High-throughput sequencing of two populations of extracellular vesicles provides an mRNA signature that can be detected in the circulation of breast cancer patients.
High-throughput sequencing of two populations of extracellular vesicles provides an mRNA signature that can be detected in the circulation of breast cancer patients.
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DOI：
10.1080/15476286.2016.1259061
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发表时间：
2017-03-04
期刊：
影响因子：
4.1
通讯作者：
Di Vizio D
中科院分区：
文献类型：
--
作者：
Conley A;Minciacchi VR;Lee DH;Knudsen BS;Karlan BY;Citrigno L;Viglietto G;Tewari M;Freeman MR;Demichelis F;Di Vizio D
细胞外囊泡（EV）含有广泛的RNA类型，其中非编码RNA的报道普遍存在。迄今为止，仍然缺乏对EV中蛋白质编码转录物的全面表征。我们对2个EV群体进行了RNA测序（RNA-Seq），并鉴定了一小部分在大的癌小体和外泌体中以显著不同的水平表达的转录物，表明它们可能介导专门的功能。然而，这2个EV群体表现出共同的mRNA特征，与其供体细胞相比，显著富集编码E2 F转录靶点和组蛋白的mRNA。这些mRNA主要在细胞周期的S期表达，这表明它们可能在S期被包装到EV中。使用来自ENCODE细胞系纲要的亚细胞区室转录组数据的计算机模拟分析显示EV mRNA来源于细胞质RNA库。通过RNA-Seq在患有乳腺癌的患者的血浆中独立地鉴定EV特征。此外，与对照相比，患者EV中差异表达的几种转录本反映了正常和乳腺癌组织之间的差异表达。总而言之，EV mRNA的这种最大的高通量分析表明EV携带肿瘤特异性改变，并且可以作为癌症衍生货物的来源进行询问。
Extracellular vesicles (EVs) contain a wide range of RNA types with a reported prevalence of non-coding RNA. To date a comprehensive characterization of the protein coding transcripts in EVs is still lacking. We performed RNA-Sequencing (RNA-Seq) of 2 EV populations and identified a small fraction of transcripts that were expressed at significantly different levels in large oncosomes and exosomes, suggesting they may mediate specialized functions. However, these 2 EV populations exhibited a common mRNA signature that, in comparison to their donor cells, was significantly enriched in mRNAs encoding E2F transcriptional targets and histone proteins. These mRNAs are primarily expressed in the S-phase of the cell cycle, suggesting that they may be packaged into EVs during S-phase. In silico analysis using subcellular compartment transcriptome data from the ENCODE cell line compendium revealed that EV mRNAs originate from a cytoplasmic RNA pool. The EV signature was independently identified in plasma of patients with breast cancer by RNA-Seq. Furthermore, several transcripts differentially expressed in EVs from patients versus controls mirrored differential expression between normal and breast cancer tissues. Altogether, this largest high-throughput profiling of EV mRNA demonstrates that EVs carry tumor-specific alterations and can be interrogated as a source of cancer-derived cargo.