Recruitment of host noncoding RNAs by HIV-1

HIV-1 招募宿主非编码 RNA

• 批准号: 9095216
• 负责人: Sandra L. Wolin
• 金额: $ 24.66万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 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/9095216
• 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 resistance; Goals; HIV; HIV-1; Health; 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; 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; Virus Replication; base; combat; deep sequencing; genomic RNA; insight; lymphoblastoid cell line; mortality; new therapeutic target; pathogen; research study; targeted treatment

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）：人类免疫缺陷病毒1型（HIV-1）是获得免疫缺陷综合征（AIDS）的原因，负责明显的发病率和死亡率。尽管靶向HIV-1蛋白的疗法的使用大大降低了艾滋病死亡率，但必须使用抗逆转录病毒病数十年。鉴于逆转录病毒蛋白的固有突变性，靶向由HIV-1劫持的新宿主途径或对病毒复制重要的宿主成分的疗法可能非常有用。该探索性项目的目的是检验以下假设：HIV-1非随机招募新合成的宿主非编码RNA（NCRNA）的倾向可以被利用，从确定潜在目标 抗病毒药。尽管首先描述了四十多年前的宿主NCRNA经历了逆转录病毒包装，但HIV-1封装的NCRNA的谱系，这些NCRNA被募集的机制以及它们对HIV-1生命周期的贡献的程度都是在很大程度上未知。在初步实验中，我们使用高通量测序来获得对HIV-1包装的NCRNA的无偏和全面评估。我们的分析表明，除了已知的包装RNA（例如信号识别粒子的7SL RNA成分）外，还有几个意外的NCRNA。我们还发现，NCRNA加工中间体通常很少，因为它们仅在细胞内瞬时存在，因此在病毒体中高度富集。这些结果在一起表明早期 在HIV-1组装中，与宿主细胞NCRNA生物发生途径相交，从而将窗户打开到这两个过程中。我们的目标是确定HIV-1有效NCRNA包装的病毒和细胞要求，并确定NCRNA对HIV-1组装和复制的贡献程度。我们的第一个目的是建立我们的发现，即HIV-1通过执行其他生物学重复和确定与病毒基因组RNA相对于病毒基因组RNA的高度代表的NCRNA的丰度来包装特定的NCRNA和NCRNA前体。我们的第二个目的是确定HIV-1募集特定宿主NCRNA的机制以及这些RNA与HIV-1生命周期的功能相关性。我们将基于我们的发现，即HIV-1优先募集新生的NCRNA，即与病毒成分（例如基因组RNA或核素）相互作用对于封装至关重要。由于HIV-1募集了仅在核中检测到的一些NCRNA，因此我们将测试HIV-1界面是否具有新的宿主细胞监测途径，在该途径中，未加工和未组装的NCRNA导出到细胞质中以进行快速衰减。由于阐明了宿主NCRNA对HIV-1生命周期的贡献可能产生抗病毒靶标，因此我们将干扰大量代表NCRNA的包装，并检查对病毒组装和感染性的影响。通过阐明包装的NCRNA集，它们的招募机制以及它们对HIV-1复制的贡献程度，这些实验可以识别出新的治疗靶标。