Role of biomolecular condensates in regulating HIV-1 viral ribonucleoprotein complex formation in the setting of substance use disorder

物质使用障碍中生物分子缩合物在调节 HIV-1 病毒核糖核蛋白复合物形成中的作用

• 批准号: 10228358
• 负责人: Leslie J Parent
• 金额: $ 18.84万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228358
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Binding; Biochemical; Biogenesis; Biological; Biology; Biophysics; Cell Nucleus; Cells; Cessation of life; Clinic; Complex; Coupling; Data; Disease; Dissociation; Drug Targeting; Drug resistance; Gene Expression; Genetic; Genetic Determinism; Genetic Transcription; Genome; Goals; HIV; HIV-1; Health; Hela Cells; Immunologic Deficiency Syndromes; In Vitro; Incidence; Infection; Integration Host Factors; Intervention; Knowledge; Lead; Light; Liquid substance; Methods; Molecular; Nuclear; Nuclear Export; Pathway interactions; Pharmacology; Phase; Phase Transition; Physical condensation; Play; Positive Transcriptional Elongation Factor B; Process; Production; Property; Proteins; Proteomics; Proviruses; Publishing; RNA; RNA Processing; RNA chemical synthesis; Regulation; Research Project Grants; Retrovirology; Ribonucleoproteins; Role; Scientist; Site; Structure; Substance Use Disorder; T-Lymphocyte; Testing; Viral; Virion; Virus; Virus Latency; Virus Replication; Virus-Cell Membrane Interaction; Work; antiretroviral therapy; base; biophysical properties; combat; drug of abuse; drug resistant virus; follow-up; gag Gene Products; genomic RNA; in vitro Assay; innovation; knock-down; live cell imaging; new therapeutic target; novel; pandemic disease; reactivation from latency; therapeutic development; transcription factor; treatment strategy; viral RNA

Abstract HIV-1/AIDS is a devastating immunodeficiency disease that has resulted in over 35 million deaths across the globe. There is a compelling ongoing need to develop novel treatment strategies to combat the continuing emergence of drug-resistant viral strains. A drug target that has not yet been successfully brought to the clinic is the interaction of the Gag protein with its RNA genome to form the viral ribonucleoprotein complex, which initiates the process of virion assembly. A deeper understanding of the biochemical and biophysical interactions that drive viral ribonucleoprotein complex formation is needed to advance further therapeutic development. Based on recent findings that viral ribonucleoprotein complexes undergo liquid-liquid phase separation to form biomolecular condensates, this proposal explores the molecular underpinnings of Gag-viral RNA interactions. We have assembled an accomplished interdisciplinary team of scientists to work at the crossroads of retrovirology, RNA biology, biophysics, and pharmacology to shed light on our understanding of viral and cellular biomolecular condensates in HIV-1 infection. Our preliminary results suggesting that HIV-1 ribonucleoprotein complexes form in the nucleus inspired this provocative proposal to investigate the interplay of virus replication machinery with nuclear bodies that form biomolecular condensates. In the R21 phase, we will use innovative methods involving biophysics, genetics, state-of-the-art live cell imaging, and targeted pharmacological interventions to examine whether HIV-1 ribonucleoprotein complexes assemble into biomolecular condensates in the nucleus. In the R33 phase, we will extend these studies to probe mechanistic questions focusing on whether nuclear BMCs play critical roles in regulating HIV-1 transcription, latency reactivation, and genomic RNA packaging. Due to the high incidence of substance use disorder in people with HIV-1/AIDS, we will also investigate whether drugs of abuse influence HIV-1 nuclear biomolecular condensates. Elucidating the genetic determinants of HIV-1 condensate formation could lead to the identification of novel drug targets to treat HIV/AIDS.

抽象的 HIV-1/艾滋病是一种毁灭性的免疫缺陷疾病，已导致超过 3500 万人死亡 遍布全球。迫切需要开发新的治疗策略来对抗 耐药病毒株不断出现。尚未成功引入的药物靶点 诊所是Gag蛋白与其RNA基因组相互作用形成病毒核糖核蛋白复合物， 这启动了病毒粒子组装的过程。对生物化学和生物物理有更深入的了解 需要驱动病毒核糖核蛋白复合物形成的相互作用来推进进一步的治疗 发展。基于最近的发现，病毒核糖核蛋白复合物经历液-液相 分离形成生物分子缩合物，该提案探讨了 Gag 病毒的分子基础 RNA 相互作用。我们组建了一支卓有成效的跨学科科学家团队，在 逆转录病毒学、RNA 生物学、生物物理学和药理学的交叉点，阐明我们对 HIV-1 感染中的病毒和细胞生物分子凝聚体。 我们的初步结果表明 HIV-1 核糖核蛋白复合物在细胞核中形成 这项研究病毒复制机制与核体相互作用的挑衅性提议 形成生物分子缩合物。在R21阶段，我们将使用涉及生物物理学的创新方法， 遗传学、最先进的活细胞成像和有针对性的药理学干预措施，以检查是否 HIV-1 核糖核蛋白复合物在细胞核中组装成生物分子凝聚物。在R33阶段， 我们将扩展这些研究来探讨机制问题，重点关注核 BMC 是否发挥关键作用 在调节 HIV-1 转录、潜伏期再激活和基因组 RNA 包装中的作用。由于高 HIV-1/AIDS 患者中物质使用障碍的发生率，我们还将调查药物是否 滥用影响HIV-1核生物分子凝聚体。阐明 HIV-1 的遗传决定因素 冷凝物的形成可能有助于识别治疗艾滋病毒/艾滋病的新药物靶点。