The viral and cellular microRNA targetome in lymphoblastoid cell lines.
The viral and cellular microRNA targetome in lymphoblastoid cell lines.
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DOI：
10.1371/journal.ppat.1002484
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发表时间：
2012-01
期刊：
影响因子：
6.7
通讯作者：
Cullen BR
中科院分区：
文献类型：
--
作者：
Skalsky RL;Corcoran DL;Gottwein E;Frank CL;Kang D;Hafner M;Nusbaum JD;Feederle R;Delecluse HJ;Luftig MA;Tuschl T;Ohler U;Cullen BR
Epstein-Barr 病毒 (EBV) 是一种普遍存在的人类疱疹病毒，与多种 B 细胞癌和淋巴增殖性疾病有关。在潜伏感染期间，EBV 表达 25 种病毒前 microRNA (miRNA)，并诱导特定宿主 miRNA（例如 miR-155 和 miR-21）的表达，这些 miRNA 可能在病毒肿瘤发生中发挥作用。迄今为止，仅确定了有限数量的 EBV miRNA 靶标；因此，EBV miRNA 在病毒发病机制和/或淋巴瘤发生中的作用尚未明确。在这里，我们使用光活化核糖核苷增强交联和免疫沉淀（PAR-CLIP）结合深度测序和计算分析来全面检查 EBV B95-8 株感染的淋巴母细胞系（LCL）中的病毒和细胞 miRNA 靶标组。我们在 3,492 个细胞 3'UTR 中鉴定了 7,827 个 miRNA 相互作用位点。其中 531 个位点包含与病毒 miRNA 匹配的种子。报告基因检测证实了 24 个 PAR-CLIP 鉴定的 miRNA:3'UTR 相互作用。我们的结果表明，EBV miRNA 在潜伏感染期间主要靶向细胞转录物，从而操纵宿主环境。此外，EBV miRNA 的靶标参与与病毒感染直接相关的多个细胞过程，包括先天免疫、细胞存活和细胞增殖。最后，我们提供证据表明来自 miR-17/92 簇的 myc 调节的宿主 miRNA 可以调节潜在病毒基因的表达。这项对 EBV 感染的 B 细胞中 miRNA 靶标组的全面调查代表了定义 EBV 编码的 miRNA 功能的关键一步，并有可能确定 EBV 相关恶性肿瘤的新治疗靶点。全球超过 90% 的成年人感染了 Epstein-Barr 病毒 (EBV)。虽然 EBV 感染通常由健康的免疫系统控制，但在免疫功能低下的个体中，EBV 可能导致严重的疾病和/或癌症。在感染过程中，EBV 表达病毒 microRNA (miRNA) 并诱导特定细胞 miRNA 的表达。一般来说，miRNA 通过与靶信使 RNA (mRNA) 上的互补区域结合来抑制靶基因表达。虽然细胞 miRNA 调节重要的生物过程，例如细胞生长和分化，并且许多 miRNA 与癌症进展有关，但 EBV miRNA 的功能很大程度上未知。为了识别 EBV 感染细胞中 EBV miRNA 和细胞 miRNA 的靶标，我们使用基于下一代测序技术的高通量方法来全面了解 miRNA 调控的基因表达。我们的分析表明，病毒 miRNA 可以调控 500 多种 mRNA，其中许多与 EBV 感染直接相关。这项研究对 B 细胞中的病毒和细胞 miRNA 靶点进行了全面调查，这是朝着确定 EBV 相关癌症的新治疗靶点迈出的积极一步。
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus linked to a number of B cell cancers and lymphoproliferative disorders. During latent infection, EBV expresses 25 viral pre-microRNAs (miRNAs) and induces the expression of specific host miRNAs, such as miR-155 and miR-21, which potentially play a role in viral oncogenesis. To date, only a limited number of EBV miRNA targets have been identified; thus, the role of EBV miRNAs in viral pathogenesis and/or lymphomagenesis is not well defined. Here, we used photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) combined with deep sequencing and computational analysis to comprehensively examine the viral and cellular miRNA targetome in EBV strain B95-8-infected lymphoblastoid cell lines (LCLs). We identified 7,827 miRNA-interaction sites in 3,492 cellular 3′UTRs. 531 of these sites contained seed matches to viral miRNAs. 24 PAR-CLIP-identified miRNA:3′UTR interactions were confirmed by reporter assays. Our results reveal that EBV miRNAs predominantly target cellular transcripts during latent infection, thereby manipulating the host environment. Furthermore, targets of EBV miRNAs are involved in multiple cellular processes that are directly relevant to viral infection, including innate immunity, cell survival, and cell proliferation. Finally, we present evidence that myc-regulated host miRNAs from the miR-17/92 cluster can regulate latent viral gene expression. This comprehensive survey of the miRNA targetome in EBV-infected B cells represents a key step towards defining the functions of EBV-encoded miRNAs, and potentially, identifying novel therapeutic targets for EBV-associated malignancies. Over 90% of adults worldwide are infected with Epstein-Barr virus (EBV). While EBV infection is normally controlled by a healthy immune system, in immuno-compromised individuals, EBV can cause serious disease and/or cancer. During infection, EBV expresses viral microRNAs (miRNAs) and induces the expression of specific cellular miRNAs. In general, miRNAs inhibit target gene expression by binding to complementary regions on target messenger RNAs (mRNA). While cellular miRNAs regulate important biological processes such as cell growth and differentiation, and many miRNAs have been linked to cancer progression, the functions of EBV miRNAs are largely unknown. To identify targets of EBV miRNAs and cellular miRNAs in EBV-infected cells, we used a high-throughput method based on next-generation sequencing technology to give a global picture of miRNA-regulated gene expression. Our analysis showed that over 500 mRNAs can be regulated by viral miRNAs, many of which are directly relevant to EBV infection. This study provides a comprehensive survey of viral and cellular miRNA targets in B cells, which is a positive step towards identifying novel therapeutic targets for EBV-associated cancers.
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影响因子：
5.4
作者：
Cameron, Jennifer E.;Yin, Qinyan;Flemington, Erik K.
通讯作者：
Flemington, Erik K.
影响因子：
5.4
作者：
Amoroso, Richard;Fitzsimmons, Leah;Bell, Andrew I.
通讯作者：
Bell, Andrew I.
爱泼斯坦 - 巴尔病毒编码的microRNA mir-bart2下调病毒DNA聚合酶BALF5。
影响因子：
14.9
作者：
Barth, Stephanie;Pfuhl, Thorsten;Mamiani, Alfredo;Ehses, Claudia;Roemer, Klaus;Kremmer, Elisabeth;Jaeker, Christoph;Hoeck, Julia;Meister, Gunter;Graesser, Friedrich A.
通讯作者：
Graesser, Friedrich A.
DOI：
10.1073/pnas.0602266103
发表时间：
2006-05-02
影响因子：
11.1
作者：
Costinean, S;Zanesi, N;Croce, CM
通讯作者：
Croce, CM
影响因子：
8
作者：
Anastasiadou, E.;Boccellato, F.;Trivedi, P.
通讯作者：
Trivedi, P.