Mechanisms of virus-induced apoptosis in neurons and CNS

描述（由申请人提供）：中枢神经系统（CNS）病毒感染是由病毒损伤大脑和脊髓内的神经元和其他非神经元细胞的能力引起的。多种嗜神经病毒损伤中枢神经系统靶细胞的常见机制是通过细胞凋亡。尽管如此，关于导致病毒诱导的神经细胞凋亡的确切机制和特定细胞途径知之甚少。在这个提议中，我们将利用呼肠孤病毒感染原代神经元培养物和呼肠孤病毒诱导脑炎的小鼠模型来研究病毒诱导的神经元凋亡的细胞通路。我们之前已经表明，死亡受体凋亡信号有助于呼肠孤病毒诱导的神经元凋亡。我们假设线粒体和内质网细胞应激凋亡信号通路也对呼肠孤病毒诱导的神经元细胞死亡至关重要。在目的1中，我们将确定内质网（ER）和细胞应激（GADD）途径在呼肠孤病毒诱导的神经元凋亡中所起的作用，并描述所涉及的因素。在Aim 2中，我们将确定线粒体和线粒体因子在呼肠孤病毒诱导的细胞凋亡中的作用。最后，在Aim 3中，我们将在体内测试操纵细胞凋亡通路对呼肠孤病毒诱导的中枢神经系统感染发病机制的影响。提出的研究的一个关键方面是能够将体外原代神经元培养研究中获得的结果转化为中枢神经系统病毒感染的体内模型。这些研究选择呼肠孤病毒感染，因为它是病毒诱导的细胞凋亡的最广泛表征的模型之一，并且提供了关于细胞凋亡的病毒决定因素和所涉及的细胞途径的大量预先存在的知识基础，以及易于获得的小鼠中枢神经系统细胞凋亡模型，可以快速测试细胞培养中获得的结果，以确定其在体内中枢神经系统病毒性疾病的发病机制

DESCRIPTION (provided by applicant): Central nervous system (CNS) viral infections result from the capacity of viruses to injure neurons and other non-neuronal cells within the brain and spinal cord. A common mechanism by which a diverse group of neurotropic viruses injure target cells in the CNS is through apoptosis. Despite this fact, little is known about the precise mechanisms and specific cellular pathways leading to virus-induced apoptotic neural cell death. In this proposal we will utilize reovirus infection of primary neuronal cultures and a murine model of reovirus- induced encephalitis to examine the cellular pathways involved in virus-induced neuronal apoptosis. We have previously shown that death receptor apoptotic signaling contributes to reovirus-indced neuronal apoptosis. We hypothesize that both mitochondrial and endoplasmic reticulum cell stress apoptotic signaling pathways are also critical to reovirus-induced neuronal cell death. In Aim 1 we will determine the role played by endoplasmic reticulum (ER) and cellular-stress (GADD) pathways on reovirus-induced neuronal apoptosis and characterize the factors involved. In Aim 2 we will determine the role of mitochondria and mitochondrial factors in reovirus-induced apoptosis. Finally, in Aim 3 we will test the effects of manipulating apoptotic pathways on the pathogenesis of reovirus-induced CNS infection in vivo. A key aspect of the proposed studies is the ability to translate results obtained in studies of primary neuronal cultures in vitro, into an in vivo model of CNS viral infection. Reovirus infection was selected for these studies because it is one of the most extensively characterized models of virus-induced apoptosis and offers the advantage of a large pre- existing base of knowledge concerning viral determinants of apoptosis and the cellular pathways involved, as well as an easily accessible in mouse model of CNS apoptosis that allows rapid testing of results obtained in cell culture for their significance on the pathogenesis of CNS viral disease in vivo