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

描述（由申请人提供）：人类肿瘤病毒，eb病毒（EBV），编码多种microrna (mirna)，这些microrna在肿瘤潜伏感染期间表达。已经确定了两组mirna。一组仅在III型潜伏期转化淋巴细胞中表达。包含两个簇的第二组是由一个复杂的转录本家族产生的，称为BARTs，来自基因组的BamHI a区域。重要的是，这些转录本在ebv相关肿瘤中大量产生，但在感染的b淋巴样细胞系中表达水平较低。一些EBV miRNAS与恒河猴EBV中鉴定的miRNAS高度保守，与EBV/KSHV编码序列具有相当大的同源性。这种序列的保守性表明mirna可能靶向细胞信息。基于这些发现，本应用将验证EBV BART mirna靶向细胞基因影响生长调节的假设。拟议的实验将使用生物信息学、微阵列和蛋白质组学的组合来识别BART mirna的细胞靶标。将产生稳定表达BART mirna的细胞系，以确认表达并确定mirna对细胞生长特性的影响。多种细胞信号通路受到EBV转化蛋白的影响，因此BART mirna对这些通路的潜在调节也将被研究。在EBV感染的细胞中，miRNA表达缺失的影响将使用安塔戈米、海绵和miRNA合成抑制剂进行评估。细胞mirna已被证明是癌症发展的主要因素。该提案将确定EBV mirna如何促进EBV介导的肿瘤发生。

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.