Molecular Neuro-pathogenesis of Congenital Cytomegalovirus Infection

项目摘要/摘要人类巨细胞病毒(HCMV)感染是主要的感染源 新生儿先天疾病的风险。先天性巨细胞病毒感染导致永久性神经和 神经认知障碍，并导致世界各地的重大健康问题。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IE1相互作用而影响的。 因此，我们建议通过Notch途径探讨其分子致病机制。 先天性巨细胞病毒感染引起的大脑发育异常。这种提议的方法从根本上说是 与以前的研究不同，因为我们应用我们独特的技术来探索 巨细胞病毒用来干扰胚胎的大脑发育。我们将应用体外人巨细胞病毒感染 小鼠巨细胞病毒(MCMV)基因宫内电穿孔系统、条件转基因小鼠 模型和胎盘内MCMV感染系统。我们的长期目标是揭示潜在的机制 巨细胞病毒诱导的先天性神经发育障碍，并开发潜在的抗巨细胞病毒疗法。我们 假设CMV和Notch信号之间的协调相互作用是导致 胚胎中被感染的神经前体细胞的发育缺陷，导致永久性 神经和智力残疾。这一假设得到了我们的初步数据和出版物的支持 从体外和体内实验来看，HCMV感染影响Notch信号转导，即Notch信号转导 参与了CMV引起的鼻咽癌增殖缺陷，即IE1是抑制鼻咽癌的必要条件和充分因素 IE1与Notch信号通路的蛋白相互作用。我们将通过以下方式测试我们的假设 目的：1.研究人巨细胞病毒IE1基因缺陷的分子机制 人NSC/NPC在细胞培养中的增殖和分化；以及目的2.检测CMV IE1 使用IE1可诱导的转基因小鼠模型和临床上的 相关胎盘内MCMV感染小鼠模型。我们相信，目前的建议在以下方面是创新的 几点：首先，这是第一次使用可诱导转基因小鼠模型引入 特定的CMV蛋白，IE1，以解决先天性CMV感染的分子机制。第二，我们 从分子水平到行为水平研究先天性巨细胞病毒感染的病理。最后，调查人员 具有不同专业背景(病毒学和神经科学)的人正在共同努力解决 具有很高医学影响的重要问题。积极影响将包括以下方面的质的进展 特别是对分子病毒学和CMV病理生物学的了解，以及对其机制的了解 用来导致神经缺陷的巨细胞病毒可能因此推动新的、选择性的抗巨细胞病毒疗法的发展 战略。

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.