Alphavirus-manganese interactions and dopaminergic neurodegeneration

甲病毒-锰相互作用和多巴胺能神经变性

基本信息

批准号：
8755333
负责人：
RONALD TJALKENS
金额：
$ 23.64万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-05 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8755333
关键词：
Adolescent Adult Afferent Neurons Age-Months Aging Alphavirus Animal Model Animals Area Autophagocytosis Basal Ganglia Behavioral Brain Cessation of life Colorado Data Defect Development Disease Environment Epidemiology Exposure to Firefly Luciferases Functional disorder Gene Expression Genes Genetic Genetic Models Health Immunotherapy In Situ Infection Inflammatory Knowledge Lead Lesion Lewy Bodies Life Link Luciferases Manganese Mediating Methods Modeling Monitor Mus Nerve Degeneration Nervous System Physiology Neurodegenerative Disorders Neuroglia Neurologic Dysfunctions Neurons Neurotoxins Neurovirology Nose Oral Oxidative Stress Parkinson Disease Parkinsonian Disorders Passive Immunotherapy Pattern Penetrance Pharmaceutical Preparations Phenotype Predisposition Reporter Research Risk Factors Severities Staging System Structure Substantia nigra structure System Testing Therapeutic Intervention Toxic Environmental Substances Toxin Transgenes Transgenic Organisms Treatment Protocols Universities Viral Virulence Virus Virus Diseases Western Equine Encephalitis Virus alpha synuclein base bioluminescence imaging disease phenotype dopaminergic neuron human disease in vivo innovation interest mouse model neurochemistry neuroinflammation neuron loss neuropathology neurotoxicology neurotropic neurotropic virus neurovirulence nonhuman primate novel pars compacta postnatal protein aggregation response tool transgene expression whole body imaging

项目摘要

DESCRIPTION (provided by applicant): Viral infection is implicated as a possible risk-factor for neurodegenerative diseases, including Parkinson's disease (PD). Infection with neurotropic alphaviruses can produce many of the same long-term degenerative effects seen in PD, including protein aggregation, increased levels of oxidative stress, autophagy/mitophagy defects, neuroinflammation, and neuronal death. Neurotropic viruses may therefore represent a better animal model for studying gene x environment interactions in PD that recapitulate more features of the human disease than drug-based lesioning models or genetic models that often lack a pronounced phenotype in the substantial nigra. Evidence suggests that viral infection may act in synergy with other recognized risk-factors, such as aging, genetic factors, and/or previous exposures to environmental neurotoxins to promote neurodegeneration. Among neurotoxins that could have a pronounced gene x environment interaction with neurotropic viruses, Manganese (Mn) is of interest because excessive exposure early in life can have lasting effects on neurological function and can also enhance the neurovirulence of alphaviruses. Here, we propose a novel method for testing gene x environment interactions in Parkinsonism using a neurotropic alphavirus expression system (AES). Following convenient intranasal inoculation, 100% of animals become infected and the progression of infection can be non-invasively monitored in situ using luciferase-expressing viruses and whole body bioluminescence imaging. Neuroinvasion occurs through olfactory sensory neurons and the infection spreads along the neuronal axis in a pattern that mimics the Braak-staging system of PD. The severity and persistence of viral infection can be tightly controlled and our preliminary data using unbiased stereology indicate that AES infection results in significant loss of dopaminergic neurons in the substantial nigra pars compacta (SNpc). It is the Central Hypothesis of this proposal that intranasal infection with luciferase- expressing alphavirus will result in neuroinvasion through olfactory and subcortical structures resulting in progressive loss of dopaminergic neurons in the substantia nigra that is associated with inflammatory activation of glial cells. Moreover, we postulate that pre-exposure to Mn during juvenile development will enhance susceptibility to viral-mediated loss of dopaminergic neurons, in part through an increased neuroinflammatory response. This hypothesis will be tested in two Specific Aims during the 2-year project period as follows: Specific Aim 1 - Determine the optimal viral titer and phenotype for inducing persistent infection and neurodegeneration in the basal ganglia following intranasal installation of luciferase-expressing Western Equine Encephalitis Virus (WEEV); Specific Aim 2 - Characterize the disease phenotype resulting from exposure to Mn pre- and post-viral infection. We expect that these studies will demonstrate that the proposed AES-based model is a powerful method for inducing parkinsonism in mice, as well as for transgene delivery into the CNS, that can be used to identify novel gene x environment interactions relevant to PD. Additionally, we expect that exposure to Mn during juvenile development will exacerbate neurodegeneration following adult infection with WEEV. The results of these studies will increase our understanding of the environmental links to neurodegenerative disease and will provide a powerful new animal model for studying virus/toxin interactions in the CNS.

描述（由申请人提供）：病毒感染被视为神经退行性疾病（包括帕金森氏病（PD））的可能风险因素。神经性α病毒感染可以产生PD中看到的许多相同的长期退化作用，包括蛋白质聚集，氧化应激水平升高，自噬/线粒体缺陷，神经炎症和神经元死亡。因此，与基于药物的病变模型或遗传模型相比，神经性病毒可能代表了研究基因X环境相互作用的更好的动物模型，该模型概括了人类疾病的更多特征或在实质性NIGRA中通常缺乏明显表型的遗传模型。有证据表明，病毒感染可能与其他公认的风险因素协同作用，例如衰老，遗传因素和/或先前对环境神经毒素的暴露以促进神经变性。在神经毒素中可能与神经性病毒具有明显的基因X环境相互作用的神经毒素中，锰（MN）引起了人们的关注，因为生命早期过度暴露会对神经功能产生持久影响，并且还可以增强α病毒的神经毒素。在这里，我们提出了一种新的方法，用于使用神经性α病毒表达系统（AES）测试帕金森氏症基因X环境相互作用。便利的鼻内接种后，使用表达荧光素酶的病毒和全身生物发光成像，可以在原位监测100％的动物感染，并且感染的进展可以进行非侵袭性监测。神经浸觉是通过嗅觉的感觉神经元发生的，并且感染以模仿PD的Braak阶段系统的模式沿着神经元轴传播。病毒感染的严重程度和持久性可以受到严格控制，我们使用无偏的立体学的初步数据表明，AES感染会导致大量NIGRA PARS CMPSTACTA（SNPC）中多巴胺能神经元的显着丧失。这一提议的中心假设是，用荧光素酶表达α病毒的鼻腔内感染将通过嗅觉和皮层下结构导致神经侵袭，从而导致质体中多巴胺能神经元的逐渐丧失，与nigra中的多巴胺能神经元逐渐丧失，这与与听觉细胞的炎症激活相关。此外，我们假设在少年发育过程中暴露于MN将增强对病毒介导的多巴胺能神经元丧失的敏感性，部分通过神经炎症反应的增加。该假设将在2年期间的两个特定目标中进行测试：具体目的1-确定最佳病毒滴度和表达荧光素酶的西马脑炎病毒（WEEV）后，用于诱导基底神经节持续感染和神经退行性的表型；具体目标2-特征是暴露于MN前感染和病毒后感染而导致的疾病表型。我们预计这些研究将证明，基于AES的模型是诱导小鼠帕金森氏症以及转基因输送到CNS中的强大方法，可用于识别与PD相关的新型基因X环境相互作用。此外，我们预计在少年发育过程中暴露于MN会加剧成年人感染WEEV后神经变性。这些研究的结果将增加我们对与神经退行性疾病的环境联系的理解，并将为研究中枢神经系统中的病毒/毒素相互作用提供强大的新动物模型。