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

项目摘要 转化的人γ疱疹病毒EBV和KSHV在B细胞中建立稳定的潜伏感染， 提供了一个终身的病毒库，可以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预防疾病的合理战略至关重要。在 人体γ疱疹病毒的体内研究受到了严重的限制， 自然宿主鼠γ疱疹病毒68（MHV 68）与EBV和KSHV相关，并引起淋巴瘤， 小鼠淋巴组织增生性疾病，为机制研究提供了易于操作的小动物模型 病毒与宿主的关系与EBV和KSHV一样，MHV 68表达lncRNA，其在治疗过程中发挥作用。 感染和发病机理在很大程度上是未知的。由于病毒编码的lncRNA在体内大量表达， 在长期潜伏期的B细胞和淋巴组织增生性疾病的增生性B细胞病变中，我们 假设这些lncRNA在潜伏期和淋巴瘤发生中起关键作用。为了支持这一点，我们的新数据 表明MHV 68 TMER 4转录物作为一种独特的lncRNA，对潜伏期和 发病机制使用尖端的高通量测序方法和一种新的 通过生物信息学管道，我们现在已经生成了经验证的MHV 68 lncRNA转录物的综合图谱。 有了这一发现阶段的结果，我们现在将测试MHV 68 lncRNA是必需的这一假设。 潜伏期和肿瘤发生的可能性我们将：1）确定高优先级MHV 68 lncRNA在体内过程中的作用 2）定义lncRNA TMER 4促进潜伏感染的分子机制，并确定 新型M3 M2 lncRNA、反义TMERs/miRNAs与潜伏期之间的三方调控关系 蛋白M2，和3）确定病毒和宿主lncRNA在淋巴瘤发生中的作用。使用新的基因组， 生物信息学和诱变技术，结合MHV 68 lncRNA的系统性体内分析 突变体，提供了一个非常强大的手段，以确定分子机制，lncRNA 有助于体内γ疱疹病毒感染和淋巴瘤发生。此外，独特的合作性质 这项计划的项目应该使我们能够确定共同的lncRNA靶向途径的贡献， 潜伏期和肿瘤发生。

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.