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

描述（由申请人提供）：人肿瘤病毒，爱泼斯坦巴尔病毒（EBV），编码在肿瘤潜伏感染期间表达的多种微小RNA（miRNA）。已经鉴定了两组miRNAs。一组仅在III型潜伏期的转化淋巴细胞中表达。包含两个簇的第二组由来自基因组的BamHI A区域的称为BART的转录物的复杂家族产生。重要的是，这些转录物在EBV相关肿瘤中高度产生，但在感染的B淋巴细胞系中以低水平表达。几种EBV miRNAS与在恒河猴EBV中鉴定的那些高度保守，具有比EBV/KSHV编码序列大得多的同源性。这种序列的保守性表明miRNAs可能靶向细胞信息。基于这些发现，本申请将测试EBV BART miRNA靶向细胞基因以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 miRNAs的细胞靶点。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。多种细胞信号传导途径受到EBV转化蛋白的影响，因此还将检查BART miRNA对这些途径的潜在调节。将使用艾司洛尔、海绵和miRNA合成抑制剂评估EBV感染细胞中miRNA表达丧失的影响。细胞miRNA已被证明是癌症发展的主要因素。该提案将确定EBV miRNAs如何促进EBV介导的肿瘤发生。 公共卫生相关性：EB病毒感染通过病毒蛋白对细胞生长调节的影响而导致几种重要的人类恶性肿瘤的发生。已经发现了一种新的生长调节形式，涉及影响细胞基因功能并导致癌症的小RNA（miRNAs）。EBV编码多种miRNA，我们已经证明这些miRNA在一些EBV相关的癌症特别是鼻咽癌中高水平表达。鉴定EBV miRNAs靶向的细胞蛋白质将增加我们对EBV如何促进癌症发展的理解。

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. PUBLIC HEALTH RELEVANCE: Infection with Epstein-Barr virus contributes to the development of several important human malignancies through the effects of viral proteins on cell growth regulation. A new form of growth regulation has been discovered that involves small RNAs (miRNAs) that affect the function of cellular genes and contribute to cancer. EBV encodes multiple miRNAs that we have shown to be expressed at high levels in some EBV-associated cancers particularly nasopharyngeal carcinoma. The identification of cellular proteins that are targeted by the EBV miRNAs will increase our understanding of how EBV contributes to the development of cancer.