Cytor lncRNA as a positive regulator of HIV gene expression and viral latency

Cytor lncRNA 作为 HIV 基因表达和病毒潜伏期的正调节因子

• 批准号: 10681321
• 负责人: Koh Fujinaga
• 金额: $ 20.48万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-10 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10681321
• 关键词: Binding; Biochemical; CD4 Positive T Lymphocytes; Cell Separation; Cell physiology; Cells; Complex; Cytoskeleton; Development; Event; Foundations; Gene Activation; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; HIV; HIV Infections; HIV tat Protein; Histones; In Vitro; Integration Host Factors; Knowledge; Light; Mediating; Molecular; Monitor; Patients; Phosphotransferases; Physiological; Play; Positive Transcriptional Elongation Factor B; Proteins; Proteomics; Protocols documentation; RNA; RNA Polymerase II; Regulation; Regulator Genes; Rest; Role; Testing; Therapeutic; Transcription Elongation; Transcriptional Elongation Factors; Untranslated RNA; Viral; Viral reservoir; Virus Latency; Virus Replication; antiretroviral therapy; chromatin isolation by RNA purification sequencing; clinically relevant; clinically significant; cofactor; cyclin T1; differential expression; effective therapy; efficacy testing; experimental study; genetic testing; genome-wide; improved; insight; new therapeutic target; promoter; reactivation from latency; recruit; therapeutic RNA; tool; transcriptome; transcriptome sequencing

ABSTRACT To successfully eliminate the HIV reservoir, it is critical to understand the molecular events that control HIV latency. While the key roles that host protein factors play in the regulation of HIV transcription and viral latency have been extensively studied, our understanding of how the non-coding transcriptome, especially long non- coding RNA (lncRNA), contribute to viral latency control is limited. To better understand the regulatory roles lncRNAs play in the control of HIV transcription and viral latency, we employed RNA-Seq analysis and compared the transcriptome in activated versus resting HIV-infected cells. We identified numerous lncRNAs that are differentially expressed, therefore are candidates for HIV latency regulation. Among them, one lncRNA, Cytoskeleton Regulator (Cytor), activates HIV transcription and viral latency. Our results also indicate that the depletion of Cytor suppresses latent HIV reactivation by reducing the occupancy of RNA Polymerase II (Pol II) and the levels of histone activation markers on the viral promoter. Additional biochemical and proteomic analyses showed that Cytor occupies the HIV promoter and associates with Positive Transcription Elongation Factor b (P- TEFb), which is an essential cellular factor for transcription elongation of HIV and cellular genes. In light of these findings, we hypothesize that by recruiting P-TEFb to the HIV promoter, Cytor activates HIV gene expression. To test this hypothesis, we will determine whether Cytor directly binds P-TEFb and recruits the cellular transcription elongation complex to the HIV promoter. Additional preliminary RNA-Seq analysis indicated that Cytor depletion results in broad changes in the host transcriptome. We will therefore identify downstream targets of Cytor and determine their indirect effects on HIV gene expression. Significantly, our results will be further confirmed in a clinically relevant context, as we will manipulate Cytor expression in CD4+ primary cells and determine the magnitude of Cytor’s therapeutic potential for HIV reactivation and latency reversal or, alternatively, to latency induction. Our study will provide new insights into the regulation of HIV transcription and viral latency by lncRNAs. Its successful completion will lay the groundwork for the development of new RNA-based therapies that will be added to current therapeutic protocols to eliminate HIV infection and the persistent viral reservoir.

抽象的 为了成功消除HIV储层，了解控制Hibi的分子事件至关重要 延迟。 已经广泛研究过，我们的理解如何 编码RNA（LNCRNA），有助于病毒潜伏期。 LNCRNA在控制HIV转录和病毒潜伏期的控制方面发挥了作用，我们采用了RNA-Seq分析并进行了比较 活化的艾滋病毒感染细胞中的转录组。 差异表达的是，艾滋病毒潜伏期调节的候选者。 细胞骨架调节剂（细胞）激活HIV转录和病毒潜伏期。 通过还原RNA聚合酶II（POL II）来抑制细胞抑制潜在的HIV再活化（POL II） 以及病毒启动子上的组蛋白激活标记水平。 结果表明，细胞分子占据了HIB启动子，并与阳性转录伸长因子B（P-）相关联 Tefb），这是鉴于这些的转录伸长的必要细胞因子。 调查结果，我们假设通过募集P-TEFB艾滋病毒启动子，细胞运动员激活了HIV基因表达。 为了检验该假设，我们将确定细胞器直接结合P-TEFB并募集细胞 转录伸长复合物对HIB启动子 细胞耗尽会导致宿主转录组的广泛变化。 细胞体并确定对HIV基因表达的间接影响。 在临床上相关的环境中确认，因为我们将操纵CD4+主细胞中的细胞表达和 确定细胞对艾滋病毒重新激活和纬度逆转或替代性替代性的治疗潜力的大幅度。 延迟指示。 通过LNCRNA，其成功的压缩将为开发新的基于RNA的疗法提供基础 这将是消除HIV感染和陶氏储层的当前治疗方案。