Role of CMV kinase in regulating infection of ESC-derived neuroprogenitor cells

CMV 激酶在调节 ESC 来源的神经祖细胞感染中的作用

• 批准号: 8256085
• 负责人: Tarin M Bigley
• 金额: $ 4.43万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256085
• 关键词: Antiviral Agents; Apoptosis; Biological Models; Birth; Brain; Cell Cycle; Cell Death; Cell Differentiation process; Cell Line; Cells; Cessation of life; Child; Chromatin; Cyclin-Dependent Kinases; Cytolysis; Cytomegalovirus; Cytomegalovirus Infections; Data; Deacetylase; Development; Diploidy; Disease; Early Promoters; Effectiveness; Embryo; Event; Exhibits; Fellowship; Fibroblasts; Future; Gene Expression; Genome; Goals; Herpesviridae; Histones; Human; Immediate-Early Genes; Infection; Infection prevention; Life; Lytic; Mediating; Modeling; Molecular; Mothers; Nervous System Trauma; Neuraxis; Neuroglia; Neurons; Nuclear Pore Complex; Pathway interactions; Pattern; Pharmacologic Substance; Phosphorylation; Phosphotransferases; Post-Translational Protein Processing; Pregnancy; Process; Protein Kinase; Protein-Serine-Threonine Kinases; Proteomics; Recruitment Activity; Regulation; Risk; Role; Sensorineural Hearing Loss; Specificity; Testing; Time; Toxic effect; United States; Viral; Virus; Virus Diseases; cell motility; congenital cytomegalovirus; congenital infection; developmental disease; embryonic stem cell; fetal; hearing impairment; histone modification; human HDAC1 protein; inhibitor/antagonist; kinase inhibitor; maribavir; neonate; nerve stem cell; neural precursor cell; neuron development; non-genetic; precursor cell; prevent; progenitor; promoter

DESCRIPTION (provided by applicant): Human cytomegalovirus (CMV) is the most common cause of non-hereditary sensorineural hearing loss in the U.S. Current therapies to treat infection during pregnancy or upon birth are unproven and risky. The experimental antiviral compound maribavir is a specific inhibitor of the CMV protein kinase, pUL97, and exhibits low toxicity. The objective of my proposal is to identify the molecular mechanism behind pUL97 kinase-mediated regulation of CMV gene expression and determine the impact of inhibiting kinase activity on CMV infected embryonic stem cell-derived neuronal progenitor cells. I have demonstrated that inhibition of the CMV kinase resulted in decreased expression of viral immediate early (IE) genes. This is a newly identified function of the CMV pUL97 kinase. Our lab previously identified an interaction between the CMV pUL97 kinase and cellular histone deacetylase 1 (HDAC1). HDAC1 acts as a repressor and is recruited to the MIE promoter at early times during infection. Therefore, I hypothesize that the kinase activity of CMV pUL97 influences the histone modification pattern at the viral major immediate early promoter by altering cellular HDAC1 activity. I propose to evaluate the pUL97-dependent changes of histone modifications at the major immediate early promoter (MIEP) during infection. These studies will be initiated using human diploid fibroblasts. In addition, I will identify pUL97-mediated changes in HDAC1 localization within the infected cells and at the MIEP. Other herpesvirus kinases phosphorylate HDAC1 and I propose several approaches to identify potential changes in HDAC1 phosphorylation by pUL97. I will determine the impact of these changes on HDAC1 deacetylase activity. I will further these studies using an embryonic stem cell-derived neural progenitor cell line (ES-NPCs) as a model system for CMV-induced neurological damage due to congenital CMV infection. Expression of IE genes has been demonstrated to alter fetal/neonate NPC differentiation. CMV IE genes regulate cell cycle and apoptosis in human diploid fibroblasts. I will test the role of pUL97 during these early events in the context of ES-NPC's by confirming the major results that were obtained in our fibroblast studies. I will determine the effectiveness of maribavir in preventing the pathogenic effects of HCMV infection on neuronal development. My studies will determine if a pUL97 kinase inhibitor is suitable for treating CMV infected neonates and possibly CMV infected mothers at risk of congenital infection. PUBLIC HEALTH RELEVANCE: Congenital cytomegalovirus (CMV) infection occurs in 1 out of 150 pregnancies each year in the U.S. Infection results in approximately 400 neonate deaths and 5,500 children with developmental disorders of the central nervous system. We will determine the molecular mechanism behind an event that occurs very early during infection and explore possible ways of blocking that event and preventing infection within neuronal progenitor cells.

描述（由申请人提供）：人巨细胞病毒（CMV）是美国非遗传性感音神经性听力损失的最常见原因。目前治疗妊娠期间或出生时感染的治疗方法未经证实且存在风险。实验性抗病毒化合物Maribavir是CMV蛋白激酶pUL 97的特异性抑制剂，毒性较低。我的建议的目的是确定pUL 97激酶介导的CMV基因表达调控背后的分子机制，并确定抑制激酶活性对CMV感染的胚胎干细胞衍生的神经元祖细胞的影响。我已经证明，抑制CMV激酶导致病毒立即早期（IE）基因的表达减少。这是CMV pUL 97激酶的新鉴定的功能。我们的实验室以前确定了CMV pUL 97激酶和细胞组蛋白脱乙酰酶1（HDAC 1）之间的相互作用。HDAC 1作为一种阻遏物，在感染早期被募集到MIE启动子。因此，我推测CMV pUL 97的激酶活性通过改变细胞HDAC 1活性来影响病毒主要立即早期启动子处的组蛋白修饰模式。我建议在感染过程中，在主要的立即早期启动子（MIEP）的组蛋白修饰的pUL 97依赖的变化进行评估。这些研究将使用人二倍体成纤维细胞启动。此外，我将确定pUL 97介导的HDAC 1在感染细胞内和MIEP定位的变化。其他疱疹病毒激酶磷酸化HDAC 1，我提出了几种方法来确定潜在的变化HDAC 1磷酸化pUL 97。我将确定这些变化对HDAC 1脱乙酰酶活性的影响。我将使用胚胎干细胞衍生的神经祖细胞系（ES-NPCs）作为CMV诱导的先天性CMV感染引起的神经损伤的模型系统来进一步这些研究。已经证明IE基因的表达改变胎儿/新生儿NPC分化。CMV IE基因调控人二倍体成纤维细胞的细胞周期和凋亡。我将通过证实我们的成纤维细胞研究中获得的主要结果来测试pUL 97在ES-NPC背景下这些早期事件中的作用。我将确定Maribavir在预防HCMV感染对神经元发育的致病作用方面的有效性。我的研究将确定pUL 97激酶抑制剂是否适用于治疗CMV感染的新生儿和可能存在先天性感染风险的CMV感染母亲。 公共卫生关系：在美国，每年每150例妊娠中就有1例发生先天性巨细胞病毒（CMV）感染。感染导致约400例新生儿死亡和5，500例中枢神经系统发育障碍儿童。我们将确定在感染过程中非常早期发生的事件背后的分子机制，并探索阻断该事件和预防神经元祖细胞内感染的可能方法。