"Project 3" MHV68 IncRNA/miRNA interaction in latency and lympomagenesis

项目总结 转化中的人类伽马疱疹病毒EBV和KSHV在B细胞中建立稳定的潜伏感染， 提供了一个终生的病毒储存库，可能有助于恶性疾病的发展。因此，定义 控制长期潜伏期的机制对于设计合理的疾病预防策略至关重要。在……里面 伽马疱疹病毒在人体内的活体研究一直受到工作困难的严重限制 自然宿主。小鼠伽马疱疹病毒68(MHV68)与EBV和KSHV有关，可引起淋巴瘤和 小鼠淋巴增生性疾病，为机制研究提供了一种易于操作的小动物模型 体内的病毒/宿主关系。与EBV和KSHV一样，MHV68表达的lncRNAs在 感染和发病机制在很大程度上尚不清楚。由于病毒编码的lncRNA在体内大量表达 在长期潜伏期的B细胞和淋巴增生性疾病期间的增生性B细胞病变中，我们 假设这些lncRNAs在潜伏期和淋巴肿大中起关键作用。为了支持这一点，我们的新数据 证明了MHV68 TMER4转录本发挥了独特的lncRNA的作用，这对于潜伏期和 发病机制。使用尖端的高通量测序方法和一种新的 在生物信息学管道中，我们现在已经生成了一个有效的MHV68 lncRNA转录本的全面图谱。 有了这一发现阶段的结果，我们现在将检验MHV68 lncRNAs是必不可少的假设 潜伏期和肿瘤形成。我们将：1)确定高优先级MHV68 lncRNAs在体内的作用 2)确定lncRNA TMER4促进潜伏感染的分子机制，并确定 新型M3M2 lncRNA、反义TMERs/miRNAs与潜伏期的三方调控关系 蛋白M2，以及3)确定病毒和宿主lncRNAs在淋巴肿瘤发生中的作用。新基因组的使用， 生物信息学和诱变技术结合MHV68 lncRNA的体内系统分析 突变体，提供了一种极其强大的手段来确定lncRNAs 有助于体内的伽马疱疹病毒感染和淋巴肿大。此外，独特的协作性 应该允许我们定义共同的以lncRNA为靶向的途径对 潜伏期与肿瘤发生。

Project summary The transforming human gammaherpesviruses EBV and KSHV establish stable latent infections in B cells, providing a lifelong reservoir of virus that can contribute to the development of malignant disease. Thus, defining the mechanisms that govern long-term latency is critical for designing rational strategies to prevent disease. In vivo studies of gammaherpesviruses in humans have been severely limited by the difficulties of working in the natural host. Murine gammaherpesvirus 68 (MHV68) is related to EBV and KSHV and causes lymphomas and lymphoproliferative disease in mice, providing a readily manipulable small animal model for mechanistic studies of the virus/host relationship in vivo. Like EBV and KSHV, MHV68 expresses lncRNAs whose functions during infection and pathogenesis are largely unknown. As virus-encoded lncRNAs are abundantly expressed in vivo in B cells during long-term latency and in hyperplastic B cell lesions during lymphoproliferative disease, we hypothesize that these lncRNAs play key roles in latency and lymphomagenesis. In support of this, our new data demonstrates that the MHV68 TMER4 transcript acts a unique lncRNA that is essential for latency and pathogenesis. Using a combination of cutting-edge high throughput sequencing approaches and a novel bioinformatic pipeline, we have now generated a comprehensive map of validated MHV68 lncRNA transcripts. With results from this discovery phase in hand, we will now test the hypothesis that MHV68 lncRNAs are essential for latency and tumorigenesis. We will: 1) Determine the roles of high priority MHV68 lncRNAs during in vivo infection; 2) Define the molecular mechanism by which lncRNA TMER4 facilitates latent infection, and determine the tripartite regulatory relationship between the novel M3M2 lncRNA, antisense TMERs/miRNAs and latency protein M2, and 3) Determine the role of virus and host lncRNAs in lymphomagenesis. The use of new genomic, bioinformatic and mutagenesis technology, in conjunction with systematic in vivo analyses of MHV68 lncRNA mutants, provides an extremely powerful means to determine the molecular mechanism by which lncRNAs contribute to gammaherpesvirus infection and lymphomagenesis in vivo. Further, the unique collaborative nature of this program project should allow us to define the contribution of common lncRNA-targeted pathways to latency and tumorigenesis.