Investigating the Role of GDE2 in Neuronal Mitotic Inhibition and Nuclear Pore Complex Defects

研究 GDE2 在神经元有丝分裂抑制和核孔复合体缺陷中的作用

• 批准号: 10231661
• 负责人: Anna Nicole Westerhaus
• 金额: $ 4.6万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-13 至 2024-04-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231661
• 关键词: ALS patients; Address; Adult; Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease brain; Amyotrophic Lateral Sclerosis; Autopsy; Brain; Cell Cycle; Cell Cycle Inhibition; Cell Cycle Progression; Cell Cycle Regulation; Cell Proliferation; Cell surface; Cellular biology; Chronology; Cleaved cell; Data; Defect; Dementia; Development; Disease; Dissection; Embryo; Embryonic Development; Enzymes; Etiology; Exhibits; Functional disorder; GPI Membrane Anchors; Genetic Epistasis; Grant; Human; Immunohistochemistry; Impairment; In Vitro; Knock-out; Knockout Mice; Knowledge; Life; Maintenance; Mediating; Membrane; Mentors; Mentorship; Mitosis; Mitotic; Mitotic Cell Cycle; Molecular; Motor Cortex; Mus; National Research Service Awards; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Neurosciences; Nuclear Envelope; Nuclear Pore Complex; Outcome; Pathogenesis; Pathologic; Pathology; Pathway interactions; Phenotype; Physiological; Primary Cell Cultures; Process; Proteins; Public Health; RNA analysis; Research; Role; Sampling; Scientist; Severities; Signal Transduction; Structure; Technical Expertise; Testing; Therapeutic Intervention; Time; Training; Transmission Electron Microscopy; Ubiquitination; Viral; Virus; WNT Signaling Pathway; Work; base; brain tissue; collaborative environment; design and construction; effective therapy; experimental study; glycerophosphodiester phosphodiesterase; human disease; human tissue; improved; in vivo; insight; knockout animal; mouse genetics; neuronal survival; new therapeutic target; novel; nucleocytoplasmic transport; prevent; skills; stem cell proliferation; teacher; therapeutic candidate; tissue preparation; transcriptome sequencing

Project Summary / Abstract Alzheimer's disease (AD) and related diseases (ADRDs) are the leading cause of dementia, affecting millions of people each year. Despite being such a prevalent public health concern, knowledge of the etiology of neurodegeneration in these diseases is limited and has stymied the development of effective treatments. Aberrant neuronal cell-cycle reentry (CCR) and nuclear pore complex (NPC)/nucleocytoplasmic transport defects have been separately identified as causal phenomena for neurodegeneration in AD and ADRDs. However, the mechanisms leading to these phenomena and the relationship between them remains unknown. We have identified a pathway involving Glycerophosphodiester phosphodiesterase 2 (GDE2 or GDPD5) that prevents CCR and NPC/nucleocytoplasmic transport defects in the adult mammalian brain and is disrupted in human postmortem AD and ALS brain tissue. GDE2 is one of three six-transmembrane enzymes that act at the cell surface to cleave the GPI-anchor that tethers some proteins to the membrane. During embryogenesis, GDE2 inhibits progenitor cell proliferation to promote neuronal differentiation. Preliminary studies show that loss of GDE2 in the adult brain leads to CCR and NPC abnormalities that precede overt neurodegeneration. Because NPC defects are coincident with CCR in neurons lacking GDE2, and the NPC normally breaks down during mitosis, we hypothesize that GDE2 is important for maintaining neurons in a postmitotic state throughout life and that aberrant CCR elicited by GDE2 loss leads to NPC breakdown and neurodegeneration. Importantly, GDE2 localization and function is disrupted in AD and ADRDs, raising the possibility that dysfunction of the GDE2 pathway could contribute to the pathogenesis of these diseases. This proposal will test our central hypothesis in three aims. Aim 1 will define the physiological requirement for GDE2 in maintaining neuronal quiescence and NPC integrity in aging mice while Aim 2 will define the relationship between CCR and NPC breakdown and identify the mechanisms leading to CCR and NPC defects. Later experiments will determine relevance of this pathway to CCR and NPC disruption in human AD and ADRDs (Aim 3). The proposed research is expected to identify a novel physiological pathway that is essential for neuronal survival and will provide new insight into causal mechanisms of neurodegeneration that are relevant to human disease. In addition, this project has immense training potential. I will have the opportunity to improve my knowledge in neuroscience and cell biology, and to acquire technical expertise in primary cell culture, viral construct design and transduction, mouse genetics, brain dissections and tissue preparation, and immunohistochemical analysis of human tissues. This research will be performed in a highly collaborative environment, where I will have numerous opportunities to receive quality mentorship and training, to develop my written and presentation skills, and to grow as a mentor and teacher to more junior scientists. Overall, the Kirschstein-NRSA grant will support both my research aimed at discovering the molecular basis of neurodegeneration and my development as an independent scientist.

项目总结/摘要 阿尔茨海默病（AD）和相关疾病（ADRD）是痴呆症的主要原因，影响数百万人。 人每年。尽管是这样一个普遍的公共卫生问题， 这些疾病中的神经变性是有限的，并且阻碍了有效治疗的发展。 异常神经元细胞周期折返（CCR）和核孔复合物（NPC）/核质转运 缺陷已被分别鉴定为AD和ADRD中神经变性的因果现象。 然而，导致这些现象的机制以及它们之间的关系仍然未知。 我们已经鉴定了涉及甘油磷酸二酯磷酸二酯酶2（GDE 2或GDPD 5）的途径， 防止成年哺乳动物脑中CCR和NPC/核质转运缺陷， 人类AD和ALS死后脑组织。GDE 2是三种六跨膜酶之一， 细胞表面切割GPI锚，GPI锚将一些蛋白质拴在膜上。在胚胎发生过程中，GDE 2 抑制祖细胞增殖以促进神经元分化。初步研究表明， 成人大脑中的GDE 2导致明显神经变性之前的CCR和NPC异常。因为 NPC缺陷与缺乏GDE 2的神经元中的CCR一致，并且NPC通常在GDE 2缺失期间分解。 有丝分裂，我们假设GDE 2是重要的维持神经元在有丝分裂后的状态在整个生命， 由GDE 2缺失引起的异常CCR导致NPC破坏和神经变性。重要的是，GDE 2 AD和ADRD的定位和功能被破坏，增加了GDE 2功能障碍的可能性， 可能参与了这些疾病的发病机制。这一提议将检验我们的中心假设， 三个目标。目的1将定义GDE 2在维持神经元静止中的生理需求， 而目标2将定义CCR和NPC分解之间的关系， 识别导致CCR和NPC缺陷的机制。稍后的实验将确定这一点的相关性 在人类AD和ADRD中CCR和NPC破坏的途径（目的3）。预计拟议的研究将 确定了一种新的生理途径，这是神经元生存所必需的，并将提供新的见解， 与人类疾病相关的神经退行性变的因果机制。此外，该项目还 巨大的训练潜力。我将有机会提高我在神经科学和细胞生物学方面的知识， 并获得原代细胞培养，病毒构建体设计和转导，小鼠遗传学， 脑解剖和组织制备，以及人体组织的免疫组织化学分析。本研究 将在一个高度合作的环境中进行，在那里我将有很多机会获得 高质量的指导和培训，发展我的写作和演讲技巧，并成长为一个导师， 更多年轻科学家的老师。总的来说，Kirschstein-NRSA基金将支持我的研究， 发现神经退行性疾病的分子基础以及我作为独立科学家的发展。