REGULATION OF HERPES SIMPLEX TYPE 1 INFECTION IN CORNEAL NEURONS

角膜神经元 1 型单纯疱疹病毒感染的调节

• 批准号: 9096804
• 负责人: TODD P. MARGOLIS
• 金额: $ 38.13万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9096804
• 关键词: Accounting; Acute; Adult; Affect; Afferent Neurons; Antiviral Agents; Biological Assay; Biological Models; Blindness; Cells; Clinical; Cornea; Corneal Diseases; Country; Cytoplasm; Data; Disease; Eye diseases; Failure; Gene Expression; Genes; Genetic Transcription; Genital system; Goals; Growth; Herpes Simplex Infections; Herpesvirus 1; Human; Immediate-Early Genes; Immediate-Early Proteins; In Vitro; Infection; Infectious Skin Diseases; Keratoplasty; Kinetics; Lip Diseases; Mediating; Morbidity - disease rate; Mucous Membrane; Mus; Nature; Nerve; Neuro-Ocular System; Neurons; Outcome; Outcome Study; Patients; Pattern; Phase; Phenotype; Play; Population; Process; Prophylactic treatment; Proteins; Recurrence; Regulation; Research; Role; Simplexvirus; Site; Spinal Ganglia; Structure of trigeminal ganglion; System; Testing; Transactivation; VP 16; Viral; Viral Genes; Viral Proteins; Viral reservoir; Virus; Virus Diseases; Virus Latency; Virus Replication; Visual impairment; abstracting; adeno-associated viral vector; base; clinical practice; corneal scar; gene product; human disease; in vitro Model; in vivo; innovation; insight; latent infection; mutant; novel; novel therapeutic intervention; novel therapeutics; permissiveness; prevent; promoter

PROJECT SUMMARY/ABSTRACT Herpes simplex virus (HSV) type 1 is a leading cause of infectious corneal blindness. It causes eye disease by reactivation from a latent viral reservoir in corneal nerves. Despite extensive research, the mechanisms that regulate HSV infection in neurons are not well characterized. A better understanding of this is critical for identifying new therapeutic strategies. We have developed a novel culture system, using dissociated adult murine trigeminal ganglia (TG), for studying HSV infection in neurons. Preliminary data from our lab indicates that, unlike its role in replicating cells, the viral immediate early (IE) gene product, ICP27, restricts productive viral infection in neurons, especially in A5+ ganglionic neurons, and promotes viral latency. In the current proposal we will further characterize this novel function for ICP27, as well as study the mechanisms by which ICP27 accomplishes these functions. In the first two specific aims of this proposal, we will characterize the role that ICP27 plays in restricting productive infection and promoting viral latency, using ICP27 null mutants, an ICP27 promoter mutant with delayed kinetics of expression, and viral mutants with deletions in different ICP27 functional domains. We will further characterize the role of ICP27 in restricting infection in neurons through the use of novel AAV vectors for the efficient transduction of sensory neurons with ICP27. Our preliminary data also suggests that VP16, a late viral protein in replicating cells, is expressed very early in TG neurons, and that ICP27 restricts transcription of both VP16 and ICP4 in cultured TG neurons. In the third specific aim we will further characterize ICP27 mediated inhibition of ICP4 and VP16 transcription and test hypotheses about the way in which this is achieved. Finally, we will test hypotheses that ICP27 restricts productive infection in A5+ neurons, in part, by restricting ICP4, VP16 and HCF1 to the cytoplasm, thus preventing transactivation of viral IE genes. These concepts and studies are innovative, and are a result of being able to directly study HSV infection in neurons, as well as being able to differentiate the outcome of infection in A5+ neurons, the major site of HSV latency. The outcome of these studies should generate new insights into the mechanisms regulating HSV infection of neurons; the first step in developing new therapy strategies.

项目总结/摘要 单纯疱疹病毒（HSV）1型是感染性角膜盲的主要原因。它会导致 由于角膜神经中潜伏的病毒库重新激活而引起的眼病。尽管进行了广泛 研究中，调节神经元中HSV感染的机制还没有得到很好的表征。一 更好地理解这一点对于确定新的治疗策略至关重要。我们有 开发了一种新的培养系统，使用分离的成年小鼠三叉神经节（TG）， 研究神经元中的HSV感染。我们实验室的初步数据表明，与其在 复制细胞中，病毒立即早期（IE）基因产物ICP 27限制了生产性病毒 感染的神经元，特别是在A5+神经节神经元，并促进病毒潜伏期。在 目前的建议，我们将进一步表征ICP 27的这种新功能，以及研究 ICP 27实现这些功能的机制。在前两个具体目标中， 建议，我们将描述ICP 27在限制生产性感染中的作用， 促进病毒潜伏期，使用ICP 27无效突变体，ICP 27启动子突变体具有延迟 表达动力学，以及在不同ICP 27功能结构域中缺失的病毒突变体。 我们将进一步表征ICP 27在限制神经元感染中的作用， 用于用ICP 27有效转导感觉神经元的新型AAV载体。我们 初步数据还表明，VP 16，一种复制细胞中的晚期病毒蛋白， 在TG神经元中非常早，并且ICP 27限制了TG神经元中VP 16和ICP 4的转录。 培养的TG神经元。在第三个具体目标中，我们将进一步表征ICP 27介导的 ICP 4和VP 16转录的抑制，并测试关于这种抑制的方式的假设。 办妥了一批最后，我们将测试ICP 27限制A5+中的生产性感染的假设。 神经元，部分通过限制ICP 4，VP 16和HCF 1到细胞质，从而防止 病毒IE基因的反式激活。这些概念和研究是创新的，是 能够直接研究神经元中的HSV感染，以及能够区分 A5+神经元感染的结果，HSV潜伏期的主要部位。成果的 研究应该对神经元HSV感染的调节机制产生新的见解; 这是开发新疗法的第一步。