Characterization of tRNA-derived RNA Fragments (tRFs) in Respiratory Syncytial Vi

呼吸合胞体 Vi 中 tRNA 衍生的 RNA 片段 (tRF) 的表征

基本信息

批准号：
8813852
负责人：
Xiaoyong Bao
金额：
$ 38.7万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8813852
关键词：
Address Antiviral Agents Attention Attenuated Attenuated Live Virus Vaccine Biochemistry and Cellular Biology Biogenesis Bioinformatics Biological Biological Markers Biological Process Biology Breathing Bronchiolitis Cells Cellular biology Child Collaborations Communities Complex Computer Analysis Cytoplasm Data Development Ectopic Expression Elements Employee Strikes Epithelial Cells Exhibits Failure Family Foundations Functional RNA Future Genes Genetic Transcription Genome Glutamic Acid-Specific tRNA Glycine decarboxylase Goals Health Human respiratory syncytial virus Immune response Immunoprecipitation In Vitro Infant Infection Information Centers Life Lower Respiratory Tract Infection Lung Maps Mediating Messenger RNA MicroRNAs Molecular Molecular Biology Molecular Profiling Molecular Virology Morbidity - disease rate Nucleotides Pattern Play Pneumonia Production Protein Biochemistry Proteins Public Health RNA RNA Biochemistry RNA-Induced Silencing Complex Regimen Regulation Research Resources Respiratory Syncytial Virus Infections Respiratory syncytial virus Respiratory syncytial virus RSV proteins Ribosomal RNA Role Small Interfering RNA Small Nucleolar RNA System Techniques Technology Testing The Sun Therapeutic Therapeutic Intervention Transfer RNA Vaccine Design Viral Virus Virus Diseases Virus Replication angiogenin apolipoprotein B mRNA editing enzyme apolipoprotein E receptor 2 base cellular targeting data integration design endonuclease experience human DICER1 protein innovation insight member mortality next generation sequencing novel novel therapeutics nuclease polypeptide positional cloning protein expression respiratory response structural biology therapeutic vaccine vaccine candidate virology virus host interaction

项目摘要

DESCRIPTION (provided by applicant): The discovery of small non-coding RNAs (sncRNAs) with regulatory functions is a recent breakthrough in biology. Among sncRNAs, microRNAs (miRNAs) and virus-derived sncRNAs have emerged as elements of critical importance in controlling viral replication and host responses to viral infection. However, the expression patterns and functional aspects of other types of sncRNAs after viral infection are completely unexplored. To define the expression patterns of sncRNAs, as well as to discover novel regulatory sncRNAs in response to viral infection, we applied next-generation sequencing (NGS) to cells infected with human respiratory syncytial virus (RSV), the most common respiratory mucosal virus that causes lung and airway infections in infants and young children, in comparison to uninfected cells. We found that RSV infection leads to abundant production of tRNA-derived RNA fragments (tRFs) that are approximately 30 nucleotides (nts) long and correspond to the 5'- half of mature tRNAs (tRF5). The induction of tRF5 is virus-specific and RSV replication-dependent. At least one member of these tRF5s, which is derived from tRNA-Glu-CTC (tRF5-GluCTC), decreases the level of target mRNAs in the cytoplasm. It also plays a critical role in RSV infection, as RSV replication is attenuated by its suppression, but promoted by its ectopic expression. The biogenesis of this tRF is also specific, and mediated by a particular endonuclease (angiogenin, ANG), and not by other nucleases. The central hypothesis of this project is that tRFs are not a random by-product of tRNA degradation, but rather are functional molecules. This central hypothesis will be addressed by identifying the targets of tRF5-GluCTC in RSV-infected airway epithelial cells (Aim 1), determining the viral factor(s) controlling the trans-silencing activity of tRFs (Aim 2), and defining the viral component(s) contributing to ANG-mediated biogenesis of tRFs (Aim 3). Preliminary data suggest that the RSV proteins N and P will be a focus of Aims 2 and 3, respectively. Of note, despite the public health importance of RSV, no effective therapeutic interventions or vaccines are available. Therefore, the overall goal of this project is to use a combination of molecular virology, protein and RNA biochemistry, cellular and structural biology techniques, to identify the molecular mechanisms underlying the regulatory effects of this novel tRF on RSV replication. The results of this project will provide an important new perspective and novel regulatory mechanisms to study the interactions between the host and RSV, and lay the foundation for the future development of biomarkers, therapeutic interventions, and vaccine designs based on the interaction of tRFs with their targets, both host and virus-derived, and the biogenic mechanisms of tRFs.

描述（由申请人提供）：具有调节功能的小非编码RNA（sncRNA）的发现是生物学领域的最新突破。在 sncRNA 中，微小 RNA (miRNA) 和病毒衍生的 sncRNA 已成为控制病毒复制和宿主对病毒感染的反应的至关重要的元素。然而，病毒感染后其他类型 sncRNA 的表达模式和功能方面尚未完全探索。为了定义 sncRNA 的表达模式，以及发现响应病毒感染的新型调节性 sncRNA，我们对感染人呼吸道合胞病毒 (RSV) 的细胞应用了下一代测序 (NGS)，RSV 是最常见的呼吸道粘膜病毒，与未感染的细胞相比，它会导致婴儿和幼儿的肺部和气道感染。我们发现RSV感染会导致tRNA衍生的RNA片段（tRF）大量产生，其长度约为30个核苷酸（nt），对应于成熟tRNA（tRF5）的5'-一半。 tRF5 的诱导是病毒特异性的且依赖于 RSV 复制。这些 tRF5 中至少有一个成员源自 tRNA-Glu-CTC (tRF5-GluCTC)，可降低细胞质中靶标 mRNA 的水平。它在 RSV 感染中也发挥着关键作用，因为 RSV 复制因其抑制而减弱，但因异位表达而促进。该 tRF 的生物发生也是特异性的，由特定的核酸内切酶（血管生成素，ANG）介导，而不是由其他核酸酶介导。该项目的中心假设是 tRF 不是 tRNA 降解的随机副产品，而是功能分子。这一中心假设将通过确定 RSV 感染的气道上皮细胞中 tRF5-GluCTC 的靶标（目标 1）、确定控制 tRF 反式沉默活性的病毒因子（目标 2）以及定义病毒的病毒因子来解决。有助于 ANG 介导的 tRF 生物合成的成分（目标 3）。初步数据表明，RSV 蛋白 N 和 P 将分别成为目标 2 和 3 的重点。值得注意的是，尽管 RSV 对公共卫生具有重要意义，但尚无有效的治疗干预措施或疫苗。因此，该项目的总体目标是结合分子病毒学、蛋白质和RNA生物化学、细胞和结构生物学技术，确定这种新型tRF对RSV复制调节作用的分子机制。该项目的结果将为研究宿主与RSV之间的相互作用提供重要的新视角和新颖的调控机制，并为未来基于tRF与其相互作用的生物标志物、治疗干预和疫苗设计奠定基础。宿主和病毒来源的靶标，以及 tRF 的生物机制。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（2）

Respiratory Syncytial Virus Utilizes a tRNA Fragment to Suppress Antiviral Responses Through a Novel Targeting Mechanism.

呼吸道合胞病毒利用 tRNA 片段通过新颖的靶向机制来抑制抗病毒反应。

DOI：
10.1038/mt.2015.124
发表时间：
2015-10-01
期刊：
Molecular therapy : the journal of the American Society of Gene Therapy
影响因子：
0
作者：
Junfang Deng;R. Ptashkin;Yu Chen;Zhi Cheng;Guangliang Liu;T. Phan;Xiaoling Deng;Jiehua Zhou;Inhan Lee;Yong Sun Lee;X. Bao
通讯作者：
X. Bao

Recent vaccine development for human metapneumovirus.

最近开发了人类偏肺病毒疫苗。