Characterizing the Molecular Mode of Action of EBV Noncoding RNA EBER1

表征 EBV 非编码 RNA EBER1 的分子作用模式

• 批准号: 10092105
• 负责人: Nara Lee
• 金额: $ 23.48万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092105
• 关键词: Adenine; Antisense Oligonucleotides; Aptamer Technology; B-Lymphocytes; Base Sequence; Binding; Binding Sites; Biochemical; Biological Assay; Capsid Proteins; Carcinoma; Cell physiology; Cells; Chromatin; Clinical; DNA; Dam methyltransferase; Deposition; Development; Enterobacteria phage MS2; Enzymes; Epithelial Cells; Epstein-Barr virus encoded RNA 1; Epstein-Barr virus encoded RNA 2; Exhibits; Gene Expression; Genes; Genetic Transcription; Genomics; Guide RNA; Human; Human Herpesvirus 4; Immune; Immunocompromised Host; Individual; Infection; Investigation; Length; Life Cycle Stages; Light; Location; Lymphocyte; Lymphoma; MS2 coat protein; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; Methyltransferase; Modification; Molecular; Nuclear; Nucleotides; Oncogenic; Organism; Pharmaceutical Preparations; Play; Population; Property; Proteins; RNA; RNA Stability; Reader; Refractory; Role; Site; Structure; Techniques; Therapeutic; Transcript; Transplant Recipients; Untranslated RNA; Viral; Viral Proteins; Virus; Virus Diseases; Virus Replication; aptamer; base; bisulfite sequencing; cancer type; capture hybridization analysis of RNA targets; cell growth regulation; design; detection limit; gammaherpesvirus; in vivo; insight; interest; next generation sequencing; novel; recruit; transcription factor; tumor; viral RNA

PROJECT SUMMARY Epstein-Barr virus (EBV) was the first human cancer-causing virus to be discovered. EBV primarily infects lymphocytes and epithelial cells, which can result in a wide range of lymphomas and carcinomas, respectively. Over 90% of the world’s population is infected with this virus, but infection is mostly asymptomatic in healthy individuals. The cancer-causing potential of EBV becomes evident particularly in immunocompromised individuals, e.g. transplant recipients who are under long-term administration of immune-suppressing medication, where reactivation of the virus can drive tumor formation. Apart from viral proteins, EBV expresses two highly abundant noncoding RNAs called EBER1 and EBER2. Both noncoding RNAs exhibit a strict nuclear localization and are expressed at high levels on par with other greatly abundant host noncoding RNAs that carry out vital cellular functions. It can be assumed based on this high copy number as well as the fact that EBERs are found in all clinical isolates of EBV that EBERs execute or interfere with a vital cellular process during infection. While we have made significant progress in elucidating the molecular mechanism by which EBER2 benefits the EBV life cycle, the molecular function of EBER1 remains enigmatic. Given the vast parallels between both EBER1 and EBER2, we hypothesize that EBER1 may also execute a similar function to EBER2, i.e. to bind to specific sites on chromatin and regulate gene expression. Here we propose to apply cutting-edge techniques combining aptamer technology and next- generation sequencing to further our insight into the molecular mechanism of EBER1. Furthermore, we have recently obtained experimental evidence for RNA modification in EBER1. Thus, we also propose to uncover the factors involved in depositing and interpreting this modification to study the significance of RNA modification in EBER1 for the EBV life cycle. In summary, we expect that the proposed investigations will yield significant insights into how EBER1 contributes to viral replication and thus tumor development. In light of the fact that EBER1 by itself harbors oncogenic properties, our proposed studies may indicate viable therapeutic avenues for targeting the cancer- causing potential of EBV.

项目概要 爱泼斯坦-巴尔病毒（EBV）是第一个被发现的人类致癌病毒。 EBV主要感染 淋巴细胞和上皮细胞，分别可导致多种淋巴瘤和癌。 世界上90%以上的人口感染了这种病毒，但健康人的感染大多无症状 个人。 EB 病毒的致癌潜力变得明显，尤其是在免疫功能低下的人群中 个人，例如长期接受免疫抑制治疗的移植受者 药物治疗，病毒的重新激活可以促进肿瘤的形成。 除了病毒蛋白外，EBV 还表达两种高度丰富的非编码 RNA，称为 EBER1 和 EBER2。 两种非编码 RNA 都表现出严格的核定位，并且与其他非编码 RNA 一样以高水平表达。 非常丰富的宿主非编码RNA，执行重要的细胞功能。基于此可以假设 高拷贝数以及在 EBER 执行的所有 EBV 临床分离株中都发现了 EBER 或干扰感染期间的重要细胞过程。虽然我们在阐明问题方面取得了重大进展 EBER2 有益于 EBV 生命周期的分子机制、EBER1 的分子功能 仍然是个谜。鉴于 EBER1 和 EBER2 之间存在巨大相似之处，我们假设 EBER1 也可能执行与 EBER2 类似的功能，即结合染色质上的特定位点并调节基因 表达。在这里，我们建议应用适体技术与下一步相结合的尖端技术—— 世代测序以进一步了解 EBER1 的分子机制。此外，我们还有 最近获得了 EBER1 中 RNA 修饰的实验证据。因此，我们还建议揭露 参与沉积和解释这种修饰的因素以研究 RNA 的重要性 针对 EBV 生命周期对 EBER1 进行修改。 总之，我们预计拟议的调查将对 EBER1 如何 有助于病毒复制，从而促进肿瘤的发展。鉴于 EBER1 本身包含 致癌特性，我们提出的研究可能表明针对癌症的可行治疗途径- 引起EBV的潜力。