EBV BART miRNAs: Identification of Targets and Characterization of the Effects o

描述(申请人提供)：人类肿瘤病毒，爱泼斯坦-巴尔病毒(EBV)，编码多种微小RNAs(MiRNAs)，在肿瘤潜伏感染期间表达。已经确定了两组miRNAs。其中一组仅在III型潜伏期转化的淋巴细胞中表达。第二组包含两个簇，是从基因组的BamHI A区域产生的一个复杂的转录本家族，称为Barts。重要的是，这些转录本在EBV相关肿瘤中高表达，但在受感染的B淋巴样细胞系中低水平表达。其中一些EBV miRNAs与恒河猴EBV中发现的miRNAs高度保守，与EBV/KSHV编码序列有相当大的同源性。这种序列的保守性表明miRNAs可能针对细胞信息。基于这些发现，这一应用将检验EBV BART miRNAs针对细胞基因影响生长调节的假设。拟议的实验将结合生物信息学、微阵列和蛋白质组学来确定BART miRNAs的细胞靶点。将建立稳定表达BART miRNAs的细胞系，以确认其表达并确定miRNAs对细胞生长特性的影响。EBV转化蛋白影响多个细胞信号通路，因此BART miRNAs对这些通路的潜在调节也将被研究。EBV感染细胞中miRNA表达缺失的影响将使用反配子、海绵和miRNA合成抑制剂进行评估。细胞miRNAs已被证明是癌症发展的主要因素。这一提议将确定EBV miRNAs在EBV介导的肿瘤发生中的作用。

DESCRIPTION (provided by applicant): The human tumor virus, Epstein Barr Virus (EBV), encodes multiple microRNAs (miRNAs) that are expressed during latent infection in tumors. Two groups of miRNAs have been identified. One group is only expressed in transformed lymphocytes in Type III latency. The second group containing two clusters is produced from a complex family of transcripts, termed BARTs, from the BamHI A region of the genome. Importantly, these transcripts are highly produced in EBV-associated tumors but are expressed at low levels in infected B-lymphoid cell lines. Several of the EBV miRNAS are highly conserved with those identified in rhesus EBV with considerably greater homology than EBV/KSHV coding sequences. This conservation of sequences suggests that the miRNAs might target cellular messages. Based on these findings, this application will test the hypothesis that the EBV BART miRNAs target cellular genes to affect growth regulation. The proposed experiments will use a combination of bioinformatic, microarray, and proteomics to identify the cellular targets of the BART miRNAs. Cell lines that stably express the BART miRNAs will be produced to confirm expression and to determine the effects of the miRNAs on cellular growth properties. Multiple cellular signaling pathways are affected by EBV transforming proteins so potential modulation of these pathways by the BART miRNAs will also be examined. The effects of loss of miRNA expression in EBV infected cells will be assessed using antagomirs, sponges, and inhibitors of miRNA synthesis. Cellular miRNAs have been shown to be major factors in the development of cancer. This proposal will determine how the EBV miRNAs contribute to EBV-mediated oncogenesis.