Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat Gan, Israel.
Departments of Ophthalmology and of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
J Virol. 2020 Jun 16;94(13). doi: 10.1128/JVI.00123-20.
Small noncoding RNAs (sncRNA), including microRNA (miR), are expressed by many viruses to provide an additional layer of gene expression regulation. Our work has shown that varicella-zoster virus (VZV; also called human herpesvirus 3 [HHV3]), the human alphaherpesvirus causing varicella and herpes zoster, expresses 24 virally encoded sncRNA (VZVsncRNA) in infected cells. Here, we demonstrate that several VZVsncRNA can modulate VZV growth, including four VZVsncRNA (VZVsncRNA10, -11, -12, and -13) that are antisense to VLT, a transcript made in lytic infections and associated with VZV latency. The influence on productive VZV growth and spread was assessed in epithelial cells transfected with locked nucleotide analog antagonists (LNAA). LNAA to the four VZVsncRNA antisense to VLT significantly reduced viral spread and progeny titers of infectious virus, suggesting that these sncRNA promoted lytic infection. The LNAA to VZVsncRNA12, encoded in the leader to ORF61, also significantly increased the levels of VLT transcripts. Conversely, overexpression of VZVsncRNA13 using adeno-associated virus consistently increased VZV spread and progeny titers. These results suggest that sncRNA antisense to VZV may regulate VZV growth, possibly by affecting VLT expression. Transfection of LNAA to VZVsncRNA14 and VZVsncRNA9 decreased and increased VZV growth, respectively, while LNAA to three other VZVsncRNA had no significant effects on replication. These data strongly support the conclusion that VZV replication is modulated by multiple virally encoded sncRNA, revealing an additional layer of complexity of VZV regulation of lytic infections. This may inform the development of novel anti-sncRNA-based therapies for treatment of VZV diseases. Varicella-zoster virus (VZV) causes herpes zoster, a major health issue in the aging and immunocompromised populations. Small noncoding RNAs (sncRNA) are recognized as important actors in modulating gene expression. This study extends our previous work and shows that four VZVsncRNA clustering in and near ORF61 and antisense to the latency-associated transcript of VZV can positively influence productive VZV infection. The ability of multiple exogenous small oligonucleotides targeting VZVsncRNA to inhibit VZV replication strengthens the possibility that they may inform development of novel treatments for painful herpes zoster.
小非编码 RNA（sncRNA），包括 microRNA（miR），是许多病毒表达的，为基因表达调控提供了一个额外的层次。我们的工作表明，水痘带状疱疹病毒（VZV；也称为人类疱疹病毒 3 [HHV3]），一种引起水痘和带状疱疹的人类α疱疹病毒，在感染细胞中表达 24 种病毒编码的 sncRNA（VZVsncRNA）。在这里，我们证明了几种 VZVsncRNA 可以调节 VZV 的生长，包括四个 VZVsncRNA（VZVsncRNA10、-11、-12 和-13），它们是反义于 VLT 的，VLT 是在裂解感染中产生的与 VZV 潜伏相关的转录本。通过转染锁核苷酸类似物拮抗剂（LNAA）评估对产生活性 VZV 生长和传播的影响。针对 VLT 反义的四个 VZVsncRNA 的 LNAA 显著降低了病毒的传播和感染性病毒的后代滴度，表明这些 sncRNA 促进了裂解感染。编码在 ORF61 启动子至 ORF61 的 VZVsncRNA12 的 LNAA 也显著增加了 VLT 转录本的水平。相反，使用腺相关病毒过表达 VZVsncRNA13 一致增加了 VZV 的传播和后代滴度。这些结果表明，反义于 VZV 的 sncRNA 可能通过影响 VLT 表达来调节 VZV 的生长。用 LNAA 转染 VZVsncRNA14 和 VZVsncRNA9 分别降低和增加了 VZV 的生长，而三种其他 VZVsncRNA 的 LNAA 对复制没有显著影响。这些数据强烈支持 VZV 复制受多种病毒编码的 sncRNA 调节的结论，揭示了 VZV 调节裂解感染的复杂性的另一个层次。这可能为基于抗 sncRNA 的治疗 VZV 疾病的新疗法的开发提供信息。水痘带状疱疹病毒（VZV）引起带状疱疹，这是老年和免疫功能低下人群的一个主要健康问题。小非编码 RNA（sncRNA）被认为是调节基因表达的重要因素。本研究扩展了我们之前的工作，表明四个 VZVsncRNA 聚类在 ORF61 内和附近，并反义于 VZV 的潜伏相关转录本，可以积极影响产生活性 VZV 感染。四种针对 VZVsncRNA 的外源性小分子寡核苷酸的抑制 VZV 复制的能力，加强了它们可能为开发治疗疼痛性带状疱疹的新疗法提供信息的可能性。
