mRNA。总的来说，每个 EV 中不到一个非 rRNA 拷贝。我们的结果表明，需要大量的 EV/exRNA 摄取才能确保转移 RNA 对脑受体细胞的功能影响，并预测在这种情况下最具影响力的 miRNA。本研究还提供了一个用于生物标志物发现的多样化 exRNA 目录，并验证了其在脑脊液中的可行性。
