The Role of RME-8 in Neuronal Health and Parkinson's Disease

RME-8 在神经元健康和帕金森病中的作用

• 批准号: 10284925
• 负责人: Sierra Swords
• 金额: $ 3.96万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10284925
• 关键词: Affect; Age; Aging; Alleles; Animals; Architecture; Auxins; Axon; Back; Behavior; Biological; Brain; Caenorhabditis elegans; Cause of Death; Cell membrane; Cells; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Confocal Microscopy; Data; Defect; Disease; Dopamine; Dynein ATPase; Elderly; Endocytosis; Endosomes; Equilibrium; Essential Tremor; Etiology; Family; Functional disorder; Gilles de la Tourette syndrome; Golgi Apparatus; Health; Human; Image; Impairment; Label; Lead; Length; Link; Longevity; Loss of Heterozygosity; Lysosomes; Membrane; Modeling; Motor; Movement; Mus; Neurodegenerative Disorders; Neuroglia; Neurons; Organelles; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Patients; Peripheral; Play; Population; Prevalence; Process; Proteins; RNA Interference; Recycling; Risk Factors; Role; Sorting - Cell Movement; Speech; Substantia nigra structure; Synapses; System; Techniques; Temperature; Testing; Travel; Variant; Vesicle; alpha synuclein; base; behavioral study; brain cell; causal variant; confocal imaging; dopaminergic neuron; dynactin; experimental study; genetic variant; in vivo; insight; knock-down; late endosome; loss of function; mutant; neuron loss; neuronal cell body; novel; overexpression; prevent; recruit; retrograde transport; trafficking

The purpose of this project is to determine how a recently discovered, Parkinson's Disease (PD)- associated, gene variant in human patients contributes to the pathogenesis of PD. The allele under study, in RME-8, is dominant and linked to late-onset PD. RME-8 is a protein localized to endosomes that our lab previously identified as a crucial regulator of transmembrane cargo sorting after endocytosis, determining whether endocytic cargo is recycled or degraded. This is the first instance in which RME-8 has been linked to human neuronal health, though its function in neurons is not yet understood. Preliminary data in vivo in C. elegans, and in primary cultured mouse cortical neurons, indicates that the RME-8 PD-associated allele interferes with retrograde trafficking of late endosomes along the axon toward the cell body of the neuron. Due to the unique architecture of neurons, an inability of cargo to reach the cell body may be exceptionally catastrophic, since the cell body is where degradation and recycling of cargo occurs via the lysosomes and Golgi, respectively. Inability of cargo to traffic back to the cell body may lead to accumulation in the axon, resulting in neuron dysfunction and death. To accomplish this project, I will utilize a dual-model approach of both cultured mouse neurons as well as an in vivo approach in C. elegans, using CRISPR, RNAi, the auxin-inducible degron system, and temperature sensitive loss-of-function alleles. I will determine the effects on retrograde trafficking of late endosomes along the axon using confocal imaging and kymograph analysis, as well determine the effects on degradation and recycling of cargo. Since Parkinson's Disease is caused by the death of dopaminergic neurons, I will analyze changes in dopamine-dependent behaviors in C. elegans and determine the effect of RME-8 perturbation on the lifespan of dopaminergic neurons. Understanding the novel role of RME-8 in maintaining neuronal health leads us closer to understanding the factors that contribute to Parkinson's Disease and therefore closer to determining how to treat patients with PD.

该项目的目的是确定最近发现的帕金森氏病 (PD)-人类患者中的相关基因变异有助于PD的发病机制。的 研究中的RME-8等位基因是显性的，与晚发型PD相关。RME-8是一种 我们的实验室以前确定它是一个重要的调节因子， 内吞后跨膜货物分选，确定内吞货物是否 回收或降解。这是RME-8与人类基因组相关的第一个例子。 神经元健康，尽管其在神经元中的功能尚未被理解。体内初步数据 C.在原代培养的小鼠皮层神经元中，表明RME-8 PD相关的 等位基因干扰晚期内体沿着轴突向 神经元的细胞体。由于神经元的独特结构，货物无法 到达细胞体可能是异常灾难性的，因为细胞体是降解的地方， 并且货物的再循环分别通过溶酶体和高尔基体发生。货物不能 回到细胞体的交通可能导致轴突中的积累，导致神经元损伤。 功能障碍和死亡。为了完成这个项目，我将使用两种模型的方法 培养的小鼠神经元以及在C.利用CRISPR，RNAi， 生长素诱导的降解决定子系统和温度敏感的功能丧失等位基因。我会 使用共聚焦显微镜确定对晚期内体沿着轴突逆行运输的影响 成像和记波分析，以及确定对降解和回收的影响， 货物.由于帕金森病是由多巴胺能神经元死亡引起的，我将 分析C. elegans和确定的影响 RME-8干扰多巴胺能神经元的寿命。理解的新颖角色 RME-8在维持神经元健康方面的作用使我们更接近于理解 有助于帕金森病，因此更接近于确定如何治疗患者， 警局