The Role of Viral and Cellular miRNAs in B-cell Lymphomagenesis

描述(申请人提供)：至少六分之一的人类癌症与病毒感染有关。其中一部分可归因于Epstein-Barr病毒(EBV)，一种普遍存在的DNA肿瘤病毒，与Burkitt‘s、Hodgkin’s和弥漫性大B细胞淋巴瘤等癌症有关。在感染过程中，EBV表达病毒microRNAs(MiRNAs)，最近的研究表明EBV miRNAs以及EBV感染上调的细胞致癌miRNAs在B细胞转化过程中起着重要作用。此外，几个EBV miRNAs与癌症中调节失调的细胞miRNAs具有序列同源性，这些病毒miRNAs可能结合并改变现有的miRNA调节网络。参与转化的miRNA靶标尚未定义，因此，系统地识别EBV感染细胞中受miRNAs调控的基因对于了解它们在病毒肿瘤发生中的作用以及它们在EBV生命周期中的作用是至关重要的。这里概述的实验结合了来自miRNA生物学、病毒学和生物信息学多学科领域的最新技术，以全面询问miRNA目标组并检查EBV感染的B细胞的转录图谱，以提取受病毒和细胞miRNAs影响的关键基因和途径。除了患者来源的EBV+B细胞肿瘤外，这些研究还将使用EBV驱动的体外B细胞转化模型进行。为了成功地开展我的研究，我需要在EBV生物学和生物信息学方面进行新的培训，并因此成立了一个科学咨询委员会来指导我建立和/或进一步发展几种基本的方法。K99/R00奖将为我在指导阶段提供新的培训资源和时间，以便学习基本的从头感染技术，并生成额外的数据和计算工具，供我自己的实验室在独立阶段使用。通过整合这些实验产生的miRNA目标组数据、转录组数据和表型数据，我希望阐明miRNAs，特别是EBV miRNAs，促进持续病毒感染和淋巴瘤发展的机制。最后，由于EBV不仅表达病毒miRNAs，而且进一步改变致癌细胞miRNAs的表达，这些miRNAs与许多与病毒感染无关的癌症相关，这些研究将潜在地提供重要的信息，可以扩展到其他癌症模型，并提供对miRNA介导的癌症基因调控的整体机制的洞察。

DESCRIPTION (provided by applicant): At least one in six human cancers is linked to viral infection. A portion of these can be attributed to Epstein-Barr virus (EBV), a ubiquitous DNA tumor virus associated with cancers such as Burkitt's, Hodgkin's, and diffuse large B cell lymphomas. During infection, EBV expresses viral microRNAs (miRNAs), and recent studies have demonstrated an important role for the EBV miRNAs as well as the cellular oncogenic miRNAs upregulated by EBV infection in the B cell transformation process. Furthermore, several EBV miRNAs share sequence homology with cellular miRNAs that are dysregulated in cancers and potentially, these viral miRNAs can tie into and alter existing miRNA-regulated networks. The miRNA targets involved in transformation are not yet defined, and thus, systemically identifying the genes regulated by miRNAs in EBV-infected cells is essential to understanding their contributions to viral oncogenesis and their roles during the EBV life cycle. Experiments outlined here combine state-of-the-art techniques from the multidisciplinary fields of miRNA biology, virology, and bioinformatics to comprehensively interrogate the miRNA targetome and examine the transcriptional landscape of EBV-infected B cells in order to extract critical genes and pathways influenced by viral and cellular miRNAs. These studies will be carried out using an EBV-driven in vitro B cell transformation model in addition to patient-derived EBV+ B cell tumors. To successfully carry out my studies, I require new training in both EBV biology and bioinformatics, and have accordingly assembled a scientific advisory committee to guide me in establishing and/or further developing several of the essential methodologies. The K99/R00 award will provide me both resources and time for new training during the mentored phase in order to learn fundamental de novo infection techniques and generate additional data and computational tools to be used in my own laboratory during the independent phase. By integrating the miRNA targetome data, transcriptome data, and phenotypic data generated through these experiments, I hope to elucidate the mechanisms by which miRNAs, particularly EBV miRNAs, contribute to persistent viral infection and the development of lymphoma. Finally, since EBV expresses not only viral miRNAs but further alters expression of oncogenic cellular miRNAs that have been linked to many cancers not associated with viral infection, these studies will potentially provide important information that can be extended to other cancer models and provide insight into the overall mechanisms governing miRNA-mediated gene regulation in cancers.