Recruitment of host noncoding RNAs by HIV-1

HIV-1 招募宿主非编码 RNA

基本信息

批准号：
8846742
负责人：
Sandra L. Wolin
金额：
$ 22.05万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8846742
关键词：
7SL RNA Acquired Immunodeficiency Syndrome Address Adverse effects Affect Antiviral Agents Binding Biogenesis Biological Biology Cell Nucleus Cells Cellular Structures Complement Cytoplasm Disease Drug Targeting Drug resistance Gagging Goals HIV HIV-1 Hepatotoxicity High-Throughput Nucleotide Sequencing Human Infection Knowledge Lactic Acidosis Learning Life Lipodystrophy Molecular Morbidity - disease rate Mutate Northern Blotting Nucleocapsid Nucleocapsid Proteins Pathway interactions Primates Process Property Proteins RNA RNA-Directed DNA Polymerase Recruitment Activity Relative (related person)Reverse Transcriptase Polymerase Chain Reaction Ribonucleases Role Signal Recognition Particle Staging Subfamily lentivirinae T-Lymphocyte Testing Total Internal Reflection Fluorescent Transfer RNA Untranslated RNA Variant Viral Viral Proteins Virion Virus Virus Assembly base combat deep sequencing genomic RNA insight lymphoblastoid cell line mortality new therapeutic target pathogen public health relevance research study targeted treatment

项目摘要

DESCRIPTION (provided by applicant): Human immunodeficiency virus type 1 (HIV-1), the cause of acquired immune deficiency syndrome (AIDS), is responsible for significant morbidity and mortality. Although the use of therapies that target HIV-1 proteins has greatly reduced AIDS mortality, anti-retrovirals must be administered for decades. Given the inherent mutability of retroviral proteins, therapies that target new host pathways hijacked by HIV-1 or host components important for viral replication could be extremely useful. The objective of this exploratory project is to test the hypothesis that the propensity of HIV-1 to non-randomly recruit newly synthesized host noncoding RNAs (ncRNAs) into virions can be exploited to obtain insights into poorly understood early steps in HIV-1 assembly and to identify potential targets for antivirals. Although host ncRNAs were first described to undergo retroviral packaging more than forty years ago, the spectrum of ncRNAs encapsidated by HIV-1, the mechanisms by which these ncRNAs are recruited, and the extent to which they contribute to the HIV-1 lifecycle are all largely unknown. In preliminary experiments, we used high-throughput sequencing to obtain an unbiased and comprehensive assessment of the ncRNAs packaged by HIV-1. Our analyses revealed that, in addition to known packaged RNAs such as the 7SL RNA component of the signal recognition particle, several unexpected ncRNAs were present. We also found that ncRNA processing intermediates, which are usually rare because they exist only transiently within cells, were highly enriched in virions. Together, these results indicate that an early stage in HIV-1 assembly intersects with host cell ncRNA biogenesis pathways, thus opening windows into both of these processes. Our goals are to identify the viral and cellular requirements for efficient ncRNA packaging by HIV-1 and to determine the extent to which the ncRNAs contribute to HIV-1 assembly and replication. Our first aim is to build on our findings that HIV-1 packages specific ncRNAs and ncRNA precursors by performing additional biological replicates and by determining the abundance of highly represented ncRNAs in virions relative to the viral genomic RNA. Our second aim is to determine the mechanism(s) by which HIV-1 recruits specific host ncRNAs and the functional relevance of these RNAs to the HIV-1 lifecycle. We will test the hypothesis, based on our findings that HIV-1 preferentially recruits nascent ncRNAs, that interactions with viral components, such as genomic RNA or nucleocapsid, are critical for encapsidation. Because HIV-1 recruits some ncRNAs that have only been detected in nuclei, we will test whether HIV-1 interfaces with a new host cell surveillance pathway in which unprocessed and unassembled ncRNAs are exported to the cytoplasm for rapid decay. Since elucidating the contributions that host ncRNAs make to the HIV-1 lifecycle could yield antiviral targets, we will interfere with packaging of abundantly represented ncRNAs and examine effects on virus assembly and infectivity. By elucidating the set of packaged ncRNAs, their mechanisms of recruitment and the extent to which they contribute to HIV-1 replication, these experiments could identify new therapeutic targets.

描述（由申请人提供）： 1 型人类免疫缺陷病毒 (HIV-1) 是获得性免疫缺陷综合征 (AIDS) 的病因，是造成显着发病率和死亡率的原因。尽管使用针对 HIV-1 蛋白的疗法已大大降低了艾滋病死亡率，但抗逆转录病毒药物必须持续使用数十年。鉴于逆转录病毒蛋白固有的可变性，针对被 HIV-1 劫持的新宿主途径或对病毒复制重要的宿主成分的治疗可能非常有用。这个探索性项目的目的是测试这样一个假设：HIV-1 非随机地将新合成的宿主非编码 RNA (ncRNA) 招募到病毒粒子中的倾向可以被利用，以深入了解 HIV-1 组装和形成过程中人们知之甚少的早期步骤。确定潜在目标抗病毒药物。尽管宿主 ncRNA 首次被描述为在四十多年前经历逆转录病毒包装，但 HIV-1 包裹的 ncRNA 谱、这些 ncRNA 的招募机制以及它们对 HIV-1 生命周期的贡献程度都是未知的。很大程度上不为人所知。在初步实验中，我们使用高通量测序对 HIV-1 包装的 ncRNA 进行公正且全面的评估。我们的分析表明，除了已知的包装 RNA（例如信号识别颗粒的 7SL RNA 成分）之外，还存在一些意想不到的 ncRNA。我们还发现，ncRNA 加工中间体通常很少见，因为它们仅短暂存在于细胞内，但在病毒粒子中高度富集。总之，这些结果表明早期阶段 HIV-1 组装中的 DNA 与宿主细胞 ncRNA 生物合成途径相交叉，从而为这两个过程打开了窗口。我们的目标是确定 HIV-1 有效包装 ncRNA 的病毒和细胞要求，并确定 ncRNA 对 HIV-1 组装和复制的贡献程度。我们的首要目标是基于 HIV-1 包装特定 ncRNA 和 ncRNA 前体的发现，通过执行额外的生物学复制并确定病毒体中高度代表性的 ncRNA 相对于病毒基因组 RNA 的丰度。我们的第二个目标是确定 HIV-1 招募特定宿主 ncRNA 的机制以及这些 RNA 与 HIV-1 生命周期的功能相关性。基于我们的发现，HIV-1 优先招募新生的 ncRNA，与基因组 RNA 或核衣壳等病毒成分的相互作用对于衣壳形成至关重要，我们将检验这一假设。由于 HIV-1 招募了一些仅在细胞核中检测到的 ncRNA，因此我们将测试 HIV-1 是否与新的宿主细胞监视途径相互作用，在该途径中，未加工和未组装的 ncRNA 被输出到细胞质进行快速衰减。由于阐明宿主 ncRNA 对 HIV-1 生命周期的贡献可能会产生抗病毒靶标，因此我们将干扰大量代表性 ncRNA 的包装，并检查对病毒组装和感染性的影响。通过阐明一组包装的 ncRNA、它们的募集机制以及它们对 HIV-1 复制的贡献程度，这些实验可以确定新的治疗靶点。