Disruption of Neural Stem Cell Homeostasis by Cytomegalovirus

巨细胞病毒破坏神经干细胞稳态

基本信息

批准号：
8570815
负责人：
DEBORAH Hye SPECTOR
金额：
$ 23.25万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-15 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8570815
关键词：
6-Phosphofructo-2-kinase Address Affect Allogenic Antioxidants Astrocytes Attention Autophagocytosis Autophagosome Biological Assay Cell Death Cell Differentiation process Cell Lineage Cell Survival Cells Cessation of life Cholecalciferol Congenital Abnormality Cytomegalovirus Cytomegalovirus Infections Data Defect Development Environment Enzymes Equilibrium Fetal Development Fibroblasts Foundations Free Radical Scavengers Fructose Future Generations Gestational Age Glucosephosphate Dehydrogenase Glutathione Glutathione Disulfide Glycolysis Glycolysis Induction Goals HIV Health Homeostasis Human Individual Infection Infection prevention Isomerase Measures Mediating Metabolism Methods Midbrain structure NADP Natural regeneration Neuraxis Neurons Oxidation-Reduction Oxidative Stress Pathway interactions Pattern Pentosephosphate Pathway Pregnant Women Protocols documentation Reactive Oxygen Species Research Role Serum Severity of illness Staging Stress Sulforaphane Testing Therapeutic Time Transplantation Viral Virus Virus Diseases analog anaphase-promoting complex base congenital infection cytotoxicity dopaminergic neuron embryonic stem cell glucose metabolism human embryonic stem cell immunosuppressed insight knock-down meetings metabolic abnormality assessment nerve stem cell neurogenesis novel therapeutic intervention oxidative damage pluripotency prevent protective effect public health relevance relating to nervous system self-renewal

项目摘要

DESCRIPTION (provided by applicant): Human cytomegalovirus (HCMV) is the major viral cause of birth defects and affects the central nervous system. It is likely that gestational age an the stage of differentiation of the susceptible neural cells at the time of infection will determin the severity of the disease. A primary goal of this proposal is to determine mechanistically how HCMV infection disrupts the homeostasis of cells of the neural lineage, and impairs survival and differentiation. The focus is on glycolysis, induction of ROS, and autophagy. A second goal is to determine whether we can interfere with the viral manipulation of these pathways, and thus inhibit the viral infection and restore cell survival and appropriate differentiation. For these studies we will use human primitive neural stem cells (pNSCs) that have been derived from approved embryonic stem cells. Established protocols will be used to differentiate the pNSCs to neurons and astrocytes. Cells will be infected with HCMV at various stages during differentiation as well as after differentiation to neurons and astrocytes. In Aim 1, we will determine how oxidative stress and altered metabolism in infected cells of the neural lineage impairs survival and differentiation. Various strategies will be employed to counter the effects of reactive oxygen species (ROS) and redirect glucose metabolism through the pentose phosphate pathway to maintain an antioxidant environment. In Aim 2, we will elucidate the effects of viral infection on autophagy. We predict that analogous to the effects of the infection in fibroblasts, HCMV will inhibit autophagy and this will contribute to altered differentiation and cell death. Since the actve form of Vitamin D3 has been shown to induce autophagy and impair HIV replication, we will determine whether raising the levels of active vitamin D3 will inhibit HCMV infection and have a positive impact on the health of the cell. We expect that these studies will provide great insight into the effects of HCMV on cellular metabolism and neurogenesis and serve as a foundation for future therapeutic efforts in preventing the birth defects due to congenital HCMV. In addition, the results will provide important information regarding how disseminated HCMV infection in immunosuppressed individuals might affect the function and differentiation of transplanted allogeneic neural stem cells, a serious potential problem that has received little attention thus far.

描述（由申请人提供）：人类巨细胞病毒（HCMV）是导致出生缺陷的主要病毒原因，影响中枢神经系统。孕龄和感染时易感神经细胞的分化阶段可能决定疾病的严重程度。该提案的主要目标是从机制上确定 HCMV 感染如何破坏神经谱系细胞的稳态，并损害存活和分化。重点是糖酵解、ROS 诱导和自噬。第二个目标是确定我们是否可以干扰病毒对这些途径的操纵，从而抑制病毒感染并恢复细胞存活和适当的分化。在这些研究中，我们将使用源自批准的胚胎干细胞的人类原始神经干细胞 (pNSC)。已建立的方案将用于将 pNSC 区分为神经元和星形胶质细胞。细胞在分化过程中以及分化成神经元和星形胶质细胞后的各个阶段都会被HCMV感染。在目标 1 中，我们将确定神经谱系受感染细胞中的氧化应激和代谢改变如何损害存活和分化。将采用各种策略来对抗活性氧（ROS）的影响，并通过戊糖磷酸途径重新定向葡萄糖代谢，以维持抗氧化环境。在目标 2 中，我们将阐明病毒感染对自噬的影响。我们预测，与成纤维细胞感染的影响类似，HCMV 将抑制自噬，这将导致分化改变和细胞死亡。由于维生素 D3 的活性形式已被证明可以诱导自噬并损害 HIV 复制，因此我们将确定提高活性维生素 D3 的水平是否会抑制 HCMV 感染并对细胞的健康产生积极影响。我们期望这些研究能够深入了解 HCMV 对细胞代谢和神经发生的影响，并为未来预防先天性 HCMV 引起的出生缺陷的治疗工作奠定基础。此外，结果将提供重要信息，说明免疫抑制个体中的播散性 HCMV 感染如何影响移植的同种异体神经干细胞的功能和分化，这是一个迄今为止很少受到关注的严重潜在问题。