Regulation of Herpes Simplex Type 1 Infection in Corneal Neurons

角膜神经元 1 型单纯疱疹感染的调节

• 批准号: 8576684
• 负责人: TODD P. MARGOLIS
• 金额: $ 39.27万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8576684
• 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 structure; 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; Virus; Virus Diseases; Virus Latency; Visual impairment; adeno-associated viral vector; base; clinical practice; corneal scar; human disease; in vitro Model; in vivo; innovation; insight; latent infection; mutant; novel; novel therapeutics; permissiveness; prevent; promoter; public health relevance

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) type 1 is a leading cause of infectious corneal blindness. It causes eye disease by reactivation from a latent viral reservoi 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 produt, 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 fo 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 i 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.

描述（由申请人提供）：1型单纯疱疹病毒（HSV）是感染性角膜失明的主要原因。它通过角膜神经中潜伏的病毒性角膜炎的再激活而引起眼病。尽管进行了广泛的研究，但调节神经元中HSV感染的机制尚未得到很好的表征。更好地理解这一点对于确定新的治疗策略至关重要。我们已经开发了一种新的培养系统，使用解离的成年小鼠三叉神经节（TG），研究HSV感染的神经元。我们实验室的初步数据表明，与其在复制细胞中的作用不同，病毒立即早期（IE）基因产物ICP 27限制了神经元中的生产性病毒感染，特别是在A5+神经节神经元中，并促进病毒潜伏期。在目前的提案中，我们将进一步表征ICP27的这种新功能，并研究ICP27实现这些功能的机制。在本提案的前两个具体目标中，我们将描述ICP27在限制生产性感染和促进病毒潜伏期方面的作用，使用ICP27无效突变体，具有延迟表达动力学的ICP27启动子突变体，以及在不同ICP27功能结构域中具有缺失的病毒突变体。我们将通过使用新的AAV载体用ICP 27有效转导感觉神经元来进一步表征ICP 27在限制感染神经元中的作用。我们的初步数据还表明，VP16，复制细胞中的晚期病毒蛋白在TG神经元中非常早地表达，并且ICP 27在培养的TG神经元中限制VP16和ICP 4的转录。在第三个具体目标中，我们将进一步表征ICP27介导的对ICP4和VP16转录的抑制，并测试关于实现这一点的方式的假设。最后，我们将测试ICP27限制A5+中的生产性感染的假设。 部分通过将ICP 4、VP 16和HCF 1限制在细胞质中，从而阻止病毒IE基因的反式激活。这些概念和研究是创新的，并且是能够直接研究神经元中HSV感染的结果，以及能够区分A5+神经元中感染的结果，A5+神经元是HSV潜伏期的主要部位。这些研究的结果应该产生新的见解调节HSV感染的神经元的机制，在开发新的治疗策略的第一步。