MicroRNAs and the HSV Lytic/Latent Balance

MicroRNA 和 HSV 裂解/潜伏平衡

• 批准号: 9102890
• 负责人: DONALD M COEN
• 金额: $ 42.12万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9102890
• 关键词: Affect; Afferent Neurons; Autophagocytosis; Biological; Cell Culture Techniques; Cells; Chromatin; Clinical; Collaborations; Dicer Enzyme; Equilibrium; Family; Gene Expression; Genes; Herpesvirus 1; Human Herpesvirus 2; In Vitro; Infection; Instruction; Knock-out; Laboratories; Lead; Lesion; Life; Lytic; Lytic Phase; Maintenance; Messenger RNA; MicroRNAs; Mus; Mutation; Natural Immunity; Neurons; Oligonucleotides; Process; Production; Property; Research; Role; Seeds; Side; Simplexvirus; Site; System; Techniques; Testing; Viral; Viral Genes; Virus; Virus Diseases; Virus Latency; biological systems; fascinate; in vivo; interest; latent infection; lytic gene expression; lytic replication; mutant; novel therapeutics; overexpression; pathogen; programs; reactivation from latency; viral DNA

The long term objective of this project is to investigate how microRNAs (miRNAs) tilt the interaction of herpes simplex virus (HSV) and neurons either towards lytic infection or towards latency. HSV latency is the most fascinating biological property ofthe virus and its most important clinical feature. Understanding HSV latency may lead to new therapies or even a cure for this widespread pathogen. MiRNAs are important regulatory molecules in many biological systems, but much remains unknown about how they impact HSV infections. The first specific aim of this project is to determine the roles of host and viral miRNAs in repressing lytic gene expression during latency. Mice with the miRNA processing enzyme Dicer knocked out in a subset of sensory neurons, and viral mutants with lesions that affect specific miRNAs or target sites on specific mRNAs will be used to test the effects of miRNAs on gene expression during establishment and maintenance of latency in vivo and, with Project 3, in vitro. Effects of these mutations on chromatin status will be examined with Project 1. Targets for an interesting miRNA, miR-H5, will be identified using multiple approaches. Oligonucleotides that antagonize miRNA function (anti-mlRs) will be tested for effects on gene expression and reactivation in vitro with Project 3, and, if results are promising, in vivo. The second specific aim is to investigate the roles of HSV-1 miRNAs that counteract innate immunity using viral mutants affected for these miRNAs, with emphasis on whether these miRNAs promote latency by abetting host survival in the face of HSV-1 infection. Effects of these miRNAs in neurons on innate immunity and autophagy will be explored in vitro with Project 3. The third specific aim focuses on the lytic side of the lytic/latent balance, and seeks to determine how HSV-1 miRNAs promote lytic replication and reactivation. How ATRX, a target of HSV-1 miR-H1, is downregulated in lytic infection and how that impacts virus production, chromatin status, and reactivation from latency will be explored with Project 1 in cell culture and in vivo and, with Project 3, in an in vitro latency system, using virus mutants and anti-mlRs. Additional targets for miR-H1 and targets for a conserved family of miRNAs expressed during lytic infection will be identified. RELEVANCE (See instructions); This project proposes research to understand how miRNAs contribute to herpes simplex virus (HSV) latency, which is the most fascinating biological and most vexing clinical aspect of this virus. Understanding HSV latency might lead to new treatments or even cures for HSV infections, which are widespread and, in some cases, life-threatening.

该项目的长期目标是研究microRNA（miRNA）如何倾斜疱疹的相互作用 单纯形病毒（HSV）和神经元朝向裂解感染或倾向潜伏期。 HSV延迟最多 该病毒的引人入胜的生物学特性及其最重要的临床特征。了解HSV潜伏期 可能导致新的疗法，甚至可以治愈这种广泛的病原体。 miRNA是重要的调节 许多生物系统中的分子，但对于它们如何影响HSV感染仍未知。 该项目的第一个具体目的是确定宿主和病毒miRNA在抑制裂解基因中的作用 延迟期间的表达。 MiRNA加工酶DICER的小鼠在一部分 感觉神经元和具有影响特定特异性miRNA或靶位点病变的病毒突变体 mRNA将用于测试miRNA在建立过程中对基因表达的影响 维持体内潜伏期的延迟，并在项目3中维持体外。这些突变对染色质状态的影响 将使用项目1进行检查。 方法。将测试拮抗miRNA功能（抗MLR）的寡核苷酸对基因的影响 在体外表达和重新激活项目3，如果结果有前途，则在体内。第二个特定 目的是研究使用影响的病毒突变体抵消先天免疫力的HSV-1 miRNA的作用 对于这些miRNA，重点是这些miRNA是否通过教宿主生存来促进潜伏期 HSV-1感染的面孔。这些miRNA在神经元中对先天免疫和自噬的影响将是 通过项目3在体外进行了探索。第三个特定目的侧重于裂解/潜在平衡的裂解侧， 试图确定HSV-1 miRNA如何促进裂解复制和重新激活。 ATRX如何，目标 HSV-1 miR-H1在裂解感染中被下调，这如何影响病毒的产生，染色质状态， 与潜伏期的重新激活将与细胞培养和体内的项目1一起探索，以及项目3，在 使用病毒突变体和抗MLR的体外潜伏系统。 miR-H1的其他目标和A的目标 将确定在裂解感染期间表达的保守家族。 相关性（请参阅说明）； 该项目提出了研究，以了解miRNA如何对单纯疱疹病毒（HSV）潜伏期做出贡献， 这是该病毒中最迷人的生物学和最烦人的临床方面。了解HSV 潜伏期可能会导致新的治疗方法，甚至可以治愈HSV感染，这是广泛的，在某些人中， 案件，威胁生命。