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

EBV BART miRNA：目标鉴定和效应表征

• 批准号: 7785008
• 负责人: NANCY JOAN RAAB-TRAUB
• 金额: $ 30.58万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7785008
• 关键词: 3' Untranslated Regions; Ablation; Affect; Antibodies; Apoptosis; Bioinformatics; Biological Assay; Cell Line; Cell physiology; Cells; Code; Complex; Development; Epithelial Cells; Epstein-Barr Virus Infections; Factor Analysis; Family; Gene Expression; Genes; Genetic Transcription; Genome; Goals; Growth; Human; Human Herpesvirus 4; Immunoblotting; Individual; Infection; Lymphocyte; Lymphoid Cell; Macaca mulatta; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; MicroRNAs; Nasopharynx Carcinoma; Nuclear Pore Complex; Oncogenic Viruses; Pathway interactions; Porifera; Process; Production; Property; Protein Array; Proteins; Proteomics; Regulation; Reporter; Retroviral Vector; Signal Pathway; Small RNA; Testing; Transcript; Tumor Cell Line; Variant; Viral; Viral Proteins; Virus; Virus Diseases; Work; anticancer research; base; cell growth; cell growth regulation; cellular targeting; established cell line; gel electrophoresis; inhibitor/antagonist; latent infection; protein expression; prototype; public health relevance; research study; transforming virus; tumor; tumorigenesis; two-dimensional

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.

描述（由申请人提供）：人类肿瘤病毒，爱泼斯坦Barr病毒（EBV），编码在肿瘤潜在感染期间表达的多个microRNA（miRNA）。已经确定了两组miRNA。仅在III型潜伏期中转化的淋巴细胞中表达一组。第二组包含两个簇是由一个复杂的转录本，称为BART，来自基因组的BAMHI A区域。重要的是，这些转录本在与EBV相关的肿瘤中高度产生，但在感染的B淋巴样细胞系中以低水平表达。与EBV/KSHV编码序列相比，几种EBV miRNA具有高度保守的EBV miRNA。序列的保存表明miRNA可能靶向细胞信息。基于这些发现，该应用将检验以下假设：EBV BART miRNA靶基因影响生长调节。提出的实验将结合生物信息学，微阵列和蛋白质组学的组合来鉴定巴特miRNA的细胞靶标。稳定表达BART miRNA的细胞系将产生以确认表达并确定miRNA对细胞生长特性的影响。多个细胞信号通路受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miRNA合成的Antagomirs，海绵和抑制剂评估miRNA表达丧失在EBV感染细胞中的影响。细胞miRNA已被证明是癌症发展的主要因素。该建议将决定EBV miRNA如何促进EBV介导的肿瘤发生。 公共卫生相关性：爱泼斯坦 - 巴尔病毒的感染通过病毒蛋白对细胞生长调节的影响有助于发展几种重要的人类恶性肿瘤。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有助于癌症功能的小RNA（miRNA）。 EBV编码多种miRNA，我们已证明在某些EBV相关的癌症（尤其是鼻咽癌）中以高水平表达。 EBV miRNA靶向的细胞蛋白的鉴定将增加我们对EBV如何促进癌症发展的理解。