Mechanisms of Neuronal Maintenance and Protection.

神经元维持和保护机制。

• 批准号: 8097985
• 负责人: Rong Grace Zhai
• 金额: $ 32.8万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8097985
• 关键词: Address; Biochemical; Biological Models; Degenerative Disorder; Disease; Drosophila genus; Environmental Risk Factor; Enzymes; Epigenetic Process; Event; Genes; Genetic; Genetic Screening; Goals; Health; Heat-Shock Response; Heating; Hippocampus (Brain); Homeostasis; Homologous Gene; Housekeeping; Human; Indium; Life; Light; Maintenance; Mediating; Messenger RNA; Modeling; Molecular; Molecular Chaperones; Mus; Mutagenesis; Mutation; Nerve Degeneration; Nerve Regeneration; Nervous system structure; Neurons; Nicotinamide-Nucleotide Adenylyltransferase; Organism; Phototransduction; Process; Property; Protein Isoforms; Proteins; Regulation; Research; Retinal Degeneration; Role; Severities; Spinal Ganglia; Stress; Structure; System; Tertiary Protein Structure; Testing; Therapeutic; Time; Toxic effect; Transcriptional Regulation; Ubiquitination; Visual system structure; Wallerian Degeneration; Work; ataxin-1; base; design; effective therapy; genetic manipulation; insight; mouse model; neurotoxicity; overexpression; polyglutamine; promoter; protective effect; protein aggregation; protein degradation; protein folding; protein misfolding; public health relevance; receptor; repaired; research study

DESCRIPTION (provided by applicant): Neurodegeneration can be triggered by a variety of genetic, epigenetic, and environmental factors. Healthy neurons are able to maintain their integrity throughout the life of an organism, suggesting the existence of a maintenance mechanism that allows neurons to sustain, mitigate or even repair damage. Recently, we have identified a neuronal maintenance factor NMNAT in a forward genetic screen in Drosophila. Loss of nmnat causes rapid and severe neurodegeneration, whereas over-expression of NMNAT protein offers protection against neurodegeneration. These findings suggest that normal level of NMNAT maintains neuronal homeostasis, and increased level offers protection. NMNAT is a highly conserved housekeeping enzyme, and the neuroprotective function of NMNAT has also been implicated in a mouse model of slow Wallerian Degeneration. Currently, the detailed mechanisms of this maintenance function and the protective capability of NMNAT in mammalian neurons are unclear. Our preliminary experiments suggest that in addition to its NAD synthesis activity, NMNAT has a chaperone function that is involved in regulating protein misfolding and degradation. We hypothesize that like other chaperones, NMNAT is up-regulated under stress, reduces protein aggregation, and thus protects neurons from degenerative conditions. In the proposed research, we will characterize the biochemical and cellular mechanisms underlying the protective process mediated by NMNAT using both Drosophila and mammalian primary neuronal models. In Specific Aim 1, we will use structure- function analysis to define the protein domains that are required for chaperone function, and characterize the transcriptional regulation of NMNAT under stress. In Specific Aim 2, we will first determine the neuroprotective activity of mammalian NMNAT isoforms in primary neurons, and then characterize the role of NMNAT in reducing protein aggregation-induced neurotoxicity. In Specific Aim 3, we will test whether NMNAT proteins can exert protective activity when their expression is induced after the onset of degeneration. For this last study, we will take advantage of the Drosophila genetic system and control the expression of NMNAT using a heat-inducible promoter. In summary, our proposed research in both Drosophila and mammalian model systems will help unmask the function of NMNAT and its regulation as a molecular chaperone, determine the neuroprotective properties of human NMNAT in primary neurons, and reveal the repair potential of NMNAT in neural regeneration after neuronal damage. PUBLIC HEALTH RELEVANCE: Neurodegenerative conditions are among the most intractable of diseases and therefore present an urgent need for developing effective treatments. Our studies on the neuronal maintenance process suggest that neurons have a self-defense system that can be augmented to protect against neurodegeneration. The experiments in this proposal will help us better understand the molecular events of protein folding and aggregation in neurodegenerative conditions, reveal potential neuroprotective mechanisms in mammalian neurons, and aid in the design of therapeutic treatments.

描述（由申请人提供）：神经变性可由多种遗传、表观遗传和环境因素引发。健康的神经元能够在生物体的整个生命周期中保持其完整性，这表明存在一种维持机制，可以使神经元维持、减轻甚至修复损伤。最近，我们在果蝇的正向遗传筛选中发现了一种神经元维持因子 NMNAT。 nmnat 的缺失会导致快速而严重的神经退行性变，而 NMNAT 蛋白的过度表达则可以防止神经退行性变。这些发现表明，正常水平的 NMNAT 可以维持神经元稳态，而升高的水平则可以提供保护。 NMNAT 是一种高度保守的管家酶，NMNAT 的神经保护功能也与缓慢沃勒变性小鼠模型有关。目前，NMNAT在哺乳动物神经元中的这种维持功能和保护能力的详细机制尚不清楚。我们的初步实验表明，除了 NAD 合成活性外，NMNAT 还具有参与调节蛋白质错误折叠和降解的伴侣功能。我们假设与其他伴侣一样，NMNAT 在压力下上调，减少蛋白质聚集，从而保护神经元免受退化状况的影响。在拟议的研究中，我们将使用果蝇和哺乳动物初级神经元模型来表征 NMNAT 介导的保护过程背后的生化和细胞机制。在具体目标 1 中，我们将使用结构功能分析来定义伴侣功能所需的蛋白质结构域，并表征 NMNAT 在压力下的转录调控。在具体目标 2 中，我们将首先确定哺乳动物 NMNAT 亚型在初级神经元中的神经保护活性，然后表征 NMNAT 在减少蛋白质聚集引起的神经毒性中的作用。在具体目标 3 中，我们将测试 NMNAT 蛋白在变性开始后诱导表达时是否可以发挥保护活性。对于最后一项研究，我们将利用果蝇遗传系统并使用热诱导启动子控制 NMNAT 的表达。总之，我们在果蝇和哺乳动物模型系统中提出的研究将有助于揭示 NMNAT 的功能及其作为分子伴侣的调节作用，确定人 NMNAT 在初级神经元中的神经保护特性，并揭示 NMNAT 在神经再生中的修复潜力。神经元损伤。公共卫生相关性：神经退行性疾病是最难治的疾病之一，因此迫切需要开发有效的治疗方法。我们对神经元维持过程的研究表明，神经元具有自我防御系统，可以增强该系统以防止神经变性。该提案中的实验将帮助我们更好地了解神经退行性疾病中蛋白质折叠和聚集的分子事件，揭示哺乳动物神经元的潜在神经保护机制，并有助于设计治疗方法。