Studying the role of KSHV-encoded microRNAs

研究 KSHV 编码的 microRNA 的作用

基本信息

批准号：
7418877
负责人：
ROLF F RENNE
金额：
$ 41.3万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-06 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7418877
关键词：
Address Angiogenesis Inhibitors Apoptosis Binding Bioinformatics Biological Biological Assay Biological Process Biology Cell Proliferation Cells Cultured Cells DNA Viruses Data Detection Endothelial Cells Environment Epithelial Cells Escherichia coli Functional RNA Gene Expression Gene Expression Profiling Gene Targeting Genes Genomics Hematopoiesis Herpesviridae Human Human Herpesvirus 4 Human Herpesvirus 8 Infection Journals Kinetics Knock-out Laboratories Length Life Cycle Stages Locales Lymphoid Lymphoma Lytic Mammals Maps MicroRNAs Northern Blotting Numbers Organism Pathogenesis Pathway interactions Plants Play Post-Transcriptional Regulation RNA Range Recombinants Reporting Reverse Transcriptase Polymerase Chain Reaction Role Sampling Sequence Analysis Simplexvirus Small RNA Technology Testing Thrombospondin 1 Tissue Sample Transcript Translations Untranslated Regions Vero Cells Viral Viral Genome Virus Work base effusion interest lytic replication mRNA Transcript Degradation novel pathogen prevent programs promoter recombinant virus transcription factor tumor tumorigenesis virology

项目摘要

DESCRIPTION (provided by applicant): MicroRNAs are small non-coding regulatory RNA molecules that bind to 3'UTRs of mRNAs to either prevent their translation or induce their degradation. Previously identified in a variety of organisms ranging from plants to mammals, miRNAs are also now known to be produced by DNA viruses. The human ?-herpesvirus Epstein-Barr Virus has been shown to encode miRNAs which potentially regulate both viral and cellular genes. To determine whether Kaposi's Sarcoma-associated herpesvirus (KSHV) encodes miRNAs, we cloned small RNAs from KSHV positive primary effusion lymphoma derived cells. Sequence analysis revealed 11 isolated RNAs of 19 to 23 bases in length that perfectly align to KSHV. Surprisingly, all candidate miRNAs mapped to a single genomic locale within the latency-associated region of KSHV (Samols et al., Journal of Virology 2005). While our work was in review, two other laboratories reported the identification of KSHV-encoded miRNAs. Hence, these data suggest that virally-encoded miRNAs represent a novel mechanism by which KSHV may regulate viral and/or cellular gene expression during both latent and lytic KSHV replication. In human cells, over 450 miRNAs have been identified. Targets and functions of only a few miRNAs have been experimentally determined thus far, yet some miRNAs such as human hsa-miR-14 and hsa-miR-181 regulate fundamental biological processes like apoptosis, cell proliferation, and hematopoiesis and very recently have been implicated in tumorigenesis. Based on our preliminary results we hypothesize that miRNAs play an important role in the KSHV life cycle and may contribute to KSHV pathogenesis. To directly address this hypothesis we propose the following specific aims: SA1: Analysis of KSHV miRNA expression during latent and lytic replication in cultured cells and tissues samples of different origins. SA2: Identification of cellular and viral targets regulated by KSHV-encoded miRNAs. SA3: Evaluate the role of miRNA expression in the context of the viral genome. In support of our hypothesis we provide new preliminary data on identifying cellular miRNA targets in stably miRNA-expressing 293 cells (Samols et al., PLoS Pathogens, 2007, May 11). In summary, the identification of virally encoded miRNAs within the latency-associated region of KSHV is novel and very exciting as it suggests an entirely new level of regulating gene expression by which KSHV can potentially reprogram the host cell environment.

描述（由申请人提供）：MicroRNA 是小的非编码调节 RNA 分子，它们与 mRNA 的 3'UTR 结合，以阻止其翻译或诱导其降解。以前在从植物到哺乳动物的多种生物体中都发现了 miRNA，现在也知道它是由 DNA 病毒产生的。人类β-疱疹病毒 Epstein-Barr 病毒已被证明编码可能调节病毒和细胞基因的 miRNA。为了确定卡波西肉瘤相关疱疹病毒 (KSHV) 是否编码 miRNA，我们从 KSHV 阳性原发性渗出性淋巴瘤衍生细胞中克隆了小 RNA。序列分析显示 11 个分离的 RNA，长度为 19 至 23 个碱基，与 KSHV 完美对齐。令人惊讶的是，所有候选 miRNA 都映射到 KSHV 潜伏相关区域内的单个基因组区域（Samols 等人，Journal of Virology 2005）。当我们的工作正在接受审查时，另外两个实验室报告了 KSHV 编码的 miRNA 的鉴定。因此，这些数据表明，病毒编码的 miRNA 代表了一种新机制，KSHV 可以通过该机制在潜伏和裂解性 KSHV 复制过程中调节病毒和/或细胞基因表达。在人类细胞中，已鉴定出超过 450 种 miRNA。迄今为止，只有少数 miRNA 的靶标和功能已通过实验确定，但一些 miRNA（例如人类 hsa-miR-14 和 hsa-miR-181）调节细胞凋亡、细胞增殖和造血等基本生物过程，并且最近已被涉及在肿瘤发生中。根据我们的初步结果，我们假设 miRNA 在 KSHV 生命周期中发挥重要作用，并可能有助于 KSHV 发病机制。为了直接解决这一假设，我们提出以下具体目标： SA1：不同来源的培养细胞和组织样本中潜伏和裂解复制过程中 KSHV miRNA 表达的分析。 SA2：鉴定由 KSHV 编码的 miRNA 调节的细胞和病毒靶标。 SA3：评估 miRNA 表达在病毒基因组背景下的作用。为了支持我们的假设，我们提供了关于在稳定表达 miRNA 的 293 细胞中识别细胞 miRNA 靶点的新初步数据（Samols 等人，PLoS Pathogens，2007 年，5 月 11 日）。总之，在 KSHV 潜伏相关区域内病毒编码的 miRNA 的鉴定是新颖且非常令人兴奋的，因为它表明 KSHV 可以通过全新的水平调节基因表达来重新编程宿主细胞环境。