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: 8196870
• 负责人: NANCY JOAN RAAB-TRAUB
• 金额: $ 29.66万
• 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/8196870
• 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; Human Herpesvirus 8; 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; Research; Retroviral Vector; Signal Pathway; Small RNA; Testing; Transcript; Tumor Cell Line; Variant; Viral; Viral Proteins; Virus; Virus Diseases; Work; 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，EB病毒（EBV），编码多种微RNA（miRNA），这些微RNA（miRNA）在肿瘤潜伏感染期间表达。已鉴定出两组 miRNA。一组仅在 III 型潜伏期的转化淋巴细胞中表达。第二组包含两个簇，由来自基因组 BamHI A 区域的复杂转录本家族（称为 BART）产生。重要的是，这些转录本在 EBV 相关肿瘤中大量产生，但在受感染的 B 淋巴细胞系中表达水平较低。一些 EBV miRNA 与恒河猴 EBV 中鉴定的 miRNA 高度保守，与 EBV/KSHV 编码序列具有显着更高的同源性。这种序列的保守性表明 miRNA 可能以细胞信息为目标。基于这些发现，该应用将测试 EBV BART miRNA 靶向细胞基因以影响生长调节的假设。拟议的实验将结合生物信息学、微阵列和蛋白质组学来识别 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 antagomir、海绵和 miRNA 合成抑制剂来评估 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 miRNA 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，导致多种重要的人类恶性肿瘤的发生。人们发现了一种新的生长调节形式，它涉及影响细胞基因功能并导致癌症的小 RNA (miRNA)。 EBV 编码多种 miRNA，我们已证明这些 miRNA 在一些 EBV 相关癌症（尤其是鼻咽癌）中高水平表达。 EBV miRNA 靶向的细胞蛋白的鉴定将加深我们对 EBV 如何促进癌症发展的理解。