检测非小细胞肺癌中的EB病毒感染
先前的研究提出了爱泼斯坦-巴尔病毒(EBV)与肺癌(LC)之间的联系,但由于样本量不足以及传统病毒筛选方法(如PCR)的固有局限性,结果引起了很大争议。与PCR不同的是,当前的下一代测序(NGS)利用无偏的方法以高灵敏度和特异性对癌症样品中的所有外源性因子进行全局评估。在我们当前的研究中,我们旨在通过利用我们无偏倚的基于NGS的信息学方法与传统分子方法相结合,来研究EBV在总共1127 LC中的作用,以解决这一长期存在的争议。对110个LC和10个正常肺样本进行原位杂交分析,在3个LC样本中检测到EBV转录本。来自1017个LC和110对配对的正常肺标本的RNA测序(RNA-seq)数据集的综合病毒学分析显示,在3例肺鳞状细胞癌和1例肺腺癌样本中,EBV转录本。在EBV覆盖率最高的样本中,来自BamHI A区域的转录本占了EBV读数的大部分。观察到EBNA-1,LMP-1和LMP-2的表达。还检测到许多病毒环状RNA候选物。因此,我们首次揭示了在体内LC环境中的II型潜伏期样病毒转录组。 LC中病毒BamHI A转录物的高水平表达表明这些转录物的功能性作用,很可能是长的非编码RNA。细胞基因表达和染色的组织切片的分析表明,与表达低水平EBV转录本的样品相比,表达高水平EBV转录本的样品中免疫细胞浸润增加。 EBV转录物水平较高的样品中还检测到了更高的免疫检查点阻断因子水平,表明诱导了免疫耐受性。最后,与高EBV转录本相比,高EBV转录本的样品中检测到免疫途径的抑制和致癌途径的激活,表明EBV直接调节了癌症途径。综上所述,我们的数据支持以下观点:EBV可能在LC的一个子集中发挥病理作用。**
Detection of Epstein-Barr Virus Infection in Non-Small Cell Lung Cancer
Previous investigations proposed a link between the Epstein-Barr virus (EBV) and lung cancer (LC), but the results are highly controversial largely due to the insufficient sample size and the inherent limitation of the traditional viral screening methods such as PCR. Unlike PCR, current next-generation sequencing (NGS) utilizes an unbiased method for the global assessment of all exogenous agents within a cancer sample with high sensitivity and specificity. In our current study, we aim to resolve this long-standing controversy by utilizing our unbiased NGS-based informatics approaches in conjunction with traditional molecular methods to investigate the role of EBV in a total of 1127 LC. In situ hybridization analysis of 110 LC and 10 normal lung samples detected EBV transcripts in 3 LC samples. Comprehensive virome analyses of RNA sequencing (RNA-seq) data sets from 1017 LC and 110 paired adjacent normal lung specimens revealed EBV transcripts in three lung squamous cell carcinoma and one lung adenocarcinoma samples. In the sample with the highest EBV coverage, transcripts from the BamHI A region accounted for the majority of EBV reads. Expression of EBNA-1, LMP-1 and LMP-2 was observed. A number of viral circular RNA candidates were also detected. Thus, we for the first time revealed a type II latency-like viral transcriptome in the setting of LC in vivo. The high-level expression of viral BamHI A transcripts in LC suggests a functional role of these transcripts, likely as long non-coding RNA. Analyses of cellular gene expression and stained tissue sections indicated an increased immune cell infiltration in the sample expressing high levels of EBV transcripts compared to samples expressing low EBV transcripts. Increased level of immune checkpoint blockade factors was also detected in the sample with higher levels of EBV transcripts, indicating an induced immune tolerance. Lastly, inhibition of immune pathways and activation of oncogenic pathways were detected in the sample with high EBV transcripts compared to the EBV-low LC indicating the direct regulation of cancer pathways by EBV. Taken together, our data support the notion that EBV likely plays a pathological role in a subset of LC.
pmid: 31159203 Cancers (Basel) 影响因子: 6.162 发表日期: 20190531 官网 免费下载
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