Investigating the Influence of Ribavirin on Human Respiratory Syncytial Virus RNA Synthesis by Using a High-Resolution Transcriptome Sequencing Approach.
Investigating the Influence of Ribavirin on Human Respiratory Syncytial Virus RNA Synthesis by Using a High-Resolution Transcriptome Sequencing Approach.
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DOI：
10.1128/jvi.02349-15
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发表时间：
2016-05-15
影响因子：
5.4
通讯作者：
Hiscox JA
中科院分区：
文献类型：
--
作者：
Aljabr W;Touzelet O;Pollakis G;Wu W;Munday DC;Hughes M;Hertz-Fowler C;Kenny J;Fearns R;Barr JN;Matthews DA;Hiscox JA
人呼吸道合胞病毒（HRSV）是引起严重呼吸道感染的主要病原。治疗方案包括给予利巴韦林，一种嘌呤类似物，尽管其抗HRSV活性的机制尚不清楚。我们使用转录组测序（RNA-seq）来研究基因组突变频率和病毒mRNA在HRSV感染的细胞中的积累，这些细胞未经处理或用利巴韦林处理。在没有利巴韦林的情况下，HRSV特异性转录物占感染细胞RNA群体总RNA读数的三分之一。利巴韦林处理导致病毒mRNA读数丰度降低>90%，而同时未检测到细胞转录物丰度的这种降低。所提供的数据表明，利巴韦林显着增加的频率HRSV特异性RNA突变，这表明直接影响的保真度的HRSV聚合酶。所提供的数据表明，转换和颠换发生在HRSV复制，这些变化发生在热点沿着HRSV基因组。病毒基因组中核苷酸取代率的检测表明，在利巴韦林存在下，转换频率增加，但颠换突变频率没有增加。此外，我们的数据表明，在所用的连续细胞类型和分析的时间点，一些HRSV mRNA的丰度并不反映mRNA转录的顺序。重要性人类呼吸道合胞病毒（HRSV）是一种主要的儿科病原体。利巴韦林可用于患有重病的儿童，以减少病毒数量并降低疾病负担。利巴韦林被用作其他病毒的实验性治疗。利巴韦林对HRSV的作用机制尚不清楚，但认为它会增加病毒复制过程中聚合酶的突变率。为了研究这一假设，我们使用了一种高分辨率的方法，使我们能够确定病毒的基因序列，以达到很大的覆盖深度。我们发现，利巴韦林没有引起病毒mRNA转录物的相对量的可检测的变化。然而，我们发现利巴韦林治疗确实会引起突变数量的增加，这与病毒产量的减少有关。
Human respiratory syncytial virus (HRSV) is a major cause of serious respiratory tract infection. Treatment options include administration of ribavirin, a purine analog, although the mechanism of its anti-HRSV activity is unknown. We used transcriptome sequencing (RNA-seq) to investigate the genome mutation frequency and viral mRNA accumulation in HRSV-infected cells that were left untreated or treated with ribavirin. In the absence of ribavirin, HRSV-specific transcripts accounted for up to one-third of total RNA reads from the infected-cell RNA population. Ribavirin treatment resulted in a >90% reduction in abundance of viral mRNA reads, while at the same time no such reduction was detected for the abundance of cellular transcripts. The presented data reveal that ribavirin significantly increases the frequency of HRSV-specific RNA mutations, suggesting a direct influence on the fidelity of the HRSV polymerase. The presented data show that transitions and transversions occur during HRSV replication and that these changes occur in hot spots along the HRSV genome. Examination of nucleotide substitution rates in the viral genome indicated an increase in the frequency of transition but not transversion mutations in the presence of ribavirin. In addition, our data indicate that in the continuous cell types used and at the time points analyzed, the abundances of some HRSV mRNAs do not reflect the order in which the mRNAs are transcribed. IMPORTANCE Human respiratory syncytial virus (HRSV) is a major pediatric pathogen. Ribavirin can be used in children who are extremely ill to reduce the amount of virus and to lower the burden of disease. Ribavirin is used as an experimental therapy with other viruses. The mechanism of action of ribavirin against HRSV is not well understood, although it is thought to increase the mutation rate of the viral polymerase during replication. To investigate this hypothesis, we used a high-resolution approach that allowed us to determine the genetic sequence of the virus to a great depth of coverage. We found that ribavirin did not cause a detectable change in the relative amounts of viral mRNA transcripts. However, we found that ribavirin treatment did indeed cause an increase in the number of mutations, which was associated with a decrease in virus production.