Molecular Neuro-pathogenesis of Congenital Cytomegalovirus Infection

人类巨细胞病毒（HCMV）感染是导致感染的主要原因 新生儿的先天性疾病。先天性HCMV感染导致永久性神经和 神经认知障碍，并导致世界范围内的重大健康问题。我们的初步调查结果显示 Notch途径受HCMV感染的影响，可能是通过与CMV编码的蛋白IE 1相互作用。 因此，我们建议通过Notch通路来探讨其致病的分子机制， 先天性巨细胞病毒感染引起的大脑发育异常。这种方法从根本上 与以前的研究不同，因为我们应用我们独特的技术来探索 CMV用于干扰胚胎的大脑发育。我们将应用体外HCMV感染 系统，用于小鼠CMV（MCMV）基因递送子宫内电穿孔系统，条件性转基因小鼠 模型和胎盘内MCMV感染系统。我们的长期目标是揭示 CMV诱导的先天性神经发育障碍和开发潜在的抗CMV治疗。我们 我假设CMV和Notch信号之间的协调相互作用是导致 胚胎中受感染的神经祖细胞（NPC）的发育缺陷，导致永久性的 神经和智力残疾。我们的初步数据和出版物支持这一假设 从HCMV感染影响Notch信号传导的体外和体内实验，Notch信号传导是 IE 1参与CMV引起的NPC增殖缺陷，IE 1是必需的，足以抑制NPC IE 1与Notch信号通路的蛋白相互作用。我们将测试我们的假设通过 具体目标：目标1。为了研究HCMV IE 1如何导致细胞缺陷的分子机制， 细胞培养中人NSC/NPC增殖和分化;和目的2.为了研究CMV IE 1 使用IE 1诱导型转基因小鼠模型和临床- 相关胎盘内MCMV感染小鼠模型。我们认为，目前的建议是创新的， 几点：首先，这是使用诱导型转基因小鼠模型的首批研究之一， 特异性CMV蛋白，IE 1，以解决先天性CMV感染的分子机制。二是 从分子水平到行为水平研究先天性CMV感染的病理。最后，调查人员 具有不同专业背景（病毒学和神经科学）的人正在共同努力， 具有高度医学影响的重要问题。积极影响将包括以下方面的质的进步： 了解分子病毒学，特别是CMV病理生物学， CMV用于引起神经缺陷可能因此推动新的选择性抗CMV治疗药物的开发 战略布局

PROJECT SUMMARY/ABSTRACT Human cytomegalovirus (HCMV) infection is the leading infectious cause of congenital disorders in newborns. Congenital HCMV infection causes permanent neurological and neurocognitive disabilities and results in significant health problems worldwide. Our preliminary findings reveal that Notch pathway is affected by HCMV infection and probably via interaction with CMV-encoded protein, IE1. Therefore, we propose to explore the molecular pathogenic mechanism via Notch pathway underlying abnormal brain development caused by congenital CMV infection. This proposed approach is fundamentally different from previous studies because we apply our unique technologies to explore the mechanisms that CMV employs to interfere with the brain development of embryos. We will apply the in vitro HCMV infection system, in utero electroporation system for mouse CMV (MCMV) gene delivery, conditional transgenic mouse model, and intraplacental MCMV infection system. Our long-term goal is to reveal the mechanisms underlying CMV-induced congenital neurodevelopmental disorders and develop potential anti-CMV therapeutics. We hypothesize that the coordinative interaction between CMV and Notch signaling is the key to causing developmental defects of infected neural progenitor cells (NPCs) in embryos, resulting in permanent neurological and intellectual disabilities. The hypothesis is supported by our preliminary data and publications from in vitro and in vivo experiments that HCMV infection affects Notch signaling, that Notch signaling is involved in CMV-caused defects of NPC proliferation, that IE1 is essential and sufficient to inhibit NPC proliferation, and that IE1 interacts with proteins of Notch signaling pathway. We will test our hypothesis via the following Specific Aims: Aim 1. To examine molecular mechanisms for how HCMV IE1 causes defects of human NSC/NPC proliferation and differentiation in cell culture; and Aim 2. To examine how the CMV IE1 causes different stages of neurogenic defect using an IE1-inducible transgenic mouse model and a clinically- relevant intraplacental MCMV infection mouse model. We believe that the present proposal is innovative in several points: First, this is one of the first studies that uses an inducible transgenic mouse model introducing a specific CMV protein, IE1, to address molecular mechanisms underlying congenital CMV infection. Second, we study the congenital CMV infection pathology from molecular levels to behavior levels. Lastly, investigators with different professional backgrounds (virology and neuroscience) are working together to address an important question with high medical impact. Positive impacts will include qualitative advances in understanding molecular virology and CMV pathobiology in particular, and that understanding the mechanisms CMV uses to cause neural defects may thus drive development of new, selective anti-CMV therapeutic strategies.