Intra- and Inter-Neuronal Viral Trafficking

描述(申请人提供)：许多嗜神经性病毒通过突触传播，包括麻疹、脊髓灰质炎、狂犬病和一些疱疹病毒。然而，控制这一过程的病毒蛋白-神经元蛋白相互作用尚不清楚。此外，虽然这些病毒会在外围引起快速、高效的感染，但大多数病毒都与大脑中的慢性感染有关，这可能是由于病毒后代的产生减少和神经元的细胞病理学所致。因此，这项建议的目标是确定病毒，如麻疹病毒，如何与神经元蛋白相互作用，影响神经元内和神经元间的运输，并确定这种传播方式与慢性中枢神经系统感染的关系。这些广泛目标将通过完成两个具体目标来实现。在第一个目标中，研究了麻疹病毒在神经元内向突触的传播，这将表征存在于突触膜上的病毒蛋白，以及这些蛋白是否需要跨突触传播。此外，还将确定与控制微管运动的细胞马达蛋白的相互作用。在第二个目标中，研究了麻疹的神经元间传播，并确定了一种可能的、新型的融合蛋白受体--神经激肽-1的作用。具体地说，我们将确定病毒神经传递是否需要神经激肽-1，麻疹融合蛋白和神经激肽-1是否相互作用，以及最终阻断这种拟议的相互作用如何影响病毒的传播和在易感转基因小鼠模型中的致病作用。成功完成这些目标将有助于我们的长期目标，其中包括：(1)确定导致病毒在神经元中持续存在的细胞因素；(2)确定病毒持续存在如何可能导致宿主受损而不导致神经元丢失；以及最终(3)开发预防或逆转慢性中枢神经系统感染威胁生命后果的治疗方法。与公共卫生相关的中枢神经系统(CNS)病毒感染通常与慢性神经系统疾病有关，而不是这些病毒在外围引起的急性疾病。在这一应用中，我们使用新的小鼠模型、原代神经细胞培养和人神经细胞系来研究麻疹病毒的神经发病机制。这项提议的广泛、长期的目标是确定麻疹病毒颗粒如何与神经元蛋白相互作用以影响神经元内和神经元间的病毒运输，并确定跨突触传播可能如何有助于神经发病。成功完成拟议的目标将为开发预防或扭转病毒引起的慢性中枢神经系统感染危及生命的后果的治疗方法奠定基础。

DESCRIPTION (provided by applicant): Many neurotropic viruses are transmitted trans-synaptically, including measles, polio, rabies and some herpesviruses. Nevertheless, the viral protein-neuronal protein interactions that govern this process are not known. Moreover, while these viruses cause rapid, highly productive infections in the periphery, most are associated with chronic infections in the brain, likely due to reduced viral progeny production and cytopathology in neurons. Thus, the objectives of this proposal are to define how viruses, such as measles virus, interact with neuronal proteins to affect both intra- and inter-neuronal transport, and to establish how this mode of spread is associated with chronic central nervous system infections. These broad objectives will be accomplished through completion of two Specific Aims. In the first aim, intra-neuronal spread of measles virus to the synapse is studied, which will characterize the viral proteins that are present at synaptic membranes, and whether these proteins are required for trans-synaptic spread. In addition, interaction with cellular motor proteins that govern movement along microtubules will be determined. In the second aim, inter-neuronal measles transmission is addressed, and the role of a putative, novel fusion protein receptor, neurokinin-1, ascertained. Specifically, we will establish if neurokinin-1 is required for viral neurotransmission, whether the measles fusion protein and neurokinin-1 interact, and, ultimately, how blockade of this proposed interaction affects viral spread and pathogenesis in a susceptible transgenic mouse model. Successful completion of these aims will contribute to our long-term goals that include: (1) identification of cellular factors that contribute to viral persistence in neurons; (2) determination of how viral persistence may lead to host impairment without neuronal loss; and eventually (3) development of treatments to prevent or reverse the life- threatening consequences of chronic CNS infections. PUBLIC HEALTH RELEVANCE Viral infections of the central nervous system (CNS) are often associated with chronic neurological diseases, rather than the acute illnesses these viruses cause in the periphery. In this application, we focus on measles virus neuropathogenesis using novel mouse models, primary neuronal cell cultures, and human neuronal cell lines. The broad, long-term objectives of this proposal are to define how measles virus particles interact with neuronal proteins to affect both intra- and inter- neuronal viral transport, and to determine how spread across the synapse may contribute to neuropathogenesis. Successful completion of the proposed aims will provide the foundation for the development of treatments to prevent or reverse the life-threatening consequences of chronic CNS infections caused by viruses.