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

描述（由申请人提供）：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eb病毒（EBV），这是一种普遍存在的DNA肿瘤病毒，与伯基特病、霍奇金病和弥漫性大B细胞淋巴瘤等癌症有关。在感染过程中，EBV表达病毒microrna (mirna)，最近的研究表明EBV microrna以及EBV感染上调的细胞致癌mirna在B细胞转化过程中发挥了重要作用。此外，一些EBV mirna与癌症中失调的细胞mirna具有序列同源性，这些病毒mirna可能结合并改变现有的mirna调节网络。参与转化的miRNA靶点尚未确定，因此，系统地识别EBV感染细胞中受miRNA调控的基因对于了解它们在病毒肿瘤发生中的作用及其在EBV生命周期中的作用至关重要。本文概述的实验结合了miRNA生物学、病毒学和生物信息学等多学科领域的最新技术，全面研究了miRNA靶组，并检查了ebv感染的B细胞的转录景观，以提取受病毒和细胞miRNA影响的关键基因和途径。这些研究将使用EBV驱动的体外B细胞转化模型以及患者来源的EBV+ B细胞肿瘤进行。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，并为此组建了一个科学咨询委员会，以指导我建立和/或进一步发展几个基本方法。K99/R00奖将为我提供资源和时间，在指导阶段进行新的培训，以学习基本的新生感染技术，并生成额外的数据和计算工具，以便在独立阶段在我自己的实验室中使用。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miRNA，特别是EBV miRNA，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mirna，还会进一步改变与许多与病毒感染无关的癌症相关的致癌细胞mirna的表达，因此这些研究将可能提供重要的信息，可以扩展到其他癌症模型，并为癌症中mirna介导的基因调控的整体机制提供见解。

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.