Viral And host mechanisms that tilt the HSV lytic/latent balance

导致 HSV 裂解/潜伏平衡倾斜的病毒和宿主机制

• 批准号: 8871671
• 负责人: DONALD M COEN
• 金额: $ 177.52万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-02 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8871671
• 关键词: Acute; Affect; Animals; Antiviral Agents; Area; Autophagocytosis; Back; Biological Assay; Chromatin; Chromatin Structure; Complex; Cultured Cells; DNA Sequence; Disease; Drug Targeting; Enzymes; Equilibrium; Gene Expression; Genes; Genital system; Genome; Growth; HIV Infections; Herpesvirus 1; Histones; Immune response; In Vitro; Infection; Investigation; Laboratories; Latent Virus; Latent virus infection phase; Life; Lytic; Lytic Phase; Maintenance; MicroRNAs; Morbidity - disease rate; Mus; Mutate; Natural Immunity; Neurons; Pharmaceutical Preparations; Proteins; RNA; Recruitment Activity; Recurrence; Research; Risk; Role; Sensory Ganglia; Simplexvirus; Site; Structure of trigeminal ganglion; System; Testing; Transcript; Untranslated RNA; Viral; Viral Genes; Viral Genome; Viral Physiology; Viral Proteins; Virus; Virus Latency; Work; chromatin remodeling; combat; combinatorial; genital herpes; histone modification; in vivo; latency associated transcript; latent infection; lytic gene expression; mortality; mucosal site; mutant; nervous system disorder; programs; promoter; response; therapeutic target; viral DNA

DESCRIPTION (provided by applicant): Herpes simplex viruses cause considerable morbidity and mortality. They undergo a lytic, productive infection at the mucosal sites and spread into sensory ganglia, where they undergo a latent infection for the life of the host. Reactivation leads to recurrent infection and disease. Antiviral drugs have been defined that inhibit the lytic infection cycle, but there are no approaches that target the latent virus. We have defined the role of viral gene products such as LAT and ICPO in modulating the chromatin structure during lytic and latent infection, but further basic information is needed about these mechanisms for discovery of therapeutics that target HSV latent infection. In this application our specific aims are: a. To test hypotheses for possible mechanisms by which the HSV latency-associated transcript reduces lytic gene expression during acute infection and during latent infection of trigeminal ganglia: LAT acts as a long noncoding RNA that recruits histone-modifying complexes to the viral genome, b. LAT leads to chromatin changes by serving as a precursor to an miRNA that reduces ICPO expression through studies of miRNA mutant viruses (with Coen lab), c. LAT regulatory DNA sequences act in a cis-acting manner to promote chromatin on the viral lytic genes, d. LAT leads to differential targeting of the viral genome through studies of genome targeting in cultured neurons with the Leib lab. 2. To define the mechanisms by which the HSV ICPO protein regulates chromatin structure during acute infection and latent infection of trigeminal ganglia. We have exciting unpublished results that ICPO mutant viruses have a different chromatin profile on their genome during latent infection. We will test the hypothesis that ICPO acts to alter the chromatin state by recruiting histone modification enzymes to viral and cellular genes. 3. To define Interactions between LAT and ICPO in regulating HSV chromatin. We will test the hypothesis that LAT forms duplex RNA with ICPO transcripts to recruit histone modifying enzymes by mutating the ICPO promoter or mutating the ICPO translational initiation site and by constructing LAT and ICPO double mutant viruses to determine their combinatorial effects on latent infection, and viral chromatin. RELEVANCE: Herpes simplex viruses cause considerable genital, ocular and nervous system disease, and genital herpes increases the risk of HIV infection. There are drugs that target the active growth of herpes simplex virus but none that target the latent infection. This research wil define basic mechanisms of herpes simplex virus latent infection and new targets for potential drugs to treat the latent infection of these viruses. Project 1: Chromatin and the lytic/latent balance Project Leader (PL): Knipe, David M. DESCRIPTION (provided by applicant): Herpes simplex viruses cause considerable morbidity and mortality. They undergo a lytic, productive infection at the mucosal sites and spread into sensory ganglia, where they undergo a latent infection for the life of the host. Reactivation leads to recurrent infection and disease. Antiviral drugs have been defined that inhibit the lytic infection cycle, but there are no approaches that target the latent virus. We have defined the role of viral gene products such as LAT and ICP0 in modulating the chromatin structure during lytic and latent infection, but further basic information is needed about these mechanisms for discovery of therapeutics that target HSV latent infection. In this application our specific aims are: a. To test hypotheses for possible mechanisms by which the HSV latency-associated transcript reduces lytic gene expression during acute infection and during latent infection of trigeminal ganglia: LAT acts as a long noncoding RNA that recruits histone-modifying complexes to the viral genome. b. LAT leads to chromatin changes by serving as a precursor to an miRNA that reduces ICP0 expression through studies of miRNA mutant viruses (with Coen lab). c. LAT regulatory DNA sequences act in a cis-acting manner to promote chromatin on the viral lytic genes. d. LAT leads to differential targeting of the viral genome through studies of genome targeting in cultured neurons with the Leib lab. 2. To define the mechanisms by which the HSV ICP0 protein regulates chromatin structure during acute infection and latent infection of trigeminal ganglia. We have exciting unpublished results that ICP0 mutant viruses have a different chromatin profile on their genome during latent infection. We will test the hypothesis that ICP0 acts to alter the chromatin state by recruiting histone modification enzymes to viral and cellular genes. 3. To define Interactions between LAT and ICP0 in regulating HSV chromatin. We will test the hypothesis that LAT forms duplex RNA ICP0 transcripts to recruit histone modifying enzymes by mutating the ICP0 promoter or mutating the ICP0 translational initiation site and by constructing LAT and ICP0 double mutant viruses to determine their combinatorial effects on latent infection, and viral chromatin. RELEVANCE: Herpes simplex viruses cause considerable genital, ocular, and nervous system disease, and genital herpes increases the risk of HIV infection. There are drugs that target the active growth of herpes simplex virus but none that target the latent infection. This research wil define basic mechanisms of herpes simplex virus latent infection and new targets for potential drugs to treat the latent infection of these viruses.

描述（由申请人提供）：单纯疱疹病毒引起了相当大的发病率和死亡率。他们在粘膜部位经历了裂解的生产性感染，并扩散到感官神经节中，在那里他们会在宿主的生命中感染潜在感染。重新激活导致 复发感染和疾病。已经定义了抑制裂解感染周期的抗病毒药物，但没有针对潜在病毒的方法。我们已经定义了病毒基因产物（例如LAT和ICPO）在调节裂解和潜在感染过程中的染色质结构中的作用，但是对于发现靶向HSV潜在感染的治疗剂，需要进一步的基本信息。在此应用中，我们的具体目的是：为了测试假设，以使HSV潜伏相关的转录本在急性感染过程和三叉神经节的潜在感染过程中降低裂解基因的表达：LAT充当长期的非编码RNA，可募集组蛋白对病毒基因组的组蛋白改性为复合物，b。 LAT通过作为miRNA的前体来导致染色质变化，该miRNA通过研究miRNA突变病毒（带有Coen Lab），c。 LAT调节DNA序列以顺式作用方式作用，以促进病毒裂解基因上的染色质。 LAT通过使用Leib Lab的培养神经元中的基因组靶向来导致病毒基因组的差异靶向。 2。定义HSV ICPO蛋白在急性感染和三叉神经节的潜在感染过程中调节染色质结构的机制。我们有令人兴奋的未发表的结果，即ICPO突变病毒在潜在感染期间的基因组具有不同的染色质特征。我们将检验以下假设：ICPO通过募集组蛋白修饰酶为病毒和细胞基因来改变染色质状态。 3。在调节HSV染色质时定义LAT和ICPO之间的相互作用。我们将测试以ICPO转录物形成双链RNA的假说，通过突变ICPO启动子或突变ICPO转化起始位点并通过构建LAT和ICPO双重突变病毒来确定其组合对拉力感染以及病毒染色质的组合作用，从而通过ICPO转录物形成招募组蛋白来修饰酶。 相关性：单纯疱疹病毒引起大量生殖器，眼和神经系统疾病，生殖器疱疹增加了HIV感染的风险。有一些药物靶向单纯疱疹病毒的积极生长，但没有针对潜在感染的药物。这项研究定义了单纯疱疹病毒潜在感染的基本机制，以及用于治疗这些病毒潜在感染的潜在药物的新靶标。 项目1：染色质和裂解/潜在平衡 项目负责人（PL）：Knipe，David M. 描述（由申请人提供）：单纯疱疹病毒引起了相当大的发病率和死亡率。他们在粘膜部位经历了裂解的生产性感染，并扩散到感官神经节中，在那里他们会在宿主的生命中感染潜在感染。重新激活导致 复发感染和疾病。已经定义了抑制裂解感染周期的抗病毒药物，但没有针对潜在病毒的方法。我们已经定义了病毒基因产物（例如LAT和ICP0）在调节裂解和潜在感染过程中的染色质结构中的作用，但是对于发现靶向HSV潜在感染的治疗剂，需要进一步的基本信息。在此应用中，我们的具体目的是：为了测试假设，以使HSV潜伏期相关的转录本在急性感染和三叉神经节的潜在感染过程中降低裂解基因的表达：LAT充当长期非编码RNA，可募集组蛋白对病毒基因组的组蛋白改性复合物募集。 b。 LAT通过作为miRNA的前体来导致染色质变化，该miRNA通过研究miRNA突变病毒（使用Coen Lab）来降低ICP0表达。 c。 LAT调节DNA序列以顺式作用方式起作用，以促进病毒裂解基因上的染色质。 d。 LAT通过使用Leib Lab的培养神经元中的基因组靶向来导致病毒基因组的差异靶向。 2。定义HSV ICP0蛋白在急性感染和三叉神经节的潜在感染过程中调节染色质结构的机制。我们有令人兴奋的未发表的结果，即在潜在感染期间，ICP0突变病毒在其基因组上具有不同的染色质特征。我们将检验以下假设：ICP0通过募集组蛋白修饰酶为病毒和细胞基因来改变染色质状态。 3。在调节HSV染色质时定义LAT和ICP0之间的相互作用。我们将检验以下假设：LAT形成双链RNA ICP0转录本来通过突变ICP0启动子或突变ICP0转化起始位点并构建LAT和ICP0双突变病毒以确定其对潜水感染的组合作用以及病毒染色质来确定其联合效应，从而通过突变ICP0启动子突变或突变ICP0转化起始位点来修饰酶。 相关性：单纯疱疹病毒会引起大量生殖器，眼和神经系统疾病，而生殖器疱疹会增加HIV感染的风险。 有一些药物靶向单纯疱疹病毒的积极生长，但没有针对潜在感染的药物。 这项研究定义了单纯疱疹病毒潜在感染的基本机制，以及用于治疗这些病毒潜在感染的潜在药物的新靶标。