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）：单纯疱疹病毒引起相当大的发病率和死亡率。它们在粘膜部位经历裂解性、生产性感染，并扩散到感觉神经节，在那里它们经历潜伏感染直至宿主死亡。再激活电极导线 复发性感染和疾病。抗病毒药物已被定义为抑制裂解感染周期，但没有靶向潜伏病毒的方法。我们已经确定了病毒基因产物如LAT和ICPO在裂解和潜伏感染过程中调节染色质结构的作用，但需要进一步的基本信息，这些机制的发现治疗HSV潜伏感染的目标。在本申请中，我们的具体目标是：为了检验HSV潜伏相关转录物在急性感染和三叉神经节潜伏感染期间降低裂解基因表达的可能机制的假设：LAT作为一种长的非编码RNA，将组蛋白修饰复合物募集到病毒基因组中，B。LAT通过作为miRNA的前体而导致染色质变化，所述miRNA通过miRNA突变病毒的研究（与Coen实验室一起）减少ICPO表达，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，d. LAT通过Leib实验室对培养的神经元中的基因组靶向研究，导致病毒基因组的差异靶向。2.明确三叉神经节急性感染和潜伏感染时HSV ICPO蛋白调节染色质结构的机制。我们有令人兴奋的未发表的结果，ICPO突变病毒在潜伏感染期间在其基因组上具有不同的染色质谱。我们将检验ICPO通过向病毒和细胞基因募集组蛋白修饰酶来改变染色质状态的假设。3.目的探讨LAT和ICPO在HSV染色质调控中的相互作用。我们将测试的假设，LAT形成双链体RNA与ICPO转录招募组蛋白修饰酶突变的ICPO启动子或突变的ICPO翻译起始位点，并通过构建LAT和ICPO双突变体病毒，以确定其对潜伏感染和病毒染色质的组合效应。 相关性：单纯疱疹病毒可引起相当多的生殖器、眼部和神经系统疾病，生殖器疱疹会增加感染艾滋病毒的风险。有针对单纯疱疹病毒活跃生长的药物，但没有针对潜伏感染的药物。本研究将明确单纯疱疹病毒潜伏感染的基本机制，并为治疗这些病毒潜伏感染的潜在药物提供新的靶点。 项目1：染色质和溶解/潜在平衡 项目负责人（PL）：Alberpe，大卫M. 描述（由申请人提供）：单纯疱疹病毒引起相当大的发病率和死亡率。它们在粘膜部位经历裂解性、生产性感染，并扩散到感觉神经节，在那里它们经历潜伏感染直至宿主死亡。再激活电极导线 复发性感染和疾病。抗病毒药物已被定义为抑制裂解感染周期，但没有靶向潜伏病毒的方法。我们已经确定了病毒基因产物如LAT和ICP 0在裂解和潜伏感染过程中调节染色质结构的作用，但需要进一步的基本信息，这些机制的发现治疗HSV潜伏感染的目标。在本申请中，我们的具体目标是：为了测试HSV潜伏相关转录物在急性感染和三叉神经节潜伏感染期间减少裂解基因表达的可能机制的假设：LAT作为一种长的非编码RNA，将组蛋白修饰复合物招募到病毒基因组中。B. LAT通过作为miRNA的前体导致染色质变化，通过对miRNA突变病毒的研究（与Coen实验室）降低ICP 0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Leib实验室对培养的神经元中的基因组靶向研究，导致病毒基因组的差异靶向。2.明确三叉神经节急性感染和潜伏感染时HSV ICP 0蛋白调节染色质结构的机制。我们有令人兴奋的未发表的结果，ICP 0突变病毒在潜伏感染期间在其基因组上具有不同的染色质谱。我们将检验ICP 0通过向病毒和细胞基因募集组蛋白修饰酶来改变染色质状态的假设。3.目的：明确LAT和ICP 0在调节HSV染色质中的相互作用。我们将测试的假设，LAT形成双链RNA ICP 0转录招募组蛋白修饰酶突变的ICP 0启动子或突变的ICP 0翻译起始位点，并通过构建LAT和ICP 0双突变病毒，以确定其潜伏感染和病毒染色质的组合效应。 相关性：单纯疱疹病毒引起相当多的生殖器，眼部和神经系统疾病，生殖器疱疹增加了艾滋病毒感染的风险。 有针对单纯疱疹病毒活跃生长的药物，但没有针对潜伏感染的药物。 本研究将明确单纯疱疹病毒潜伏感染的基本机制，并为治疗这些病毒潜伏感染的潜在药物提供新的靶点。