Intra- and Inter-Neuronal Viral Trafficking

描述（由申请人提供）：许多嗜神经病毒通过突触传播，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，虽然这些病毒在外周引起快速、高效的感染，但大多数与大脑的慢性感染有关，可能是由于神经元中病毒子代产生和细胞病理学减少。因此，本提案的目标是确定病毒（如麻疹病毒）如何与神经元蛋白相互作用以影响神经元内和神经元间的转运，并确定这种传播模式如何与慢性中枢神经系统感染相关。这些广泛的目标将通过完成两个具体目标来实现。第一个目的是研究麻疹病毒向突触的神经元内传播，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，将确定与控制微管运动的细胞运动蛋白的相互作用。在第二个目标中，神经间麻疹传播得到解决，并确定了一种假定的新型融合蛋白受体，神经激肽-1的作用。具体而言，我们将在易感转基因小鼠模型中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，以及最终阻断这种相互作用如何影响病毒传播和发病机制。这些目标的成功完成将有助于我们的长期目标，包括：(1)鉴定有助于病毒在神经元中持续存在的细胞因子；(2)确定病毒持久性如何导致宿主损伤而不造成神经元损失；最终(3)开发治疗方法，以预防或逆转慢性中枢神经系统感染危及生命的后果。中枢神经系统（CNS）的病毒感染通常与慢性神经系统疾病有关，而不是这些病毒在周围引起的急性疾病。在本应用中，我们主要使用新型小鼠模型、原代神经细胞培养和人类神经细胞系来研究麻疹病毒的神经发病机制。该建议的广泛、长期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，并确定如何通过突触传播可能有助于神经发病机制。上述目标的成功完成将为开发预防或逆转由病毒引起的慢性中枢神经系统感染的危及生命的后果的治疗方法提供基础。

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.